AIM:To investigate the value of combined detection of circulating cell-free DNA(cfDNA),a-fetal protein(AFP) and a L-fucosidase(AFU) for diagnosis of hepatocellular carcinoma(HCC).METHODS:Serum samples from 39 HCC pati...AIM:To investigate the value of combined detection of circulating cell-free DNA(cfDNA),a-fetal protein(AFP) and a L-fucosidase(AFU) for diagnosis of hepatocellular carcinoma(HCC).METHODS:Serum samples from 39 HCC patients and 45 normal controls were collected.Branched DNA(bDNA) was used to detect the level of cfDNA,and a receiver operating characteristic curve was employed to evaluate the diagnostic sensitivity,specificity,accuracy,positive predictive value,negative predictive value,positive likelihood ratio,negative likelihood ratio and Youden index,and to assess the diagnostic efficiency and their correlations with the clinicopathological features.AFP and AFU were detected by chemiluminescence and colorimetry,respectively.The significance of combined detection of the three biomarkers was discussed.RESULTS:cfDNA level was increased in 22 of the 39 HCC samples and in 2 of the 45 normal controls.cfDNA level in HCC samples was significantly higher than that in normal controls(P < 0.05).There were significant differences in sex and extra-and intrahepatic metastasis(P < 0.05).There was no significant correlation between cfDNA,AFP and AFU in the detection of HCC.The sensitivity of combined detection of cfDNA with one marker(AFP or AFU) and cfDNA with two markers(AFP and AFU) was 71.8%,87.2% and 89.7% vs 56.4%,53.8% and 66.7% for cfDNA,AFP and AFU used alone,respectively,the difference being statistically significant(P < 0.05).CONCLUSION:Quantitative analysis of cfDNA is sensitive and feasible,and the combined detection of cfDNA with AFP or AFU or both could improve the diagnostic sensitivity for HCC.展开更多
BACKGROUND Biliary diseases are common digestive system disorders which may combine with biliary tract infection such as cholecystitis or cholangitis.Thus,rapid identification of the bacteria and their antibiotic susc...BACKGROUND Biliary diseases are common digestive system disorders which may combine with biliary tract infection such as cholecystitis or cholangitis.Thus,rapid identification of the bacteria and their antibiotic susceptibility profiles are crucial for reducing the mortality of patients with biliary tract infection.AIM To identify bacterial species and antibiotic susceptibility for antibacterial therapy and analyze bile cultivation risk factors for increasing detection rates.METHODS This retrospective study was conducted from July 2008 to July 2017.In total,1339 bile samples which were collected during therapeutic endoscopic retrograde cholangiopan-creatography or percutaneous transhepatic cholangiodrainage or other biliary surgeries or biliary drainage were obtained to characterize pathogen spectra,antibiotic susceptibility,and clinical features.Clinical data including age,sex,comorbidities,clinical symptoms,protopathies,and history of biliary tract diseases and surgeries were collated from hospital medical records.Species identification and initial drug susceptibility were further identified by biochemical characterization using the VITEK 2 Compact test.RESULTS Positive microbiological findings were observed in 738 samples.The most frequently encountered strains were gram-negative bacteria(74.94%),including Escherichia coli(37.78%),Pseudomonas aeruginosa(8.96%),and Klebsiella pneumoniae(10.29%).Bile bacteria were largely sensitive to carbapenems,piperacillin/tazobactam,and gentamicin.Gram-negative strains had low susceptibility to ceftriaxone,quinolones and ampicillin.Almost the same microorganisms were present in patients with malignant and benign diseases.The number of samples with Klebsiella pneumoniae in the bile culture were significantly different between patients with malignant and benign diseases(55 vs 30;P=0.019).Age(P<0.001),fever(P<0.001),history of biliary tract diseases and surgeries(both P<0.001),benign disease(P=0.002),and the comorbidity chronic renal insufficiency(P=0.007)affected the positive rates of the bile samples.CONCLUSION Gram-negative bacteria were the most commonly isolated biliary bacteria.We determined the major factors associated with positive detection rates.Microbiological analysis of bile samples allowed accurate antibiotic treatments.展开更多
Live poultry markets（LPMs） are crucial places for human infection of influenza A（H7N9 virus）.In Yangtze River Delta,LPMs were closed after the outbreak of human infection with avian influenza A（H7N9） virus,and t...Live poultry markets（LPMs） are crucial places for human infection of influenza A（H7N9 virus）.In Yangtze River Delta,LPMs were closed after the outbreak of human infection with avian influenza A（H7N9） virus,and then reopened when no case was found.Our purpose was to quantify the effect of LPMs＇ operations in this region on the transmission of influenza A（H7N9） virus.We obtained information about dates of symptom onset and locations for all human influenza A（H7N9） cases reported from Shanghai,Jiangsu and Zhejiang provinces by May 31,2014,and acquired dates of closures and reopening of LPMs from official media.A two-phase Bayesian model was fitted by Markov Chain Monte Carlo methods to process the spatial and temporal influence of human cases.A total of 235 cases of influenza A（H7N9） were confirmed in Shanghai,Jiangsu and Zhejiang by May 31,2014.Using these data,our analysis showed that,after LPM closures,the influenza A（H7N9） outbreak disappeared within two weeks in Shanghai,one week in Jiangsu,and one week in Zhejiang,respectively.Local authorities reopened LPMs when there was no outbreak of influenza A（H7N9）,which did not lead to reemergence of human influenza A（H7N9）.LPM closures were effective in controlling the H7N9 outbreak.Reopening of LPM in summer did not increase the risk of human infection with H7N9.Our findings showed that LPMs should be closed immediately in areas where the H7N9 virus is confirmed in LPM.When there is no outbreak of H7N9 virus,LPMs can be reopened to satisfy the Chinese traditional culture of buying live poultry.In the long term,local authorities should take a cautious attitude in permanent LPM closure.展开更多
Objective： To investigate the effects of sleep electroencephalogram-modulated repetitive transcranial magnetic stimulation （SEM-rTMS） and conventional rTMS （C-rTMS） on the activity of hypothalamic-pituitary-adren...Objective： To investigate the effects of sleep electroencephalogram-modulated repetitive transcranial magnetic stimulation （SEM-rTMS） and conventional rTMS （C-rTMS） on the activity of hypothalamic-pituitary-adrenal （HPA） axis in patients with depression. Methods： In a double-blind, randomized controlled trial, 164 patients diagnosed with depression were randomized to treatment with SEM-rTMS （n=57）, C-rTMS （n=55） or sham rTMS （n=52） for 30 rain every day for 10 d. Before and after treatment plasma concentrations of adrenocorticotropic hormone （ACTH） and cortisol （CORT） were measured, and the 24-item Hamilton Depression Rating Scale （HAMD-24） was used for assessment. Results： The HAMD-24 scores and plasma ACTH and CORT concentrations of these depressive patients before treatment were significantly different from those of the normal control group （P〈0.05）. The HAMD-24 scores and plasma ACTH and CORT concentrations in the SEM-rTMS group and conventional rTMS group were decreased significantly （P〈0.05）. There was a significant positive correlation between the HAMD-24 scores and plasma ACTH （n=240, r=0.105, P=0.048） and CORT concentrations （n-240, r=0.126, P=0.023） in the patients with depression before and after treatment. Conclusion： The antidepressant effect of rTMS, including SEM-rTMS, may be related to its decreasing HPA axis activity. （This trail was registered. No： ChiCTR-TRC-00000465）展开更多
By using the density functional theory （B3LYP） and four highly accurate complete basis set （CBS-Q, CBS-QB3, CBS-Lq, and CBS-4M）ab initio methods, the X（C, N, O）-NO2 bond dissociation energies （BDEs） for CH3NO2...By using the density functional theory （B3LYP） and four highly accurate complete basis set （CBS-Q, CBS-QB3, CBS-Lq, and CBS-4M）ab initio methods, the X（C, N, O）-NO2 bond dissociation energies （BDEs） for CH3NO2, C2H3NO2, C2H5NO2, HONO2, CH3ONO2, C2H5ONO2, NH2NO2 （CH3）2NNO2 are computed. By comparing the computed BDEs and experimental results, it is found that the B3LYP method is unable to predict satisfactorily the results of bond dissociation energy （BDE）; however, all four CBS models are generally able to give reliable predication of the X（C, N, O）-NO2 BDEs for these nitro compounds. Moreover, the CBS-4M calculation is the least computationally demanding among the four CBS methods considered, Therefore, we recommend CBS-4M method as a reliable method of computing the BDEs for this nitro compound system.展开更多
