Objective To preliminarily explore the early process of hepatitis B virus (HBV) infection in HepG2 cells induced by dimethyl sulfoxide(DMSO) ,and provide cytological bases for mechanism study of HBV infection in vitro .Methods HepG cells were divided into the DMSO inducing group and control group ,and were cultured 4 days by DMEM containing 1 .5% DMSO and normal DMEM respectively ;changes of cellular morphology were observed .In addition ,selected ECV304 cells as the negative con‐trol group and treated with DMSO .Cells in the three groups were incubated 2 hours with HBV positive serum after culturing 24 hours ,then trypsin digestive solution ,HepG2 cells and ECV304 cells were collected and polymerase chain reaction (PCR) was used for the determination of HBV DNA .Simultaneously ,the blank control group was set ,and the position of HBsAg on HepG2 cells were detected by using indirect immunofluorescence .Results HepG2 cell volume in the DMSO inducing group was obviously in‐creased .HBV DNA was found in HepG2 cells both in DMSO inducing group and control group ,and DMSO inducing group ex‐pressed much stronger .The results of IIF shown that green fluorescent signals of cell membrane and cytoplasm of HepG 2 cells in the DMSO inducing group were increased obviously ,while the results of HBV DNA and IIF both were negative in the negative con‐trol group .Conclusion DMSO could facilitate adsorption of HBsAg in some extent ,which are benefit for completing the process of early infection .

Objective To explore the underlying clinical factors and precautionary measures of fluid extravasation in patients with calyceal calculi treated by ureteroscopic holmium laser lithotripsy.Methods A retrospective review was made on clinical records of 138 patients with calyceal calculi receiving retrograde ureteroscopic holmium laser lithotripsy from May 2005 to March 2009. The relevance was studied between the occurance of fluid extravasation complications and various clinical factors using x2 test and binary Logistic regression. The clinical factors included patients' sexes, age groups (＜30 years, 30-50 years, ＞50 years), history of treatment (ESWL or open surgery) for upper urinary tract calculi, preoperative upper urinary tract infection, intraoperative placement of ureteral catheter and the length of procedure duration (＜ 50 min, 50-80 min, ＞ 80 min). Results Fluid extravasation complications occurred in 24 patients. The sexes and age groups were irrelevant to the occurance of fluid extravasation complications; while history of ESWL or open surgery and preoperative infection in upper urinary tract, without intraoperative ureteral catheter placement and long duration of procedure were responsible for the higher rates of the fluid extravasation complications.Conclusion Reasonable selection of patients and timing of operation, regular intraoperative ureteral catheter placement and control the length of procedure duration help to reduce fluid extravasation during retrograde ureteroscopic lithotripsy.

Objective To detect the mouse testicular gene expression pattern differences be-tween spermatogonial stem cell (SSC) proliferative and differential stages and study the molecular reg-ulation mechanism in SSC proliferation and differentiation. Methods With the interval of 24 days, male Kunming mice were injected intraperitoneally with two doses of busulfan (10 mg/kg) to establish spermatogenesis regeneration models. 36 k Mouse Genome Array was used to detect the differential gene expression profiles between the stages of SSC proliferation and differentiation. Bioinforrnsties analysis was conducted in GO (gene ontology) and KEGG (Kyoto Encyclopedia of Genes and Ge-nomes) pathway to describe the potential roles that may play in spermatogonial stern cells behavior regulation. Results Nine hundred and eleven differential expression genes were identified by gene arrays in mice testes, consisting of 608 up-regulated and 303 down-regulated in SSC proliferation stage and SSC differentiation stage. The differential expression genes were classified by their biological process, molecular function and cellular component, respectively. Alterations with statistical signifi-cance (P<0.05)appeared in 84 KEG(;signal pathways, including Notch and Wnt signaling pathways which had been proved to be important for stem cell maintenance. Fifty-six differential expression genes were selected as genes related to stem cells, among which 40 genes were up-regulated, including some stem cell biomarkers(such as Cd9, StraS, hgbl-, Oct4 and Thyl)and some growth factors(such as Fgf2, Pdgfa and Csfl). Conclustion The regulation of SSC proliferation and differentiation involves inmany differentially expressed genes in various signal pathways. This study provides a molecular basis for the elucidation of the molecular mechanism behind self-renewal and differentiation of spermatogonial stem cells.

