Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnosti...Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.展开更多
BACKGROUND The diagnosis of sepsis combined with acute respiratory distress syndrome(ARDS)has increased owing to the enhanced awareness among medical profes-sionals and the continuous development of modern medical tec...BACKGROUND The diagnosis of sepsis combined with acute respiratory distress syndrome(ARDS)has increased owing to the enhanced awareness among medical profes-sionals and the continuous development of modern medical technologies,while early diagnosis of ARDS still lacks specific biomarkers.One of the main patho-genic mechanisms of sepsis-associated ARDS involves the actions of various pathological injuries and inflammatory factors,such as platelet and white blood cells activation,leading to an increase of surface adhesion molecules.These adhesion molecules further form platelet-white blood cell aggregates,including platelet-mononuclear cell aggregates(PMAs).PMAs has been identified as one of the markers of platelet activation,here we hypothesize that PMAs might play a potential biomarker for the early diagnosis of this complication.METHODS We selected 72 hospitalized patients diagnosed with sepsis as the study population between March 2019 and March 2022.Among them,30 patients with sepsis and ARDS formed the study group,while 42 sepsis patients without ARDS comprised the control group.After diagnosis,venous blood samples were imme-diately collected from all patients.Flow cytometry was employed to analyze the expression of PMAs,platelet neutrophil aggregates(PNAs),and platelet aggregates(PLyAs)in the serum.Additionally,the Acute Physiology and Chronic Health Evaluation(APACHE)II score was calculated for each patient,and receiver operating characteristic curves were generated to assess diagnostic value.RESULTS The study found that the levels of PNAs and PLyAs in the serum of the study group were higher than those in the control group,but the difference was not statistically significant(P>0.05).However,the expression of PMAs in the serum of the study group was significantly upregulated(P<0.05)and positively correlated with the APACHE II score(r=0.671,P<0.05).When using PMAs as a diagnostic indicator,the area under the curve value was 0.957,indicating a high diagnostic value(P<0.05).Furthermore,the optimal cutoff value was 8.418%,with a diagnostic sensitivity of 0.819 and specificity of 0.947.CONCLUSION In summary,the serum levels of PMAs significantly increase in patients with sepsis and ARDS.Therefore,serum PMAs have the potential to become a new biomarker for clinically diagnosing sepsis complicated by ARDS.展开更多
Introduction: Vascular endothelial growth factor-C (VEGF-C) is the primary lymphangiogenic factor that stimulates lymphangiogenesis by signaling via specific receptor, vascular endothelial growth factor receptor 3 (VE...Introduction: Vascular endothelial growth factor-C (VEGF-C) is the primary lymphangiogenic factor that stimulates lymphangiogenesis by signaling via specific receptor, vascular endothelial growth factor receptor 3 (VEGFR3). This study was conducted to evaluate the change in the level of VEGF-C before and after autologous bone marrow mononuclear cell transplantation for treatment of Lower limb lymphedema. Patient and methods: Forty patients with lower limb lymphedema were divided into two groups. Group I included 20 patients with chronic lower limb lymphedema who underwent autologous bone marrow mononuclear cell transplantation. Group II included 20 patients with chronic lower limb lymphedema who were exposed only to compression therapy as a control group. VEGF-C level in the diseased limbs was measured in both groups at the beginning of the study then 3 and 6 months respectively. Results: Group I included 20 patients, 8 patients were male (40%) and 12 patients were females (60%) with mean age 29.5 ± 12.15 while group II included 20, 10 patients were male (50%) and 10 patients were females (50%) with mean age 39.5 ± 11.5. In group I, the specimens were taken at 3 and 6 months after transplantation showed a marked decrease in the VEGF-C level with statistically significant p value, 0.02 and 0.001 respectively. In group II the level of VEGF-C after compression therapy alone at 3 and 6 months interval showed fluctuation with statistically non-significant p value, 0.64 and 0.55 respectively. Conclusion: VEGF-C is essential for regulation of lymphangiogenesis. The level of VEGF-C was found elevated in patients with lymphedema and decrease after autologous mononuclear bone marrow cells, however these results were statically non-significant.展开更多
Objective:To compare the effect of human chorionic gonadotropin(hCG)-producing peripheral blood mononuclear cells(PBMCs)and PBMCs activated by hCG in vitro and expressions of related immune genes in mouse implantation...Objective:To compare the effect of human chorionic gonadotropin(hCG)-producing peripheral blood mononuclear cells(PBMCs)and PBMCs activated by hCG in vitro and expressions of related immune genes in mouse implantation.Methods:hCG-producing PBMCs(transfected PBMC)and PBMCs activated by hCG in vitro were introduced into isolated mouse endometrial cells,while cell cultures were divided into four groups:the control,PBMC,transfected,and activated PBMC groups.The expression of studied genes(IL-1β,IL-6,Lif,and Vegf)was evaluated and blastocyst attachment on the cocultured cells(isolated endometrial cells and PBMC cells)was monitored in all four groups.Results:Data showed that expression decreased in the PBMC group compared to the treated PBMC(transfected and activated PBMCs)and increased in transfected PBMC compared to the activated PBMC.Attachment and migration of blastocysts were dramatically enhanced in the transfected PBMC group compared to the activated PBMC group(P<0.05).Conclusions:Use of hCG-producing PBMCs(transfected PBMC)has more influence on endometrial receptivity.展开更多
BACKGROUND: Interleukin-18 (IL-18), a pro-inflamma- tory cytokine that induces interferon-γ (IFN-γ) production in T cells and natural killer cells, plays a critical role in the T-lymphocyte helper type 1 ( Th1) resp...BACKGROUND: Interleukin-18 (IL-18), a pro-inflamma- tory cytokine that induces interferon-γ (IFN-γ) production in T cells and natural killer cells, plays a critical role in the T-lymphocyte helper type 1 ( Th1) response. This study was designed to explore the effect of IL-18 on peripheral blood mononuclear cells ( PBMCs) derived from chronic hepatitis B (CHB) and on hepatitis B virus (HBV) DNA released by HepG2.2.15 cell lines, which were transfected with hepatitis B virus gene in vitro. METHODS: PBMCs isolated from 25 healthy people and 25 patients with CHB were stimulated with HBcAg and IL-18 of various concentrations for 72 hours. The levels of IFN-γ in the supernatants of cultured PBMCs were determined by ELISA. After the stimulation of IL-18 of various concentra- tions, PBMCs derived from one patient were co-cultured for 96 hours with HepG2. 2. 15 cells which had been cul- tured for 24 hours, and then the supernatants were collected by centrifugation and used for HBV DNA quantitative as- say. RESULTS: When PBMCs were stimulated by HBcAg and IL-18 at various concentrations, the levels of IFN-γ in the supernatants of CHB groups were much higher than those in normal control groups, at 0.2 ng/ml: t =11.70, P< 0.01; at 1.0 ng/ml: t =16.19, P<0.01; and at5.0 ng/ml: t =20.12, P <0.01. In the CHB groups, the levels of IFN-γ in the supernatants of PBMCs stimulated by HBcAg alone were lower than both those stimulated by HBcAg and EL-18 at various concentrations and those stimulated by HBcAg and EL-18 (5.0 ng/ml) together with EL-12 (mild: t = 2.20, P<0.05; moderate; t=2.97, P<0.05; severe; t = 0.66, P >0.05). The content of HBV DNA in the superna- tant of co-cultivation of HepG2. 2. 15 cells and PBMCs without stimulated materials was higher than that stimula-ted by HBcAg and EL-18 at various concentrations of HBc- Ag and IL-18 together with IL-12/IFN-α1lb. CONCLUSION: DL-18 can induce IFN-γ secretion and pro- bably play a key role in the modulation of both innate and adaptive immunity. It has implications in improving im- munoregulatory effect and increasing the ability of immune cells to kill cells infected by virus.展开更多