Objective： To survey the special killing activity of CD3AK on anti-CEA-positive tumor enhanced by umbilical cord blood dendritic cell （DC） loaded with CEA recombinant vaccinia virus （CEA-rV）. Methods： Freshly is...Objective： To survey the special killing activity of CD3AK on anti-CEA-positive tumor enhanced by umbilical cord blood dendritic cell （DC） loaded with CEA recombinant vaccinia virus （CEA-rV）. Methods： Freshly isolated umbilical blood mononuclear calls （UBMC） were cultivated for 3 h. Suspension cells and attached calls were used to induce CD3AK calls and DC separately. DC was loaded with CEA-rV on the 3rd day to prepare CEA-rV＋DC. CD3AK cells were co-cultured with CEA- rV＋DC on the 8th day, to prepare CEA-rV＋DC＋CD3AK. The killing activity of each effector＇s cell, which included UBMC, CD3AK, DC＋CD3AK and CEA-rV＋DC＋CD3AK, was measured respectively by MTT reduction assay. Results： （1） 4 target cells were con- firmed by CEA monoclonal antibody of rabbit anti-human. Lovo and A549 were really CEA positive cell lines, while Bel-7402 and K562 were CEA negative cell lines. （2） It was showed by flow-cytometry that the mature DC cultured at 10th day expressed MHC I, II molecules such as CD86, CDS0, CD83 and CD40 highly, but CD123 lowly. The expression rates of CD86, CDS0, CD83 and CD40 was 82.7%, 51.1%, 57.5% and 69.4%, respectively. The appearances and intra-cellular structures of DC were observed through light and electron microscope. The diameter of mature DC was 15-20 μm presented the irregular morphologic appearanca, much prominences and pseudopodium. There were abundant mitochondria and endoplasmic reticulum in DC endochylema. （3） The rates of CD3, CD4, CD8 and CD28 in CD3AK cells group were 2 folds higher than that in UBMC group by FACS. It was said that the numbers of the mature T lymphocyte in CD3AK cells group were much greater than that in UBMC group. （4） The killing activities to 4 target cells of 3 effector＇s cells, which included CEA-rV＋DC＋CD3AK, DC＋CD3AK and CD3AK, were much greater than that of UBMC （P〈0.01）. Moreover, comparing with the killing activities of 4 effector＇s： CEA-rV＋DC＋CD3AK group 〉 DC＋CD3AK group 〉 CD3AK group 〉 UBMC group. It showed that, cytokine, DC and CEA-rV could efficiently elevate the killing activity of UBMC on broad-spectrum tumor cells. （5） Comparing with the killing activities of CEA-rV＋DC＋CD3AK and CD3AK cells to CEA positive and negative cells, the killing activities of CEA-rV＋DC＋CD3AK to CEA positive tumor calls, Lovo and A549 calls （P〈0.01） were remarkably better than that to CEA negative tumor cells BEL-7402 and K562 cells （P〈0.05）. It was said that the CEA-rV＋DC could obviously enhance the killing activity of CD3AK on CEA positive tumor cells. Comparing with the killing activities of CEA-rV＋DC＋CD3AK and DC＋CD3AK cells, the killing activity of CD3AK on CEA negative tumor cells was no statistical difference （P〉0.05）. However, the killing activity to CEA positive cells of CEA-rV＋DC＋CD3AK group was notably higher than that of DC＋CD3AK group. Namely, CEA-rV could distinctly promote the special killing activity to CEA positive tumor cells of CD3AK, but could not do it to CEA negative tumor cells. Conclusion： CEA-rV＋DC could obviously enhance the special killing activity of CD3AK on CEA positive tumor cell lines, while the DC only couldn＇t. The results indicated that the CEA-rV played an important role during the special killing activity of CD3AK cells to CEA positive tumor cells.展开更多
Background: In the previous study, we established an ischemia-prone gerbil population(IG), which was selectively bred to increase the incidence of unilateral carotid arterial occlusion(UCO)-induced ischemia in Mongoli...Background: In the previous study, we established an ischemia-prone gerbil population(IG), which was selectively bred to increase the incidence of unilateral carotid arterial occlusion(UCO)-induced ischemia in Mongolian gerbils. However, if the characteristics of ischemia model in IG are the same as those in general gerbils(GG), and if the neurological symptoms are associated with the neurological insults in IG is still unclear.Methods: In the present study, we evaluated the UCO model in IG by analyzing neurological symptoms, neurological injury in the hippocampal CA1 region and compared with GG.Results: The data showed that the ratios of neurological symptom scores ≥ 2 in the IG and GG groups were 65.0% vs 30.0%, respectively, and were significantly different(P <.01).The neuronal damage following a UCO ischemic insult in the IG group was more severe compared to the GG group. There was a high correlation between the neurological insults' scale and the neurological symptom score in the IG and GG groups(r =.979 and.943 in the IG and GG groups, respectively). In animals with mild neurological symptom scores(2 and 3), the neuronal insults were significantly different between female and male gerbils in both IG and GG.Conclusion: Our findings suggest that IG population would likely be more advantageous to establish an ischemic model.展开更多
Objective：To study the genetic variation of mitochondrial DNA （mtDNA） among common laboratory strains of inbred mice. Methods： The genetic polymorphism of mtDNA among 4 classical laboratory strains of inbred mice ...Objective：To study the genetic variation of mitochondrial DNA （mtDNA） among common laboratory strains of inbred mice. Methods： The genetic polymorphism of mtDNA among 4 classical laboratory strains of inbred mice and 3 inbred strains of mice established in China was analyzed by polymerase chain reaction coupled with restriction fragment length polymorphism（PCR-RFLP） and PCR coupled with single-stranded conformation polymorphism（PCR-SSCP）. Results： With regard to the D-loop （Displacement loop, D-loop）, tRNA^ Met＋Glu＋Ile, and ND3 （NADH dehydrogenase subunit 3, ND3） gene fragments of mtDNA from these mice,no variation was revealed by PCR-RFLP at 46 restriction enzyme sites. Further analyzed by PCR-SSCP,the D-loop 5＇fragment and 3＇end fragment of mtDNA from these mice also showed no genetic variation. Conclusion： Owing to maternal mode of inheritance of mtDNA,the results indicate that these common inbred strains of mice share the same maternal lineage.展开更多
The nonlineat third order optical susceptibility of C_(60) films on roughed glass plate has been studied by using a inode locked Nd:YAG laser emitting 8ps pulsed at 1.064 μm of 0.3 J energy.The large|χ^(3)(-3ω,ω,...The nonlineat third order optical susceptibility of C_(60) films on roughed glass plate has been studied by using a inode locked Nd:YAG laser emitting 8ps pulsed at 1.064 μm of 0.3 J energy.The large|χ^(3)(-3ω,ω,ω,ω)|=2 ×10^(-8) esu for C_(60) films of 15μm thickness has been observed,which is 1O^(3) larger than the value in the bulk C_(60).This is due to surface enhanced effect.展开更多
Staphylococcus epidermidis is one of the leading pathogens of nosocomial infections. Twenty- eight strains of biofilm-negative Staphylococcus epidermidis, of which thirteen strains considered as the clinical strains w...Staphylococcus epidermidis is one of the leading pathogens of nosocomial infections. Twenty- eight strains of biofilm-negative Staphylococcus epidermidis, of which thirteen strains considered as the clinical strains were isolated from catheters, blood and urine of patients in Huashan Hospital （Shanghai, China） and fifteen strains considered as commensal strains were isolated from the skin of healthy students who had no contact with hospitals in recent half year, were investigated for the adherence to human umbilical vein endothelial cell line ECV304 and human cervical adenocarcinoma cell line Hela, respectively. Our results showed that the clinical strains were much more adhesive to both cells than commensal ones. The endothelial adherence may be a virulence factor associated with this bacterial pathogenesis.展开更多
