Objective To investigate the expression of sphingosine kinase 1(SphK1)and NF-κB in colon carcinoma tissues and their correlation with clinicopathologic features.Methods Sixty-six paraffinembedded colon carcinoma samples and 66 fresh colon carcinoma samples were tested using immunohistochemistry,RT-PCR and Western blot,respectively.Results In 66 fresh colon carcinoma samples,the positive rate of SphK1 and NF-κB mRNA expression were 84.85％(56/66)and 74.24％(49/66),while the positive rate of SphK1 and NF-κB protein detected by Western blot were 78.79％(52/66)and 69.70％(46/66).The positive rates were higher than those in the adjacent tissues[mRNA:63.64％(42/66),48.49％(32/66);protein:57.58％(38/66),45.45％(30/66)]and the normal mucosa [mRNA:42.42％(28/66),25.76％(17/66); protein:36.36％(24/66),24.24％(16/66)],with statistical significances(all P values ＜ 0.05).The mean expressive levels of SphK1 and NF-kB mRNA and protein in colon carcinoma were both significantly higher than those in the adjacent tissues and the normal mucosa(mRNA:0.55±0.06 vs0.35 ±0.05 vs0.25±0.05,0.75 ±0.06 vs0.43±0.05 vs0.30±0.04 ; protein:0.77 ± 0.05 vs 0.38 ± 0.06 vs 0.12 ± 0.03,0.45 ± 0.08 vs 0.23 ± 0.05 vs 0.13 ± 0.03 ;all P values ＜ 0.05).There was a close correlation between SphK1 and NF-kB expression levels (r =0.459,P =0.036).The results of immunohistochemistry were similar to those of RT-PCR and Western blot.Overexpression of SphK1 and NF-κB in colon carcinoma was related with depth of invasion,distant and lymph node metastasis and Dukes'stages(all P values ＜0.05).The expression of SphK1 was also related with differentiation(P ＜ 0.05).Conclusions Overexpression of SphK1 and NF-κB may be involved in the pathogenesis and progression of colon carcinoma.Moreover,SphK1 and NF-κB may be correlated with the invasion and metastasis of colon carcinoma.

Objective To investigate clinical effect of in vitro maturation(IVM)of immature oocytes transferred from conventional in vitro fertilization embryo transfer(IVF-ET) cycles.Methods From January 2008 to June 2009.medical documents of 155 infertile patients underwent IVF-ET in the Reproductive Medical Center of Fimt Affiliated Hospital of Wenzhou Medical College were analyzed retrospectively.If more than 20 oocytes were monitored after 5-7 days of ovulation induction or follicular developmental retardation were confirmed after 8-13 days of ovulation induction.according to patients' wish,IVM were transferred in 60 cycles(group A).In the mean time.IVF was continued in 95 cycles (group B).The mean dosage of gonadotropin,the cancel lation rate of cycles,the mean numbers of oocytes retrieved and maturation,the rate of fertilization and excellent quality embryos.pregnancy outcome and the incidence rate of ovarian hyperstimulation syndrome(OHSS)were compared and analyzed.Results The rates of embryo transfer were 92%(55/60)in group A and 63%(60/95)in group B,which showed significant differences (P<0.05).In group A,the mean dosage of the gonadotropin,the mean number of oocytes retrieved,the cleavage rate and OHSS rate were(1030±468)U,10±6,82.2%(231/281)and O,and were(1544±338)U,14±4,94.0%(502/534)and 35%(21/60)in group B,respectively,all data abeve exhibited statistieal difference between two groups(P<0.05).However,the rates of fertilization and excellent quality embryos had no significant differences between two groups(P>0.05).In group A,the rate of clinical pregnancy per transfer was 53%(29/55)and multiple pregnancy was 14%(4/29),and were 47%(28/60) and 32%(9/28)in group B,they all had no significant differences (P>0.05).Conclusion IVM of immature oocytes used in conventional IVF cycles not only obtained a high clinical pregnancy rate,but also reduced gonadotropin using dosage and avoided OHSS completely.