The effect of Sinomenine on IL-8, IL-6, IL-2 and mIL-2R produced by peripheral blood mononuclear cells was investigated by using cell culture, radioimmunoassay and flow cytometry. It was showed that production of IL-8...The effect of Sinomenine on IL-8, IL-6, IL-2 and mIL-2R produced by peripheral blood mononuclear cells was investigated by using cell culture, radioimmunoassay and flow cytometry. It was showed that production of IL-8 and mIL-2R was inhibited, but the levels of IL-6 were enhanced by Sinomenine. Our results also demonstrated that Sinomenine did not have any effect on the production of IL-2. The study demonstrated that Sinomenine was able to regulate the production of cytokines. This may be one of the mechanisms by which Sinomenine works on rheumatoid arthritis.展开更多
BACKGROUND: The resolution of hepatitis C, evidenced by normalization of liver function and disappearance of hepatitis C virus RNA from serum as determined by conventional laboratory assays, reflects virus eradication...BACKGROUND: The resolution of hepatitis C, evidenced by normalization of liver function and disappearance of hepatitis C virus RNA from serum as determined by conventional laboratory assays, reflects virus eradication. But in interferon treated patients the HCV RNA in serum sometimes could not show the virus in cells. Such factors as virus genotype, HCV RNA contents in serum, HCV specific cellular immunities after treatment were reported to predict the response to interferon therapy. In most patients, HCV RNA could detect the virus in peripheral blood mononucle-ar cell. The aim of this study was to investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C after interferon treatment. METHODS: Sixteen patients with chronic hepatitis C were treated with interferon for 24 weeks, and they all get complete responses at 12 weeks of treatment. At the end of treatment, the HCV RNA in PBMC and serum were detected by RT-PCR, and after stopping treatment, HCV RNA in serum was monitored continually. RESULTS: In 9 patients who were HCV RNA positive in their PBMC at the end of treatment, 8 showed serum HCV RNA positive after 24 weeks and another 1 after 1 year. In 7 patients with negative HCV RNA in their PBMC, only 2 patients relapsed in serum HCV RNA after 1-year follow-up, and others remained viral response after 3.5 years. CONCLUSION: HCV RNA in PBMC at the end of IFN treatment is a predictor of durable response to antiviral therapy in patients with chronic hepatitis C.展开更多
BACKGROUND:The ischemic-type biliary lesion(ITBL)is one of the most serious biliary complications of liver transplantation. This study aimed to investigate the effects of autologous bone marrow mononuclear cell(BM-MNC...BACKGROUND:The ischemic-type biliary lesion(ITBL)is one of the most serious biliary complications of liver transplantation. This study aimed to investigate the effects of autologous bone marrow mononuclear cell(BM-MNC)implantation on neovascularization and the prevention of intrahepatic ITBL in a rabbit model. METHODS:The rabbits were divided into control,experimental model,and cell implantation groups,with 10 in each group. The model of intrahepatic ITBL was established by clamping the hepatic artery and common bile duct.Autologous BM-MNCs were isolated from the tibial plateau by density gradient centrifugation and were implanted through the common hepatic artery.Changes in such biochemical markers as aspartate aminotransferase,alanine aminotransferase,alkaline phosphatase,gamma-glutamyltranspeptidase,total bilirubin and direct bilirubin were measured.Four weeks after operation, cholangiography,histopathological manifestations,differentiation of BM-MNCs,microvessel density and the expression of vascular endothelial growth factor were assessed. RESULTS:Compared with the experimental model group, the BM-MNC implantation group showed superiority in the time to recover normal biochemistry.The microvessel density and vascular endothelial growth factor expression of the implantation group were significantly higher than those of the control and experimental model groups.The ITBL in the experimental model group was more severe than that in the implantation group and fewer new capillary blood vessels occurred around it.CONCLUSIONS:Implanted autologous BM-MNCs can differentiate into vascular endothelial cells,promote neovascu-larization and improve the blood supply to the ischemic bile duct,and this provides a new way to diminish or prevent intrahepatic ITBL after liver transplantation.展开更多
AIM:To determine the long-term efficacy of autologous bone marrow mononuclear cells(BM-MNCs)transplantation in terms of improving liver function and reducing complications in patients with decompensated cirrhosis.METH...AIM:To determine the long-term efficacy of autologous bone marrow mononuclear cells(BM-MNCs)transplantation in terms of improving liver function and reducing complications in patients with decompensated cirrhosis.METHODS:A total of 47 inpatients with decompensated liver cirrhosis were enrolled in this trial,including32 patients undergoing a single BM-MNCs transplantation plus routine medical treatment,and 15 patients receiving medical treatment only as controls.Fortythree of 47 patients were infected with hepatitis B virus.Bone marrow of 80-100 mL was obtained from each patient and the BM-MNCs suspension was transfused into the liver via the hepatic artery.The efficacy of BM-MNCs transplantation was monitored during a24-mo follow-up period.RESULTS:Liver function parameters in the two groups were observed at 1 mo after BM-MNCs transfusion.Prealbumin level was 118.3±25.3 mg/L vs 101.4±28.7 mg/L(P=0.047);albumin level was 33.5±3.6g/L vs 30.3±2.2 g/L(P=0.002);total bilirubin 36.9±9.7 mmol/L vs 45.6±19.9 mmol/L(P=0.048);prothrombin time 14.4±2.3 s vs 15.9±2.8 s(P=0.046);prothrombin activity 84.3%±14.3%vs 74.4%±17.8%(P=0.046);fibrinogen 2.28±0.53 g/L vs1.89±0.44 g/L(P=0.017);and platelet count 74.5±15.7×109/L vs 63.3±15.7×109/L(P=0.027)in the treatment group and control group,respectively.Differences were statistically significant.The efficacy of BM-MNCs transplantation lasted 3-12 mo as compared with the control group.Serious complications such as hepatic encephalopathy and spontaneous bacterial peritonitis were also significantly reduced in BM-MNCs transfused patients compared with the controls.However,these improvements disappeared 24 mo after transplantation.CONCLUSION:BM-MNCs transplantation is safe and effective in patients with decompensated cirrhosis.It also decreases the incidence of serious complications.展开更多
Objective: To determine the role of mononuclearmacrophages in the pathogenesis of acute lung injuryduring acute obstructive cholangitis.Methods: Sixty Wistar rats were used to study thecorrelation between the behavior...Objective: To determine the role of mononuclearmacrophages in the pathogenesis of acute lung injuryduring acute obstructive cholangitis.Methods: Sixty Wistar rats were used to study thecorrelation between the behavior of mononuclearmacrophages and acute pulmonary injury during a-cute obstructive cholangitis (AOC). Animal modelof AOC was made according to the method that thecommon bile duct was injected with Escherichia coliand ligated. The rats were killed at 6 h, 12 h, 24 hand 48 h after operation. The phagocytic function ofKupffer cells (KCs), the number of alveolar macro-phages (AMs) in bronchoalveolar lavage liquid, andthe extravascular water content of lung tissue weremeasured. The levels of lipid peroxide (LPO) andsupperoxide dismutase (SOD) were determined too.Pathological alterations of liver and lung tissue wereobserved under light and electron microscopes.Results: KCs phagocytic function was significantly el-evated at the 6th hour but markedly decreased fromthe 24th hour to the 48th hour in the AOC group ascompared with the control (P【0. 05). From the12th to the 48th hour, the number of AMs, the ex-travascular water content of lung tissue, and the con-tent of LPO significantly increased, but the SOD lev-el of lung tissue decreased greatly (P【0. 05). Mor-phologically, KCs proliferated diffusely in the earlyperiod in livers of the AOC group, but decreasedmarkedly in the late period. Mitochondria of KCswere swollen or even vacuolated; focal cytoplasmicdegeneration and many myeli like figures could beseen in the cytoplasm. The changes of injury such asdisturbance of pulmonary capillary blood circula-tion, degeneration and/or necrosis of the lung tissueand endothelium, and inflammatory reactions couldbe observed. In other two groups, no evident mor-phological changes were observed.Conclusions: KCs phagocytic function is decreased,whereas AM is activated by the invading bacteria torelease such inflammatory mediators as free radicals,resulting in acute pulmonary injury. It seems thatthere is a close relationship between the functionalstatus of mononuclear macrophages and the develop-ment of acute lung injury. The dysfunction of mono-nuclear macrophages may play an important role inthe pathogenesis of multiple organ damage, especial-ly acute pulmonary injury.展开更多