The CRISPR/Cas9 system has shown great potential for treating human genetic diseases through gene therapy.However,there are concerns about the safety of this system,specifically related to the use of guide-free Cas9.P...The CRISPR/Cas9 system has shown great potential for treating human genetic diseases through gene therapy.However,there are concerns about the safety of this system,specifically related to the use of guide-free Cas9.Previous studies have shown that guidefree Cas9 can induce genomic instability in vitro.However,the in vivo safety risks associated with guide-free Cas9 have not been evaluated,which is necessary for the development of gene therapy in clinical settings.In this study,we used doxycycline-inducible Cas9-expressing pigs to evaluate the safety risks of guide-free Cas9 in vivo.Our findings demonstrated that expression of guide-free Cas9 could induce genomic damages and transcriptome changes in vivo.The severity of the genomic damages and transcriptome changes were correlate with the expression levels of Cas9 protein.Moreover,prolonged expression of Cas9 in pigs led to abnormal phenotypes,including a significant decrease in body weight,which may be attributable to genomic damage-induced nutritional absorption and metabolic dysfunction.Furthermore,we observed an increase in whole-genome and tumor driver gene mutations in pigs with long-term Cas9 expression,raising the risk of tumor occurrence.Our in vivo evaluation of guide-free Cas9 in pigs highlights the necessity of considering and monitoring the detrimental effects of Cas9 alone as genome editing via the CRISPR/Cas9 system is implemented in clinical gene therapy.This research emphasizes the importance of further study and implementation of safety measures to ensure the successful and safe application of the CRiSPR/Cas9 system in clinical practice.展开更多
Background and objective Heparanase has been thought to be a good molecular marker of tumor,and the heparanase expression level was correlated closely with tumor metastasis. In this study,we investigate the effects of...Background and objective Heparanase has been thought to be a good molecular marker of tumor,and the heparanase expression level was correlated closely with tumor metastasis. In this study,we investigate the effects of heparanase on angiogenesis and lymphangiogenesis of lung cancer and the relationship between heparanase expression and vascular endothelial growth factor (VEGF),vascular endothelial growth factor-C (VEGF-C). Methods Immunohistochemistry was used to detect the expression of heparanase,VEGF,VEGF-C protein and microvascular density (MVD),lymphatic vessel density (LVD) in 115 cases of non-small cell lung cancer (NSCLC) and 45 cases of adjacent normal tissue samples. Results Our results showed that heparanase expression was significantly increased in 91 (79.13%) of the 115 cases and correlated with lymph node metastasis (node positive rate 87.0%; node negative rate 36.8%; P=0.003). Heparanase positive expression cases have significantly higher concentration of microvascular density (MVD) and lymphatic vessel density (LVD) as compared with heparanase negative expression cases (P<0.01,P<0.01,respectively),heparanase expression was significantly correlated with VEGF,VEGF-C expression in NSCLC. Conclusion Heparanase overexpression was associated with angiogenesis and lymphangiogenesis of lung cancer,targeting of heparanase may represent a significant therapeutic potential for lung cancer.展开更多
Thoracic aortic aneurysms(TAAs)develop asymptomatically and are characterized by dilatation of the aorta.This is considered a life-threating vascular disease due to the risk of aortic rupture and without effective tre...Thoracic aortic aneurysms(TAAs)develop asymptomatically and are characterized by dilatation of the aorta.This is considered a life-threating vascular disease due to the risk of aortic rupture and without effective treatments.The current understanding of the pathogenesis of TAA is still limited,especially for sporadic TAAs without known genetic mutation.Sirtuin 6(SIRT6)expression was significantly decreased in the tunica media of sporadic human TAA tissues.Genetic knockout of Sirt6 in mouse vascular smooth muscle cells accelerated TAA formation and rupture,reduced survival,and increased vascular inflammation and senescence after angiotensin II infusion.Transcriptome analysis identified interleukin(IL)-1βas a pivotal target of SIRT6,and increased IL-1βlevels correlated with vascular inflammation and senescence in human and mouse TAA samples.Chromatin immunoprecipitation revealed that SIRT6 bound to the Il1b promoter to repress expression partly by reducing the H3K9 and H3K56 acetylation.Genetic knockout of Il1b or pharmacological inhibition of IL-1βsignaling with the receptor antagonist anakinra rescued Sirt6 deficiency mediated aggravation of vascular inflammation,senescence,TAA formation and survival in mice.The findings reveal that SIRT6 protects against TAA by epigenetically inhibiting vascular inflammation and senescence,providing insight into potential epigenetic strategies for TAA treatment.展开更多
AIM: To investigate whether enteroviral infection might trigger acute pancreatitis in patients made susceptible due to high alcohol consumption. METHODS: Patients with alcohol-induced acute pan-creatitis were analyzed...AIM: To investigate whether enteroviral infection might trigger acute pancreatitis in patients made susceptible due to high alcohol consumption. METHODS: Patients with alcohol-induced acute pan-creatitis were analyzed for signs of simultaneous or preceding enteroviral infection. We studied the serum samples of 40 patients hospitalized for alcohol-induced acute pancreatitis and 40 controls recruited from an alcohol detoxification center. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect enterovirus RNA and diagnose acute viremia. Immuno-globulin G (IgG), immunoglobulin A (IgA) and immuno-globulin M (IgM) enteroviral antibodies were measured using enzyme immunoassay to detect subacute andprevious infections. The samples were considered posi-tive when the antibody titers were≥15 IU. Further-more, using RT-PCR, we studied pancreatic biopsy sam-ples obtained during surgery from nine patients with chronic pancreatitis, one patient with acute pancreatitis and ten control patients with pancreatic carcinoma for evidence of persisting enteroviral RNA in the pancreatic tissue. RESULTS: No enterovirus RNA indicating acute viremia was detected by RT-PCR in the serum samples of any patient or control. A high incidence of positive antibody titers was observed in both study groups: IgM antibod-ies had positive titers in 5/40 (13%) vs 4/40 (10%), P=0.723; IgG in 15/40 (38%) vs 19/40 (48%), P=0.366; and IgA in 25/40 (63%) vs 33/40 (83%), P=0.045, patients and controls, respectively. Ten (25%) patients had severe pancreatitis and two (5%) required treatment in intensive care. The median length of hos-pitalization was 7 d (range: 3-47 d). The severity of acute pancreatitis or the length of hospitalization was not associated with enteroviral IgM, IgG or IgA anti-bodies. Five pancreatic biopsy samples tested positive with RT-PCR, three (8%) in the control group and two (5%) in the patient group (P=0.64). CONCLUSION: The rate of enteroviral infection is not increased in patients with alcohol-induced acute pan-creatitis when compared to alcoholics with similar high alcohol use.展开更多
Xenotransplantation,involving animal organ transplantation into humans to address the human organ shortage,has been studied since the 17th century.Early attempts to obtain organs from animals such as goats,dogs,and no...Xenotransplantation,involving animal organ transplantation into humans to address the human organ shortage,has been studied since the 17th century.Early attempts to obtain organs from animals such as goats,dogs,and non-human primates proved unsuccessful.In the 1990s,scientists agreed that pigs were the most suitable donor animals for xenotransplantation.However,immune rejection between pig and human has hindered the application.To overcome these challenges,researchers developed genetically modified pigs that deactivate xenoreactive antigen genes and express human protective genes.These advances extended xenograft survival from days to years in non-human primates,resulting in the first human heart xenotransplant trial.Using genetically engineered pigs for the organ shortage is promising.This review provides an overview of potential incompatibilities of immunogenicity and functional proteins related to xenotransplantation between humans and pigs.Furthermore,it elucidates possible approaches for multiplex gene modification to breed better-humanized pigs for clinical xenotransplantation.展开更多
Background It remains almost a helpless situation for the recurrent implantation failure and pregnancy loss caused by endometrial injury at present. The purpose of this study was to develop a rabbit model of endometri...Background It remains almost a helpless situation for the recurrent implantation failure and pregnancy loss caused by endometrial injury at present. The purpose of this study was to develop a rabbit model of endometrial mechanical injury that could provide a research platform for this difficult clinical predicament. Methods Three experiments were conducted. Experiment 1： Curettages in both uterus horns and copper wire inserting after curettage （double-injury） in one horn. The histological changes were monitored at 0, 24, 48, 72 hours, as well as in 1 and 2 weeks after operation. Experiment 2： Direct copper wire inserting in one horn and double-injury in other horn. The wires in both horns were removed after 2 weeks.展开更多
