Objective To study the gene expression of metallothionein 1 (MT-1) isoforms in human peripheral blood lymphocytes (HPBLs). Methods The expression of mRNA representing the seven active MT-1 genes was determined in HPBL...Objective To study the gene expression of metallothionein 1 (MT-1) isoforms in human peripheral blood lymphocytes (HPBLs). Methods The expression of mRNA representing the seven active MT-1 genes was determined in HPBLs by quantitative RT-PCR before and after exposure to cadmium. Results Basal expressions of MT-1X, and MT-1A in HPBLs were similar to expression of housekeeping gene. In contrast, the basal gene expressions of MT-1H, 1F, 1E, and 1G were a little transcripts in human HPBLs. No signal was detected for MT-1B. There was a sex difference (P<0.05). in basal gene expression of MT-1E. The levels of gene expression of MT-1A, 1E, 1F, 1G, 1H, and 1X increased, but the level of MT-1B did not increase after exposure to cadmium. Conclusions Gene expressions of MT-1G, MT-1H, MT-1F, and MT-1X in HPBLs can be used as a potential biomarker of cadmium exposure.展开更多
OBJECTIVE:Ligusticum porteri is a traditional Native American herb.The roots of L.porteri are traditionally used in the treatment of many diseases,however,its cytotoxicity,antioxidative and immunemodulatory effects ne...OBJECTIVE:Ligusticum porteri is a traditional Native American herb.The roots of L.porteri are traditionally used in the treatment of many diseases,however,its cytotoxicity,antioxidative and immunemodulatory effects need to be investigated.In this study,we evaluated the effects of the root extract at different doses on human peripheral blood lymphocytes(PBLs).METHODS:The lymphocytes were incubated with different concentrations of the root extracts(0,50,100,200,and 400 ug/mL) and harvested every 6 h for 2 d(P<0.05).The protective effect of the herb against oxidative damage was determined by inducing oxidative stress with the administration of50 pmol/L of hydrogen peroxide(H_2O_2).RESULTS:Treatments with L.porteri at 200 and 400 pg/mL increased the viability of PBLs.The deleterious effect of H_2O_2 was ameliorated by 400 ug/mL L.porteri treatment.Addition of 400 pg/mL L.porteri reduced lipid peroxidation in stressed PBLs by 94%(P<0.05).Treatment with 400 μg/mL of L.porteri resulted in a 26.4%increase of reduced glutathione levels.Activities of superoxide dismutase and catalase increased by 17.5%and 55.2%respectively,when stressed PBLs were treated with 400 μg/mL L.porteri for 2 d(P<0.05).Treatment with 400 pg/mL L.porteri increased interferon-γ and interleukin-2expressions in H_2O_2-challenged PBLs(P<0.05),however,the root extract did not cause a significant difference in interleukin-10 levels compared to the control(P>0.05).CONCLUSION:The findings suggest that L.porteri might be a potential immune-modulating agent involving protective effects against oxidative damage.展开更多
2,4-dinitrophenol(DNP),an organic compound which frequently used in industry,is considered to have high toxicity.This study aimed to investigate the early changes of lymphocyte subpopulations in patients with occupati...2,4-dinitrophenol(DNP),an organic compound which frequently used in industry,is considered to have high toxicity.This study aimed to investigate the early changes of lymphocyte subpopulations in patients with occupational 2,4-DNP poisoning.Totally 9 patients with acute occupational 2,4-DNP poisoning and 30 healthy volunteers as展开更多
Chronic obstructive pulmonary disease (COPD) is a chronic, progressive respiratory disease and the third leading cause of respiratory disease mortality. The diagnosis of COPD is changed to acute exacerbation of COPD (...Chronic obstructive pulmonary disease (COPD) is a chronic, progressive respiratory disease and the third leading cause of respiratory disease mortality. The diagnosis of COPD is changed to acute exacerbation of COPD (AECOPD) when respiratory symptoms become worse, beyond normal day-to-day variations and severely enough that changes in medication are required. Both neutrophils to lymphocyte ratio (NLR) and peripheral blood eosinophilia (PBE) are rapid and relatively inexpensive tests that can be easily applied in the clinical practice for the diagnosis and treatment of AECOPD patients. Furthermore, current studies found that NLR and PBE had a higher accuracy rate than other traditional markers (Leukocyte count and C-reactive protein) for the diagnosis and management of AECOPD. Besides, recent studies determined that NLR and PBE can be used for prediction of future exacerbations in COPD patients. This review aims to explore the current knowledge about the significance of NLR and PBE in AECOPD patients.展开更多
DNA damage response (DDR) in different cell cycle status of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin (PH...DNA damage response (DDR) in different cell cycle status of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA). The apoptotic ratio and the phosphorylation H2AX (S139) were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray. The expressions of γH2AX, Bcl-2, caspase-3 and caspase-9 were detected by Western blotting. DDR in 293T cells was detected after H2AX was silenced by RNAi method. Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment. The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P<0.05). The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased. No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls. It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates. γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.展开更多