The expression of Toll-like receptor 4 (TLR4) in neonatal cord blood mononuclear cells (MNCs) and serum TNF-α were investigated in order to explore the roles of TLR4 in the pathogenesis of preeclampsia.The study enro...The expression of Toll-like receptor 4 (TLR4) in neonatal cord blood mononuclear cells (MNCs) and serum TNF-α were investigated in order to explore the roles of TLR4 in the pathogenesis of preeclampsia.The study enrolled 27 patients suffering from preeclampsia (experimental group) and 21 normal pregnancy patients (control group).After MNCs were separated, the expression of TLR4 mRNA and protein was detected by using real-time quantitative PCR and Western blotting respectively, and the expression of TNF-α by using ELISA.The results showed the TLR4 mRNA level in cord blood MNCs (2-CT:0.07±0.17), TLR4 protein expression level (absorbance ratio:0.81%±0.15%) and TNF-α level (9.5±1.73 pg/mL) were all increased in experimental group as compared with control group with the differences being statistically significant (P<0.05).There was a positive correlation between the expression of TLR4 mRNA and TNF-α in both experimental group and control group (r=0.54 and 0.53, respectively, P<0.05).It was concluded that TLR4 expression in the experimental group of cord blood MNCs was increased and there was a positive correlation between the expression of TLR4 mRNA and TNF-α in both groups.TLR4-mediated release of inflammatory cytokines may be one of the important reasons leading to preeclampsia.展开更多
During malaria infection,elevated levels of pro-inflammatory mediators and nitric;oxide production have been associated with pathogenesis and disease severity.Previous in vitro and in vivo studies have proposed that b...During malaria infection,elevated levels of pro-inflammatory mediators and nitric;oxide production have been associated with pathogenesis and disease severity.Previous in vitro and in vivo studies have proposed that both Plasmodium falciparum hemozoin and glycosylphosphatidylinositols are able to modulate blood mononuclear cells,contributing to stimulation of signal transduction and downstream regulation of the NF-KB signaling pathway,and subsequently leading to the production of pro-inflammatory cytokines,chemokines,and nitric oxide.The present review summarizes the published in vitro and in vivo studies that have investigated the mechanism of intracellular signal transduction and activation of the NF-KB signaling pathway in blood mononuclear cells after being inducted by Plasmodium falciparum malaria components.Particular attention is paid to hemozoin and glycosylphosphatidylinositols which reflect the important mechanism of signaling pathways involved in immune response.展开更多
AIM: To identify peripheral blood mononuclear cell (PBMC) gene expression profiles of ulcerative colitis (UC) patients, using oligonucleotide microarrays, to gain insights into UC molecular mechanisms. METHODS: The Hu...AIM: To identify peripheral blood mononuclear cell (PBMC) gene expression profiles of ulcerative colitis (UC) patients, using oligonucleotide microarrays, to gain insights into UC molecular mechanisms. METHODS: The Human OneArray microarrays were used for a complete genome-wide transcript profiling of PBMCs from 12 UC patients and 6 controls. Differential analysis per gene was performed with a random variance model; t test and P values were adjusted to control the false discovery rate (5%). Gene ontology (GO) was deployed to analyze differentially expressed genes at significant levels between patients and controls to identify the biological processes involved in UC. RESULTS: Comparative analysis revealed that 4438 probes (4188 genes) were differentially expressed between the two groups, of which 3689 probes (3590 genes) were down-regulated whereas 749 probes (598 genes) were up-regulated. Many disregulated genes in our data have been reported by previous microarray studies carried out on intestinal mucosa samples, such as S100A8 , CEACAM1 and S100A9 . GO enrichment analysis revealed 67 high enrichment up-regulated categories and one significant down-regulated category. The up-regulated genes were mainly involved in immune and inflammatory response, cell cycle and proliferation, DNA metabolism and repair. CONCLUSION: Gene expression profiling of PBMCs from patients with UC has highlighted several novel gene categories that could contribute to the pathogenesis of UC.展开更多
Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C(IL-17RC),a phenomenon observed in peripheral blood and chorioretinal tissues with age-relate...Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C(IL-17RC),a phenomenon observed in peripheral blood and chorioretinal tissues with age-related macular degeneration(AMD),was associated with altered activation of phosphatidylinositide 3-kinase(PI3K),Akt,and glycogen synthase kinase 3(GSK3).We wondered whether or not altered PI3 K,Akt,and GSK3 activities could be detected in peripheral blood mononuclear cells(PBMC) obtained from AMD patients.In the patients' PBMC,absent or reduced serine-phosphorylation of GSK3α or GSK3β was observed,which was accompanied with increased phosphorylation of GSK3 substrates(e.g.CCAAT enhancer binding protein a,insulin receptor substrate 1,and TAU),indicative of enhanced GSK3 activation.In addition,decreased protein mass of PI3K85α and tyrosinephosphorylation of PI3K50α was present in PBMC of the AMD patients,suggesting impaired PI3 K activation.Moreover,abnormally lowered molecular weight forms of Akt and GSK3 were detected in PBMC of the AMD patients.These data demonstrate that despite the presence of high levels of IL-17 RC,Wnt-3a and vascular endothelial growth factor,the PI3K/Akt/GSK3 signaling pathway is insensitive to these stimuli in PBMC of the AMD patients.Thus,measurement of PI3K/Akt/GSK3 expression and activity in PBMC may serve as a surrogate biomarker for AMD.展开更多
Objective:To evaluate retrospectively the effect of general anesthesia on DNA damage in the blood mononuclear cells(PBMCs) of surgical patients in order to provide evidence for a better nursing care during the procedu...Objective:To evaluate retrospectively the effect of general anesthesia on DNA damage in the blood mononuclear cells(PBMCs) of surgical patients in order to provide evidence for a better nursing care during the procedure.Methods:Clinical charts of 76 patients who underwent operation under general anesthesia and 76 healthy control subjects with documented results of DNA damage extent in PBMCs from the single-cell gel electrophoresis(SCGE) or cornel assay and scrum contents of superoxide dismutase(SOD) and malondialdehyde(MDA) from biochemical analyses were reviewed.The percentage of comet PBMCs and tail DNA and semm contents of SOD and MAD were analyzed by student t-test.Results:Compared with health) control subjects, generally anesthetized surgical patients had significantly higher%comet PBMCs and%tail DNA(P【0.05) and significantly lower serum concentrations of SOD(P【0.05) and significantly higher serum concentrations of MAD(P【0.05).Compared with levels before general anesthesia in surgical patients,%comet PBMCs,%tail DNA.and serum lexels of MAD were significantly higher(P【0.05 or 0.01).and semm levels of SOD were significantly lower(P【0.05).alter general anesthesia.Conclusions:General anesthesia during surgery causes a certain degree of hypoxia and PBMC damage.Particular attention should be paid to monitoring and maintenance of blood oxygen saturation in patients undergoing surgery under general anesthesia.展开更多