The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interestingcandidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic...The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interestingcandidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cellnuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs expressapproximately 15–25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was alsomarkedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs withinfluenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profounddecrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-foldreduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found thatthe Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These resultsdemonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate thatthe Mx1 transgene can be harnessed to develop disease-resistant pigs.展开更多
Background： Measuring total serum calcium is important for the diagnosis of diseases. Currently, results from commercial kits for calcium measurement are variable. Generally, the performance of serum calcium measurem...Background： Measuring total serum calcium is important for the diagnosis of diseases. Currently, results from commercial kits for calcium measurement are variable. Generally, the performance of serum calcium measurements is monitored by external quality assessment （EQA） or proficiency testing schemes. However, the commutability of the EQA samples and calibrators is often unknown, which limits the effectiveness of EQA schemes. The aim of this study was to evaluate the bias of serum calcium measurements and the commutability of processed materials. Methods： Inductively coupled plasma mass spectrometry was applied as a comparative method, and 14 routine methods were chosen as test methods. Forty-eight serum samples from individual patients and 25 processed materials were quantified. A scatter plot was generated from patient samples, and 95% prediction intervals were calculated to evaluate the commutability of the processed materials and measurement bias at three concentration levels was used to determine the accuracy of routine assays. Results： All assays showed high precision （total coefficient of variation [CV] 〈2.26%） and correlation coefficients （r 〉 0.99）. For all assays, the mean bias for the 48 patient samples ranged from 0.13 mmol/L to 0.00 mmol/L （-5.61 0.01%）, and the ranges for the three concentrations were 0.10-0.04 mmoUL （-5.71-2.35%）, -0.14-0.01 mmol/L （-5.80-0.30%）, and -0.19-0.04 mmol/L （-6.24-1.22%）. The EQA samples, calibrators, and animal sera exhibited matrix effects in some assays; human serum pools were commutable in all assays; certificate reference materials were commutable in most assays, and only GBW09152 exhibited a matrix effect in one assay： and aqueous reference materials exhibited matrix effects in most assays. Conclusions： Biases for most assays were within the acceptable range, although the accuracy of some assays needs improvement. Human serum pools prepared from patient samples were commutable, and the other tested materials exhibited a matrix effect.展开更多
Background： Hemoglobin A 1 c （HbA1 c） measurement is of great value for the diagnosis and monitoring of diabetes. Many manufacturers have developed various experiments to determine the HbAlc concentration. However,...Background： Hemoglobin A 1 c （HbA1 c） measurement is of great value for the diagnosis and monitoring of diabetes. Many manufacturers have developed various experiments to determine the HbAlc concentration. However, the longitudinal use of these tests requires strict quality management. This study aimed to analyze the quality of HbAlc measurement systems in China using six sigma techniques to help improve their performances. Methods： A total of 135 laboratories were involved in this investigation in 2015. Bias values and coefficients of variation were collected from an HbA 1 c trueness verification external quality assessment program and an internal quality control program organized by the National Center of Clinical Laboratories in China. The sigma （σ） values and the quality goal index （QGI） were used to evaluate the performances of different groups, which were divided according to principles and instruments. Results： The majority of participants （88, 65.2%） were scored as ＂improvement needed （σ 〈 3）＂, suggesting that the laboratories needed to improve their measurement performance. Only 8.2% （11/135） of the laboratories were scored as ＂world class （σ≥ 6）＂. Among all the 88 laboratories whose σ values were below 3, 52 （59.1%） and 23 （26.1%） laboratories needed to improve measurement precision （QGI 〈8.0） and trueness （QGI 〉 1.2）, respectively; the remaining laboratories （13, 14.8%） needed to improve both measurement precision and trueness. In addition, 16.1% （5/31） and 15.0% （3/20） of the laboratories in ＂TO SOH＂ and ＂ARKRAY＂ groups, respectively, were scored as ＂world class＂, whereas none of the laboratories in ＂BIO-RAD＂ group were scored as ＂world class＂. Conclusions： This study indicated that, although participating laboratories were laboratories with better performance in China, the performances were still unsatisfactory. Actions should be taken to improve HbAlc measurement performance before we can include HbAlc assays in diabetes diagnosis in China.展开更多
Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological process.Binary Tet-On system has been widely employed to regulate transgenic expression by doxycycline.Previous...Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological process.Binary Tet-On system has been widely employed to regulate transgenic expression by doxycycline.Previous pig models with tetracycline regulatory elements were generated through random integration.This process often resulted in uncertain expression and unpredictable phenotypes,thus hindering their applications.Here,by precise knock-in of binary Tet-On 3G elements into Rosa26 and Hipp11 locus,respectively,a double knock-in reporter pig model was generated.We characterized excellent properties of this system for controllable transgenic expression both in vitro and in vivo.Two att P sites were arranged to flank the td Tomato to switch reporter gene.Single or multiple gene replacement was efficiently and faithfully achieved in fetal fibroblasts and nuclear transfer embryos.To display the flexible application of this system,we generated a pig strain with Dox-inducing h KRASexpression through phiC31 integrase-mediated cassette exchange.After eight months of Dox administration,squamous cell carcinoma developed in the nose,mouth,and scrotum,which indicated this pig strain could serve as an ideal large animal model to study tumorigenesis.Overall,the established pig models with controllable and switchable transgene expression system will provide a facilitating platform for transgenic and biomedical research.展开更多
文摘AIM:To investigate the value of combined detection of circulating cell-free DNA(cfDNA),a-fetal protein(AFP) and a L-fucosidase(AFU) for diagnosis of hepatocellular carcinoma(HCC).METHODS:Serum samples from 39 HCC patients and 45 normal controls were collected.Branched DNA(bDNA) was used to detect the level of cfDNA,and a receiver operating characteristic curve was employed to evaluate the diagnostic sensitivity,specificity,accuracy,positive predictive value,negative predictive value,positive likelihood ratio,negative likelihood ratio and Youden index,and to assess the diagnostic efficiency and their correlations with the clinicopathological features.AFP and AFU were detected by chemiluminescence and colorimetry,respectively.The significance of combined detection of the three biomarkers was discussed.RESULTS:cfDNA level was increased in 22 of the 39 HCC samples and in 2 of the 45 normal controls.cfDNA level in HCC samples was significantly higher than that in normal controls(P < 0.05).There were significant differences in sex and extra-and intrahepatic metastasis(P < 0.05).There was no significant correlation between cfDNA,AFP and AFU in the detection of HCC.The sensitivity of combined detection of cfDNA with one marker(AFP or AFU) and cfDNA with two markers(AFP and AFU) was 71.8%,87.2% and 89.7% vs 56.4%,53.8% and 66.7% for cfDNA,AFP and AFU used alone,respectively,the difference being statistically significant(P < 0.05).CONCLUSION:Quantitative analysis of cfDNA is sensitive and feasible,and the combined detection of cfDNA with AFP or AFU or both could improve the diagnostic sensitivity for HCC.
