BACKGROUND:Bone marrow is the main source of mesenchymal stem cells(MSCs)at present,but its application has been limited,because of some reasons such as inconvenience of isolation,and the quantity of cells decreases with human increased age.Umbilical cord as a new source of MSCs has been widespread concerned recently.OBJECTIVE:To explore the approach of isolating and culturing MSCs from Wharton's jelly of human umbilical cord,and the methods of identifying the surface antigens and the differentiation potential.METHODS:MSCs were isolated and amplified via tissue-cultivation,and cultured by FasGrow medium.Morphology of MSCs from Wharton's jelly of human umbilical cord was observed under the optical microscope.Its immunophenotypes were detected using immunohistochemistry.The differentiation of MSCs into the osteoblasts was determined utilizing Gomori calcium-cobalt alkaline phosphatase staining,von Kossa calcium node staining,and tetracyclinefluorescence labeling.The differentiation of MSCs into the adipocytes was detected using oil red O staining.RESULTS AND CONCLUSION:MSCs were easily obtained from Wharton's jelly of human umbilical cord via the proposed approach.The primary cells grew up to 70%-80% confluence after 12-16 days of culture,and meanwhile the undifferentiated state was maintained and proliferation was stabilized after passage.The cell cycle of double increase was about 2 days,and proliferation in vitro reached twenty generation above.Surface antigen analysis showed that CD44,CD105,CD133,MHC-I were positively expressed,while CD34,CD45 were negatively expressed.Experiments of differentiation in vitro indicated that the obtained cells were capable of differentiating into fat,osteoblast and nerve-like cells.

Objective To research the removal of the low concentration of formaldehyde in the indoor air by using copper sulfate.Methods The low concentration of formaldehyde(10.0 mg/L)in the indoor air was determined by the way of MBTH spectrophotometry.The influence of pH,chelon and concentration on the removal of different concentration formaldehyde was investigated by the way of chemisorption.Results When pH was 11.99,12.86,13.08 and 13.42,using copper sulfate,the removal rate of 10.0 mg/L formaldehyde was 43.82%,62.75%,69.21% and 73.40% respectively.When the concentration of copper sulfate was at 3.0 g/L,5.0 g/L,7.0 g/L and 10.0 g/L,the removal rate was 51.43%,73.40%,66.36% and 62.18% respectively in the condition of pH=13.42.When used potassium sodium tartrate and EDTA as the ehelon,pH=13.42,concentration of copper sulfate was 5.0 g/L,the removal rate of 2.0 mg/L formaldehyde was 77.21% and 62.51% respectively,that of 10.0 mg/L formaldehyde was 86.54% and 73.40% respectively,that of 100.0mg/L formaldehyde was 96.71% and 91.32% respectively.Conclusion Using potassium sodium tartrate as the chelon,at pH=13.42,5.0 g/L copper sulfate can produce a good removal efficiency for indoor low level formaldehyde.

20?10 9/L) was observed (mean 41 days, range 12 to 118 days). There w as the association between the number of CD34 +CD44 + cells infusion and time to neutrophic recovery (?= -0.657 , P

Objective To explore the chromosome aberration of cervical cancers induced by human papillomavirus(HPV16)virus.Methods The change of change of chromosomes of Hela cells induced by HPV-16 infection HPV-16 in fection was detected by FISH.Results The integration sites of HPV-16 were not observed in Hela cells.But HPV16 virus was integrated on chromosomes 3.Conclusion HPV16 could cause chromosome 3 aberration in Hela cells and result in the occurance of cervical cancers.