Objective To evaluate the therapeutic effect of endoscopic management of plastic stents of post-liver transplantation anastomotic biliary stricture. Methods From January 2010 to October 2015, clinical data of patients with post-liver transplantation anastomotic biliary stricture and received endoscopic retrograde cholangiopancreatog﹣raphy and plastic stents management was collected. The technical success rate, ERCP-related complications, clinical remission rate and long-term complications were main outcome measurements to compare the efficacy and safety of different number of stents in managing post-liver transplantation anastomotic biliary stricture. Results Among the 18 patients (0.5 ~ 60.0 months) with post-liver transplantation ABSs, seven patients received less plastic stents treat﹣ment (< 3 stents), nine patients with persistent anastomotic or recurrent stricture received multiple plastic stents treatment (≥ 3 stents), two patients received multiple plastic stents treatment once suffered with post-liver trans﹣plantation ABSs. The endoscopic technical success rate was seventy-six over eighty (95.0 %). Among the seven pa﹣tients received less plastic stents treatment, one loss to follow-up, two were still under treatment, one died of acute hepatic failure, one died of septic stock, one combined with biliary fistula resulted in treatment failure, one achieved clinical remission, the clinical remission rate was one third (33.3 %). Among the eleven patients received multiple plastic stents treatment, two loss to follow-up, one was still under treatment, two received surgery because of failed treatment, six achieved clinical remission, the clinical remission rate was 75.0 % (6/8). The average diameters and stent durations of management of 1 stent, 2 stents, 3 stents, 4 stents, 5 stents, 7 stents were 8.5 F, 17.0 F, 24.0 F, 28.0 F, 36.0 F, 50.0 F. Among the six early postoperative complications, five cases occurred in less stent manage﹣ment and one occurred in MPSs management, the early postoperative complication rate was 7.5 %(6/80). No severe ERCP-related complications and procedure-related deaths. Conclusions Endoscopic management of plastic stents is safe and effective for post-liver transplantation ABSs. Providing larger biliary support, the multiple plastic stents treatment was superior to less plastic stents treatment in view of clinical remission rate, especially for refractory one. Multiple plastic stents did not increase the incidence of complications, it could be used as the first-line treatment of post-liver transplantation duct-to-duct biliary anastomosis for its safety and effectivity.

Objective To investigate the correlation between sperm-nucleporotein transition and sperm parameters and embryo development,also to evaluate the influence of pregnancy out comes of assisted reproductive technology (ART).Methods Sperm-nucleoprotein transition assay of a total of 676 patients underwent ART treatment were detected by aniline blue staining,and the correlation analysis between spermnucleoprotein transition and sperm parameters,DNA damage,acrosin activity,fertilization rate,cleavage rate,quality of early embryo development as well as blastocyst formation rate was performed.Results The sperm concentration,(a+b) ％ sperm,sperm count and acrosin activity was (66.5±4.6) × 109/L,(149.2±9.9)×109/L,(51.2±1.3)％ and (72.2±3.3) mU/106 sperm in abnormal group,and (91.9±2.7) ×109/L,(240.0±8.0) ×109/L,(57.3±0.8)％ and (85.7±1.9) mU/106 sperm in normal group,which reached significant difference (P＜0.01).DNA fragmentation index (DFI) was (17.3± 1.0)％ in abnormal group,which was significantly higher than (14.6±0.5)％ in normal group.The cleavage rate of 95.0％,D3/D5 high quality embryo rates of 34.2％ and 1.28％,D5 blastocyst formation rate and the total rate of blastocyst formation rate of 22.4％ and 38.6％ in abnormal group,which were significantly lower than that in normal group (96.9％,38.2％,2.70％,27.9％ and 46.4％) (P＜0.01).The rate of spontaneous abortion was 12.3％ in abnormal group,which was significantly higher than that in normal group (4.7％) (P＜0.01).However,there was no significant difference in biochemical pregnancy rate and ectopic pregnancy rate between the 2 groups (P＞0.05).Conclusions Sperm-nucleoprotein transition was positively related with sperm parameters,DNA damage,acrosin activity,and also has an adverse effect on embryo development and the outcomes of ART.It is suggested that the sperm-nucleoprotein transition should be detected before ART.

