Objective To investigate the relationships and clinicopathologic features of the expressions of cytochrome P450 2E1 (CYP2E1) and PC cell-derived growth factor (PCDGF) in esophageal squamous cell carcinoma (ESCC),and to determine the role of CYP2E1 and PCDGF in angiogenesis.Methods The expression levels of CYP2E1 and PCDGF in 42 surgical cancer specimens and 20 adjacent normal esophageal mucosa specimens from patients with ESCC were detected by immunohistochemistry.Results The positive expression rates of CYP2E1 and PCDGF in ESCC were 83.3 ％ (35/42) and 88.1％ (37/42),respectively,which were obviously higher than those of normal mucosa (P ＜ 0.05).Expression of PCDGF was correlated with the degree of histological differentiation (r =0.444,P ＜ 0.05),depth of tumor invasion (r =0.332,P ＜ 0.05),lymph node metastasis (r =0.476,P ＜ 0.05),and TNM classification (r =0.450,P ＜ 0.05).The expression of CYP2E1 was negatively correlated with tumor tissue differentiation (r =-0.518,P ＜ 0.05),and positively correlated with depth of invasion in ESCC (r =0.388,P ＜ 0.05).The expression of CYP2E1 was related to that of PCDGF in the tumor (r =0.483,P ＜ 0.05).Conclusion The expressions of CYP2E1 and PCDGF are synergistically involved in tumor growth,infiltration andmetastasis.Overexpression of CYP2E1 and PCDGF can be used as the important predictors for evaluating the biological behavior of ESCC and predicting prognosis of patients.

Objective To establish an easier and better method for primary culture of rabbit fibroblast-like synoviocytes(FLS) by modified tissue cell culture.Method Healthy rabbit joint synovial layers were cut into small pieces and siphoned off water attached on the tissue with sterile filter paper and then cultured.The growth status and morphology were observed.The growth curve of the 3rd passage cells was measured by CCK-8 assay, and the expression of vimentin was detected by immunocytochemistry.Results The FLS cultured in vitro exhibited a spindle-shaped appearance and could rapidly expand.The cell growth curve was typical of S type, and the cells highly expressed vimentin.Conclusions Primary culture of rabbit FLS by the improved explant culture method is successfully established.The method is simple and highly efficient.It provideds a new method for isolation of FLS.

BACKGROUND:It is unclear whether serial cel passage in vitro influences the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s.
OBJECTIVE:To investigate the effect of cel passage on the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s.
METHODS:Rat bone marrow mesenchymal stem cel s were isolated and cultured by the whole bone marrow adherence method. Bone marrow mesenchymal stem cel s at passages 3, 6, 9, 12 were incubated in serum-free medium. After culture for 7 and 14 days, cel biological characterization was observed and differenitaiton ability into neural stem cel s was observed by detecting Nestin expression in cel s using flow cytometry. Then, the cel s were further induced to differentiate and cel multipotential differentiation capacity was detected by measurement of nerve enolase and glial acidic protein expression.
RESULTS AND CONCLUSION:Under induction, bone marrow mesenchymal stem cel s at different passages were al differentiated into Nestin-positive neural stem cel s. However, there was a significant difference in differentiation proportion of cel s at different passages (P<0.05). Strongest differentiation ability was found in the passage 6 cel s, with the Nestin expression up to (93.7±2.3)%at 7 days of induction and (96.2±1.8)%at 14 days of induction. The proportion of differentiated cel s at passages 6 and 9 was signfi cantly higher than that at passages 3 and 12. Moreover, adherent cel s were positive for nerve enolase and glial acidic protein. Al these findings indicate that the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s is correlated with cel passage. Cel s at lower or higher passages are both detrimental to cel differentiation.

Objective:To isolate rabbit fibroblast-like synoviocytes(FLSs) by suspended explant culture method. Methods:The healthy rabbit joint synovial layers were obtained. The cells were cultured by suspended explant culture method and compared with the explants adherent culture method. The growth status and morphology were observed. The growth curve of the 3rd passage cells was measured by CCK-8 assay,and the expression of vimentin was tested by immunocytochemistry. Results: The FLS obtained by the two methods exhibited a spindle-shaped appearance and could rapidly expand. The cell growth curve was typical of S type, and the cells highly expressed vimentin. Conclusion:Primary culture of rabbit fibroblast-like synoviocytes by suspended explant culture method were successfully established. The method was simple and highly efficient. It provided a new method for the isolation of FLS in vitro.

BACKGROUND:Currently, fibroblast-like synoviocytes from the synovial tissue of patients with rheumatoid arthritis or adjuvant arthritis rats are isolated and cultured mainly using enzyme digestion method, but tissue explant adherent method is rarely reported in primary culture. OBJECTIVE:To establish a culture method of fibroblast-like synoviocytes from adjuvant arthritis rabbits and to investigate cell biological characteristics. METHODS:Rabbit models of adjuvant arthritis were prepared by injected with adjuvant at multiple sites and times, the knee synovial tissues were obtained, and fibroblast-like synoviocytes were separated and cultured by the tissue explant adherent method. Furthermore, the cell morphology was observed. The cell proliferation was measured using cell counting kit-8 assay, and the expression of Vimentin was detected by immunofluorescence.RESULTS AND CONCLUSION:The primary cultured rabbit synoviocytes were long spindle fiber-like and coincided with the fibroblast-like synoviocytes. Moreover, cells were positive for Vimentin. In conclusion, fibroblast-like synoviocytes from rabbits with adjuvant arthritis are successfully isolated and cultured by the tissue explant method.

