Objective To evaluate the significance of immediate early 1 (IE) mRNA detection by nucleic acid sequence-based amplification (NASBA) in the diagnosis of human cytomegalovirus (HCMV) infection after renal transplantation.Methods The expression of IE-mRNA and pp67-(mRNA) was detected by NASBA in 55 patients after renal transplantation, and pp65 antigenemia (assays) were done for all cases.Results Twenty cases were positive for IE-mRNA. Thirteen cases were suffered from CMV disease with symptoms. The sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of IE-mRNA by NASBA were 92.3 %, 80.9 %, 60.0 % and 97.1 %, respectively. Time to first IE-mRNA positive detection after transplantation was shortest, compared with pp67 and antigenemia (P

Gastric cancer is one of the most common malignant tumors in China, and its pathogenesis is complex. At present, there is no good method for the diagnosis and treatment of gastric cancer. However, studies have shown that microRNA (miRNA) has abnormal expression in gastric cancer, which participates in the regulation of gastric cancer related genes, and has an impact on the occurrence, development, diagnosis and treatment of gastric cancer. This paper aims to review the relationship between miRNA and the diagnosis and treatment of gastric cancer, as well as the drug resistance in the treatment, so as to lay the foundation for the follow-up research and clinical diagnosis and treatment.

Objective To evaluate the curative effects and safty of contra-aperture dissection and skin bridge preservation in the treatment of low transphincteric perianal fistula.Methods Sixty subjects of low transphincteric perianal fistula were randomly divided into two groups.Thirty cases of treatment group were treated by contra-aperture dissection and skin bridge preservation,and 30 cases of control group were treated by anal fistulectomy.On the second,seventh and fourteenth day after operation,the postoperative pain,exudate and fever were recorded and scored.The curative time was observed.Anorectal dynamic changes were compared simultaneously to evaluate the curative effects and safety of contra-aperture dissection and skin bridge preservation.Results The scores of pain and exudate were significantly different between the treatment and the control group(P＜0.01).The wound-healing time was shorter in the treatment group than that in the control group((27.37±8.93) d vs.(32.73±9.45) d,P=0.000).There were significant differences in the anal resting pressure,anal maximal contraction pressure and active systolic pressure between the two groups (t =13.12,10.21,12.10;P＜0.01).There was no significant difference of total effect between the two groups(100％ and 93.3％;x2 =2.07,P＞0.05).Conclusion Contra-aperture dissection and skin bridge preservation can reduce postoperative pain,exudate and fever,shorten wound-healing time and protect anal functions in the treatment of low transphincteric perianal fistula.

Two isolation methods for sorting of endothelial progenitor cells (EPCs): from peripheral blood mononuclear cells (PBMCs) and CD133(+) enriched cells were compared, by defining the cell morphology, phenotype, reproductive activities and function in vitro, to provide a reference for clinical application of EPCs. PBMCs from healthy subjects were used either directly for cell culture or for CD133(+) sorting. The two groups of cells were cultured in complete medium 199 (M199) for 7 to 14 days and the phenotypes of EPCs were analyzed by FACS. The proliferation of differentiated EPCs was studied by MTT assay, and the VEGF concentration was measured using an ELISA kit. ECM gel experiment and migration assay were performed in vivo. The results showed that PBMCs produced more colony-forming units (CFU) than CD133(+) enriched cells from the same volume of blood (P<0.01). From day 7 to 14, the two groups showed decreased expression of hematopoietic stem cell markers and increased level of endothelial markers, but CD144(+) cells in CD133(+) group were less than in PBMCs group (P<0.01). PBMCs group secreted more VEGF than CD133(+) group on the day 7 (P<0.01). As compared with CD133(+) group, PBMCs group had more potent potential of proliferation and vascularization in vitro. It was concluded that CD133(+) sorted cells showed a lower capacity of differentiation, secretion, proliferation and vascularization in vitro, suggesting that CD133-negative cells may be a preferential way to get EPCs for clinical therapy.

AIM: To compare the methods of two currently employed isolation methods for endothelial progenitor cells (EPCs): from total peripheral blood mononuclear cells (PBMCs) and from enriched CD133~+ cells, by defining the cell morphology, phenotype, reproductive activities and function in vitro, providing a reference for clinic application. METHODS: PBMCs from the healthy subjects were used for CD133~+ sorting or not. The two groups of isolated cells were suspended in complete medium M199 for 7 d to 14 d. EPCs phenotype were characterized by FACS. The proliferation of differentiated EPCs was studied by MTT assay, and VEGF concentration was measured using an ELISA kit. Matrigel experiment and migration assay were imitated vascularization in vivo. RESULTS: PBMCs produced more colony-forming units (CFU) than CD133~+ cells from the same volume of blood (P<0.01). From 7 d to 14 d, the two groups show decreased expression of hematopoietic stem cell markers and increased level of endothelial markers, but CD144~+ cells in CD133~+ group were lower than those in PBMCs groups (P<0.01). Cells in PBMCs group secreted more VEGF than that in CD133~+ group on 7 d (P<0.01). Compared to CD133~+ group, PBMCs group showed more potential of proliferation and vascularization in vitro. CONCLUSION: CD133~+ sorted cells show a lower capacity of differentiation, secretion, proliferation and vascularization in vitro, which is unable to differentiate to mature endothelial cells, indicating that it's not a preferential way to obtain EPCs for clinic therapy.