[ ABSTRACT] AIM:To investigate the effects of sphingosine kinase l ( SphK1) and focal adhesion kinase ( FAK) on the epithelial-mesenchymal transition ( EMT) of human colon cancer HCT 116 cells.METHODS:Human colon cancer HCT116 cells were divided into 3 groups.N, N-dimethylsphingosine (DMS) was used to suppress the activity of SphK1. PF573228 was used to suppress the activation of FAK .The cells treated with equal volume of culture medium severed as control group.The cell viability was measured by MTT assay .The protein expression of SphK1, FAK and the EMT relative protein E-cadherin, N-cadherin, vimentin and matrix metalloproteinase (MMP) 2 was analyzed by Western blot.The mR-NA expression of SphK1, sphingosine-1-phosphate (S1P), FAK, E-cadherin and vimentin was detected by real-time PCR. The ability of tumor cell migration was measured by wound-healing assay.RESULTS:The cell viability of HCT116 cells was suppressed by DMS and PF 573228 in dose and time dependent manners .DMS significantly suppressed the expression of SphK1, FAK, N-cadherin, vimentin and MMP2, meanwhile enhanced the expression of E-cadherin.PF573228 reduced the expression of FAK , SphK1, N-cadherin, vimentin and MMP2, meanwhile increased the expression of E-cadherin (P<0.01).In addition, the migration ability of HCT116 cells was significantly decreased by treating with DMS and PF573228 (P<0.01).Compared with control group , the mRNA expression of FAK, SphK1, S1P and vimentin was de-creased, while the expression of E-cadherin was increased significantly in PF573228 group and DMS group (P<0.05). CONCLUSION:SphK1 and FAK signaling pathways may play an important role in the occurrence of EMT in the colon cancer HCT116 cells.

Objective To construct an eukaryotic expression vector of human telomerase reverso transcriptase (hTERT) gene specific shRNA, and investigate the effect of pshRNA-hTERT combined with γ-irradiation on telomerase activity and DNA damage. Methods The recombinant expression plasmid pshRNA-hTERT was constructed and transfected into Hep-2 cells. The telomerase activity was examined by the PCR-hased telomeric repeat amplification protocol (TRAP). DNA single-stranded break (SSB) and the DNA double-stranded break (DSB) were detected by Comet assay. The xenograft model of human laryngeal carcinoma with the same genetic background and different radiosensitivity (Hep-2 and Hep-2R) was established in nude mice. The mixture of pshRNA-hTERT and liposome was injected to the transplanted tumor to observe the inhibition of the tumor growth. The cell apoptosis was detected by TUNEL. The hTERT protein expression was determined by streptavidin-peroxidase conjugated method (AP). Results Recombinant expression plasmid pshRNA-hTERT was successfully constructed and transfected into Hep-2 cells. The hTERT expression inhibition rate reached 60.78 %. pshRNA-hTERT not only inhibited telomerase activity of Hep-2 inehiding the increase of telomerase activity induced by γ-irradiation, but also inhibited the repair of the SSB and DSB induced by irradiation in the human laryngeal carcinoma xenograft in nude mice with the same genetic background and different radiosensitivity. The pshRNA-hTERT combined with γ-irradiation could inhibit the growth of transplanted tumor (Hep-2: EPO = 1.79; Hep-2R: EPO = 2.01) with reduced telomerase activity and hTERT protein expression. Conclusions The eukaryotic expression vector pshRNA-hTERT could enhance the radiosensitivity of Hep-2 cells in vitro and the human laryngeal carcinoma xenograft in nude mice which had the same genetic background with different radiosensitivity.

