Objective To investigate protective effects of dexmedetomidine on oxygen-glucose deprivation/reperfusion(OGD/R)-induced neuronal apoptosis.Methods SH-SY5Y cells were differentiated to neurons with ATRA and followed by TPA.According to the results of preliminary experiment, OGD/R modle was constructed by oxygen-glucose deprivation(OGD) for 12 h and reperfusion(R) for another 12 h.During the start of the OGD, neurons were immediately divided into six groups: group D0(0 μmol/L dexmedetomidine), group D1(0.1 μmol/L dexmedetomidine), group D2 (1 μmol/L dexmedetomidine), group D3 (10 μmol/L dexmedetomidine), group D4(100 μmol/L dexmedetomidine), group D5 (1 000 μmol/L dexmedetomidine).After reperfusion 12 h, the cell viability was evaluated by the method of MTT.The cellular apoptosis was observed by flow cytometry method.The protective effects of different concentration dexmedetomidine on OGD/R-induced neuronal apoptosis were investigated.Then in chosen the exact group having protective effects, endoplasmic reticulum stress specific protein mesencephalic astrocyte-derived neurotrophic factor (MANF) and pro-apoptotic protein Caspase-3 and CHOP were detected by Westernblot method.Results Compared with group D0, there was no difference on the cell viability and cellular apoptosis induced by OGD/R in groups D1 and D2, but a significant decrease and increase in groups D4 and D5 (P<0.01 or P<0.05).And only group D3 had a neuroprotective effect, significantly increased the cell viability and inhibited the apoptosis (P<0.01).Further studys found that group D3 significantly up-regulated ER stress specific protein MANF (P<0.01) and inhibited up-regulation of Caspase-3 and CHOP (P<0.01).Conclusion These data suggest that 10 μmol/L dexmedetomidine had neuroprotective effect on OGD/R-induced neuronal apoptosis and significantly increased cell viability.Our results also indicate that up-regulation of ER stress specific protein MANF and inhibition of CHOP and Caspase-3 by MANF are involved in the neuroprotective effects of Dexmedetomidine.

OBJECTIVEThis study was designed to explore the incidence rate of occult HBV infection in patients with anti-HBc positive alone and analyze the possible reasons of occult infection.

METHODSSera of 183 patients carrying anti-HBc alone(A < or = 0.1) were collected and real-time PCR was used to select samples with HBV DNA positive. HBV pre-S/S amplification products were obtained by PCR, and clonal sequencing were then used for these samples with HBV DNA positive.

RESULTSDNA quantitative results of three samples were greater than 10(3) copies/ml in 183 samples, with a fraction of 1.6%. Pre-S/S sequencing results of two samples from these three samples were obtained. Point mutations within "a" determinant with Q129R/P mutations and co-existence of the mutant type and wild type were found in the two samples.

CONCLUSIONSOccult HBV infection existed in samples with anti-HBc alone. Factors contributing to the loss of HBsAg detection by immunoassays include S gene mutations and low levels of circulating antigen which are below the assay limit of detection. Occult HBV infection not only can lead to a false clinical diagnosis, but also can result in hematological pollution due to such occult infection of blood donors.

Base Sequence ; Blood Donors ; DNA, Viral ; analysis ; Genotype ; Hepatitis B ; diagnosis ; immunology ; Hepatitis B Antibodies ; Hepatitis B Core Antigens ; isolation & purification ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; genetics ; immunology ; Humans ; Polymerase Chain Reaction ; Protein Precursors ; genetics

Objective To compare the effect between intra-arterial and intra-venous neoadjuvant chemotherapy(NACT)in stage Ⅰb2-Ⅱ b cervical carcinoma.Methods A retrospective analysis Was done on 52 cases of intra-venous NACT and 95 eases of intm-arterial NACT for stage Ⅰ b2-Ⅱ b cervical carcinoma treatad in Peking Union Medical College Hospital from 1999.ResulIs The response rate of intraveHous NACT and intra-arterial NACT was 88%(46/52)and 79%(75/95).and the operative rate after NACT Was 81%(42/52)and 72%(68/95)respectively(P>0.05).There were no significant differences in surgery time,blood loss and pest-operative morbidity between these two groups.Pathological parametrial positive rate after NACT in arterial group(6%)Was significantly lower than that of venous group (50%,P>0.05).The venous group had very similar recurrence rates(13%vs 17%)and death rates (9%VS 12%)when compared with the arterial group(P>0.05).Conclusions The intra-arterial and intra-venous NACT for stage Ⅰ b2-Ⅱb cervical carcinoma show similar response rate.operative rate and surgical difficulties.Arterial NACT shows a better effect on parametrial infiltration.