基金National Natural Science Foundation of China,No.81802071.
文摘BACKGROUND Biliary diseases are common digestive system disorders which may combine with biliary tract infection such as cholecystitis or cholangitis.Thus,rapid identification of the bacteria and their antibiotic susceptibility profiles are crucial for reducing the mortality of patients with biliary tract infection.AIM To identify bacterial species and antibiotic susceptibility for antibacterial therapy and analyze bile cultivation risk factors for increasing detection rates.METHODS This retrospective study was conducted from July 2008 to July 2017.In total,1339 bile samples which were collected during therapeutic endoscopic retrograde cholangiopan-creatography or percutaneous transhepatic cholangiodrainage or other biliary surgeries or biliary drainage were obtained to characterize pathogen spectra,antibiotic susceptibility,and clinical features.Clinical data including age,sex,comorbidities,clinical symptoms,protopathies,and history of biliary tract diseases and surgeries were collated from hospital medical records.Species identification and initial drug susceptibility were further identified by biochemical characterization using the VITEK 2 Compact test.RESULTS Positive microbiological findings were observed in 738 samples.The most frequently encountered strains were gram-negative bacteria(74.94%),including Escherichia coli(37.78%),Pseudomonas aeruginosa(8.96%),and Klebsiella pneumoniae(10.29%).Bile bacteria were largely sensitive to carbapenems,piperacillin/tazobactam,and gentamicin.Gram-negative strains had low susceptibility to ceftriaxone,quinolones and ampicillin.Almost the same microorganisms were present in patients with malignant and benign diseases.The number of samples with Klebsiella pneumoniae in the bile culture were significantly different between patients with malignant and benign diseases(55 vs 30;P=0.019).Age(P<0.001),fever(P<0.001),history of biliary tract diseases and surgeries(both P<0.001),benign disease(P=0.002),and the comorbidity chronic renal insufficiency(P=0.007)affected the positive rates of the bile samples.CONCLUSION Gram-negative bacteria were the most commonly isolated biliary bacteria.We determined the major factors associated with positive detection rates.Microbiological analysis of bile samples allowed accurate antibiotic treatments.
文摘Live poultry markets（LPMs） are crucial places for human infection of influenza A（H7N9 virus）.In Yangtze River Delta,LPMs were closed after the outbreak of human infection with avian influenza A（H7N9） virus,and then reopened when no case was found.Our purpose was to quantify the effect of LPMs＇ operations in this region on the transmission of influenza A（H7N9） virus.We obtained information about dates of symptom onset and locations for all human influenza A（H7N9） cases reported from Shanghai,Jiangsu and Zhejiang provinces by May 31,2014,and acquired dates of closures and reopening of LPMs from official media.A two-phase Bayesian model was fitted by Markov Chain Monte Carlo methods to process the spatial and temporal influence of human cases.A total of 235 cases of influenza A（H7N9） were confirmed in Shanghai,Jiangsu and Zhejiang by May 31,2014.Using these data,our analysis showed that,after LPM closures,the influenza A（H7N9） outbreak disappeared within two weeks in Shanghai,one week in Jiangsu,and one week in Zhejiang,respectively.Local authorities reopened LPMs when there was no outbreak of influenza A（H7N9）,which did not lead to reemergence of human influenza A（H7N9）.LPM closures were effective in controlling the H7N9 outbreak.Reopening of LPM in summer did not increase the risk of human infection with H7N9.Our findings showed that LPMs should be closed immediately in areas where the H7N9 virus is confirmed in LPM.When there is no outbreak of H7N9 virus,LPMs can be reopened to satisfy the Chinese traditional culture of buying live poultry.In the long term,local authorities should take a cautious attitude in permanent LPM closure.
基金Supported by the Social Development Guide Projects of Science and Technology Committee of Jiangsu Province in 2005 (BS2005629)"The Six Major Expert Peaks" Projects of Jiangsu Provincial Government in 2006 [Surentong (2006) 174]
文摘Objective： To investigate the effects of sleep electroencephalogram-modulated repetitive transcranial magnetic stimulation （SEM-rTMS） and conventional rTMS （C-rTMS） on the activity of hypothalamic-pituitary-adrenal （HPA） axis in patients with depression. Methods： In a double-blind, randomized controlled trial, 164 patients diagnosed with depression were randomized to treatment with SEM-rTMS （n=57）, C-rTMS （n=55） or sham rTMS （n=52） for 30 rain every day for 10 d. Before and after treatment plasma concentrations of adrenocorticotropic hormone （ACTH） and cortisol （CORT） were measured, and the 24-item Hamilton Depression Rating Scale （HAMD-24） was used for assessment. Results： The HAMD-24 scores and plasma ACTH and CORT concentrations of these depressive patients before treatment were significantly different from those of the normal control group （P〈0.05）. The HAMD-24 scores and plasma ACTH and CORT concentrations in the SEM-rTMS group and conventional rTMS group were decreased significantly （P〈0.05）. There was a significant positive correlation between the HAMD-24 scores and plasma ACTH （n=240, r=0.105, P=0.048） and CORT concentrations （n-240, r=0.126, P=0.023） in the patients with depression before and after treatment. Conclusion： The antidepressant effect of rTMS, including SEM-rTMS, may be related to its decreasing HPA axis activity. （This trail was registered. No： ChiCTR-TRC-00000465）
基金Project supported by the National Natural Science Foundation of China and China Academy of Engineering Physics （Grant Nos 10376021, 10274055）.
文摘By using the density functional theory （B3LYP） and four highly accurate complete basis set （CBS-Q, CBS-QB3, CBS-Lq, and CBS-4M）ab initio methods, the X（C, N, O）-NO2 bond dissociation energies （BDEs） for CH3NO2, C2H3NO2, C2H5NO2, HONO2, CH3ONO2, C2H5ONO2, NH2NO2 （CH3）2NNO2 are computed. By comparing the computed BDEs and experimental results, it is found that the B3LYP method is unable to predict satisfactorily the results of bond dissociation energy （BDE）; however, all four CBS models are generally able to give reliable predication of the X（C, N, O）-NO2 BDEs for these nitro compounds. Moreover, the CBS-4M calculation is the least computationally demanding among the four CBS methods considered, Therefore, we recommend CBS-4M method as a reliable method of computing the BDEs for this nitro compound system.
基金Supported by a grant from Science & Technique Department of Guang-dong Province of China (No. 2002C30305).
