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.

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.

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.

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.

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

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.

To analyze the structural characteristics of the Rap2B gene from Clonorchis sinensis by bioinformatics and to clone and express this gene through genetic engineering methods in order to obtain the corresponding recombinant protein.The structural and functional characteristics of the Rap2B protein were analyzed by using the related bioinformatic softwares. Using the full-length cDNA plasmid clone (Noc009e03) as template , the coding region of Rap2B gene sequence was amplified with PCR and cloned into the prokaryotic expression vector pET-28a(+) and then expressed in E.coli BL21 through IPIG induction. The recombinant protein expressed was purified by Ni-IDA affinity chromatography and detected by SDS-PAGE and Western blotting. It was demonstrated that the size of this gene was 847 bp in length, encoding 141 amino acids and its theoretical molecular weight was 15851.1 daltons. As demonstrated by PCR, double enzyme digestion and DNA sequencing, the recombinant expression plasmid pET-28a(+)-Rap2B was successively constructed and the Rap2B-like gene was highly expressed in E.coli BL21. The recombinant protein was obtained through purification with His affinity chromatography and could react specifically with the serum of rats infected with C.sinensis. Through these investigations, the Rap2B protein may be predicted as a member of the Ras oncogene family protein and is potential in the carcinogenic process of C.sinensis.

AIM: To examine the relationship between corneal lymphangiogenesis and hemangiogenesis after keratoplasty.METHODS: Nineteen human corneas were obtained from 19 patients undergoing a second corneal transplantation in Zhongshan Ophthalmic Center in 2005. Blood and lymphatic vessels in human transplanted corneas were identified by lymphatic vessel endothelial receptor(LYVE-1) and platelet endothelial cell adhesion modecule-1(PECAM-1) immunohistochemistry, and double enzyme-histochemistry; then the association of corneal blood vessel counting (BVC) with lymphatic vessel counting (LVC) was examined.RESULTS: Corneal hemangiogenesis was present in 12 cases (63%), and lymphangiogenesis occurred in 5 cases (26%) of human transplanted corneas. In addition, corneal lymphangiogenesis was only present in vascularized corneas. LVC was strongly and positively correlated with BVC(r=0.725;P<0.01).CONCLUSION: Corneal lymphangiogenesis develops after keratoplasty, and strongly associates with hemangiogenesis.

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.