Two new compounds,1([Cu(IDB_)2]Cl_2·2CH_3CH_2OH·2H_2O(IDB = N,N-di(2-benzimidazolylmethyl)imine]) and 2([Cu(EDTB)]·2[C_6H_4(OH)COO]·6H_2O(EDTB = N,N,N',N'-tetrakis-[(2-benzimidazolyl)methyl...Two new compounds,1([Cu(IDB_)2]Cl_2·2CH_3CH_2OH·2H_2O(IDB = N,N-di(2-benzimidazolylmethyl)imine]) and 2([Cu(EDTB)]·2[C_6H_4(OH)COO]·6H_2O(EDTB = N,N,N',N'-tetrakis-[(2-benzimidazolyl)methyl]-1,2-ethanediamine]),have been synthesized and their crystal structures were determined by single-crystal X-ray diffraction. Crystal 1 belongs to the triclinic system,space group P1 with a = 9.565(2),b = 9.863(2),c = 10.252(3) ?,α = 81.915,β = 88.330,γ = 87.347°,V = 956.28(40) ?3,Z = 1,F(000) = 427,D_c = 1.419 g/cm3,Mr = 817.27 and μ(MoKα) = 0.764 mm^(-1). The final R indices(I > 2σ(I)): R = 0.0505,wR = 0.1417; R indices(all data): R = 0.0591,wR = 0.1525. Crystal 2 is of triclinic system,space group P1 with a = 11.487(3),b = 13.396(4),c = 17.977(5) ?,α = 73.899(5),β = 86.629(5),γ = 65.018(4)°,V = 2403.8(12) ?3,Z = 2,F(000) = 1072,D3c = 1.417 g/cm,Mr = 1025.54,μ(MoKα) = 0.528 mm^(-1). The final R indices(I > 2σ(I)): R = 0.0769,w R = 0.163 2; R indices(all data): R = 0.0769,wR = 0.1632. The two compounds exhibit mononuclear structures. The copper(Ⅱ) atom of crystal 1 is six-coordinated by four nitrogen atoms of benzimidazole groups and two amine nitrogen atoms from the ligand IDB to adopt a distorted octahedral coordination geometry. The copper(Ⅱ) atom of crystal 2 is six-coordinated by four nitrogen atoms of benzimidazole groups and two amine nitrogen atoms from the ligand EDTB to show a distorted octahedral coordination geometry. Particularly,the two compounds exhibit fluorescent properties at room temperature in ethanol.展开更多
The mononuclear phagocyte system(MPS)consists of monocytes,dendritic cells and macrophages,which play vital roles in innate immune defense against cancer.Hepatocellular carcinoma(HCC)is a complex disease that is affec...The mononuclear phagocyte system(MPS)consists of monocytes,dendritic cells and macrophages,which play vital roles in innate immune defense against cancer.Hepatocellular carcinoma(HCC)is a complex disease that is affected or initiated by many factors,including chronic hepatitis B virus infection,hepatitis C virus infection,metabolic disorders or alcohol consumption.Liver function,tumor stage and the performance status of patients affect HCC clinical outcomes.Studies have shown that targeted treatment of tumor microenvironment disorders may improve the efficacy of HCC treatments.Cytokines derived from the innate immune response can regulate T-cell differentiation,thereby shaping adaptive immunity,which is associated with the prognosis of HCC.Therefore,it is important to elucidate the function of the MPS in the progression of HCC.In this review,we outline the impact of HCC on the MPS.We illustrate how HCC reshapes MPS cell phenotype remodeling and the production of associated cytokines and characterize the function and impairment of the MPS in HCC.展开更多
AIM To investigate disease-specific gene expression profiles of peripheral blood mononuclear cells(PBMCs) from Crohn's disease(CD) patients in clinical remission.METHODS Patients with CD in clinical remission or w...AIM To investigate disease-specific gene expression profiles of peripheral blood mononuclear cells(PBMCs) from Crohn's disease(CD) patients in clinical remission.METHODS Patients with CD in clinical remission or with very low disease activity according to the Crohn's disease activity index were genotyped regarding nucleotidebinding oligomerization domain 2(NOD2),and PBMCs from wild-type(WT)-NOD2 patients,patients with homozygous or heterozygous NOD2 mutations and healthy donors were isolated for further analysis.The cells were cultured with vitamin D,peptidoglycan(PGN) and lipopolysaccharide(LPS) for defined periods of time before RNA was isolated and subjected to microarray analysis using Clariom S assays and quantitative realtime PCR.NOD2-and disease-specific gene expression profiles were evaluated with repeated measure ANOVA by a general linear model.RESULTS Employing microarray assays,a total of 267 genes were identified that were significantly up-or downregulated in PBMCs of WT-NOD2 patients,compared to healthy donors after challenge with vitamin D and/or a combination of LPS and PGN(P < 0.05;threshold:≥ 2-fold change).For further analysis by real-time PCR,genes with known impact on inflammation and immunity were selected that fulfilled predefined expression criteria.In a larger cohort of patients and controls,a disease-associated expression pattern,with higher transcript levels in vitamin D-treated PBMCs from patients,was observed for three of these genes,CLEC5 A(P < 0.030),lysozyme(LYZ;P < 0.047) and TREM1(P < 0.023).Six genes were found to be expressed in a NOD2-dependent manner(CD101,P < 0.002;CLEC5 A,P < 0.020;CXCL5,P < 0.009;IL-24,P < 0.044;ITGB2,P < 0.041;LYZ,P < 0.042).Interestingly,the highest transcript levels were observed in patients with heterozygous NOD2 mutations.CONCLUSION Our data identify CLEC5 A and LYZ as CD-and NOD2-associated genes of PBMCs and encourage further studies on their pathomechanistic roles.展开更多
BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human gra...BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1(2×108 L-1,5×108 L-1,1×109 L-1)], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=(1-A experimental well-A effector cell well/A target cell well)×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was (43.12±5.83)%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[(31.00 ±4.41)%,(30.92±5.27)%,(33.57±5.35)%,(26.23±5.20)%, t=3.51,2.98,4.24, P < 0.01); But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 (t=2.01,2.36, P < 0.05), and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1(t=0.06,P > 0.05). CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.展开更多
基金This study was supported by grants from the Key Project of the Chinese Ministry of Science and Technology(2017ZX102022022)National Key Research and Development Program of China(2021YFC2301801).
文摘Background: Glycine dehydrogenase(GLDC) plays an important role in the initiation and proliferation of several human cancers. In this study, we aimed to detect the methylation status of GLDC promoter and its diagnostic value for hepatitis B virus-associated hepatocellular carcinoma(HBV-HCC). Methods: We enrolled 197 patients, 111 with HBV-HCC, 51 with chronic hepatitis B(CHB), and 35 healthy controls(HCs). The methylation status of GLDC promoter in peripheral mononuclear cells(PBMCs) was identified by methylation specific polymerase chain reaction(MSP). The mRNA expression was examined using real-time quantitative polymerase chain reaction(q PCR). Results: The methylation frequency of the GLDC promoter was significantly lower in HBV-HCC patients(27.0%) compared to that in CHB patients(68.6%) and HCs(74.3%)( P < 0.001). The methylated group had lower alanine aminotransferase level( P = 0.035) and lower rates of tumor node metastasis(TNM) Ⅲ/Ⅳ( P = 0.043) and T3/T4( P = 0.026). TNM stage was identified to be an independent factor for GLDC promoter methylation. GLDC mRNA levels in CHB patients and HCs were significantly lower than those in HBV-HCC patients( P = 0.022 and P < 0.001, respectively). GLDC mRNA levels were significantly higher in HBV-HCC patients with unmethylated GLDC promoters than those with methylated GLDC promoters( P = 0.003). The diagnostic accuracy of alpha-fetoprotein(AFP) combined with GLDC promoter methylation for HBV-HCC was improved compared with that of AFP alone(AUC: 0.782 vs. 0.630, P < 0.001). In addition, GLDC promoter methylation was an independent predictor for overall survival of HBV-HCC patients( P = 0.038). Conclusions: The methylation frequency of GLDC promoter was lower in PBMCs from HBV-HCC patients than that from patients with CHB and HCs. The combination of AFP and GLDC promoter hypomethylation significantly improved the diagnostic accuracy of HBV-HCC.