Human-induced neural stem cells(iNSCs)transplantation is a potential treatment of neurodegeneration diseases.However,whether the reprogrammed cells have the same characterizations as human fetal neural stem cells need...Human-induced neural stem cells(iNSCs)transplantation is a potential treatment of neurodegeneration diseases.However,whether the reprogrammed cells have the same characterizations as human fetal neural stem cells needs further exploration.Here we isolated human fetal neural stem cells from aborted 12-week fetal brains and compared with iNSCs reprogrammed from human peripheral blood mononuclear cells in gene expression,proliferation ability,differentiation capacity,and the responses to tumor necrosis factor-α.We found that iNSCs and NSCs both expressed neural stem cell markers Nestin,SOX1,and SOX2.However,only iNSCs can be patterned into dopaminergic neurons and motor neurons.Furthermore,both iNSCs and NSCs can differentiate into oligodendrocyte progenitor cells.In addition,a low dose of tumor necrosis factor-αdid not inhibit the proliferation and differentiation of iNSCs and NSCs.In conclusion,iNSCs have properties similar to,and even better than,fetal neural stem cells and may be suitable for disease modeling and transplantation.展开更多
Day 100 prognostic factors post-autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT) to predict clinical outcomes in diffuse large B-cell lymphoma (DLBCL) patients have not been studied. Thus,...Day 100 prognostic factors post-autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT) to predict clinical outcomes in diffuse large B-cell lymphoma (DLBCL) patients have not been studied. Thus, we retrospectively examined if day 100 absolute monocyte/lymphocyte prognostic score (AMLPS-100) affects clinical outcomes by landmark analysis from day 100 post-APBHSCT in DLBCL. Only DLBCL patients in complete remission at day 100 post-APBHSCT were evaluated. From 2000 to 2007, 134 consecutive DLBCL patients are qualified for the study. Patients with a day 100 absolute monocyte count (AMC-100) ≥ 630 cells/μL and day 100 absolute lymphocyte count (ALC-100) ≤ 1000 cells/μL experienced inferior overall survival (OS) and progression free survival (PFS). On multivariate analysis, the AMC-100 and ALC-100 remained independent predictors of OS and PFS. Combining both values into the AMLPS-100, the 5-year OS rates for low, intermediate, and high AMLPS-100 risk groups were 94% (95% CI, 83.0% - 98.1%), 70% (95% CI, 58.6% - 80.1%), and 13% (95% CI, 3.4% - 40.5%), respectively;and the 5-year PFS rates were 87% (95% CI, 74.0% - 94.1%), 68% (95% CI, 56.0% - 77.8%), and 13% (95% CI, 3.4% - 40.5%), respectively. The AMLPS-100 is a simple biomarker score that can stratify clinical outcomes from day 100 post-APBHSCT in DLBCL patients.展开更多
In order to provide pertinent evidence of ion channel with immune response in the fish,whole cell patch-clamp technique was employed for potassium ion channel study in turbot(Scophthalmus maximus) . Lymphocytes were i...In order to provide pertinent evidence of ion channel with immune response in the fish,whole cell patch-clamp technique was employed for potassium ion channel study in turbot(Scophthalmus maximus) . Lymphocytes were isolated by Percoll density gradient centrifugation from peripheral blood samples,and electrophysiological characters of the channel were analyzed. In the recorded cells,activated voltage of the channels was-42.5 ± 3.7 mV and the average peak current was 313.12 ± 28.2 pA. The channel was identified as voltage dependent,the current was outward and it could be inhibited by 10 mmol/dm 3 TEA or 5 mmol/dm 3 4-AP,a specific potassium channel inhibitor,identifying the existence of potassium channel in peripheral lymphocytes of the turbot.展开更多
Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into th...Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.展开更多
Objective: To study expression of membrane receptors of interleukin-2 (CD25) on the peripheral blood lymphocytes (PBL) of patients with colorectal cancer and its clinical significance. Methods: CD25 percentages (CD25%...Objective: To study expression of membrane receptors of interleukin-2 (CD25) on the peripheral blood lymphocytes (PBL) of patients with colorectal cancer and its clinical significance. Methods: CD25 percentages (CD25%) in PBL of 105 colorectal cancer patients before operation and 100 normal individuals were examined by flow cytometer, and the results were clinically and pathologically analyzed. Results: The mean of CD25% in PBL of the normal individuals was 17.24(5.33, it was significantly lower (p<0.01) than that of the colon cancer patients (21.29(7.95) or rectal cancer patients (21.62(6.11). In contrast to the normal individuals, the means of CD25% in PBL in ulcer type (20.53(6.50) or protruded type (21.56(6.16) colorectal cancer patients were notably elevated (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was observed between the normal individuals and patients with less than 4 cm mass (22.10(5.43) or 4cm(8cm mass (20.90(6.96). The significant difference (P<0.05) of means of CD25% in PBL was also observed between the normal individuals and patients with greater than 8 cm mass (21.56(5.41). The mean of CD25% in PBL in patients with well differentiation colorectal cancer was 22.20(5.50, it was significantly higher than that in normal individuals (P<0.05). The means of CD25% in PBL in patients with middle or poor differentiation colorectal cancer were 21.30(6.89 and 22.15(5.71 respectively, they were obviously higher than that in normal individuals (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was present between the colorectal cancer patients without metastatic lymph nodes (22.06(6.90) and normal individuals. The significant difference (P<0.05) of means of CD25% in PBL was present between the colorectal cancer patients with metastatic lymph nodes (20.73(6.40) and normal individuals. The means of CD25% in PBL in colorectal cancer patients in various clinic stages were significantly higher than that in the health subjects (p<0.01). The significant difference (p<0.01) of means of CD25% in PBL was present between the patients whose ages were equal to or less than 60 (21.00(5.76) and normal individuals in the same age group. The significant difference (p<0.05) of means of CD25% in PBL was also present between the patients whose ages were greater than 60 (22.54(7.75) and normal individuals in the same age group. The significant difference (P<0.01) of means of CD25% in PBL was present between the male patients (22.55(7.05) and normal men. The significant difference (p<0.05) of means of CD25% in PBL was also present between the female patients (20.09(5.48) and normal women. Conclusion: The mean of CD25% in PBL of colorectal cancer patients was significantly higher than that in health subjects. Abnormally elevated CD25% were correlative with site of tumor growth, macropathology type of tumor, the degree of tumor differentiation, clinical stage and patient’s age and sex. It may be helpful to detect CD25% in PBL of colorectal cancer patients before operation for diagnosis, immune treatment and judging prognosis.展开更多