文摘Objective： To survey the special killing activity of CD3AK on anti-CEA-positive tumor enhanced by umbilical cord blood dendritic cell （DC） loaded with CEA recombinant vaccinia virus （CEA-rV）. Methods： Freshly isolated umbilical blood mononuclear calls （UBMC） were cultivated for 3 h. Suspension cells and attached calls were used to induce CD3AK calls and DC separately. DC was loaded with CEA-rV on the 3rd day to prepare CEA-rV＋DC. CD3AK cells were co-cultured with CEA- rV＋DC on the 8th day, to prepare CEA-rV＋DC＋CD3AK. The killing activity of each effector＇s cell, which included UBMC, CD3AK, DC＋CD3AK and CEA-rV＋DC＋CD3AK, was measured respectively by MTT reduction assay. Results： （1） 4 target cells were con- firmed by CEA monoclonal antibody of rabbit anti-human. Lovo and A549 were really CEA positive cell lines, while Bel-7402 and K562 were CEA negative cell lines. （2） It was showed by flow-cytometry that the mature DC cultured at 10th day expressed MHC I, II molecules such as CD86, CDS0, CD83 and CD40 highly, but CD123 lowly. The expression rates of CD86, CDS0, CD83 and CD40 was 82.7%, 51.1%, 57.5% and 69.4%, respectively. The appearances and intra-cellular structures of DC were observed through light and electron microscope. The diameter of mature DC was 15-20 μm presented the irregular morphologic appearanca, much prominences and pseudopodium. There were abundant mitochondria and endoplasmic reticulum in DC endochylema. （3） The rates of CD3, CD4, CD8 and CD28 in CD3AK cells group were 2 folds higher than that in UBMC group by FACS. It was said that the numbers of the mature T lymphocyte in CD3AK cells group were much greater than that in UBMC group. （4） The killing activities to 4 target cells of 3 effector＇s cells, which included CEA-rV＋DC＋CD3AK, DC＋CD3AK and CD3AK, were much greater than that of UBMC （P〈0.01）. Moreover, comparing with the killing activities of 4 effector＇s： CEA-rV＋DC＋CD3AK group 〉 DC＋CD3AK group 〉 CD3AK group 〉 UBMC group. It showed that, cytokine, DC and CEA-rV could efficiently elevate the killing activity of UBMC on broad-spectrum tumor cells. （5） Comparing with the killing activities of CEA-rV＋DC＋CD3AK and CD3AK cells to CEA positive and negative cells, the killing activities of CEA-rV＋DC＋CD3AK to CEA positive tumor calls, Lovo and A549 calls （P〈0.01） were remarkably better than that to CEA negative tumor cells BEL-7402 and K562 cells （P〈0.05）. It was said that the CEA-rV＋DC could obviously enhance the killing activity of CD3AK on CEA positive tumor cells. Comparing with the killing activities of CEA-rV＋DC＋CD3AK and DC＋CD3AK cells, the killing activity of CD3AK on CEA negative tumor cells was no statistical difference （P〉0.05）. However, the killing activity to CEA positive cells of CEA-rV＋DC＋CD3AK group was notably higher than that of DC＋CD3AK group. Namely, CEA-rV could distinctly promote the special killing activity to CEA positive tumor cells of CD3AK, but could not do it to CEA negative tumor cells. Conclusion： CEA-rV＋DC could obviously enhance the special killing activity of CD3AK on CEA positive tumor cell lines, while the DC only couldn＇t. The results indicated that the CEA-rV played an important role during the special killing activity of CD3AK cells to CEA positive tumor cells.
基金Key Projects in the National Science&Technology Pillar Program,Grant/Award Number:2015BAI09B01
文摘Background: In the previous study, we established an ischemia-prone gerbil population(IG), which was selectively bred to increase the incidence of unilateral carotid arterial occlusion(UCO)-induced ischemia in Mongolian gerbils. However, if the characteristics of ischemia model in IG are the same as those in general gerbils(GG), and if the neurological symptoms are associated with the neurological insults in IG is still unclear.Methods: In the present study, we evaluated the UCO model in IG by analyzing neurological symptoms, neurological injury in the hippocampal CA1 region and compared with GG.Results: The data showed that the ratios of neurological symptom scores ≥ 2 in the IG and GG groups were 65.0% vs 30.0%, respectively, and were significantly different(P <.01).The neuronal damage following a UCO ischemic insult in the IG group was more severe compared to the GG group. There was a high correlation between the neurological insults' scale and the neurological symptom score in the IG and GG groups(r =.979 and.943 in the IG and GG groups, respectively). In animals with mild neurological symptom scores(2 and 3), the neuronal insults were significantly different between female and male gerbils in both IG and GG.Conclusion: Our findings suggest that IG population would likely be more advantageous to establish an ischemic model.
基金Supported by Developmental Programming Item of National Keystone Basic Research (G2000016106) and National Natural Science Foundation of China (No. 39600079)
文摘Objective：To study the genetic variation of mitochondrial DNA （mtDNA） among common laboratory strains of inbred mice. Methods： The genetic polymorphism of mtDNA among 4 classical laboratory strains of inbred mice and 3 inbred strains of mice established in China was analyzed by polymerase chain reaction coupled with restriction fragment length polymorphism（PCR-RFLP） and PCR coupled with single-stranded conformation polymorphism（PCR-SSCP）. Results： With regard to the D-loop （Displacement loop, D-loop）, tRNA^ Met＋Glu＋Ile, and ND3 （NADH dehydrogenase subunit 3, ND3） gene fragments of mtDNA from these mice,no variation was revealed by PCR-RFLP at 46 restriction enzyme sites. Further analyzed by PCR-SSCP,the D-loop 5＇fragment and 3＇end fragment of mtDNA from these mice also showed no genetic variation. Conclusion： Owing to maternal mode of inheritance of mtDNA,the results indicate that these common inbred strains of mice share the same maternal lineage.
基金Supported by the National Natuiil Science Foundation of China,S.F.of Zhejiang Province,P.R.C.and Doctoral Foundation of China.
文摘The nonlineat third order optical susceptibility of C_(60) films on roughed glass plate has been studied by using a inode locked Nd:YAG laser emitting 8ps pulsed at 1.064 μm of 0.3 J energy.The large|χ^(3)(-3ω,ω,ω,ω)|=2 ×10^(-8) esu for C_(60) films of 15μm thickness has been observed,which is 1O^(3) larger than the value in the bulk C_(60).This is due to surface enhanced effect.
基金This work was supported by the Chinese National Natural Science Foundation(30670108)Shanghai Municipal Science and Technology Commission(05PJ14025).
文摘Staphylococcus epidermidis is one of the leading pathogens of nosocomial infections. Twenty- eight strains of biofilm-negative Staphylococcus epidermidis, of which thirteen strains considered as the clinical strains were isolated from catheters, blood and urine of patients in Huashan Hospital （Shanghai, China） and fifteen strains considered as commensal strains were isolated from the skin of healthy students who had no contact with hospitals in recent half year, were investigated for the adherence to human umbilical vein endothelial cell line ECV304 and human cervical adenocarcinoma cell line Hela, respectively. Our results showed that the clinical strains were much more adhesive to both cells than commensal ones. The endothelial adherence may be a virulence factor associated with this bacterial pathogenesis.
基金This work was financially supported by National Key Research and Development Program of China(2022YFA1105403,2022YFA1105402,2021YFA0805903,2023YFF0724703,2021YFF0702601)Research Unit of Generation of Large Animal Disease Models,Chinese Academy of Medical Sciences(2019-12M-5-025)+4 种基金National Natural Science Foundation of China(32170542,32300426)Major Science and Technology Projects of Hainan Province(ZDKJ2021030)Science and Technology Planning ProjectofGuangdong ProvinceC,hina(2023B1212060050,2021B1212040016,2021A1515110909)Hainan Provincial Joint Project of Sanya Yazhou Bay Science and Technology City(2021JJLH0085,2021JJLH0096)the Youth Innovation Promotion Association of the Chinese Academy of Sciences(Y2023096).
