Objective To clone and express Schistosoma j aponicum cyclophilin B (SjCyPB) gene in E.coli,and to identify and analyze the immunity of recombinant proteins.Methods A pair of specific primers was designed according to GenBank of Schistosoma japonicum sequence.SjCyPB gene was amplify by PCR and then connected to pET28 vector.The recombinant plasmid pET28a (+)-SjCyPB was constructed and transformed into E.coli BL21 cell line,the recombinant plasmid was identified by double enzyme digestion and sequence analysis.After induced by isopropyl-B-D-thiogalaetoside (IPTG),the expressed recombinant proteinwas purified by affinity-chromatography,and then verified by Western blotting.Rats were immunized recombinant SjCyPB,andthe SjCyPB-specific IgG was detected by ELISA.Results SjCyPB gene was successfully inserted into pET28a(-) vecter which identified by double enzyme digestion and sequence analysis.Recombinant SjCyPB protein was highly expressed in E.coli.The Western Blotting analysis confirmed that the recombinant protein could specifically combine to S.japonicum-infected rabbit serum.Using recombinant protein to immunize rats,the SjCyPB-specific IgG antibody titer was 1 ∶ 51 200 detected by ELISA.Conclusion The recombinant SjCyPB is successfully constructed,and recombinant SjCyPB has immunogenicity and antigenicity.

BACKGROUND: Proximal humeral fractures are one of the most common types of upper limb fractures In adults. With the aging of the population, the Incidence of proximal humeral fractures has been increasing in recent years, and its treatment is still controversial. OBJECTIVE: To compare the clinical efficacy of locking plate and intramedullary nail in the treatment of proximal humeral fractures using meta-analysis. METHODS: We retrieved PubMed, the Cochrane Library, Embase, CNKI, Wanfang, and VIP database to collect studies on intramedullary nail and locking plate in the treatment of proximal humeral fractures. In accordance with the inclusion and exclusion criteria, studies were included. Data were extracted from the included studies. Continuous variables (Constant score, operation time, blood loss) and binary classification variables (total complications, postoperative infection, screw cutout and internal fixation failure, humerus head necrosis, secondary surgery, and acromial impact) were analyzed, and considered as evaluation indexes for the meta-analysis. Statistical software Stata12.0 was used to analyze the data. RESULTS AND CONCLUSION: (1) A total of 8 articles and 891 patients were included, and 9 data were analyzed. (2) The amount of blood loss (SMD=-1.82, 95%C/: -2.77—0.87, P < 0.001) and operation time (SMD=-1.47, 95%C/: -2.13—0.80, P < 0.001) in the intramedullary nail group were superior to those in the locking plate group. The remaining results were not statistically significant between the two groups (P > 0.05). (3) These results confirmed that intramedullary nailing is superior to locking plate in operation time and intraoperative blood loss in the treatment of proximal humeral fractures. The results were similar in total complication rate, common complication rate and postoperative recovery of shoulder joint function between the two groups.

Pdx-1, an important transcription factor highlighting in the early pancreatic development, islet functions and pancreatic disorders, needs to be more investigated in zebrafish, and siRNA is still seldom applied in zebrafish embryo-related research. Our aim was to explore the role of pdx-1 in pancreatic development of zebrafish embryos by using siRNA approach. Microinjection, reverse transcriptase-PCR (RT-PCR), in situ hybridization and immunofluorescent staining were used in this research, and the morphology of the islet in normal zebrafish embryos, and in those treated with the siRNA specific to pdx-1 (siPDX-1) or siGFP was observed and compared. The expression of pdx-1 was detected in the stages of 1-cell, 2-cell, 4-cell, 8-cell, 16-cell, 16-hour by RT-PCT. The in situ hybridization and immunofluorescent staining results showed that siPDX-1 disturbed the formation of the islet in zebrafish embryos. Pdx-1 played multiple roles in maintaining the phenotype of the islet during embryogenesis in zebrafish.
Embryo, Nonmammalian ; Homeodomain Proteins/genetics ; Homeodomain Proteins/*metabolism ; Islets of Langerhans/cytology ; Islets of Langerhans/*embryology ; Islets of Langerhans/metabolism ; RNA Interference ; RNA, Small Interfering/*genetics ; Trans-Activators/genetics ; Trans-Activators/*metabolism ; Zebrafish