In this study,sterlet( Acipenser ruthenus) was chosen as the model species of sturgeon,different solutions were used to isolate the sturgeon peripheral blood lymphocytes and study their optimal proliferate response co...In this study,sterlet( Acipenser ruthenus) was chosen as the model species of sturgeon,different solutions were used to isolate the sturgeon peripheral blood lymphocytes and study their optimal proliferate response condition. Phytohemagglutinin( PHA),concanavalin A( Con A) and lipopolysaccharide( LPS) were used as lymphocyte proliferation mitogen,respectively. According to L25( 56) five-factor five-level orthogonal experimental design,the conditions for sturgeons proliferation response of peripheral blood lymphocytes were optimized using enhanced cell counting Kit-8( enhanced CCK-8 or WST-8). Five factors were selected to explore the optimal response conditions,including culture time,culture temperature,cell concentration,fetal bovine serum( FBS) concentration,and mitogen concentration. The results showed that 70% Percoll( 1. 092 g/ml) used as the sturgeon lymphocyte separation solution had the best separating effect. The optimal proliferation conditions were as follows: 3. 625 × 10~6 initial cells,20 μg/ml PHA or 50 μg/ml Con A or 10 μg/ml LPS as mitogen,10%-20% FBS,the temperature at 20-25 ℃,and the culture time of 2 d.展开更多
Alterations of mitochondria DNA(mtDNA)4977 bp common deletion(CD)and mtDNA copy number induced by ionizing radiation were observed in human different cell lines and total body irradiation patients.However,only few exp...Alterations of mitochondria DNA(mtDNA)4977 bp common deletion(CD)and mtDNA copy number induced by ionizing radiation were observed in human different cell lines and total body irradiation patients.However,only few experiments have evaluated the levels of the CD and mtDNA copy number in human peripheral blood exposed to ionizing radiation till now.The aim of this study is to analyze the mtDNA alterations in irradiated human peripheral blood from healthy donors as well as to explore their feasibility as biomarkers for constructing new biodosimeter.Peripheral blood samples were collected from six healthy donors,and exposed to 60Co gamma ray with the doses of 0 Gy,1 Gy,2 Gy,3 Gy,4 Gy and 5 Gy.Levels of the CD and mtDNA copy number in irradiated samples after 2h or 24h incubation were detected using TaqMan real-time PCR,and the CD ratio was calculated.The results showed that the mean of the CD ratio and the CD copy number exhibited a dose-dependent increase 2 h in the dose range from 0–5 Gy,and of the mtDNA copy number significantly increased 24 h in irradiated groups compared with 0 Gy group after irradiation.It indicates that the parameters in human peripheral blood may be considered as molecular biomarkers to applying construction of new biodosimeter.展开更多
Background:Cancer-targeted T-cell receptor T(TCR-T)cells hold promise in treating cancers such as hematological malignancies and breast cancers.However,approaches to obtain cancer-reactive TCR-T cells have been unsucc...Background:Cancer-targeted T-cell receptor T(TCR-T)cells hold promise in treating cancers such as hematological malignancies and breast cancers.However,approaches to obtain cancer-reactive TCR-T cells have been unsuccessful.Methods:Here,we developed a novel strategy to screen for cancer-targeted TCR-T cells using a special humanized mouse model with person-specific immune fingerprints.Rare steady-state circulating hematopoietic stem and progenitor cells were expanded via three-dimensional culture of steady-state peripheral blood mononuclear cells,and then the expanded cells were applied to establish humanized mice.The human immune system was evaluated according to the kinetics of dendritic cells,monocytes,T-cell subsets,and cytokines.To fully stimulate the immune response and to obtain B-cell precursor NAML-6-and triple-negative breast cancer MDA-MB-231-targeted TCR-T cells,we used the inactivated cells above to treat humanized mice twice a day every 7 days.Then,human T cells were processed for TCRβ-chain(TRB)sequencing analysis.After the repertoires had been constructed,features such as the fraction,diversity,and immune signature were investigated.Results:The results demonstrated an increase in diversity and clonality of T cells after treatment.The preferential usage and features of TRBV,TRBJ,and the V–J combination were also changed.The stress also induced highly clonal Science and Technology,Grant/Award Number:2021C03010;Zhejiang Provincial Natural Science Foundation of China,Grant/Award Numbers:LTGY24H080003,LY21H080004 expansion.Tumor burden and survival analysis demonstrated that stress induction could significantly inhibit the growth of subsequently transfused live tumor cells and prolong the survival of the humanized mice.Conclusions:We constructed a personalized humanized mouse model to screen cancer-targeted TCR-T pools.Our platform provides an effective source of cancer-targeted TCR-T cells and allows for the design of patient-specific engineered T cells.It therefore has the potential to greatly benefit cancer treatment.展开更多