Objective To observe the efficacy of Chinese herbal fumigant sitz bath for different exposure durations on patients for the complicatiors after mixed hemorrhoidectomy.Methods One hundred and twenty patients with mixed hemorrhoidectomy were randomized into four groups(2 treatment groups and 2 control groups) with 30 cases in each group.The treatment groups were treated with the Chinese Herbal fumigant sitz bath for 10 minutes and 15 minutes respectively;while the control groups were treated with the hemorrhoids lotion for 10 minutes and 15 minutes respectively.The severity of pain,edema,bleeding,constipation and urinary disturbance were scored on the second,seventh and fourteenth days after treatment.Data were analyzed statistically.Results Chinese herbal fumigant Sitz baths for 10 minutes could significantly reduce postoperative pain,edema and bleeding symptoms[(1.72±1.23) points,(0.72±1.25) points,(0.91±1.37) points,respectively,P ＜0.05],followed sequentially by groups of Chinese herbal fumigant Sitz baths for 15 minutes [(1.79±1.21)points,(0.89±1.28) points,(1.03±1.24) points]; The hemorrhoids lotion for 10 minutes ((1.86±1.25) points,(1.14±1.47) points,(1.49±1.56)) and 15 minutes[(2.03±1.48) points,(1.05±1.29) points,(1.39±1.47) points,respectively,P ＜ 0.05]With the successful cases of 27,21,18,and 6 in the groups,respectively.Though the overall response rates of for the groups of 10 min's treatment,15min's treatment and 10 min's control were not significantly different between each other(P =0.0634,P =0.2560),they showed a superior efficacy in the overall response rate to the 15 min's control group(P =0.0064,P =0.0365,P =0.0089),respectively.Conclusion Chinese herbal fumigant Sitz baths for 10minutes have a success rate of 100％ on alleviating complications after mixed hemorrhoidectomy.It has a prominent efficacy on reducing postoperative edema and bleeding symptoms than other treatments studied in this program.

Objective:To explore the effect of Wnt/β-catenin signaling pathway on growth plate development of tibial growth plate in young chronic renal failure (CRF) rats.Methods:Four-week-old male SD rats were randomly divided into control group and CRF group ( n=20/per group). Control group was intragastric administration with distilled water, and CRF group was given adenine suspension (150 mg·kg -1·d -1). All the young rats were sacrificed after continuous gavages for 6 weeks. The full length of tibia was compared between the two groups. The width of tibia proximal growth plates was measured by micro-CT scanning, and the width of the growth plate was also measured in histological sections. Chondrocytes isolated from growth plate in two groups were cultured in vitro to P3 generation. Immunohistochemical staining was used to detect the expression of collagen Ⅱ, matrix metalloproteinase 13 (MMP-13) and β-catenin in chondrocytes. Western blotting was used to detect the protein expressions of collagen Ⅱ, MMP-13 and β-catenin. Results:Compared with the control group, the tibial length of rats in the CRF group was shorter [(27.32±5.81) mm vs (35.43±3.61) mm, t=5.226, P<0.001], the width of growth plate in micro-CT picture was more narrow [(0.72±0.22) mm vs (1.13±0.27) mm, t=5.096, P<0.001], and the relative width of the growth plate was also more narrow ( t=6.744, P<0.001) in histological sections. The results of immunohistochemistry and Western blotting showed the expressions of collagen Ⅱ in the CRF group decreased significantly ( t=8.212, P<0.001), MMP-13 ( t=13.091, P<0.001) and β-catenin ( t=7.534, P<0.001) increased significantly compared the control group in chondrocytes. Conclusion:The Wnt/β-catenin signaling pathway is highly expressed in the tibial growth plate of young rats with chronic renal failure, which leads to accelerated degeneration and differentiation of chondrocytes and a closure tendency of growth plate.

Two isolation methods for sorting of endothelial progenitor cells(EPCs): from peripheral blood mononuclear cells(PBMCs)and CD133+ enriched cells were compared,by defining the cell morphology,phenotype,reproductive activities and function in vitro,to provide a reference for clinical application of EPCs.PBMCs from healthy subjects were used either directly for cell culture or for CD133+ sorting.The two groups of cells were cultured in complete medium 199(M199)for 7 to 14 days and the phenotypes of EPCs were analyzed by FACS.The proliferation of differentiated EPCs was studied by MTT assay,and the VEGF concentration was measured using an ELISA kit.ECM gel experiment and migration assay were performed in vivo.The results showed that PBMCs produced more colony-forming units(CFU)than CD133+enriched cells from the same volume of blood(P<0.01).From day 7 to 14,the two groups showed decreased expression of hematopoietic stem cell markers and increased level of endothelial markers,but CD144+cells in CD133+ group were less than in PBMCs group(P<0.01).PBMCs group secreted more VEGF than CD133+group on the day 7(P<0.01).As compared with CD133+ group,PBMCs group had more potent potential of proliferation and vascularization in vitro.It was concluded that CD133+sorted cells showed a lower capacity of differentiation,secretion,proliferation and vascularization in vitro,suggesting that CD133-negative cells may be a preferential way to get EPCs for clinical therapy.