文摘The CRISPR/Cas9 system has shown great potential for treating human genetic diseases through gene therapy.However,there are concerns about the safety of this system,specifically related to the use of guide-free Cas9.Previous studies have shown that guidefree Cas9 can induce genomic instability in vitro.However,the in vivo safety risks associated with guide-free Cas9 have not been evaluated,which is necessary for the development of gene therapy in clinical settings.In this study,we used doxycycline-inducible Cas9-expressing pigs to evaluate the safety risks of guide-free Cas9 in vivo.Our findings demonstrated that expression of guide-free Cas9 could induce genomic damages and transcriptome changes in vivo.The severity of the genomic damages and transcriptome changes were correlate with the expression levels of Cas9 protein.Moreover,prolonged expression of Cas9 in pigs led to abnormal phenotypes,including a significant decrease in body weight,which may be attributable to genomic damage-induced nutritional absorption and metabolic dysfunction.Furthermore,we observed an increase in whole-genome and tumor driver gene mutations in pigs with long-term Cas9 expression,raising the risk of tumor occurrence.Our in vivo evaluation of guide-free Cas9 in pigs highlights the necessity of considering and monitoring the detrimental effects of Cas9 alone as genome editing via the CRISPR/Cas9 system is implemented in clinical gene therapy.This research emphasizes the importance of further study and implementation of safety measures to ensure the successful and safe application of the CRiSPR/Cas9 system in clinical practice.
文摘Background and objective Heparanase has been thought to be a good molecular marker of tumor,and the heparanase expression level was correlated closely with tumor metastasis. In this study,we investigate the effects of heparanase on angiogenesis and lymphangiogenesis of lung cancer and the relationship between heparanase expression and vascular endothelial growth factor (VEGF),vascular endothelial growth factor-C (VEGF-C). Methods Immunohistochemistry was used to detect the expression of heparanase,VEGF,VEGF-C protein and microvascular density (MVD),lymphatic vessel density (LVD) in 115 cases of non-small cell lung cancer (NSCLC) and 45 cases of adjacent normal tissue samples. Results Our results showed that heparanase expression was significantly increased in 91 (79.13%) of the 115 cases and correlated with lymph node metastasis (node positive rate 87.0%; node negative rate 36.8%; P=0.003). Heparanase positive expression cases have significantly higher concentration of microvascular density (MVD) and lymphatic vessel density (LVD) as compared with heparanase negative expression cases (P<0.01,P<0.01,respectively),heparanase expression was significantly correlated with VEGF,VEGF-C expression in NSCLC. Conclusion Heparanase overexpression was associated with angiogenesis and lymphangiogenesis of lung cancer,targeting of heparanase may represent a significant therapeutic potential for lung cancer.
基金This work was supported by grants from the National Key Research and Development Project of China(grant numbers:2020YFC2008003,2021YFA0804900 and 2019YFA0801500)the National Natural Science Foundation of China(grant numbers:92149305,8222500782030017 and 81801627)the Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences(grant numbers:2021-I2M-1-016,2022-RC180-03,2022-I2M-JB-006 and 2022-I2M-2-002).
文摘Thoracic aortic aneurysms(TAAs)develop asymptomatically and are characterized by dilatation of the aorta.This is considered a life-threating vascular disease due to the risk of aortic rupture and without effective treatments.The current understanding of the pathogenesis of TAA is still limited,especially for sporadic TAAs without known genetic mutation.Sirtuin 6(SIRT6)expression was significantly decreased in the tunica media of sporadic human TAA tissues.Genetic knockout of Sirt6 in mouse vascular smooth muscle cells accelerated TAA formation and rupture,reduced survival,and increased vascular inflammation and senescence after angiotensin II infusion.Transcriptome analysis identified interleukin(IL)-1βas a pivotal target of SIRT6,and increased IL-1βlevels correlated with vascular inflammation and senescence in human and mouse TAA samples.Chromatin immunoprecipitation revealed that SIRT6 bound to the Il1b promoter to repress expression partly by reducing the H3K9 and H3K56 acetylation.Genetic knockout of Il1b or pharmacological inhibition of IL-1βsignaling with the receptor antagonist anakinra rescued Sirt6 deficiency mediated aggravation of vascular inflammation,senescence,TAA formation and survival in mice.The findings reveal that SIRT6 protects against TAA by epigenetically inhibiting vascular inflammation and senescence,providing insight into potential epigenetic strategies for TAA treatment.
文摘AIM: To investigate whether enteroviral infection might trigger acute pancreatitis in patients made susceptible due to high alcohol consumption. METHODS: Patients with alcohol-induced acute pan-creatitis were analyzed for signs of simultaneous or preceding enteroviral infection. We studied the serum samples of 40 patients hospitalized for alcohol-induced acute pancreatitis and 40 controls recruited from an alcohol detoxification center. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect enterovirus RNA and diagnose acute viremia. Immuno-globulin G (IgG), immunoglobulin A (IgA) and immuno-globulin M (IgM) enteroviral antibodies were measured using enzyme immunoassay to detect subacute andprevious infections. The samples were considered posi-tive when the antibody titers were≥15 IU. Further-more, using RT-PCR, we studied pancreatic biopsy sam-ples obtained during surgery from nine patients with chronic pancreatitis, one patient with acute pancreatitis and ten control patients with pancreatic carcinoma for evidence of persisting enteroviral RNA in the pancreatic tissue. RESULTS: No enterovirus RNA indicating acute viremia was detected by RT-PCR in the serum samples of any patient or control. A high incidence of positive antibody titers was observed in both study groups: IgM antibod-ies had positive titers in 5/40 (13%) vs 4/40 (10%), P=0.723; IgG in 15/40 (38%) vs 19/40 (48%), P=0.366; and IgA in 25/40 (63%) vs 33/40 (83%), P=0.045, patients and controls, respectively. Ten (25%) patients had severe pancreatitis and two (5%) required treatment in intensive care. The median length of hos-pitalization was 7 d (range: 3-47 d). The severity of acute pancreatitis or the length of hospitalization was not associated with enteroviral IgM, IgG or IgA anti-bodies. Five pancreatic biopsy samples tested positive with RT-PCR, three (8%) in the control group and two (5%) in the patient group (P=0.64). CONCLUSION: The rate of enteroviral infection is not increased in patients with alcohol-induced acute pan-creatitis when compared to alcoholics with similar high alcohol use.
基金supported by the National Key Research and Development Program of China(2022YFA1105404,2021YFF0702601 and 2021YFA0805300)the Research Unit of Generation of Large Animal Disease Models,Chinese Academy of Medical Sciences(2019I2M-5-025)+3 种基金the Science and Technology Program of Guangzhou(202201010409)the Key Research&Development Program of Hainan Province(ZDYF2021SHFZ052)the Major Science and Technology Project of Hainan Province(ZDKJ2021030)the 2020 Research Program of Sanya Yazhou Bay Science and Technology City(202002011)
文摘Xenotransplantation,involving animal organ transplantation into humans to address the human organ shortage,has been studied since the 17th century.Early attempts to obtain organs from animals such as goats,dogs,and non-human primates proved unsuccessful.In the 1990s,scientists agreed that pigs were the most suitable donor animals for xenotransplantation.However,immune rejection between pig and human has hindered the application.To overcome these challenges,researchers developed genetically modified pigs that deactivate xenoreactive antigen genes and express human protective genes.These advances extended xenograft survival from days to years in non-human primates,resulting in the first human heart xenotransplant trial.Using genetically engineered pigs for the organ shortage is promising.This review provides an overview of potential incompatibilities of immunogenicity and functional proteins related to xenotransplantation between humans and pigs.Furthermore,it elucidates possible approaches for multiplex gene modification to breed better-humanized pigs for clinical xenotransplantation.