Objective:Chimeric antigen receptor-modified T(CAR-T)cells have shown impressive results against relapsed/refractory B cell malignancies.However,the traditional manufacture of CAR-T cells requires leukapheresis to iso...Objective:Chimeric antigen receptor-modified T(CAR-T)cells have shown impressive results against relapsed/refractory B cell malignancies.However,the traditional manufacture of CAR-T cells requires leukapheresis to isolate large amounts of peripheral blood T cells,thus making some patients ineligible for the procedure.Methods:We developed a simple method for CAR-T cell preparation requiring small volumes of peripheral blood.First,CD3+T cells isolated from 50 mL peripheral blood from patients(B-cell malignancies)were stimulated with immobilized anti-CD3/RetroNectin in 6-well plates and then transduced with CAR-expressing lentiviral vector.After 4 d,the T cells were transferred to culture bags for large-scale CAR-T cell expansion.In vitro and animal experiments were performed to evaluate the activity of the manufactured CAR-T cells.Finally,29 patients with B-cell acute lymphoblastic leukemia(B-ALL)and 9 patients with B-cell lymphoma were treated with the CAR-T cells.Results:The CAR-T cells were expanded to 1–3×10^(8) cells in 8–10 d and successfully killed B cell-derived malignant tumor cells in vitro and in vivo.For patients with B-ALL,the complete remission rate was 93%1 month after CAR-T cell infusion;after 12 months,the overall survival(OS)and leukemia-free survival rates were 69%and 31%,respectively.For patients with lymphoma,the objective response rate(including complete and partial remission)was 78%2 months after CAR-T cell infusion,and after 12 months,the OS and progression-free survival rates were 71%and 43%,respectively.Cytokine-release syndrome(CRS)occurred in 65.51%and 55.56%of patients with B-ALL and B-cell lymphoma,respectively;severe CRS developed in 20.69%of patients with B-ALL and in no patients with lymphoma.Conclusions:Our novel method can generate sufficient numbers of CAR-T cells for clinical use from 50–100 mL peripheral blood,thus providing an alternative means of CAR-T cell generation for patients ineligible for leukapheresis.展开更多
Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-d...Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells in guiding peripheral nerve regeneration remain poorly understood. This study was designed to investigate the effects of human umbilical cord blood-derived mesenchymal stem cells on neural regeneration using a rat sciatic nerve crush injury model. Human umbilical cord blood-derived mesenchymal stem cells (1 × 10 6 ) or a PBS control were injected into the crush-injured segment of the sciatic nerve. Four weeks after cell injection, brain-derived neurotrophic factor and tyrosine kinase receptor B mRNA expression at the lesion site was increased in comparison to control. Furthermore, sciatic function index, Fluoro Gold-labeled neuron counts and axon density were also significantly increased when compared with control. Our results indicate that human umbilical cord blood-derived mesenchymal stem cells promote the functional recovery of crush-injured sciatic nerves.展开更多
Opportunistic bacteremia in adult HIV-infected patients is a normal co-infectious condition caused by Gram-negative bacilli. Respiratory infections, including cough, shortness of breath, and chest pain and skin infect...Opportunistic bacteremia in adult HIV-infected patients is a normal co-infectious condition caused by Gram-negative bacilli. Respiratory infections, including cough, shortness of breath, and chest pain and skin infection with eruptions, pustules and itchiness, are common complaints in the setting of late HIV infection. The variety of infections ranges from mild, self-limited viral, bacteremia and fungal infections to severe, life-threatening demanding urgent care and hospitalization. Varicella pneumonia, for instance, is the most severe complication of chickenpox in HIV infected adults, potentially refractory, fulminant respiratory failure can ensue. Patients with impaired immune status and chronic lung disease are at an increased risk. In the United States as well as in Vietnam, bacterial/viral pneumonia and skin infection are the two most common HIV-associated conditions. While globally the incidence of opportunistic infection has decreased since the introduction of highly active antiretroviral therapy during the last 3 decades, HIV-associated diseases remain a significant source of mortality, thus any manifestation must be taken seriously. This study will present the most common HIV-related pulmonary and skin infections and provide an overview of the epidemiology, characteristic clinical and chest radiograph findings, diagnosis, treatment, and prevention globally as well in Vietnam. Though the extensive efforts of the Vietnamese Government during last decade contributed to a valuable decrease, yet epidemic in Vietnam still remains high, ranking Vietnam 5th in the South-East region. The second part of the study focuses on a unique and severe HIV case report of a 35-year-old man, with a rare form of pneumonia caused by Acitenobacter spp. concomitant with a prolonged and disseminating skin infection. The case has been treated with a combination of conventional anti-retroviral medication and autologous peripheral blood stem cells, the results showed that within 5 months there was an impressive amelioration of HIV viral activity together with a total recovery from pneumonia and skin infection.展开更多
基金This work was supported by the National 973 Program of China 2002 CB 512905
文摘Objective To study the gene expression of metallothionein 1 (MT-1) isoforms in human peripheral blood lymphocytes (HPBLs). Methods The expression of mRNA representing the seven active MT-1 genes was determined in HPBLs by quantitative RT-PCR before and after exposure to cadmium. Results Basal expressions of MT-1X, and MT-1A in HPBLs were similar to expression of housekeeping gene. In contrast, the basal gene expressions of MT-1H, 1F, 1E, and 1G were a little transcripts in human HPBLs. No signal was detected for MT-1B. There was a sex difference (P<0.05). in basal gene expression of MT-1E. The levels of gene expression of MT-1A, 1E, 1F, 1G, 1H, and 1X increased, but the level of MT-1B did not increase after exposure to cadmium. Conclusions Gene expressions of MT-1G, MT-1H, MT-1F, and MT-1X in HPBLs can be used as a potential biomarker of cadmium exposure.