文摘BACKGROUND The diagnosis of sepsis combined with acute respiratory distress syndrome(ARDS)has increased owing to the enhanced awareness among medical profes-sionals and the continuous development of modern medical technologies,while early diagnosis of ARDS still lacks specific biomarkers.One of the main patho-genic mechanisms of sepsis-associated ARDS involves the actions of various pathological injuries and inflammatory factors,such as platelet and white blood cells activation,leading to an increase of surface adhesion molecules.These adhesion molecules further form platelet-white blood cell aggregates,including platelet-mononuclear cell aggregates(PMAs).PMAs has been identified as one of the markers of platelet activation,here we hypothesize that PMAs might play a potential biomarker for the early diagnosis of this complication.METHODS We selected 72 hospitalized patients diagnosed with sepsis as the study population between March 2019 and March 2022.Among them,30 patients with sepsis and ARDS formed the study group,while 42 sepsis patients without ARDS comprised the control group.After diagnosis,venous blood samples were imme-diately collected from all patients.Flow cytometry was employed to analyze the expression of PMAs,platelet neutrophil aggregates(PNAs),and platelet aggregates(PLyAs)in the serum.Additionally,the Acute Physiology and Chronic Health Evaluation(APACHE)II score was calculated for each patient,and receiver operating characteristic curves were generated to assess diagnostic value.RESULTS The study found that the levels of PNAs and PLyAs in the serum of the study group were higher than those in the control group,but the difference was not statistically significant(P>0.05).However,the expression of PMAs in the serum of the study group was significantly upregulated(P<0.05)and positively correlated with the APACHE II score(r=0.671,P<0.05).When using PMAs as a diagnostic indicator,the area under the curve value was 0.957,indicating a high diagnostic value(P<0.05).Furthermore,the optimal cutoff value was 8.418%,with a diagnostic sensitivity of 0.819 and specificity of 0.947.CONCLUSION In summary,the serum levels of PMAs significantly increase in patients with sepsis and ARDS.Therefore,serum PMAs have the potential to become a new biomarker for clinically diagnosing sepsis complicated by ARDS.
文摘Introduction: Vascular endothelial growth factor-C (VEGF-C) is the primary lymphangiogenic factor that stimulates lymphangiogenesis by signaling via specific receptor, vascular endothelial growth factor receptor 3 (VEGFR3). This study was conducted to evaluate the change in the level of VEGF-C before and after autologous bone marrow mononuclear cell transplantation for treatment of Lower limb lymphedema. Patient and methods: Forty patients with lower limb lymphedema were divided into two groups. Group I included 20 patients with chronic lower limb lymphedema who underwent autologous bone marrow mononuclear cell transplantation. Group II included 20 patients with chronic lower limb lymphedema who were exposed only to compression therapy as a control group. VEGF-C level in the diseased limbs was measured in both groups at the beginning of the study then 3 and 6 months respectively. Results: Group I included 20 patients, 8 patients were male (40%) and 12 patients were females (60%) with mean age 29.5 ± 12.15 while group II included 20, 10 patients were male (50%) and 10 patients were females (50%) with mean age 39.5 ± 11.5. In group I, the specimens were taken at 3 and 6 months after transplantation showed a marked decrease in the VEGF-C level with statistically significant p value, 0.02 and 0.001 respectively. In group II the level of VEGF-C after compression therapy alone at 3 and 6 months interval showed fluctuation with statistically non-significant p value, 0.64 and 0.55 respectively. Conclusion: VEGF-C is essential for regulation of lymphangiogenesis. The level of VEGF-C was found elevated in patients with lymphedema and decrease after autologous mononuclear bone marrow cells, however these results were statically non-significant.
文摘Objective:To compare the effect of human chorionic gonadotropin(hCG)-producing peripheral blood mononuclear cells(PBMCs)and PBMCs activated by hCG in vitro and expressions of related immune genes in mouse implantation.Methods:hCG-producing PBMCs(transfected PBMC)and PBMCs activated by hCG in vitro were introduced into isolated mouse endometrial cells,while cell cultures were divided into four groups:the control,PBMC,transfected,and activated PBMC groups.The expression of studied genes(IL-1β,IL-6,Lif,and Vegf)was evaluated and blastocyst attachment on the cocultured cells(isolated endometrial cells and PBMC cells)was monitored in all four groups.Results:Data showed that expression decreased in the PBMC group compared to the treated PBMC(transfected and activated PBMCs)and increased in transfected PBMC compared to the activated PBMC.Attachment and migration of blastocysts were dramatically enhanced in the transfected PBMC group compared to the activated PBMC group(P<0.05).Conclusions:Use of hCG-producing PBMCs(transfected PBMC)has more influence on endometrial receptivity.
文摘BACKGROUND: Interleukin-18 (IL-18), a pro-inflamma- tory cytokine that induces interferon-γ (IFN-γ) production in T cells and natural killer cells, plays a critical role in the T-lymphocyte helper type 1 ( Th1) response. This study was designed to explore the effect of IL-18 on peripheral blood mononuclear cells ( PBMCs) derived from chronic hepatitis B (CHB) and on hepatitis B virus (HBV) DNA released by HepG2.2.15 cell lines, which were transfected with hepatitis B virus gene in vitro. METHODS: PBMCs isolated from 25 healthy people and 25 patients with CHB were stimulated with HBcAg and IL-18 of various concentrations for 72 hours. The levels of IFN-γ in the supernatants of cultured PBMCs were determined by ELISA. After the stimulation of IL-18 of various concentra- tions, PBMCs derived from one patient were co-cultured for 96 hours with HepG2. 2. 15 cells which had been cul- tured for 24 hours, and then the supernatants were collected by centrifugation and used for HBV DNA quantitative as- say. RESULTS: When PBMCs were stimulated by HBcAg and IL-18 at various concentrations, the levels of IFN-γ in the supernatants of CHB groups were much higher than those in normal control groups, at 0.2 ng/ml: t =11.70, P< 0.01; at 1.0 ng/ml: t =16.19, P<0.01; and at5.0 ng/ml: t =20.12, P <0.01. In the CHB groups, the levels of IFN-γ in the supernatants of PBMCs stimulated by HBcAg alone were lower than both those stimulated by HBcAg and EL-18 at various concentrations and those stimulated by HBcAg and EL-18 (5.0 ng/ml) together with EL-12 (mild: t = 2.20, P<0.05; moderate; t=2.97, P<0.05; severe; t = 0.66, P >0.05). The content of HBV DNA in the superna- tant of co-cultivation of HepG2. 2. 15 cells and PBMCs without stimulated materials was higher than that stimula-ted by HBcAg and EL-18 at various concentrations of HBc- Ag and IL-18 together with IL-12/IFN-α1lb. CONCLUSION: DL-18 can induce IFN-γ secretion and pro- bably play a key role in the modulation of both innate and adaptive immunity. It has implications in improving im- munoregulatory effect and increasing the ability of immune cells to kill cells infected by virus.
文摘The effect of Sinomenine on IL-8, IL-6, IL-2 and mIL-2R produced by peripheral blood mononuclear cells was investigated by using cell culture, radioimmunoassay and flow cytometry. It was showed that production of IL-8 and mIL-2R was inhibited, but the levels of IL-6 were enhanced by Sinomenine. Our results also demonstrated that Sinomenine did not have any effect on the production of IL-2. The study demonstrated that Sinomenine was able to regulate the production of cytokines. This may be one of the mechanisms by which Sinomenine works on rheumatoid arthritis.