Objective To do screening acute lymphoblastic leukemia patients scFv antibody single chain variable region to cre-ate conditions for the expression and obtain further specificity of antibody fragments.Methods In this study,patients with newly diagnosed acute lymphoblastic leukemia serum as coating antigen using phage display technology,screening phage an-tibody specificity from the semi-synthetic human phage antibody libraries,the first to target the immune antigen-coated tab-let,phage library was added,so that with the target antigen-specific binding phage antibody was immobilized on plates immu-nization,could not be specifically bound phages were rinsed.The eluted specific binding phage,E.coli infection.Could get the specific antibody gene containing phagemid.Results After three “adsorption-elution-amplification”screening process,got stronger leukemia patient antigen-specific phage antibody variable region fragment and identification.Conclusion Got better strain affinity antibody fragments,to create the conditions for the next fragment expression,identification and clinical appli-cation.

Objective To screen permethrin human single-chain variable region (scFv) antibody for aims of developing rapid detection kit. Methods Phage display technology was used in this study. Permethrin was solid phase coated on Nunc plate as antigen. Semi-synthetic single-chain variable region of human antibody library technology was applied, and single chain variable region was screened from phage antibody library after 3 rounds "adsorption - elution - amplification" of the selection process. 100 clones were random selected as resistance to permethrin clones , enzyme-linked immunosorbent assay (ELISA), crossreactivity and competitive inhibition experiments were used to validate permethrin binding activity with strong scFv clones from the selected phage antibody clones plasmid. The plasmid was digested with restriction enzyme Sfi Ⅰ / Not Ⅰ and subcloned into pCANTAB5E vector. After transformed into E. coli XL1BIue, the plasmid was identified by restriction enzyme analysis. Results After screening in 100 clones, 18 clones had high ELISA absorbance values ( A value) at 490nm wavelength ( A490nm), then bovine serum albumin (BSA) cross-reactions identified five weak cross-reaction. Combined with the triplicate ELISA and competitive inhibition experiment results, one positive clone was acquired at last. And this clone was subcloned into pCANTAB5E vector and transformed into competent cells XL1-Blue. Conclusion Plasmid fragment was consistent with the purpose, which provided the foundation for further study of its specific affinity.

Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.

Objective To evaluate the prognosis and fertility outcomes of patients with early stage of cervical cancer treated by vaginal radical trachelectomy ( VRT ) in combination with laparoscopic pelvic lymphadenectomy.Methods The surgical data , disease recurrences and fertility outcomes were analyzed retrospectively for 51 patients who received VRT in Peking Union Medical College Hospital from Dec.2003 to Nov.2013.Results Forty-eight patients succeeded in preserving fertility.The median age was 29 years.International Federation of Gynecology and Obstetrics ( FIGO ) stage: 5 cases Ⅰa1 with lymph vascular space invasion (LVSI),4 cases Ⅰa2 and 39 cases in stage Ⅰb1.Tumor size: 20 cases with no visible lesion, 20 cases with tumor size ≤2 cm, 8 cases with tumor size >2 cm.Histological type:42 cases with squamous carcinoma , 6 cases with adenocarcinoma or adeno-squamous carcinoma.The mean excised cervical length and parametrial width was ( 2.6 ±0.6 ) cm and ( 1.9 ±0.5 ) cm, respectively.Six recurrences ( 12%) were observed after following up for a mean duration of ( 35 ±21 ) months.The recurrent rate in patients with tumor size >2 cm was 3/8, which was significantly higher than that of the
patients with tumor size ≤2 cm (8%, 3/40;P<0.01).Of the 35 patients who desired to conceive after the surgery, 13 women had 17 pregnancies and the pregnant rate was 37% (13/35).Nine women obtained 10 healthy live birth babies.The fertility rate was 26%( 9/35 ).Conclusions VRT in combination with laparoscopic pelvic lymphadenectomy could preserve the fertility of patients with early stage of cervical cancer with acceptable oncologic and fertility outcomes.Tumor size ≤2 cm should be emphasized as the indication of VRT in considering of the higher recurrent rate in patients with tumor size >2 cm.