基金supported by a grant from Texas A&M University-Corpus Christi
文摘OBJECTIVE:Ligusticum porteri is a traditional Native American herb.The roots of L.porteri are traditionally used in the treatment of many diseases,however,its cytotoxicity,antioxidative and immunemodulatory effects need to be investigated.In this study,we evaluated the effects of the root extract at different doses on human peripheral blood lymphocytes(PBLs).METHODS:The lymphocytes were incubated with different concentrations of the root extracts(0,50,100,200,and 400 ug/mL) and harvested every 6 h for 2 d(P<0.05).The protective effect of the herb against oxidative damage was determined by inducing oxidative stress with the administration of50 pmol/L of hydrogen peroxide(H_2O_2).RESULTS:Treatments with L.porteri at 200 and 400 pg/mL increased the viability of PBLs.The deleterious effect of H_2O_2 was ameliorated by 400 ug/mL L.porteri treatment.Addition of 400 pg/mL L.porteri reduced lipid peroxidation in stressed PBLs by 94%(P<0.05).Treatment with 400 μg/mL of L.porteri resulted in a 26.4%increase of reduced glutathione levels.Activities of superoxide dismutase and catalase increased by 17.5%and 55.2%respectively,when stressed PBLs were treated with 400 μg/mL L.porteri for 2 d(P<0.05).Treatment with 400 pg/mL L.porteri increased interferon-γ and interleukin-2expressions in H_2O_2-challenged PBLs(P<0.05),however,the root extract did not cause a significant difference in interleukin-10 levels compared to the control(P>0.05).CONCLUSION:The findings suggest that L.porteri might be a potential immune-modulating agent involving protective effects against oxidative damage.
基金supported by the grants from the Foundation of Science and Technology Department of Zhejiang Province for Beneficial Technology Research of Social Development(2011C23013)
文摘2,4-dinitrophenol(DNP),an organic compound which frequently used in industry,is considered to have high toxicity.This study aimed to investigate the early changes of lymphocyte subpopulations in patients with occupational 2,4-DNP poisoning.Totally 9 patients with acute occupational 2,4-DNP poisoning and 30 healthy volunteers as
文摘Chronic obstructive pulmonary disease (COPD) is a chronic, progressive respiratory disease and the third leading cause of respiratory disease mortality. The diagnosis of COPD is changed to acute exacerbation of COPD (AECOPD) when respiratory symptoms become worse, beyond normal day-to-day variations and severely enough that changes in medication are required. Both neutrophils to lymphocyte ratio (NLR) and peripheral blood eosinophilia (PBE) are rapid and relatively inexpensive tests that can be easily applied in the clinical practice for the diagnosis and treatment of AECOPD patients. Furthermore, current studies found that NLR and PBE had a higher accuracy rate than other traditional markers (Leukocyte count and C-reactive protein) for the diagnosis and management of AECOPD. Besides, recent studies determined that NLR and PBE can be used for prediction of future exacerbations in COPD patients. This review aims to explore the current knowledge about the significance of NLR and PBE in AECOPD patients.
基金supported by grants from Program 973 from Ministry of ScienceTechnology of China (Nos. 2004CB518705, 2009CB5218702)the National Natural Sciences Foundation of China (Nos. 30872472, 30800569)
文摘DNA damage response (DDR) in different cell cycle status of human peripheral blood lymphocytes (PBLs) and the role of H2AX in DDR were investigated. The PBLs were stimulated into cell cycle with phytohemagglutinin (PHA). The apoptotic ratio and the phosphorylation H2AX (S139) were flow cytometrically measured in resting and proliferating PBLs after treatment with camptothecin (CPT) or X-ray. The expressions of γH2AX, Bcl-2, caspase-3 and caspase-9 were detected by Western blotting. DDR in 293T cells was detected after H2AX was silenced by RNAi method. Our results showed that DNA double strand breaks (DSBs) were both induced in quiescent and proliferating PBLs after CPT or X-ray treatment. The phosphorylation of H2AX and apoptosis were more sensitive in proliferating PBLs compared with quiescent lymphocytes (P<0.05). The expression levels of anti-apoptotic proteins Bcl-2 were reduced and cleaved caspase-3 and caspase-9 were increased. No significant changes were observed in CPT-induced apoptosis in 293T cells between H2AX knocking down group and controls. It was concluded that proliferating PBLs were more vulnerable to DNA damage compared to non-stimulated lymphocytes and had higher apoptosis rates. γH2AX may only serve as a marker of DNA damage but exert no effect on apoptosis regulation.
文摘Human-induced neural stem cells(iNSCs)transplantation is a potential treatment of neurodegeneration diseases.However,whether the reprogrammed cells have the same characterizations as human fetal neural stem cells needs further exploration.Here we isolated human fetal neural stem cells from aborted 12-week fetal brains and compared with iNSCs reprogrammed from human peripheral blood mononuclear cells in gene expression,proliferation ability,differentiation capacity,and the responses to tumor necrosis factor-α.We found that iNSCs and NSCs both expressed neural stem cell markers Nestin,SOX1,and SOX2.However,only iNSCs can be patterned into dopaminergic neurons and motor neurons.Furthermore,both iNSCs and NSCs can differentiate into oligodendrocyte progenitor cells.In addition,a low dose of tumor necrosis factor-αdid not inhibit the proliferation and differentiation of iNSCs and NSCs.In conclusion,iNSCs have properties similar to,and even better than,fetal neural stem cells and may be suitable for disease modeling and transplantation.