文摘BACKGROUND: The resolution of hepatitis C, evidenced by normalization of liver function and disappearance of hepatitis C virus RNA from serum as determined by conventional laboratory assays, reflects virus eradication. But in interferon treated patients the HCV RNA in serum sometimes could not show the virus in cells. Such factors as virus genotype, HCV RNA contents in serum, HCV specific cellular immunities after treatment were reported to predict the response to interferon therapy. In most patients, HCV RNA could detect the virus in peripheral blood mononucle-ar cell. The aim of this study was to investigate the predictive value of HCV RNA in PBMC of patients with chronic hepatitis C after interferon treatment. METHODS: Sixteen patients with chronic hepatitis C were treated with interferon for 24 weeks, and they all get complete responses at 12 weeks of treatment. At the end of treatment, the HCV RNA in PBMC and serum were detected by RT-PCR, and after stopping treatment, HCV RNA in serum was monitored continually. RESULTS: In 9 patients who were HCV RNA positive in their PBMC at the end of treatment, 8 showed serum HCV RNA positive after 24 weeks and another 1 after 1 year. In 7 patients with negative HCV RNA in their PBMC, only 2 patients relapsed in serum HCV RNA after 1-year follow-up, and others remained viral response after 3.5 years. CONCLUSION: HCV RNA in PBMC at the end of IFN treatment is a predictor of durable response to antiviral therapy in patients with chronic hepatitis C.
文摘BACKGROUND:The ischemic-type biliary lesion(ITBL)is one of the most serious biliary complications of liver transplantation. This study aimed to investigate the effects of autologous bone marrow mononuclear cell(BM-MNC)implantation on neovascularization and the prevention of intrahepatic ITBL in a rabbit model. METHODS:The rabbits were divided into control,experimental model,and cell implantation groups,with 10 in each group. The model of intrahepatic ITBL was established by clamping the hepatic artery and common bile duct.Autologous BM-MNCs were isolated from the tibial plateau by density gradient centrifugation and were implanted through the common hepatic artery.Changes in such biochemical markers as aspartate aminotransferase,alanine aminotransferase,alkaline phosphatase,gamma-glutamyltranspeptidase,total bilirubin and direct bilirubin were measured.Four weeks after operation, cholangiography,histopathological manifestations,differentiation of BM-MNCs,microvessel density and the expression of vascular endothelial growth factor were assessed. RESULTS:Compared with the experimental model group, the BM-MNC implantation group showed superiority in the time to recover normal biochemistry.The microvessel density and vascular endothelial growth factor expression of the implantation group were significantly higher than those of the control and experimental model groups.The ITBL in the experimental model group was more severe than that in the implantation group and fewer new capillary blood vessels occurred around it.CONCLUSIONS:Implanted autologous BM-MNCs can differentiate into vascular endothelial cells,promote neovascu-larization and improve the blood supply to the ischemic bile duct,and this provides a new way to diminish or prevent intrahepatic ITBL after liver transplantation.
基金Supported by Grants for Key Bio-Medical Research Projects in Henan Province,China,No.2011020122,No.112102310251
文摘AIM:To determine the long-term efficacy of autologous bone marrow mononuclear cells(BM-MNCs)transplantation in terms of improving liver function and reducing complications in patients with decompensated cirrhosis.METHODS:A total of 47 inpatients with decompensated liver cirrhosis were enrolled in this trial,including32 patients undergoing a single BM-MNCs transplantation plus routine medical treatment,and 15 patients receiving medical treatment only as controls.Fortythree of 47 patients were infected with hepatitis B virus.Bone marrow of 80-100 mL was obtained from each patient and the BM-MNCs suspension was transfused into the liver via the hepatic artery.The efficacy of BM-MNCs transplantation was monitored during a24-mo follow-up period.RESULTS:Liver function parameters in the two groups were observed at 1 mo after BM-MNCs transfusion.Prealbumin level was 118.3±25.3 mg/L vs 101.4±28.7 mg/L(P=0.047);albumin level was 33.5±3.6g/L vs 30.3±2.2 g/L(P=0.002);total bilirubin 36.9±9.7 mmol/L vs 45.6±19.9 mmol/L(P=0.048);prothrombin time 14.4±2.3 s vs 15.9±2.8 s(P=0.046);prothrombin activity 84.3%±14.3%vs 74.4%±17.8%(P=0.046);fibrinogen 2.28±0.53 g/L vs1.89±0.44 g/L(P=0.017);and platelet count 74.5±15.7×109/L vs 63.3±15.7×109/L(P=0.027)in the treatment group and control group,respectively.Differences were statistically significant.The efficacy of BM-MNCs transplantation lasted 3-12 mo as compared with the control group.Serious complications such as hepatic encephalopathy and spontaneous bacterial peritonitis were also significantly reduced in BM-MNCs transfused patients compared with the controls.However,these improvements disappeared 24 mo after transplantation.CONCLUSION:BM-MNCs transplantation is safe and effective in patients with decompensated cirrhosis.It also decreases the incidence of serious complications.
基金This study was supported by the National Natural Science Foundation of China (No. 39970719, 30170919).
文摘Objective: To determine the role of mononuclearmacrophages in the pathogenesis of acute lung injuryduring acute obstructive cholangitis.Methods: Sixty Wistar rats were used to study thecorrelation between the behavior of mononuclearmacrophages and acute pulmonary injury during a-cute obstructive cholangitis (AOC). Animal modelof AOC was made according to the method that thecommon bile duct was injected with Escherichia coliand ligated. The rats were killed at 6 h, 12 h, 24 hand 48 h after operation. The phagocytic function ofKupffer cells (KCs), the number of alveolar macro-phages (AMs) in bronchoalveolar lavage liquid, andthe extravascular water content of lung tissue weremeasured. The levels of lipid peroxide (LPO) andsupperoxide dismutase (SOD) were determined too.Pathological alterations of liver and lung tissue wereobserved under light and electron microscopes.Results: KCs phagocytic function was significantly el-evated at the 6th hour but markedly decreased fromthe 24th hour to the 48th hour in the AOC group ascompared with the control (P【0. 05). From the12th to the 48th hour, the number of AMs, the ex-travascular water content of lung tissue, and the con-tent of LPO significantly increased, but the SOD lev-el of lung tissue decreased greatly (P【0. 05). Mor-phologically, KCs proliferated diffusely in the earlyperiod in livers of the AOC group, but decreasedmarkedly in the late period. Mitochondria of KCswere swollen or even vacuolated; focal cytoplasmicdegeneration and many myeli like figures could beseen in the cytoplasm. The changes of injury such asdisturbance of pulmonary capillary blood circula-tion, degeneration and/or necrosis of the lung tissueand endothelium, and inflammatory reactions couldbe observed. In other two groups, no evident mor-phological changes were observed.Conclusions: KCs phagocytic function is decreased,whereas AM is activated by the invading bacteria torelease such inflammatory mediators as free radicals,resulting in acute pulmonary injury. It seems thatthere is a close relationship between the functionalstatus of mononuclear macrophages and the develop-ment of acute lung injury. The dysfunction of mono-nuclear macrophages may play an important role inthe pathogenesis of multiple organ damage, especial-ly acute pulmonary injury.
文摘The expression of Toll-like receptor 4 (TLR4) in neonatal cord blood mononuclear cells (MNCs) and serum TNF-α were investigated in order to explore the roles of TLR4 in the pathogenesis of preeclampsia.The study enrolled 27 patients suffering from preeclampsia (experimental group) and 21 normal pregnancy patients (control group).After MNCs were separated, the expression of TLR4 mRNA and protein was detected by using real-time quantitative PCR and Western blotting respectively, and the expression of TNF-α by using ELISA.The results showed the TLR4 mRNA level in cord blood MNCs (2-CT:0.07±0.17), TLR4 protein expression level (absorbance ratio:0.81%±0.15%) and TNF-α level (9.5±1.73 pg/mL) were all increased in experimental group as compared with control group with the differences being statistically significant (P<0.05).There was a positive correlation between the expression of TLR4 mRNA and TNF-α in both experimental group and control group (r=0.54 and 0.53, respectively, P<0.05).It was concluded that TLR4 expression in the experimental group of cord blood MNCs was increased and there was a positive correlation between the expression of TLR4 mRNA and TNF-α in both groups.TLR4-mediated release of inflammatory cytokines may be one of the important reasons leading to preeclampsia.