Objective To evaluate the possible association among serum folate,polymorphisms related reduced folate carrier gene (RFC-1) AS0G,methionine synthase reductase gene (MTRR) A66G,and cervical cancer,and to provide clues for the etiology of cervical cancer.Methods Based on a hospital-based case-control study,107 cases diagnosed as cervical cancer pathematologically and 107 controls with hysteromyoma,were selected by frequency,matched with age and habitation.Serum folate concentration was detected by RIA and polymorphism RFC-1 A80G and MTRR A66G was examed by RFLP-PCR.Results Serum folate concentration in patient group [(1.86±0.60) ng/rml] was significantly lower than that in control group [(2.30 ± 1.14) ng/ml],and risk of cervical cancer increased with the decreased serum folate levels (x2trend =12.57,P =0.001).Risks to catch cervical cancer for women with RFC-1 80 GG were 2.42 times (95 ％ CI 1.01-5.81) as much as for those with RFC-1 80 AA,and 1.65 times (95 ％ CI 0.77-3.53) for those with RFC-1 80 AA and RFC-1 80 AG,risks to catch cervical cancer for women with MTRR 66 GG were 1.35 times (95 ％ CI 0.40-4.56) as much as for those with MTRR 66 AA and 1.26 times (95 ％ CI 0.38-4.16) for those with RFC-1 80 AA and RFC-1 80 AG.Conclusion Serum folate deficiency to a certain degree can increase the risk of cervical cancer.RFC-1 A80G mutation may be a risk factor for cervical cancer and homozygous (GG) gene may increase the susceptibility of cervical cancer.MTRR A66G gene mutation may have nothing to do with cervical cancer.

Purpose To study the clinic-pathologic features, differential diagnosis and prognosis of lymphoepithelioma-like carcinoma of cervix. Methods Eight cases of lymphoepithelioma-like carcinoma of cervix were retrieved from our institution. HE staining, im-munohistochemistry and in situ hybridization were perform on those cases. Results Of the 8 cases of lymphoepithelioma-like carcino-ma,the median age was 44 years old ( ranged from 29-67 years) and all of the clinical stage belonged toⅠB. Grossly, 5 cases presen-ted a cauliflower-like appearance, 1 case grew as a polyp protrusion, and the other 2 cases showed a non-specified thicken change of cervix. Histologically, the tumors were composed of undifferentiated large cells, either loosely or closely-packed, with the nested and trabecular growth pattern. The tumor cells were generally oval or polygonal in shape, while the demarcation between the cells was indis-tinct and form the syncytial appearance. The nuclei were vesicular and had one or more prominent nucleoli. There were abundant infil-trates of lymphocyte and plasma cells among the tumor nests. The tumor cells showed positive expression of CK in 100% of cases, p63 in 37. 5% of cases, p16 in 62. 5% of cases, CK5/6 in 75% of cases, Ki-67 in 10% ~30% of cases. The stromal lymphocytes mainly expressed CD3 and CD8. EBER was negative in the entire tumor cells while was weakly positive in the stromal lymphocytes in one case. The follow-up data, which terminated at June 2013, demonstrate 7 cases were alive and 1 case died of the disease. Conclusions Lymphoepithelioma-like carcinoma occurred of cervix is rare. They have distinctive morphologic features and overall favorable prog-nosis. The accurate diagnosis is dependant on the combination of the histologic and immunohistochemical examination.

Objective To investigate the hand hygiene status of urban and rural residents in Hangzhou City of Zhejiang Province to provide scientific base for health promotion.Methods Urban residents,rural residents,personnel in government institutions,urban students,rural students and enterprise workers were enrolled in this investigation.By using constant volume sampling method,fifty individuals in each group were recruited from 13 districts of Hangzhou City,and a total of 3 337 people filled in the questionnaire,which included demographic characteristics,knowledge of hand hygiene and hand washing.Logistic regression analysis and x2 test were used for data analysis.Results Correct hand-washing was found in 35.84％ urban and rural residents (urban vs.rural:41.45％ vs.27.06％).Awareness rate of hand hygiene was 35.06％.The influencing factors of proper hand washing were location,gender,age,educational level and understanding of hand hygiene.The influencing factors of awareness of hand hygiene were location,gender,age and educational level.Conclusion Hangzhou citizens had a relatively lower rate of proper hand washing,and significant difference was found between urban and rural residents; the study suggested that para-urben residents need to improve their knowledge and behavior of hand hygiene.Meanwhile,the students urgenty need to improve their hand hygiene