文摘Day 100 prognostic factors post-autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT) to predict clinical outcomes in diffuse large B-cell lymphoma (DLBCL) patients have not been studied. Thus, we retrospectively examined if day 100 absolute monocyte/lymphocyte prognostic score (AMLPS-100) affects clinical outcomes by landmark analysis from day 100 post-APBHSCT in DLBCL. Only DLBCL patients in complete remission at day 100 post-APBHSCT were evaluated. From 2000 to 2007, 134 consecutive DLBCL patients are qualified for the study. Patients with a day 100 absolute monocyte count (AMC-100) ≥ 630 cells/μL and day 100 absolute lymphocyte count (ALC-100) ≤ 1000 cells/μL experienced inferior overall survival (OS) and progression free survival (PFS). On multivariate analysis, the AMC-100 and ALC-100 remained independent predictors of OS and PFS. Combining both values into the AMLPS-100, the 5-year OS rates for low, intermediate, and high AMLPS-100 risk groups were 94% (95% CI, 83.0% - 98.1%), 70% (95% CI, 58.6% - 80.1%), and 13% (95% CI, 3.4% - 40.5%), respectively;and the 5-year PFS rates were 87% (95% CI, 74.0% - 94.1%), 68% (95% CI, 56.0% - 77.8%), and 13% (95% CI, 3.4% - 40.5%), respectively. The AMLPS-100 is a simple biomarker score that can stratify clinical outcomes from day 100 post-APBHSCT in DLBCL patients.
基金The National Natural Science Foundation of China under contract Nos 40476060 and 40876089the State Key Basic Research Program under contract No. 2006CB101805
文摘In order to provide pertinent evidence of ion channel with immune response in the fish,whole cell patch-clamp technique was employed for potassium ion channel study in turbot(Scophthalmus maximus) . Lymphocytes were isolated by Percoll density gradient centrifugation from peripheral blood samples,and electrophysiological characters of the channel were analyzed. In the recorded cells,activated voltage of the channels was-42.5 ± 3.7 mV and the average peak current was 313.12 ± 28.2 pA. The channel was identified as voltage dependent,the current was outward and it could be inhibited by 10 mmol/dm 3 TEA or 5 mmol/dm 3 4-AP,a specific potassium channel inhibitor,identifying the existence of potassium channel in peripheral lymphocytes of the turbot.
文摘Stem cells are commonly classified based on the developmental stage from which they are isolated, although this has been a source of debate amongst stem cell scientists. A common approach classifies stem cells into three different groupings: Embryonic Stem Cells (ESCs), Umbilical Cord Stem Cells (UCBSCs) and Adult Stem Cells (ASCs), which include stem cells from bone marrow (BM), fat tissue (FT), engineered induced pluripotent (IP) and peripheral blood (PB). By definition stem cells are progenitor cells capable of self-renewal and differentiation hypothetically “ab infinitum” into more specialized cells and tissue. The main intent of this study was to determine and characterize the different sub-groups of stem cells present within the human PB-SCs that may represent a valid opportunity in the field of clinical regenerative medicine. Stem cells in the isolated mononucleated cells were characterized using a multidisciplinary approach that was based on morphology, the expression of stem cell markers by flowcytometry and fluorescence analysis, RT-PCR and the capacity to self-renew or proliferate and differentiate into specialized cells. This approach was used to identify the expression of hematopoietic, mesenchymal, embryonic and neural stem cell markers. Both isolated adherent and suspension mononucleated cells were able to maintain their stem cell properties during in-vitro culture by holding their capacity for proliferation and differentiation into osteoblast cells, respectively, when exposed to the appropriate induction medium.
文摘Objective: To study expression of membrane receptors of interleukin-2 (CD25) on the peripheral blood lymphocytes (PBL) of patients with colorectal cancer and its clinical significance. Methods: CD25 percentages (CD25%) in PBL of 105 colorectal cancer patients before operation and 100 normal individuals were examined by flow cytometer, and the results were clinically and pathologically analyzed. Results: The mean of CD25% in PBL of the normal individuals was 17.24(5.33, it was significantly lower (p<0.01) than that of the colon cancer patients (21.29(7.95) or rectal cancer patients (21.62(6.11). In contrast to the normal individuals, the means of CD25% in PBL in ulcer type (20.53(6.50) or protruded type (21.56(6.16) colorectal cancer patients were notably elevated (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was observed between the normal individuals and patients with less than 4 cm mass (22.10(5.43) or 4cm(8cm mass (20.90(6.96). The significant difference (P<0.05) of means of CD25% in PBL was also observed between the normal individuals and patients with greater than 8 cm mass (21.56(5.41). The mean of CD25% in PBL in patients with well differentiation colorectal cancer was 22.20(5.50, it was significantly higher than that in normal individuals (P<0.05). The means of CD25% in PBL in patients with middle or poor differentiation colorectal cancer were 21.30(6.89 and 22.15(5.71 respectively, they were obviously higher than that in normal individuals (P<0.01). The significant difference (P<0.01) of means of CD25% in PBL was present between the colorectal cancer patients without metastatic lymph nodes (22.06(6.90) and normal individuals. The significant difference (P<0.05) of means of CD25% in PBL was present between the colorectal cancer patients with metastatic lymph nodes (20.73(6.40) and normal individuals. The means of CD25% in PBL in colorectal cancer patients in various clinic stages were significantly higher than that in the health subjects (p<0.01). The significant difference (p<0.01) of means of CD25% in PBL was present between the patients whose ages were equal to or less than 60 (21.00(5.76) and normal individuals in the same age group. The significant difference (p<0.05) of means of CD25% in PBL was also present between the patients whose ages were greater than 60 (22.54(7.75) and normal individuals in the same age group. The significant difference (P<0.01) of means of CD25% in PBL was present between the male patients (22.55(7.05) and normal men. The significant difference (p<0.05) of means of CD25% in PBL was also present between the female patients (20.09(5.48) and normal women. Conclusion: The mean of CD25% in PBL of colorectal cancer patients was significantly higher than that in health subjects. Abnormally elevated CD25% were correlative with site of tumor growth, macropathology type of tumor, the degree of tumor differentiation, clinical stage and patient’s age and sex. It may be helpful to detect CD25% in PBL of colorectal cancer patients before operation for diagnosis, immune treatment and judging prognosis.