文摘Background It remains almost a helpless situation for the recurrent implantation failure and pregnancy loss caused by endometrial injury at present. The purpose of this study was to develop a rabbit model of endometrial mechanical injury that could provide a research platform for this difficult clinical predicament. Methods Three experiments were conducted. Experiment 1： Curettages in both uterus horns and copper wire inserting after curettage （double-injury） in one horn. The histological changes were monitored at 0, 24, 48, 72 hours, as well as in 1 and 2 weeks after operation. Experiment 2： Direct copper wire inserting in one horn and double-injury in other horn. The wires in both horns were removed after 2 weeks.
基金This work was supported by grants from National Basic Research Program of China(973 program)(2011CB944203)ZNGI-2011-010 from the Guangzhou Municipality and the Chinese Academy and Sciences to L.L.
文摘The myxovirus resistance gene (Mx1) has a broad spectrum of antiviral activities. It is therefore an interestingcandidate gene to improve disease resistance in farm animals. In this study, we report the use of somatic cellnuclear transfer (SCNT) to produce transgenic pigs over-expressing the Mx1 gene. These transgenic pigs expressapproximately 15–25 times more Mx1 mRNA than non-transgenic pigs, and the protein level of Mx1 was alsomarkedly enhanced. We challenged fibroblast cells isolated from the ear skin of transgenic and control pigs withinfluenza A virus and classical swine fever virus (CFSV). Indirect immunofluorescence assay (IFA) revealed a profounddecrease of influenza A proliferation in Mx1 transgenic cells. Growth kinetics showed an approximately 10-foldreduction of viral copies in the transgenic cells compared to non-transgenic controls. Additionally, we found thatthe Mx1 transgenic cells were more resistant to CSFV infection in comparison to non-transgenic cells. These resultsdemonstrate that the Mx1 transgene can protect against viral infection in cells of transgenic pigs and indicate thatthe Mx1 transgene can be harnessed to develop disease-resistant pigs.
文摘Background： Measuring total serum calcium is important for the diagnosis of diseases. Currently, results from commercial kits for calcium measurement are variable. Generally, the performance of serum calcium measurements is monitored by external quality assessment （EQA） or proficiency testing schemes. However, the commutability of the EQA samples and calibrators is often unknown, which limits the effectiveness of EQA schemes. The aim of this study was to evaluate the bias of serum calcium measurements and the commutability of processed materials. Methods： Inductively coupled plasma mass spectrometry was applied as a comparative method, and 14 routine methods were chosen as test methods. Forty-eight serum samples from individual patients and 25 processed materials were quantified. A scatter plot was generated from patient samples, and 95% prediction intervals were calculated to evaluate the commutability of the processed materials and measurement bias at three concentration levels was used to determine the accuracy of routine assays. Results： All assays showed high precision （total coefficient of variation [CV] 〈2.26%） and correlation coefficients （r 〉 0.99）. For all assays, the mean bias for the 48 patient samples ranged from 0.13 mmol/L to 0.00 mmol/L （-5.61 0.01%）, and the ranges for the three concentrations were 0.10-0.04 mmoUL （-5.71-2.35%）, -0.14-0.01 mmol/L （-5.80-0.30%）, and -0.19-0.04 mmol/L （-6.24-1.22%）. The EQA samples, calibrators, and animal sera exhibited matrix effects in some assays; human serum pools were commutable in all assays; certificate reference materials were commutable in most assays, and only GBW09152 exhibited a matrix effect in one assay： and aqueous reference materials exhibited matrix effects in most assays. Conclusions： Biases for most assays were within the acceptable range, although the accuracy of some assays needs improvement. Human serum pools prepared from patient samples were commutable, and the other tested materials exhibited a matrix effect.
文摘Background： Hemoglobin A 1 c （HbA1 c） measurement is of great value for the diagnosis and monitoring of diabetes. Many manufacturers have developed various experiments to determine the HbAlc concentration. However, the longitudinal use of these tests requires strict quality management. This study aimed to analyze the quality of HbAlc measurement systems in China using six sigma techniques to help improve their performances. Methods： A total of 135 laboratories were involved in this investigation in 2015. Bias values and coefficients of variation were collected from an HbA 1 c trueness verification external quality assessment program and an internal quality control program organized by the National Center of Clinical Laboratories in China. The sigma （σ） values and the quality goal index （QGI） were used to evaluate the performances of different groups, which were divided according to principles and instruments. Results： The majority of participants （88, 65.2%） were scored as ＂improvement needed （σ 〈 3）＂, suggesting that the laboratories needed to improve their measurement performance. Only 8.2% （11/135） of the laboratories were scored as ＂world class （σ≥ 6）＂. Among all the 88 laboratories whose σ values were below 3, 52 （59.1%） and 23 （26.1%） laboratories needed to improve measurement precision （QGI 〈8.0） and trueness （QGI 〉 1.2）, respectively; the remaining laboratories （13, 14.8%） needed to improve both measurement precision and trueness. In addition, 16.1% （5/31） and 15.0% （3/20） of the laboratories in ＂TO SOH＂ and ＂ARKRAY＂ groups, respectively, were scored as ＂world class＂, whereas none of the laboratories in ＂BIO-RAD＂ group were scored as ＂world class＂. Conclusions： This study indicated that, although participating laboratories were laboratories with better performance in China, the performances were still unsatisfactory. Actions should be taken to improve HbAlc measurement performance before we can include HbAlc assays in diabetes diagnosis in China.
基金the National Key Research and Development Program of China(2017YFA0105103,2021YFA0805903)the National Natural Science Foundation of China(81941004,32170542)+10 种基金2020 Research Program of Sanya Yazhou Bay Science and Technology City(202002011)Major Science and Technology Projects of Hainan Province(ZDKJ2021030)Key Research&Development Program of Hainan Province(ZDYF2021SHFZ052)Youth Innovation Promotion Association of the Chinese Academy of Sciences(2019347)Young Elite Scientist Sponsorship Program by CAST(YESS20200024)Biological Resources Progaramme,Chinese Academy of Sciences(KFJBRP-017-57)Key Research&Development Program of Bioland Laboratory(Guangzhou Regenerative Medicine and Health Guangdong Laboratory)(2018GZR110104004)China Postdoctoral Science Foundation(2020M682943)Science and Technology Planning Project of Guangdong Province,China(2019A030317010,2020B1212060052,2021B1212040016,2021A1515011110)Science and Technology Program of Guangzhou,China(202007030003)Research Unit of Generation of Large Animal Disease Models,Chinese Academy of Medical Sciences(2019-I2M-5-025)。
文摘Inducible expression systems are indispensable for precise regulation and in-depth analysis of biological process.Binary Tet-On system has been widely employed to regulate transgenic expression by doxycycline.Previous pig models with tetracycline regulatory elements were generated through random integration.This process often resulted in uncertain expression and unpredictable phenotypes,thus hindering their applications.Here,by precise knock-in of binary Tet-On 3G elements into Rosa26 and Hipp11 locus,respectively,a double knock-in reporter pig model was generated.We characterized excellent properties of this system for controllable transgenic expression both in vitro and in vivo.Two att P sites were arranged to flank the td Tomato to switch reporter gene.Single or multiple gene replacement was efficiently and faithfully achieved in fetal fibroblasts and nuclear transfer embryos.To display the flexible application of this system,we generated a pig strain with Dox-inducing h KRASexpression through phiC31 integrase-mediated cassette exchange.After eight months of Dox administration,squamous cell carcinoma developed in the nose,mouth,and scrotum,which indicated this pig strain could serve as an ideal large animal model to study tumorigenesis.Overall,the established pig models with controllable and switchable transgene expression system will provide a facilitating platform for transgenic and biomedical research.