BACKGROUND: At present, the transplantation of bone marrow-derived endothelial progenitor cells (BM-EPCs) or bone marrow-derived hepatocyte stem cells (BDHSCs) is common in the treatment of liver fibrosis, but the combined treatment for liver fibrosis is rarely reported. Combined transplantation of BM-EPCs possessing the function of angiogenesis and BDHSCs possessing the function of hepatocyte regeneration might play a dual anti-fibrosis role. OBJECTIVE: To evaluate the reversal effect on liver fibrosis by the combined transplantation of BM-EPCs and BDHSCs in rats. METHODS: The liver fibrosis rat models were induced with CCl4 subcutaneous injections for 6 weeks. BM-EPCs of rats with liver fibrosis were obtained by culture induction in vitro.BDHSCs of rats with liver fibrosis were obtained by magnetic bead cell sorting.BM-EPCs and/or BDHSCs were transplanted into liver fibrosis rats via the tail vein and branch of the portal vein,and then the effects of BDHSCs transplantatiron on liver fibrosis and liver function were observed. RESULTS AND CONCLUSION: (1) Masson staining results showed transplantations of BDHSCs and BM-EPCs, alone or both, could suppress the formation of collagen fibers. However, the staging scores of liver fibrosis showed that only the combined transplantation of BM-EPCs and BDHSCs could significantly improve liver fibrosis,which was significantly different from the model group(1.75±0.25 vs. 3.00±0.19, P < 0.05). (2) The liver biochemical assay in the blood showed that the levels of all five parameters of alanine aminotransferase, aspartate aminotransferase, total bilirubin, prothrombin time, and activated partial thromboplastin time in the BM-EPCs/BDHSCs group were significantly improved to be equivalent to normal levels, compared with those in the model group (P < 0.05). To conclude, it is an effective treatment for liver fibrosis by the co-transplantation of BM-EPCs and BDHSCs.

Objective To up-regulate the expression of Glil gene in periodontal ligament stem cells ( PDLSCs) and to explore the effect of Glil gene on PDLSCs proliferation and osteogenesis differentiation by establishing Glil gene adenovirus vectors. Methods Subcloned Glil to viral backbone vector Adtrack-CMV and transfered the established vector to 293T cells, which was to acquire the virus particles. Trans-fected aim cells,namely PDLSCs,with these virus. Detected its effect on PDLSCs proliferation with CCK-8 assay, and detected the expression of Glil and the bone-related markers ALP and Runx2 through Western blot. Results An adenovirus vector, which were over expressed Glil gene, was successfully constructed and transfected to PDLSCs. Compared with the empty vector group and normal group, the over expressed one had a much slower proliferation rate in CCK-8 assay (P=0. 003). Western blot showed that ALP and Runx2 can be overexpressing os-teogenic differentiated after PDLSCs successfully transfected with the Glil gene. Conclusion Over expressing Glil gene would lead to a much slower proliferation rate in the PDLSCs and an increase of the bone-related markers. It is concluded that Glil can enhance the osteogenic dif-ferentiation capacity in PDLSCs.

Objective To explore anti-EV71 effects of Ganlu Xiaodudan in vitro. Methods Ribavirin was taken as control drug, with the help of cell culture to observe anti-EV71 inhibition rate of Ganlu Xiaodudan in inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment, preventive-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment. Results In inhibiting-virus-directly experiment, therapeutic-inhibiting-virus experiment and preventive-therapeutic-inhibiting-virus experiment, virus inhibition rate of Ganlu Xiaodudan was higher than ribavirin. In preventive-inhibiting-virus experiment, virus inhibition rates of Ganlu Xiaodudan and ribavirin both were almost zero. Conclusion Ganlu Xiaodudan has better antiviral effects on EV71 than ribavirin, and it can affect more than one link of multiplication of EV71.

AIM:To study the chemical constituents of Erigeron Injection by HPLC. METHODS: Under the guidance of HPLC analysis,two constituents were separated and purified by the aid of column chromatography with silica gel,and identified by UV、 IR、 MS、 NMR respectively and documentary data. RESULTS: Two compounds from semi-finished products of Erigeron Injection were identified as caffeic acid(Ⅰ) and chlorogenic acid(Ⅱ). CONCLUSION: Two main peaks in HPLC chromatogram of Erigeron Injection are Ⅰ and Ⅱ.

