AIM: To study whether mesenchymal stem cells (MSCs) from fetal liver can differentiate into skeletal muscle-like cells. METHODS: MSCs were isolated from C57BL/6J mouse fetal liver and were induced by 5-azacytidine and amphotericin B. Myf5 and myogenin were tested by RT-PCR. Desmin and ?-actin was examined by immunocytochemistry. RESULTS: RT-PCR showed that the treated cells expressed Myf5 and myogenin orderly from 6 hours to 72 hours, while the untreated cells did not. Immunocytochemistry confirmed that these cells were positive for desmin and ?-actin and the positive rate was higher in 7 days than that in 14 days. CONCLUSION: MSCs derived from fetal liver can be induced into skeletal muscle-like cells in vitro.

Purpose:To investigate the relationship between HELICOBACTER PYLORI (Hp) and gastric carcinoma, as well as the possible carcinogenic mechanism by Hp. Methods: DNEL technique and immunohistochemical staining were used in the research to study the state of apoptosis, proliferation and p53 gene expression. Total 100 gastric mucosal biopsy specimens, including normal mucosa, Hp negative and Hp positive gastric precancerous lesions as well as gastric carcinomas were studied. Results:There were several apoptotic cells in the superficial epithelium and a few proliferative cells within neck of gastric glands, and no p53 protein expression in normal mucosa. In gastric carcinoma, apoptotic cells accounted for 1.62%, while proliferative cells 4.99%, and P53 protein significantly increased. in metaplasia, the apoptotic index (AI), proliferative index (PI) and positivity of p53 expression in Hp positive group were higher than those in normal mucosa ( P

Objective:To explore potential differentiation from antigen-1 positive stem cells (Sca-1 +cells) in murine fetal liver tissue into cardiomyocytes in vivo. Methods: Sca-1 + cells from the murine fetal liver of 14.5 d were separated by Magnetic cell sorting (MACS).The sequence of the sex determination region of the Y chromosome was determined by PCR.Sca-1 + cells (2?10 3) of male mice were transfused to female mice receiving 8-12 weeks irradiation at a lethal dose of ?-ray(10 Gy) from a 60Co source.Two months later,the mice were killed and the hearts were removed to make slices. Fluorescence in situ hybridisation (FISH) tests with a Y chromosome probe were used to detect sex of the cells.Immunohistochemistry with antibodies of desmin,Flt-1,white blood cell common antigen CD45 and macrophage F 4/80 were used to detect tissue characteristics of cardiomyocytes. Results: Cells with Y chromosome were found in the cardiomyocytes of female mice transplanted with Sca-1 + cells,and showed phenotypic characteristics of Desmin +/Flt-1-/CD45-/F- 4/80 at the same time.Conclusion: Sca-1 + cells from the murine fetal liver are mostly hematopoietic stem cells,having the potential of differentiating into cardiomyocytes.

AIM: To establish HPLC for determination of 2,3,5,4′tetrahydroxystilbene-?-O-?-D-glucoside in Radix polygoni multiflori preparata. METHODS: The column was diamonsil C_(18)(5 ?m,4.6 mm?250 mm).The mobile phase was CH_3CN-CH_3OH-H_2O(10∶20∶70) and ?=320 nm.The flow rate was 1.5 ml/min. RESULTS: The linearity was in the range of 0.0253-0.3542 ?g.The average recovery and RSD were 99.34% and 0.49%,respectively. CONCLUSION: The method is simple,precise and reproduciable and can be used for determination of 2,3,5,4′-tetrahydroxystilbene-?-O-?-D-glucoside in Radix polygoni multiflori preparata effectively.

AIM: To study the effect of acute renal failure (ARF) on the differentiated frequency of Sca-1+ cells from murine fetal liver in irradiated mice. METHODS: The Sca-1+ cells from murine fetal liver were isolated with magnetic cell sorting (MACS) technique, the sex of which was identified by PCR. The 2?104 Sca-1+ cells were transplanted into a lethally irradiated ([ 60Co], 8 Gy) inbred female mouse. After 8 weeks, these recipient mice were divided to A, B, and C groups at random (A group: irradiated; B group: ARF; C group: ARF and Sca-1+). The mice in B and C groups were induced to ARF with 50% (V/V) glycerin (11.6 mL/kg). 72 hours later, the mice in C group were injected with the fresh prepared Sca-1+ cells again. 8 weeks later, mice were sacrificed, and their kidneys were taken out, fixed and slices were prepared. Fluorescence in situ hybridization (FISH) of renal slices was performed and the pictures of them were taken and analyzed. RESULTS: The cells containing Y chromosome were found in renal slices from the mice in A, B and C groups, which located in epithelial cells of renal tubules, interstitium, glomeruli, and glomerular margin and increased gradually. The double and encircle zone of Y chromosome cells were found in the slices from the mice in B and C groups separately, which was consist of new renal tubules. The differentiation frequency of Sca-1+ cells in kidney in A, B and C groups were (1.65?0.18)%, (8.58?1.34)% and (18.13?1.91)%, respectively, which showed significant difference between former group and later group (P

liver.Combined informational analysis showed that the presenting frequency of the cells containing Y chromosome was consistent with the irradiative sensitivity of the organ.CONCLUSION: These findings suggest that the capability of differentiation of Sca-1 positive cells from murine fetal liver was potentially connected to the extent of damage in these organs when transferred in vivo.

