This study is to establish a cell-based model targeting to neuraminidase (NA) of the 2009 H1N1 influenza A virus. NA is an influenza virus structural protein with enzymatic activity of the cleavage of HA-sialic acid interaction to release new viral particles from cells. A model of HIV-1 (pNL4-3.Luc.R(-)E(-)) based pseudovirions packed with HA [hemagglutinin, A/VietNam/1203/2004 (H5N1)] and NA [A/California/04/2009 (H1N1)] was established to evaluate compounds activities on NA function. The viral release can be blocked by neuraminidase inhibitors, oseltamivir and oseltamivir carboxylate, with IC50 of (61 +/- 31) nmol L(-1) and (5.5 +/- 2.9) nmol L(-1) respectively. A point mutation of H275Y on NA leads oseltamivir-resistance. This corresponding mutation was introduced into the system which was also confirmed by oseltamivir and oseltamivir carboxylate.

Objective To identify the imaging performance and differences between type] and type Ⅱ papillary renal cell carcinoma (PRCC).Methods Data of 21 lesions of type Ⅰ,27 lesions of type Ⅱ (1 patient had 2 lesions) in 47 patients was retrospectively analyxed.All patients with pathologically proven PRCC were examined by contrast CT or MRI preoperatively.The morphological features,outside invasion signs and performance on contrast-enhanced CT were compared by qualitative and quantitative studies.The maximum diameter of tumors and CT values,△CT values in corticomedullary and nephrographic phase were analyzed by two-sample t-test,classified variable were compared by the Pearson X2 test or the Fisher exact test.Results On morphological behaviors,type Ⅱ PRCC were significantly larger than type Ⅰ PRCC (t =-2.604,P =0.013),more heterogeneous (X2 =14.928,P =0.000),greater probability to show cystic degeneration or necrosis (X2 =5.598,P =0.018) with more severity (X2 =4.769,P =0.029).There was no significant difference in hemorrhage and calcification between the two types observed by contrast-enhanced CT.Respectively,66.7 ％ of type Ⅱ PRCC and 23.8％ of type Ⅰ PRCC had papillary nodule,with obviously significant difference (X2 =8.694,P =0.003).In outside invasion signs,except for margins,type Ⅱ had more easily invaded peripheral fat,renal sinus and distant metastasis compared with type Ⅰ (P＜0.05).On contrast enhanced CT,there were significant differences in CT values and △CT values in corticomedullary phase between the two types (t =-2.674,P =0.012;t =-3.109,P =0.005).And there were no significant difference in unenhanced and nephrographic phase.Conclusions There were certain difference in morphological features,outside invasion signs and enhancement degree between type Ⅰ and type Ⅱ PRCC,and part of type Ⅱ PRCC had aggressive biological behaviors with worse prognosis.

This study is to investigate the mechanism and action characteristics of 6-chloro-3-methyl-4-(2-methyoxycarbonylthiophene-3-sulfonyl)-3, 4-dihydroquinoxa-lin-2-(1 H)-one (XU07011) against HIV-1 replication. XU07011 anti-HIV activity was tested by using VSVG/HIV pseudotype viral system and confirmed by HIV-1 live viruses' infectious assay. Time of addition was used to test HIV-1 reverse transcription process. RNA-dependent DNA polymerase activity and RNase H activity were tested by using enzyme linked immunoabsorbent assay and fluorescence method. Wild type and nine NNRTIs-resistant reverse transcriptase enzymatic models and cell-based pharmacological models were used to evaluate XU07011 bio-characteristics. The results showed that XU07011 inhibited HIV-1 replication with IC50 of (0.057 +/- 0.01) micromol x L(-1) which was comparable to nevirapine [IC50: (0.046 +/- 0.01) micromol x L(-1)]. Mechanism study data indicated that XU07011 blocked HIV-1 reverse transcription process through acting on reverse transcriptase RNA-dependent DNA polymerase with IC 50 of (1.1 +/- 0.3) micromol x L(-1). The compound showed no effect on RNase H activity. XU07011 exhibited better activities comparing with nevirapine on K103N mutated NNRTIs-resistant HIV-1 strains. This study could provide a theoretical basis for novel anti-HIV reagents development.