Purpose To study the expression of HK2 in human prostate cancer (PCa) tissues and its effect on malignant phenotype of prostate cancer cells.Methods HK2 expression in PCa tissues was determined by microarray database and immunohistochemical staining.Subsequently,the change of cellular phenotype was detected by glycometabolism kit,CCK-8 kit,and flow cytometry after HK2 knockdown.Results HK2 expression was elevated followed by prostate cancer development.HK2 depletion inhibited cellular proliferation and aerobic glycolysis,and increased the ratio of early apoptosis.Conclusion HK2 expression increases in the process of PCa malignant progression.It plays a critical role in cellular proliferation,glycometabolism,and apoptosis,the mechanism of which needs further exploration.

Objective To further investigate the accuracy of the real-time three-dimensional echocardiography(RT-3DE) in evaluating right ventricular(RV) systolic function.Methods RT-3DE,MRI and SPECT were used to study a total 30 patients including 10 with coronary artery disease,7 with hypertension,5 with cardiomyopathy,5 with atrial septal defect,and 3 with pulmonary hypertension.The RV end-diastolic volume(RVEDV),RV end-systolic volume(RVESV),RV stroke volume(RVSV) and RV ejection fraction(RVEF) were measured.The correlation and the difference between different modalities were compared.Results RT-3DE had a good correlation with MRI in RVEDV,RVESV,RVSV and RVEF (rEDV =0.811,rESV =0.837,rSV =0.818,and rEF =0.701).Also RT-3DE had a correlation with SPECT in RVEDV,RVESV,and RVSV(rEDV =0.526,rESV =0.493,rSV =0.514),but there was no correlation between RT-3DE and SPECT in RVEF (rEF =0.235).Conclusions RT-3DE can assess RV systolic function accurately.

Objective To investigate the role of pre-chemotherapy hemoglobin and platelet levels in the effect of chemotherapy and prognostic outcome in patients with International Federation of Gynecology and Obstetrics(FIGO) stage Ⅰ b2 - Ⅱb cervical cancer treated with neoadjuvant chemotherapy followed by radical hysterectomy.Methods From January 1999 to December 2010,111 patients with FIGO stage Ⅰ b2 - Ⅱ b who underwent chemosurgical treatment at the department of obstetrics and gynecology in Peking Union Medical College Hospital were reviewed.The median age of patients was 42 years (range:21 -68 years).The median level of prechemotherapy hemoglobin and platelet levels was 127 g/L and 266 ×109/L,respectively.Chemotherapy response was evaluated according to the WHO criteria,including complete response (CR),partial response (PR),arable disease (SD) and progressive disease (PD).Patients who achieved CR or PR were defined as responder.Rates of clinical response were compared with the clinicalpathological variables using chi-square test.Multiple logistic regression was carried out to evaluate the relationship among the probability of achieving an optimal clinical response and the variables.The log-rank test was used to compare the homogeneity of progression-free survival and overall survival functions across strata defined by categories of prognostic variables.The Cox proportional hazard model was used to assess the significance of potential prognostic factors for progression-free survival and overall survival.Results All patients received one to three cycles of chemotherapy.After the neoadjuvant chemotherapy,9 patients achieved CR,77 patients PR,23 patients SD, 2 patients PD.The overall response rate was 77.5％(86/111).By univariate analysis,the clinical response rate was associated with tumor grade( P =0.026),deep cervical stromal invasion ( P =0.029 ) and positive lymph nodes ( P =0.048 ).By multiple logistic regression,deep cervical stromal invasion ( P =0.015 ) and positive lymph nodes ( P =0.031 ) were independent predictors of optimal clinical response.By log-rank test,5-year overall survival rate and 5-year progression-free survival rate were associated with lymph nodes metastases status and lymphovascular invasion ( P =0.000),but not with hemoglobin and platelet levels( P ＞ 0.05 ).By Cox regression model,lymph nodes metastases status and lymph-vascular space involvement ( P ＜ 0.01 ) were independently prognostic factors of 5-year overall survival rate and 5-year progression-free survival rate.Conclusion Pretreatment hemoglobin and platelet levels were neither predictors of clinical response to chemotherapy nor prognostic factors.