基金Supported by Foundation of Beijing Municipal Science and Technology Project(Z161100004516003)Innovation Team of Sturgeon and Salmonid of Beijing(BAIC08-2018)Innovation Team of Sturgeon and Salmonid of Baafs(JNKST201611)
文摘In this study,sterlet( Acipenser ruthenus) was chosen as the model species of sturgeon,different solutions were used to isolate the sturgeon peripheral blood lymphocytes and study their optimal proliferate response condition. Phytohemagglutinin( PHA),concanavalin A( Con A) and lipopolysaccharide( LPS) were used as lymphocyte proliferation mitogen,respectively. According to L25( 56) five-factor five-level orthogonal experimental design,the conditions for sturgeons proliferation response of peripheral blood lymphocytes were optimized using enhanced cell counting Kit-8( enhanced CCK-8 or WST-8). Five factors were selected to explore the optimal response conditions,including culture time,culture temperature,cell concentration,fetal bovine serum( FBS) concentration,and mitogen concentration. The results showed that 70% Percoll( 1. 092 g/ml) used as the sturgeon lymphocyte separation solution had the best separating effect. The optimal proliferation conditions were as follows: 3. 625 × 10~6 initial cells,20 μg/ml PHA or 50 μg/ml Con A or 10 μg/ml LPS as mitogen,10%-20% FBS,the temperature at 20-25 ℃,and the culture time of 2 d.
基金Supported by grants from the building project for the National Key Clinical Special Department of China(No.2011-17)the Medical Science and Technology Foundation of Henan Province(No.201204123)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘Alterations of mitochondria DNA(mtDNA)4977 bp common deletion(CD)and mtDNA copy number induced by ionizing radiation were observed in human different cell lines and total body irradiation patients.However,only few experiments have evaluated the levels of the CD and mtDNA copy number in human peripheral blood exposed to ionizing radiation till now.The aim of this study is to analyze the mtDNA alterations in irradiated human peripheral blood from healthy donors as well as to explore their feasibility as biomarkers for constructing new biodosimeter.Peripheral blood samples were collected from six healthy donors,and exposed to 60Co gamma ray with the doses of 0 Gy,1 Gy,2 Gy,3 Gy,4 Gy and 5 Gy.Levels of the CD and mtDNA copy number in irradiated samples after 2h or 24h incubation were detected using TaqMan real-time PCR,and the CD ratio was calculated.The results showed that the mean of the CD ratio and the CD copy number exhibited a dose-dependent increase 2 h in the dose range from 0–5 Gy,and of the mtDNA copy number significantly increased 24 h in irradiated groups compared with 0 Gy group after irradiation.It indicates that the parameters in human peripheral blood may be considered as molecular biomarkers to applying construction of new biodosimeter.
基金National Natural Science Foundation of China,Grant/Award Numbers:82130003,81970158,82000180Zhejiang Provincial Key R&D Projects of Department of Science and Technology,Grant/Award Number:2021C03010Zhejiang Provincial Natural Science Foundation of China,Grant/Award Numbers:LTGY24H080003,LY21H080004。
文摘Background:Cancer-targeted T-cell receptor T(TCR-T)cells hold promise in treating cancers such as hematological malignancies and breast cancers.However,approaches to obtain cancer-reactive TCR-T cells have been unsuccessful.Methods:Here,we developed a novel strategy to screen for cancer-targeted TCR-T cells using a special humanized mouse model with person-specific immune fingerprints.Rare steady-state circulating hematopoietic stem and progenitor cells were expanded via three-dimensional culture of steady-state peripheral blood mononuclear cells,and then the expanded cells were applied to establish humanized mice.The human immune system was evaluated according to the kinetics of dendritic cells,monocytes,T-cell subsets,and cytokines.To fully stimulate the immune response and to obtain B-cell precursor NAML-6-and triple-negative breast cancer MDA-MB-231-targeted TCR-T cells,we used the inactivated cells above to treat humanized mice twice a day every 7 days.Then,human T cells were processed for TCRβ-chain(TRB)sequencing analysis.After the repertoires had been constructed,features such as the fraction,diversity,and immune signature were investigated.Results:The results demonstrated an increase in diversity and clonality of T cells after treatment.The preferential usage and features of TRBV,TRBJ,and the V–J combination were also changed.The stress also induced highly clonal Science and Technology,Grant/Award Number:2021C03010;Zhejiang Provincial Natural Science Foundation of China,Grant/Award Numbers:LTGY24H080003,LY21H080004 expansion.Tumor burden and survival analysis demonstrated that stress induction could significantly inhibit the growth of subsequently transfused live tumor cells and prolong the survival of the humanized mice.Conclusions:We constructed a personalized humanized mouse model to screen cancer-targeted TCR-T pools.Our platform provides an effective source of cancer-targeted TCR-T cells and allows for the design of patient-specific engineered T cells.It therefore has the potential to greatly benefit cancer treatment.