Objective To investigate the possibility of differentiation and spermiogenesis of spermatocytes under in vitro condition. Methods Testis biopsy was done in 11 cases with non-obstructive azoospermia.Cells were prepared from 9 samples with spermatocytes and cultured in medium containing follicle stimulating hormone 50 U/L and testosterone 1 μmol/L.Sperms and cells of other types were counted and the proportion of every cell type was calculated before and 24 hours after culture.Flow cytometry was conducted before and 24 hours after culture in 2 cases to analyze the ploidy of the cells. Results The proportion of sperm in 9 samples was ( 17.7 ± 8.9 ) ％ before culture and ( 25.6 ± 10.3 ) ％ after culture ( P =0.004).Sperm increased significantly after culture.Flow cytometry demonstrated that the diagram of 4 n,2 n and 1 n converted to 2 n and broader 1 n. Conclusion Meiosis of spermatocytes and the transformation of spermatid into sperm could arise in in vitro culture.

Objective To explore the mediating effect of attentional control between negative automatic thoughts and social anxiety.methods A total of 1 504 college freshmen of medical university were surveyed with the Interaction Anxiety Scale (IAS),the Automatic Thoughts questionnaire (ATQ) and the Attentional Control Scale (ACS).Result sThe scores of IAS,ACS and ATQ of medical freshmen were (39.36±8.15),(53.50±7.48) and (48.93±19.72) respectively.Correlation analysis showed that ATQ total score was significantly positively correlated with IAS total score(r=0.313,P<0.01),ATQ total score was significantly negatively correlated with ACS total score(r=-0.275,P<0.01),and ACS score was significantly negatively correlated with IAS total score(r=-0.455,P<0.01).Mediating effect analysis showed that attentional control partly-mediated the relationship between negative automatic thoughts and social anxiety among medical freshmen (x2/df=1.650,GFI=0.998,AGFI=0.993,RMSEA=0.021),and accounted for 50.8% of the mediating effect.Conclusion Negative automatic thoughts is an important factor which plays direct and indirect roles in social anxiety.Attentional control is the intermediate link between negative automatic thoughts and social anxiety.

Objective To analyze quantitatively intravoxel incoherent motion diffusion weighted imaging (IVIM-DWI)of normal adult kidney and to evaluate the effects of the location of the kidney,gender and age on IVIM-DWI parameters.Methods Thirty healthy adult volunteers were recruited to undergo IVIM-DWI examination.Two radiologists measured the D ,D? and f values of renal parenchyma in both the upper pole,middle part and lower pole of the kidneys separately.Results The D ,D? and f values of the middle part of kidneys in healthy adult were(1.61±0.1 6)×10 -3 mm2/s,(1 7.45 ±3.78)×10 -3 mm2/s and (26.88 ±5.1 9)%, respectively.The D values of right kidney were higher than that of left kidney (P <0.05).The f values of normal kidney in male volunteers were higher than that of female (t = 3.321,P =0.001).The D values of normal kidney in > 50 years group were lower than that of ≤50 years group (t = 3.548,P=0.001).D value of the kidney and age was negatively correlated (r=-0.406).Intraclass correlation coefficient of D,D? and f values between two observers were 0.881,0.56 and 0.741,respectively.The consistency of two observers in measurement of IVIM-DWI parameters in the middle part of kidneys was higher than that of the upper pole and lower pole of the kidneys.Conclusion The IVIM-DWI parameters of adult normal kidneys are influenced by different parts of the kidney,gender and age.

Aim To investigate the efficacy and mech-anism of FW-04-806 against HER2-positive gastric cancer cell lines,and the combination effect of FW-04-806 with lapatinib. Methods MTT assay was used to assess cell proliferous inhibition of FW-04-806 . The in-hibitory effect of colony formation was tested by colony formation. The protein expression, apoptotic induction and cell cycle arrest were detected by flow cytometry. Co-immunoprecipitation was used to investigate protein-protein interactions. The expression change of proteins was showed by immunohistochemistry. Western blot was applied to reveal the protein expression of related pro-liferous and apoptotic signaling pathway. The tumor growth inhibition was evaluated in tumor xenograft model. Results FW-04-806 obviously inhibited cell proliferation and colony formation in HER2 positive gastric cancer cell lines NCI-N87, OE19, with IC50 of (24. 17 ± 0. 02 ) , ( 29. 61 ± 0. 03 ) μmol · L-1 , re-spectively;FW-04-806 induced G2-M arrest and apop-tosis in a dose-dependent manner;200 mg · kg-1 of FW-04-806 showed tumor growth inhibition of 48. 0%( P < 0. 01 ) . In addition, FW-04-806 dissociated Hsp90/CDC37 complex, followed by degradation of HER2 and Akt,inhibiting the phosporylation of HER2, Akt and ERK, and increasing expression of apoptotic proteins,such as cleaved caspase-3 and cleaved parp. Furthermore,the combination of FW-04-806 with lapa-tinib in vitro was synergistic in NCI-N87 , which en-hanced the inhibition of cell proliferation and increased apoptotic rates. Conclusions FW-04-806 shows po-tent efficacy against HER2-positive gastric cancer cell lines in vitro and in vivo;FW-04-806 is synergistic with lapatinib.