Objective To analyze the correlation between the epithelial‐mesenchymal transition and the clinicopathologic features of IgA nephropathy .Methods A total of 168 patients diagnosed as IgA nephropathy by renal biopsy in Xinqiao hospital from Janu‐ary 2011 to December 2013 were divided into high expression of dual‐positive Snail and a‐SMA group (Group A ,117 cases) and low expression of dual‐positive Snail and a‐SMA group (Group B ,51 cases) according to results of immunohistochemical method .The clinical parameters (age ,gender ,course of disease ,BMI and chemical indicators) and renal pathology grade were compared by statis‐tical analysis .Results There were difference between group A and group B in the course of disease and BMI(P<0 .05) .There were differences between group A and group B in the incidences of creatinine ,blood urea nitrogen ,serum triglycerides and 24‐hour urine protein amount (P<0 .05) .The percentage of Lee′s grade Ⅰ ,Ⅱ ,Ⅲ in group B was significant ,while percentage of Lee′s gradeⅣ + Ⅴin group A was significant .The expression of Snail and a‐SMA in grade Ⅳ + Ⅴ was more than that in grade Ⅰ + Ⅱ (40 .2%vs .9 .4% ) ,and the difference between two groups was statistically significant (P< 0 .05) .Conclusion The expression of Snail and‐SMA were related with 24‐hour urine protein amount and kidney function in IgA nephropathy ;and Lee′s grade was severe in patients with high expression of Snail and‐SMA .

[Objective]To examine the effects of fast setting calcium phosphate scaffold on human mesenchymal stem cells(MSCs) adhesion and proliferation.[Method]Scaffolds were self-hardened in molds.Three ml human bone marrow aspirates,taken form the iliac crest of normal donors were suspended in low glucose DMEM supplemented with 10% fetal bovine serum.Atfer 72 hours the nonadherent cells were removed by changing medium,and the adherent layer cultured until it reached 80% conference.Cultured cells at passage 3 were used for the experiments.MSCs were seeded on calcium phosphate scaffold.The cell-scaffold constructs were cultured with complete media for 1 week.Scanning electron microscope(SEM)and the terazolium-based colorimetric assay(MTT test)examinations were performed.[Result]When cells were cultured in scaffolds in vitro,the number of cells on scaffold increased with time.SEM showed that the scaffold had a three-dimensional interconnected pore structure.The cells appeared to infiltrate into the macropores of the scaffold and the extracellular matrix components were observed.[Conclusion]This study demonstrates that the fast setting calcium phosphate scaffold has good biocompatibility,cells are able to adhere and proliferate on scaffold specimens,and it is a good substrate candidate for tissue engineered bone substitute.

Objective To observe the instant reactions and relationship of astrocytes(ASs) and neurons(Ns) in rat forebrain after the unilateral tibia and fibula bone fracture. Methods With immunohistochemical triple staining method,the expression of Fos-protein,glial fibrillary acidic protein(GFAP) and tyrosine hydroxydase(TH) were observed. Results Here we showed 1.After the nociception of the treatment,GFAP-like immunoreactive (-LI) ASs exhibited clear character of nuclei distribution in the lateralmedial habenular nucleus(LHb),paraventricular nucleus of the hypothalamus(Pa),supraoptic nucleus(SON),suprachiasmatic nucleus(SCh),bed nucleus of of stria terminalis(BST),central amygdaloid nucleus(Ce) medial amygdaloid nucleus(Me) and cortex;2.The distribution of Fos-LI Ns and GFAP-LI ASs in above nuclei were similar,there were close relationship between Fos-LI Ns and GFAP-LI ASs.3.There were many double-labelled Fos/TH-LI Ns that were surrounded by the GFAP-LI ASs,and formed the neuron-astrocyte complex(N-ASC).Conclusion The ASs as well as Ns of the above nuclei or regions may be involved in instant response and adjustment of the lower extremity bone fracture nociception simultanneously.

Phytoene synthase is a rate-limiting enzyme in the pathway of carotenoid biosynthesis.To aim at establishing a transformation of Ginseng callus cells, elevating the nutritive value through encouraging the composition of corresponding carotenoid,taking Ginseng callus cells as acceptor,psy gene were transformed into cells via Agrobacterium-mediated. The factors affecting genetict ransformation were also investigated seperately from concentrations of A.tumefaciens, age of Ginseng callus cells, infection time and co-culture time. PCR,PCR-Southern and RT-PCR analysis from the transferred gene plants proved that psy gene has been transferred into Ginseng callus cells and can be expressed. The content of ?-carotene was increased by an average of 26 times.A base of improving and raising the contents of carotenoid in the Ginseng callus cell was established.