基金This work was supported by grants from Henan Medical Science and Technique Foundation(Grant Nos.LHGJ2020173 and SBGJ20180850)the Natural Science Foundation of Henan(Grant No.182300410344).
文摘Objective:Chimeric antigen receptor-modified T(CAR-T)cells have shown impressive results against relapsed/refractory B cell malignancies.However,the traditional manufacture of CAR-T cells requires leukapheresis to isolate large amounts of peripheral blood T cells,thus making some patients ineligible for the procedure.Methods:We developed a simple method for CAR-T cell preparation requiring small volumes of peripheral blood.First,CD3+T cells isolated from 50 mL peripheral blood from patients(B-cell malignancies)were stimulated with immobilized anti-CD3/RetroNectin in 6-well plates and then transduced with CAR-expressing lentiviral vector.After 4 d,the T cells were transferred to culture bags for large-scale CAR-T cell expansion.In vitro and animal experiments were performed to evaluate the activity of the manufactured CAR-T cells.Finally,29 patients with B-cell acute lymphoblastic leukemia(B-ALL)and 9 patients with B-cell lymphoma were treated with the CAR-T cells.Results:The CAR-T cells were expanded to 1–3×10^(8) cells in 8–10 d and successfully killed B cell-derived malignant tumor cells in vitro and in vivo.For patients with B-ALL,the complete remission rate was 93%1 month after CAR-T cell infusion;after 12 months,the overall survival(OS)and leukemia-free survival rates were 69%and 31%,respectively.For patients with lymphoma,the objective response rate(including complete and partial remission)was 78%2 months after CAR-T cell infusion,and after 12 months,the OS and progression-free survival rates were 71%and 43%,respectively.Cytokine-release syndrome(CRS)occurred in 65.51%and 55.56%of patients with B-ALL and B-cell lymphoma,respectively;severe CRS developed in 20.69%of patients with B-ALL and in no patients with lymphoma.Conclusions:Our novel method can generate sufficient numbers of CAR-T cells for clinical use from 50–100 mL peripheral blood,thus providing an alternative means of CAR-T cell generation for patients ineligible for leukapheresis.
基金supported by a grant of the Seoul National University Dental Hospital,Republic of Korea,No.03-2010-0020
文摘Several studies have demonstrated that human umbilical cord blood-derived mesenchymal stem cells can promote neural regeneration following brain injury. However, the therapeutic effects of human umbilical cord blood-derived mesenchymal stem cells in guiding peripheral nerve regeneration remain poorly understood. This study was designed to investigate the effects of human umbilical cord blood-derived mesenchymal stem cells on neural regeneration using a rat sciatic nerve crush injury model. Human umbilical cord blood-derived mesenchymal stem cells (1 × 10 6 ) or a PBS control were injected into the crush-injured segment of the sciatic nerve. Four weeks after cell injection, brain-derived neurotrophic factor and tyrosine kinase receptor B mRNA expression at the lesion site was increased in comparison to control. Furthermore, sciatic function index, Fluoro Gold-labeled neuron counts and axon density were also significantly increased when compared with control. Our results indicate that human umbilical cord blood-derived mesenchymal stem cells promote the functional recovery of crush-injured sciatic nerves.
文摘Opportunistic bacteremia in adult HIV-infected patients is a normal co-infectious condition caused by Gram-negative bacilli. Respiratory infections, including cough, shortness of breath, and chest pain and skin infection with eruptions, pustules and itchiness, are common complaints in the setting of late HIV infection. The variety of infections ranges from mild, self-limited viral, bacteremia and fungal infections to severe, life-threatening demanding urgent care and hospitalization. Varicella pneumonia, for instance, is the most severe complication of chickenpox in HIV infected adults, potentially refractory, fulminant respiratory failure can ensue. Patients with impaired immune status and chronic lung disease are at an increased risk. In the United States as well as in Vietnam, bacterial/viral pneumonia and skin infection are the two most common HIV-associated conditions. While globally the incidence of opportunistic infection has decreased since the introduction of highly active antiretroviral therapy during the last 3 decades, HIV-associated diseases remain a significant source of mortality, thus any manifestation must be taken seriously. This study will present the most common HIV-related pulmonary and skin infections and provide an overview of the epidemiology, characteristic clinical and chest radiograph findings, diagnosis, treatment, and prevention globally as well in Vietnam. Though the extensive efforts of the Vietnamese Government during last decade contributed to a valuable decrease, yet epidemic in Vietnam still remains high, ranking Vietnam 5th in the South-East region. The second part of the study focuses on a unique and severe HIV case report of a 35-year-old man, with a rare form of pneumonia caused by Acitenobacter spp. concomitant with a prolonged and disseminating skin infection. The case has been treated with a combination of conventional anti-retroviral medication and autologous peripheral blood stem cells, the results showed that within 5 months there was an impressive amelioration of HIV viral activity together with a total recovery from pneumonia and skin infection.