Objective To investigate high risk human papillomavirus(HR-HPV)prevalence among married women in Beijing and to study the high risk flactors.nethods During March 2007 to September 2008.a total of 6185 married women sampled from 137 communities in 12 districts were screened bv HR-HPV DNA test and cytogical test.The interview was carried out with unified questionnaires.The databage was set up and twice entered in EpiDam 3.0.After checked up,the data were analyzed in SPSS 15.0.Results (1) The HR-HPV infection rate was 9.89%.The HR-HPV infection rate of the city zone,the suburb and the exurb were 9.34%,10.51% and 9.51% (P>0.05).The HR-HPV infection rate of the native and the oudander were 9.53%,11.30% (P<0.05).(2) The age distribution of HR-HPV infection was that the rate was around 10% among 25 to 44 age groups,which was the highest(11.21%) in 30 to 34 age group;then the rate was descended as the age raising,the rate of 50 to 54 age group was the lowest(7.78%).(3) Multiple logistic regression showed that the related risk factors of HR-HPV infection mainly included 1000 RMB and above of family income per person per month.possessing more than 1 sexual partner of her husband,outlander and hish levels of education.(4) The prevalence of cervical intraepithelial neoplasia(CIN)in HR-HPV positive group wag significantly higher than that in HR-HPV negative group(29.76% vs 3.32%,P<0.01).Conclusions(1)The HR-HPV infection rate among aged 25 to 54 years was 9.9% and there was no significant difference in area distribution.(2)The hish risk population which should strengthen screening was the married bearing-age women with high level of family income,outlander,high levels of education and her husband possessing more than 1 sexual partner.(3)HR-HPV infection is the main risk factor for CIN and cervical cancer.while does not provide a causal relationship with them.The high risk population should be checked regularly to understand the development of HR-HPV infection and CIN incidence.

OBJECTIVE：To provid e reference for the medicinal resources and clinical application of Tibetan medicine “Dida”. METHODS：Delphi method was adopted. By reviewing literatures ，confirming consultation scope ，inviting experts engaged in clinical，scientific research ，teaching and production of Tibetan medicine. Two methods ，online inquiry and on-site questionnaire ， were used for expert consultation to evaluate the differences and problems existing in the utilization of “Dida”medicinal resources until a consensus was reached ，consensus on the medicinal resources and clinical application of Tibetan medicine “Dida”was determined finally. RESULTS & CONCLUSIONS ：A total of 33 experts participated in the two rounds of consultation. According to the results of literature research ，the first round set up 16 inquiry indicators ；and then according to expert opinions to modify the index system ，the second round set up 18 inquiry indicators. After two rounds of inquiry ，a consensus was finally reached on 16 items on the original name ，quality standards and clinical application of Tibetan medicine “Dida”，and 2 items related to the characteristics and compatibilities of “Dida”had not reach common views. The consensuses of 16 items mainly cover the original name of “Dida”and the evolution of geographic information ，the rational selection of “Dida”medicinal materials ，the effectiveness and safety of clinical use of “Dida”.

Acute pancreatitis (AP) is a severe disease with an increasing incidence rate in clinical practice. Although most patients have mild pancreatitis, the fatality rate of severe pancreatitis remains at a relatively high level, and therefore, early-stage, simple, and accurate clinical scoring systems are urgently needed to determine the severity of AP, so as to facilitate effective disease management and symptomatic treatment and reduce the fatality rate of patients. At present, a large number of studies have demonstrated that the scoring systems such as Ranson score, APACHE Ⅱ score, BISAP score, CTSI score, and some serological markers have been used to evaluate the severity and prognosis of AP, but all of them have certain limitations. This article reviews the research advances in the existing scoring systems, single serological markers, and related modified scoring systems in recent years. Through a literature review, it is concluded that there is no a single scoring system or a single indicator that can cover the whole process of AP diagnosis and treatment and accurately judge the severity of AP, and therefore, it is necessary to develop a new scoring system or combine various indicators for comprehensive evaluation.