Objective To investigate the effects of retrograde cardioplegia on serum soluble inter cellular adhesion molecule (sICAM)-1,soluble P-selectin (sPs) and cardiac troponin Ⅰ(cTnI) in the process of heart valve replacement operation.Methods Fifty-six cases undergoing heart valve replacement with cardiopulmonary bypass (CPB) were divided into antegrade cardioplegia (AC) group and retrograde cardioplegia (RC) group.Serum levels of sICAM-1,sPs and cTnI were measured using enzyme-linked immunosorbent assayd (ELISA) at six times.Results After the CPB,the levels of sICAM-1,sPs and cTnI ascended quickly,at the time of 8-hour after operation achieved the peak value.The peak value of RC group were lower than those of AC group[sICAM-1 (817.9±133.2)μg/L vs(901.1±132.0)μg/L,sPs(83.46±16.24)μg/L vs(107.82±19.68) μg/L,cTnI(10.50±2.03) μg/L vs (14.45±2.26)μg/L],and the dedined degree of RC group were faster than those of AC group.Conclusion Retroperfusion has a better myocardial protection to the patients suffering heart valve replacement operation through active suppressed expression of sICAM-1,sPs and cTnI palliated inflammatory reaction.

Objective To evaluate the effect of lithium chloride (LiCl) pretreatment on isoflurane-induced cognitive dysfunction and inflammatory response in hippocampus in aged rats.Methods Eighty 20-month-old male Sprague-Dawley rats,weighing 350-400 g,were randomly assigned into 4 groups (n =20 each):control group (group C),1.4％ isoflurane group (group I),100 mg/kg LiCI + 1.4％ isoflurane group (group L+ I),and 100 mg/kg LiC1 group (group L).Group I was exposed to 1.4％ isoflurane in 30％ O2-70％ N2 for 6 h,while group C was exposed to 30％ O2-70％ N2 only.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and isoflurane anesthesia was performed on 4th day in group L + I.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and then the rats inhaled 30％ O2-70％ N2 for 6 h on 4th day in group L.Blood samples were taken immediately after the end of anesthesia for blood gas analysis.Hippocampi were isolated 24 h after the end of anesthesia for determination of the expression of glycogen synthase kinase-3β (GSK-3β) and acetyl-NF-κB (Lys310) (by Western blot) and TNF-α,IL-1β and IL-6 mRNA (by RT-PCR).The levels of TNF-α,IL-1β and IL-6 were determined by ELISA and the contents of TNF-α,IL-1β and IL-6 were calculated.The cognitive function was assessed on 2nd day after the end of anesthesia.Results Compared with group C,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly up-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were increased,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened in group I (P ＜ 0.05).Compared with group I,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly down-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were decreased,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in group L + I (P ＜ 0.05).Conclusion LiC1 pretreatment can improve isoflurane-induced cognitive dysfunction and inhibition of inflammatory response in hippocampus is involved in the mechanism in aged rats.

The HPLC-MS/MS method was developed to profile the dynamics of abscisic acid (ABA) and ABA-glucose ester (ABA-GE) after cloning glycosyltransferase enzyme family gene AtUGT71C5 into Arabidopsis thaliana. By constructing over-expression lines (OE) and down-expression lines (DN), we acquired mutant strains to analyze the function of AtUGT71C5. The multiple-reaction monitoring (MRM) was used for quantitative determination in negative mode. The transition was m/z 263.1-153.0 for ABA ([M-H]t), m/z 425.1-263.0 for ABA-GE ([M-H]t), and m/z 321.0-152.0 for chloramphe-nicol. The linear range was 0.8684-217.1 ng/mL for ABA and 0.3920-196.0 ng/mL for ABA-GE. The accuracy was 88.0-109.0% for ABA and 86.6-113.0% for ABA-GE; the inter-day and intra-day precisions were less than 5.4%for ABA and 8.9%for ABA-GE, respectively. This method is simple and sensitive enough for determination of ABA and ABA-GE in A. thaliana leaves. All the evidence confirmed the speculation that AtUGT71C5 can mediate abscisic acid homeostasis.

Pdx-1, an important transcription factor highlighting in the early pancreatic development,islet functions and pancreatic disorders, needs to be more investigated in zebrafish, and siRNA is still seldom applied in zebrafish embryo-related research.Our aim was to explore the role of pdx-1 in pan-creatic development of zebrafish embryos by using siRNA approach. Microinjection, reverse tran-scriptase-PCR (RT-PCR), in situ hybridization and immunofluorescent staining were used in this re-search, and the morphology of the islet in normal zebrafish embryos, and in those treated with the siRNA specific to pdx-1 (siPDX-1) or siGFP was observed and compared. The expression of pdx-1 was detected in the stages of 1-cell, 2-cell, 4-cell, 8-cell, 16-cell, 16-hour by RT-PCT. The in situ hy-bridization and immunofluorescent staining results showed that siPDX-I disturbed the formation of the islet in zebrafish embryos. Pdx-1 played multiple roles in maintaining the phenotype of the islet during embryogenesis in zebrafish.