Objective:To evaluate the therapeutic effect of CXCL1 monoclonal antibody on dextra sulfate sodium (DSS)-induced acute ulcerative colitis (UC) in mice,and to elucidate its effect on the expressions ofTNF-α,IFN-γ,,IL-17 and IL-10 as well as neutrophil infiltration.Methods:Female BALB/c mice were randomly divided into a normal group (DSS-),a disease group (DSS+saline),an anti-CXCL1 antibody group (DSS+anti-CXCL1 Ab) and a treatment control group (DSS+IgG Ab).The DSS+saline,DSS+anti-CXCL1 Ab and DSS+anti-CXCL1 Ab groups were given 3.5％ DSS solution as drinking water to induce acute intestinal inflammation,while the normal control was given distilled water freely.The DSS+anti-CXCL1 Ab mice were intraperitoneal injected with anti-CXCL1 Ab (4 mg/kg) on the 3rd and 6th day.Same amount of rat IgG Ab was given in the DSS+IgG Ab group.The normal group and the disease group were injected with 0.9％ sodium chloride solution.The value of disease activity index (DAI) and the injury of colorectal tissue were measured.The levels of TNF-α,IFN-γ,IL-10 and IL-17 in colonic tissues of mice were detected by RT-PCR.Myeloperoxidase (MPO),a specific marker of neutrophils was measured by immunohistochemistry.Results:Compared with the normal control group,DAI score and colorectal injury score in the disease group were significantly increased,but the DAI and colorectal in the mice with acute ulcerative colitis tissue damage score were significantly reduced after anti-CXCL1 Ab intervention.Compared with the normal control group,mRNA levels of TNF-α,IFN-γ and IL-17 in the colorectal tissues were significantly elevated (P＜0.05) in the disease group while the IL-10 was decreased;these effects were attenuated by anti-CXCL1 Ab intervention (P＜0.05).Immunohistochemistry showed that the infiltration of neutrophils (MPO+) in the colon tissue was significantly increased in the disease group,while the anti-CXCL1 Ab treatment could significantly reduce the neutrophil infiltration in colon tissue (P＜0.05).Conclusion:Anti-CXCL1 Ab relieves the progression of DSS-induced acute ulcerative colitis by suppressing proinflammatory expression and neutrophil infiltration.

Danshen, the dried roots and rhizomes of Salvia miltiorrhiza Bunge (S. miltiorrhiza), is widely used in the treatment of cardiovascular and cerebrovascular diseases. Tanshinones, the bioactive compounds from Danshen, exhibit a wide spectrum of pharmacological properties, suggesting their potential for future therapeutic applications. Tanshinone biosynthesis is a complex process involving at least six P450 enzymes that have been identified and characterized, most of which belong to the CYP76 and CYP71 families. In this study, CYP81C16, a member of the CYP71 clan, was identified in S. miltiorrhiza. An in vitro assay revealed that it could catalyze the hydroxylation of four para-quinone-type tanshinones, namely neocryptotanshinone, deoxyneocryptotanshinone, and danshenxinkuns A and B. SmCYP81C16 emerged as a potential broad-spectrum oxidase targeting the C-18 position of para-quinone-type tanshinones with an impressive relative conversion rate exceeding 90%. Kinetic evaluations andin vivo assays underscored its highest affinity towards neocryptotanshinone among the tested substrates. The overexpression of SmCYP81C16 promoted the accumulation of (iso)tanshinone in hairy root lines. The characterization of SmCYP81C16 in this study accentuates its potential as a pivotal tool in the biotechnological production of tanshinones, either through microbial or plant metabolic engineering.
Humans ; Salvia miltiorrhiza/metabolism* ; Biosynthetic Pathways ; Quinones/metabolism* ; Plant Roots/metabolism* ; Gene Expression Regulation, Plant