AIM: To explore the relationship between methylation status of promoter region 5′CpG island and the biological phenotype in human colorectal cancer RKO cell lines. METHODS: RKO cells were treated with selective DNA methyltransferase (DNMTs) inhibitor, 5-Aza-2′-deoxycytidine (5-Aza-CdR), for 72 h. Methylation-specific PCR (MSP), T-A clone and DNA sequence analysis were used to detect 5′CpG island methylation status of p16/CDKN2 tumor suppresor gene. Cell growth, cell cycle arrest and apoptosis were analyzed by MTT, flow cytometry (FCM), fluorescent dye staining and transmission electron microscope. RESULTS: DNMTs inhibitor (5-Aza-CdR) effectively reversed the hypermethylation status of 5′CpG island. The effects of 5-Aza-CdR on cell growth inhibition (P

Objective To determine the expression of human protection of telomeres 1 (hPOT1) and its relationship with Helicobacter pylori (HP) infection in gastric carcinoma.Methods The expressions of hPOT1 protein and hPOT mRNA were detected in 53 gastric carcinoma specimens (observed group) and 20 normal gastric mucosa tissues (control group) by SP immunohistochemical method and in situ hybridization (ISH), respectively.HP infection was examined by Warthin-Starry method in observed group.Results The positive expression rate of hPOT1 protein was 84.91％ (45/53) in observed group, higher than that in control group [30.00 ％ (6/20)] (P ＜ 0.01).The positive expression rate of hPOT1 mRNA was 58.49 ％ (31/53) in observed group, higher than that in control group [10.00 ％ (2/20)] (P ＜ 0.05).The positive co-expression rate of hPOT1 protein and mRNA was 56.60 ％ (30/53), both had positive relationship in gastric carcinoma (r =0.394, P ＜ 0.05).The rate of HP infection in 53 cases of gastric carcinoma was 52.83 ％ (28/53).The positive expression rates of hPOTI protein and mRNA in observed group with HP infection were significantly higher than those in observed group without HP infection[protein: 96.43 ％ (27/28) vs 72.00 ％ (18/25), P ＜0.05;mRNA: 85.75 ％ (24/28) vs 28.00 ％ (7/25), P ＜ 0.01].Conclusions hPOT1 may be associated with occurrence of gastric carcinoma.Combined detection of hPOT1 protein and mRNA can be used for the diagnosis of gastric carcinoma.HP infection may be associated with abnormal expression of hPOT1 in occurrence of gastric carcinoma.

Objective To explore the efficacy of tepronone and folic acid in the treatment of chronic atrophic gastritis (CAG) evaluated by the marking targeting biopsy (MTB).Methods A total of 224 H.pylori negative CAG patients were selected and divided into group A (n 96,tepronone 50 mg/time,folic acid 10 mg/time,three times/day),group B (n=23,tepronone 50 mg/time,three times/day),group C (n=74,unspecific treatment) and group D (n=31,no treatment).The treatment course lasted for one year.The clinical symptoms improvement of each group was observed before and after treatment.The pathological improvement of gastric mucosa by MTB was inspected before and after treatment.The chi square test was performed for the comparison between groups.Results The total efficacy rates of group A,B,C and D were 43.8％ (42/96),39.1％ (9/23),33.8％(25/74) and 32.3％ (10/31) respectively,there was no significant difference between groups (x2 =2.328,P =0.507).For the significant efficacy rate of gastric mucosa pathological improvement,group A was compared with group D,group A was compared with group C and group B was comparedwith group D,the differences were significant (x2 =14.520,14.628 and 8.995,all P＜0.01).In the total efficacy rate of gastric mucosa pathological improvement,group A (49.8％,131/263) was compared with group D (24.2％,16/66),group A was compared with group C (35.9％,66/184)and group B (44.7％,21/47) was compared with group D,the differences were significant (x2 =13.953,8.535 and 5.207,all P＜0.05).Conclusion Teprenone alone or teprenone and folic acid combination can obviously improve pathological changes of CAG patients.

YAEH.But ET was no statistical significance discrepancy among DBQY,DBYY and QSBS.Conclusion: In clinic to aim directly at left ventricular diastolic dysfunction of senile diabetics,no matter syndrome differentiation belonging to DBQY,DBYY,QSBS,all can benefit after treatment with supplementing QI and activating blood circulation method;but syndrome differentiation belonging to YAEH advocates not to use supplementing QI and activating blood circulation method.

Aim To investigate the effects of diallyl di-sulfide( DADS) on G2/M arrest in Chk1/MGC803 and Chk2/MGC803 cells so as to establish stable human gastric cancer MGC803 cells with overexpression of Chk1/2 gene. Methods The colony formation, flow cytometry, RT-PCR and Western blot were used to de-tect the proliferation, cell cycle, and expression of Chk1/2 mRNA and protein, p-Chk1/2, CDC25C and cyclinB1, respectively. Results The colony formation showed that the colony forming efficiency in Chk1/MGC803 and Chk2/MGC803 cells treated by 30 mg· L-1 DADS was lower than in control group and vector group ( P <0. 05 ) . Flow cytometry demonstrated that 41. 3%, 57. 4%, 68. 9% and 42. 9% of G2/M cells in Chk1/MGC803 were increased than in MGC803 and Chk2/MGC803 , respectively after treated by DADS in 12,24, 36 and 48 h(P <0. 05). At the same time, RT-PCR disclosed that expression of Chk1 and Chk2 mRNA had no marked change. Western blot showed that total proteins of Chk1 and Chk2 and p-Chk2 had invisible change, but expression of p-Chk1 was up-reg-ulated, and CDC25C and cyclinB1 were down-regula-ted time-dependently in Chk1/MGC803 cells ( P <0. 05 ) . Conclusion DADS arrests MGC803 cells at G2/M by increasing p-Chk1 expression to cause down-regulation of CDC25C and cyclinB1 simultaneously.