Disease-centered problem-based learning (PBL) integrated course inspires the thinking of medical students in the case scenario to stimulate students' motivation of active learning. In this paper, the study of diabetes cases was taken as an example. Through the design of PBL cases, the scenario was reconstructed and information was provided step by step, so as to induce the students to discuss and learn the related knowledge of glucose metabolism and understand the predisposing factors of diabetes. Furthermore, students' critical thinking could be inspired through the information of the misdiagnose and mistreatment to recognize the clinical presentation and inducement of diabetic ketoacidosis. This teaching model is conducive to the cultivation of medical students' questioning spirit and critical thinking, laying a foundation for the cultivation of innovative medical talents.

Objective:To identify the risk factors for early acute lung injury (ALI) after living-related liver transplantation in pediatric patients and evaluate the value of the risk factors in prediction of ALI.Methods:Perioperative data of patients were obtained through the electronic medical records system. Patients were divided into non-ALI group and ALI group according to whether ALI occurred within the first week after surgery. The risk factors of which P values were less than 0.05 would enter the multiple logistic regression analysis to stratify ALI-related risk factors. Area under the ROC curve was used to analyze the value of the risk factors in prediction of postoperative ALI. Results:A total of 67 patients were enrolled, including 45 cases in non-ALI group and 22 cases in ALI group. Increased intraoperative blood transfusion volume and up-regulated expression of miR-122 and miR-21 were independent risk factors for the occurrence of postoperative ALI ( P<0.05), and the area under the ROC curve (95% confidence interval) of serum miR-122 and miR-21 expression was 0.946 (0.875 to 1.00), with sensitivity and specificity of 95% and 90%, respectively. Conclusions:Increased intraoperative blood transfusion volume and up-regulated expression of serum miR-122 and miR-21 are independent risk factors for early postoperative ALI, and serum miR-122 and miR-21 levels have a high value in prediction of the development of postoperative ALI in pediatric patients undergoing living-related liver transplantation.

Objective To study the effect of miR-146a on hepatic INSR gene expression in F1 offspring of paternal rats with high-glucose-high-fat diet ( HGFD) . Methods 5-week-old male SD rats were randomly divided into normal diet ( ND) and HGFD groups. Male rats with ND or HGFD feeding for three months mated with normal female ones. Blood glucose concentration, glucose tolerance, and insulin tolerance in newborn male offspring rats were detected respectively. Differential miRNAs between ND and HGFD groups were compared using next generation sequencing and were then confirmed by real-time quantitative PCR ( qPCR) . The relative expression of INSR mRNA and the methylation level of INSR promoter in liver tissues of the offspring newborn rat were detected and the correlations between them and the relative expression of differential microRNA were analyzed respectively. In vitro, effects of miR-146a on expression and methylation of INSR gene in BRL-3A cells were detected using Western-blot assay and qPCR respectively. Results The fasting glucose concentration of different groups were without significant difference, but glucose tolerance and insulin tolerance of neonatal male rats in HGFD group were decreased significantly . Next generation sequencing has revealed 45 up-regulated miRNAs and 15 down-regulated miRNAs in HGFD group. Among them, differences of 8 miRNAs expression in the enlarged samples were confirmed by qPCR. miR-146a was up-regulated for more than 10 times in the liver of the offspring of HGFD group. Expression level of miRNA-146a was negatively correlated with the relative expression of INSR and positively correlated with the methylation level of INSR in livers of neonatal rats in HGFD group. In vitro, miR-146a ( mimics ) promoted the methylation of INSR gene and inhibited the expression of INSR in BRL-3A cells. Conclusion HGFD feeding to male SD rats leads to the inhibition of hepatic INSR gene expression in neonatal offspring via upregulating miR-146a.