Carcinoma, Hepatocellular ; metabolism ; pathology ; Hep G2 Cells ; Humans ; Interleukin-6 ; metabolism ; Liver Neoplasms ; metabolism ; pathology

Objective To develop a new method for simultaneous quantifying and genotyping of HBV in a single reaction based on dual molecular beacon real-time PCR.Methods Genotype B and C recombinant plasmids were constructed as the standards and genotype-specific primers and molecular beacons were designed for each genotype.The molecular beacons of genotype B and C were labeled with FAM and Hex respectively.In this way,a simultaneous qualification and genotyping method for HBV DNA in a single real-time PCR reaction system was developed.Firstly,10-fold gradient dilution of genotype B and C standard plasmids (103-1011 kIU/L) were utilized to evaluate the linear ranges and sensitivity of this approach.The clinical specificity was tested with twenty different serum specimens (5 cases with hepatitis C virus,5 cases with herpes simplex virus and 5 cases with human papilloma virus as well as 5 healthy volunteers) ; the reproducibility was assessed by intra-assay and inter-assay coefficient of variation (CV) of cycle threshold (Ct) value through 10 repeated detections within a batch and between batches of the B,C standard plasmids (108,106 and 104 kIU/L).Then the accuracy of qualifying and genotyping of the self-built method was evaluated by a parallel examination with 132 HBV infected patients by use of two commercial kits as the references.Finally,these HBV-positive patients were divided into 4 groups:asymptomatic carrier (n =21),chronic hepatitis (n =77),liver cirrhosis (n =25) and hepatocellular carcinoma (n =9) to investigate the relationship of genotypes,stages of disease progression and HBV DNA load.Results A simultaneous qualification and genotyping assay was successfully built and its genotyping sensitivity was 103 kIU/L and the linear range was 103-1011 kIU/L.The intra-assay CV of B genotyping was 1.51％ to 1.80％ and the interassay CV was 2.11％ to 3.03％,while the intra-assay CV of C genotyping was 1.79％ to 1.95％ and the inter-assay CV was 2.53％ to 2.91％.The results of non HBV infected cases and healthy volunteers showed negative.In the test of 132 HBV infected patients,the general coincident rate of genotyping results comparing our assay and HBV DNA genotyping kit was 90.9％ (120/132,Kappa =0.832,P ＜ 0.05).The HBV DNA quatitive results between the assay[5.07 (3.89-6.33)] and HBV DNA quatitive kit [5.19 (4.15-6.32) lg kIU/L] were well correlative (R2 =0.8477,P ＜ 0.05).69 genotype B cases,51 genotype C cases and 12 B/C mixed-genotype cases were detected by dual molecular beacon real-time PCR method and their HBV DNA load were 4.54 (3.83-6.17),5.53 (4.02-6.55),4.58 (3.68-4.98) lg kIU/L respectively.Where the patients with genotype C had higher DNA load than the patients with other two genotypes (Z =-2.195and-2.162,P ＜ 0.05).The HBV DNA load of asymptomatic group,chronic hepatitis group,liver cirrhosis group and hepatocellular carcinoma group were 7.02 (6.35-7.84),4.94 (4.16-6.25),4.37(3.50-5.17) and 3.45 (3.25-4.92) lg kIU/L,respectively.Among them,the asymptomatic group was significantly higher than those of other three groups (Z =-4.244,-4.568 and-3.489,P ＜0.001) and DNA load comparing with the chronic hepatitis group,liver cirrhosis group and hepatocellular carcinoma group also showed statistically different (Z =-2.894 and-2.413,P ＜ 0.05).However,compared with the liver cirrhosis group and hepatocellular carcinoma group there was no significant difference (Z =-0.995,P =0.335).Conclusion A dual molecular beacon real-time PCR assay which can simultaneously quantifying and genotyping HBV DNA with highly accuracy,sensitive and specificity is successfully developed.(Chin J Lab Med,2013,36:333-338)