基金Supported by the Institute of Research and Development,Walailak University,Nakhon Si Thammarat,Thailand
文摘During malaria infection,elevated levels of pro-inflammatory mediators and nitric;oxide production have been associated with pathogenesis and disease severity.Previous in vitro and in vivo studies have proposed that both Plasmodium falciparum hemozoin and glycosylphosphatidylinositols are able to modulate blood mononuclear cells,contributing to stimulation of signal transduction and downstream regulation of the NF-KB signaling pathway,and subsequently leading to the production of pro-inflammatory cytokines,chemokines,and nitric oxide.The present review summarizes the published in vitro and in vivo studies that have investigated the mechanism of intracellular signal transduction and activation of the NF-KB signaling pathway in blood mononuclear cells after being inducted by Plasmodium falciparum malaria components.Particular attention is paid to hemozoin and glycosylphosphatidylinositols which reflect the important mechanism of signaling pathways involved in immune response.
基金Supported by Grants from National Natural Science Foundation of China, No. 81260074/H0310 and 81160055/H0310Confederative Special Foundation of Science and Technology, Department of Yunnan Province and Kunming Medical College, No.2011FB183 and 2007C0010RMedical Academic Leader of Yunnan Provincial Bureau of Health, No. D-201215
文摘AIM: To identify peripheral blood mononuclear cell (PBMC) gene expression profiles of ulcerative colitis (UC) patients, using oligonucleotide microarrays, to gain insights into UC molecular mechanisms. METHODS: The Human OneArray microarrays were used for a complete genome-wide transcript profiling of PBMCs from 12 UC patients and 6 controls. Differential analysis per gene was performed with a random variance model; t test and P values were adjusted to control the false discovery rate (5%). Gene ontology (GO) was deployed to analyze differentially expressed genes at significant levels between patients and controls to identify the biological processes involved in UC. RESULTS: Comparative analysis revealed that 4438 probes (4188 genes) were differentially expressed between the two groups, of which 3689 probes (3590 genes) were down-regulated whereas 749 probes (598 genes) were up-regulated. Many disregulated genes in our data have been reported by previous microarray studies carried out on intestinal mucosa samples, such as S100A8 , CEACAM1 and S100A9 . GO enrichment analysis revealed 67 high enrichment up-regulated categories and one significant down-regulated category. The up-regulated genes were mainly involved in immune and inflammatory response, cell cycle and proliferation, DNA metabolism and repair. CONCLUSION: Gene expression profiling of PBMCs from patients with UC has highlighted several novel gene categories that could contribute to the pathogenesis of UC.
基金supported by intramural research funding of National Center for Complementary and Alternative Medicine(now is National Center for Complementary and Integrative Health),NIH,the US Department of Health and Human Services(to X.L.)and an operating grant(MOP 123279)from Canadian Institutes for Health Research(to Z.Y.)
文摘Our recent studies with cultured retinal pigment epithelium cells suggested that overexpression of interleukin 17 receptor C(IL-17RC),a phenomenon observed in peripheral blood and chorioretinal tissues with age-related macular degeneration(AMD),was associated with altered activation of phosphatidylinositide 3-kinase(PI3K),Akt,and glycogen synthase kinase 3(GSK3).We wondered whether or not altered PI3 K,Akt,and GSK3 activities could be detected in peripheral blood mononuclear cells(PBMC) obtained from AMD patients.In the patients' PBMC,absent or reduced serine-phosphorylation of GSK3α or GSK3β was observed,which was accompanied with increased phosphorylation of GSK3 substrates(e.g.CCAAT enhancer binding protein a,insulin receptor substrate 1,and TAU),indicative of enhanced GSK3 activation.In addition,decreased protein mass of PI3K85α and tyrosinephosphorylation of PI3K50α was present in PBMC of the AMD patients,suggesting impaired PI3 K activation.Moreover,abnormally lowered molecular weight forms of Akt and GSK3 were detected in PBMC of the AMD patients.These data demonstrate that despite the presence of high levels of IL-17 RC,Wnt-3a and vascular endothelial growth factor,the PI3K/Akt/GSK3 signaling pathway is insensitive to these stimuli in PBMC of the AMD patients.Thus,measurement of PI3K/Akt/GSK3 expression and activity in PBMC may serve as a surrogate biomarker for AMD.
文摘Objective:To evaluate retrospectively the effect of general anesthesia on DNA damage in the blood mononuclear cells(PBMCs) of surgical patients in order to provide evidence for a better nursing care during the procedure.Methods:Clinical charts of 76 patients who underwent operation under general anesthesia and 76 healthy control subjects with documented results of DNA damage extent in PBMCs from the single-cell gel electrophoresis(SCGE) or cornel assay and scrum contents of superoxide dismutase(SOD) and malondialdehyde(MDA) from biochemical analyses were reviewed.The percentage of comet PBMCs and tail DNA and semm contents of SOD and MAD were analyzed by student t-test.Results:Compared with health) control subjects, generally anesthetized surgical patients had significantly higher%comet PBMCs and%tail DNA(P【0.05) and significantly lower serum concentrations of SOD(P【0.05) and significantly higher serum concentrations of MAD(P【0.05).Compared with levels before general anesthesia in surgical patients,%comet PBMCs,%tail DNA.and serum lexels of MAD were significantly higher(P【0.05 or 0.01).and semm levels of SOD were significantly lower(P【0.05).alter general anesthesia.Conclusions:General anesthesia during surgery causes a certain degree of hypoxia and PBMC damage.Particular attention should be paid to monitoring and maintenance of blood oxygen saturation in patients undergoing surgery under general anesthesia.
基金supported by Research Fund for the Doctor Program of Hubei University of Technology(No.BSQD12139)Ministry of Science and Technology Small and Medium-sized Enterprise Venture Funds(No.12C26214204453)+1 种基金Natural Foundation of Hubei Province(No.2010CDA020)the Key Foundation of Industry-University-Research Cooperation of Education Department,Hubei Province(No.CXY2009A010)
文摘Two new compounds,1([Cu(IDB_)2]Cl_2·2CH_3CH_2OH·2H_2O(IDB = N,N-di(2-benzimidazolylmethyl)imine]) and 2([Cu(EDTB)]·2[C_6H_4(OH)COO]·6H_2O(EDTB = N,N,N',N'-tetrakis-[(2-benzimidazolyl)methyl]-1,2-ethanediamine]),have been synthesized and their crystal structures were determined by single-crystal X-ray diffraction. Crystal 1 belongs to the triclinic system,space group P1 with a = 9.565(2),b = 9.863(2),c = 10.252(3) ?,α = 81.915,β = 88.330,γ = 87.347°,V = 956.28(40) ?3,Z = 1,F(000) = 427,D_c = 1.419 g/cm3,Mr = 817.27 and μ(MoKα) = 0.764 mm^(-1). The final R indices(I > 2σ(I)): R = 0.0505,wR = 0.1417; R indices(all data): R = 0.0591,wR = 0.1525. Crystal 2 is of triclinic system,space group P1 with a = 11.487(3),b = 13.396(4),c = 17.977(5) ?,α = 73.899(5),β = 86.629(5),γ = 65.018(4)°,V = 2403.8(12) ?3,Z = 2,F(000) = 1072,D3c = 1.417 g/cm,Mr = 1025.54,μ(MoKα) = 0.528 mm^(-1). The final R indices(I > 2σ(I)): R = 0.0769,w R = 0.163 2; R indices(all data): R = 0.0769,wR = 0.1632. The two compounds exhibit mononuclear structures. The copper(Ⅱ) atom of crystal 1 is six-coordinated by four nitrogen atoms of benzimidazole groups and two amine nitrogen atoms from the ligand IDB to adopt a distorted octahedral coordination geometry. The copper(Ⅱ) atom of crystal 2 is six-coordinated by four nitrogen atoms of benzimidazole groups and two amine nitrogen atoms from the ligand EDTB to show a distorted octahedral coordination geometry. Particularly,the two compounds exhibit fluorescent properties at room temperature in ethanol.
基金National Natural Science Foundation of China,No 81970529The Natural Science Foundation of Jilin Province,No.20200201387JC.
文摘The mononuclear phagocyte system(MPS)consists of monocytes,dendritic cells and macrophages,which play vital roles in innate immune defense against cancer.Hepatocellular carcinoma(HCC)is a complex disease that is affected or initiated by many factors,including chronic hepatitis B virus infection,hepatitis C virus infection,metabolic disorders or alcohol consumption.Liver function,tumor stage and the performance status of patients affect HCC clinical outcomes.Studies have shown that targeted treatment of tumor microenvironment disorders may improve the efficacy of HCC treatments.Cytokines derived from the innate immune response can regulate T-cell differentiation,thereby shaping adaptive immunity,which is associated with the prognosis of HCC.Therefore,it is important to elucidate the function of the MPS in the progression of HCC.In this review,we outline the impact of HCC on the MPS.We illustrate how HCC reshapes MPS cell phenotype remodeling and the production of associated cytokines and characterize the function and impairment of the MPS in HCC.
基金Supported by a grant from the Damp-Foundation(2016-04) to Schaffler H and Rohde S
文摘AIM To investigate disease-specific gene expression profiles of peripheral blood mononuclear cells(PBMCs) from Crohn's disease(CD) patients in clinical remission.METHODS Patients with CD in clinical remission or with very low disease activity according to the Crohn's disease activity index were genotyped regarding nucleotidebinding oligomerization domain 2(NOD2),and PBMCs from wild-type(WT)-NOD2 patients,patients with homozygous or heterozygous NOD2 mutations and healthy donors were isolated for further analysis.The cells were cultured with vitamin D,peptidoglycan(PGN) and lipopolysaccharide(LPS) for defined periods of time before RNA was isolated and subjected to microarray analysis using Clariom S assays and quantitative realtime PCR.NOD2-and disease-specific gene expression profiles were evaluated with repeated measure ANOVA by a general linear model.RESULTS Employing microarray assays,a total of 267 genes were identified that were significantly up-or downregulated in PBMCs of WT-NOD2 patients,compared to healthy donors after challenge with vitamin D and/or a combination of LPS and PGN(P < 0.05;threshold:≥ 2-fold change).For further analysis by real-time PCR,genes with known impact on inflammation and immunity were selected that fulfilled predefined expression criteria.In a larger cohort of patients and controls,a disease-associated expression pattern,with higher transcript levels in vitamin D-treated PBMCs from patients,was observed for three of these genes,CLEC5 A(P < 0.030),lysozyme(LYZ;P < 0.047) and TREM1(P < 0.023).Six genes were found to be expressed in a NOD2-dependent manner(CD101,P < 0.002;CLEC5 A,P < 0.020;CXCL5,P < 0.009;IL-24,P < 0.044;ITGB2,P < 0.041;LYZ,P < 0.042).Interestingly,the highest transcript levels were observed in patients with heterozygous NOD2 mutations.CONCLUSION Our data identify CLEC5 A and LYZ as CD-and NOD2-associated genes of PBMCs and encourage further studies on their pathomechanistic roles.
文摘BACKGROUND: Dendritic cell is the most major antigen presenting cell of organism. It is proved in recent studies that human umbilical cord blood mononuclear cells induced and cultured in vitro by recombinant human granulocyte-macrophage colony stimulating factor (rhG-MCSF) and recombinant human interleukin-4(rhIL-4) can generate a great many dendritic cells and promote the lethal effect of T cells on human neuroblastoma, but it is unclear that whether the lethal effect is associated with the most proper concentration of dendritic cells. OBJECTIVE: To investigate the lethal effect of human umbilical cord blood mononuclear cells induced in vitro by cytokines differentiating into dendritic cells on human neuroblastoma, and its best concentration range. DESIGN: Open experiment. SETTING: Department of Pediatrics, the Medical School Hospital of Qingdao University. MATERIALS: The study was carried out in the Shandong Provincial Key Laboratory (Laboratory for the Department of Pediatrics of the Medical School Hospital of Qingdao University) during September 2005 to May 2006. Human umbilical cord blood samples were taken from the healthy newborn infants of full-term normal delivery during October to November 2005 in the Medical School Hospital of Qingdao University, and were voluntarily donated by the puerperas. Main instruments: type 3111 CO2 incubator (Forma Scientific, USA), type 550 ELISA Reader (Bio-Rad, USA). Main reagents: neuroblastoma cell line SK-N-SH (Shanghai Institute of Life Science, Chinese Academy of Sciences), RPMI-1640 culture fluid and fetal bovine serum (Hyclone), rhIL-4 (Promega, USA), rhG-MCSF (Harbin Pharmaceutic Group Bioengineering Co.Ltd), rat anti-human CD1a monoclonal antibody and FITC-labeled rabbit anti-rat IgG (Xiehe Stem cell Gene Engineering Co.Ltd). METHODS: ① Human umbilical cord blood mononuclear cells obtained with attachment methods differentiated into human umbilical cord blood dendritic cells, presenting typical morphology of dendritic cells after in vitro induction by rhG-MCSF and rhIL-4. ② Different concentrations of dendritic cells[ dendritic cells: neuroblastoma cells=20∶1,50∶1,100∶1(2×108 L-1,5×108 L-1,1×109 L-1)], 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the experimental group. 1×109 L-1 T cells and 1×107 L-1 neuroblastoma cells were added in the control group. ③ Main surface marker CD1a molecules of dendritic cells were detected with indirect immunofluorescence, and the percent rate of dendritic cells was counted with ultraviolet light and expressed as the expression rate of CD1a+ cells. ④ Single effector cells and target cells were respectively set in the experimental group and control group to obtain the lethal effect. The lethal effect of dendritic cells on neuroblastoma cells was indirectly evaluated by detecting cellular survival with MTT assay. The lethal effect(%)=(1-A experimental well-A effector cell well/A target cell well)×100%.⑤The experimental data were presented as Mean ±SD, and paired t test was used. MAIN OUTCOME MEASURES: ① Morphological characters of dendritic cells in the process of induction and differentiation. ②CD1a+ cellular expression rate. ③Lethal effect of dendritic cells on neuroblastoma cells. RESULTS: ①Morphological characters of dendritic cells in the process of induction and differentiation: On the 15th day after human umbilical cord blood mononuclear cells were induced by rhG-MCSF and rhIL-4, typical morphology of dendritic cells could be seen under an inverted microscope. ②Expression rate of CD1a+ cells was (43.12±5.83)%. ③Lethal effect of dendritic cells on neuroblastoma cells: Lethal effect of dendritic cells stimulated T cells in each experimental group ( dendritic cells: neuroblastoma cells=100∶1,50∶1,20∶1 respectively) on neuroblastoma cells was significantly higher than that in control group[(31.00 ±4.41)%,(30.92±5.27)%,(33.57±5.35)%,(26.23±5.20)%, t=3.51,2.98,4.24, P < 0.01); But the lethal effect of dendritic cells on neuroblastoma was significantly lower when their ratio was 100∶1 and 50∶1 in comparison with 20:1 (t=2.01,2.36, P < 0.05), and no significant difference in lethal effect existed between the ratio at 100∶1 and 50∶1(t=0.06,P > 0.05). CONCLUSION: Dendritic cells differentiated from human umbilical cord blood mononuclear cells after in vitro induction of cytokines can promote the lethal effect of T cells on neuroblastoma cells. The lethal effect is associated with the concentration of dendritic cells within some range.
