Objective:To investigate the effect and mechanism of targeted B7-H3 gene silencing on the tumorigenesis and metastasis of human hematological malignancy xenograft tumor in nude mice.Methods: Real-time fluorogentic quantitative PCR (qPCR) and flow cytometry (FCM) were used to detect the expression of B7-H3 in 13 strains of malignant hematologic cells.Then,U937,Maver and Z138 cells which expressed high level of B7-H3 were screened out.Targeted B7-H3 knockdown in U937,Ma-ver and Z138 was performed by lentivirus transduction and the effect of B7-H3 silencing in stable cell lines was tested by qPCR and FCM.Injecting the nine groups subcutaneously into the nude mice to establish xenograft models after dividing the U937,Maver and Z138 into non-infected control group (CON),B7-H3 knockdown group (KD) and negative non-targeted control infected group (NC),respectively,for detecting the tumorigenicity and metastasis in vivo.Furthermore,the expression of Ki-67 in xenograft tumors was detected by immunohistochemistry (IHC).The expression of metalloproteinase 2 (MMP-2) was detected by western blot.Results: The stable B7-H3 silencing cell lines of U937,Maver and Z138 were successfully established.Compared with the NC group,the KD groups of U937,Maver and Z138 had an obviously slower tumor growth.The average tumor inhibition rates at the end of observation period were 61.83％ (F=43.78,P<0.05),59.12％ (F=36.51,P<0.05) and 67.37％ (F=40.29,P<0.05);there was no significant difference in tumor volume growth between the NC group and the CON group (P>0.05).The liver distant metastasis of all the xenograft tumor models in nude mice was the most common and the rates of distant metastasis in KD groups were significantly lower than that of the corresponding NC groups.The Ki-67 indexes of the KD groups were significantly lower than those of the relative NC groups in three cell lines (U937: 40.3％±5.2％ vs.79.1％±6.3％,q=30.31,P<0.05,Maver: 35.2％±6.4％ vs.69.6％±5.1％,q=24.82,P<0.05;Z138: 38.4％±7.1％ vs.75.7％±4.8％,q=28.07,P<0.05);there was no significant difference in the expression of Ki-67 between the NC group and the CON group (P>0.05).The expressions of MMP-2 were also significantly lower in the KD groups than in the NC groups (U937: q=14.59,P<0.05;Maver: q=9.25,P<0.05;Z138: q=11.04,P<0.05);there was no significant difference in the expression of MMP-2 between the NC group and the CON group (P>0.05).Conclusion: Targeted B7-H3 gene silencing could inhibit the tumorigenesis and metastasis of human hematological malignancy xenograft tumor in nude mice.The mechanism may be related to the down-regulation of Ki-67 and MMP-2.

Objective To investigate radiation-induced alternations of phospholipids in epithelial cells,and to provide experimental evidence for understanding the mechanism of radiation-induced intestinal injury.Methods The intestinal epithelial cells(IEC-6)in rats were divided into three groups:normal control group,8 Gy X-ray irradiation group and 12 Gy X-ray irradiation group.Phospholipids were extracted at 6 h or 24 h after radiation and then measured by high-performance liquid chromatography and mass spectrometry(HPLC-MS).Results At 6 h after radiation,the phospholipids in 8 Gy irradiation group didn't vary significantly,while those in 12 Gy irradiation group changed.The PG,PI and Lyso PC were significantly up-regulated(F=5.37,9.60,9.88,P＜0.05).However,at 24 h after radiation,many PE and PG species in both irradiation groups declined(F=5.15-99.77,P＜0.05)and SM species increased in 12 Gy irradiation group(F=4.35-7.92,P＜0.05).Conclusions The ionizing radiation could disorder phospholipid metabolism in IEC-6 cells with a dose-dependent manner.

Objective To establish a HPLC method for glycine assay in the human thrombin. Methods The sample was derivatized with 2,4-dinitrofluorobenzene (DNFB). HPLC was performed on an ODS-C₁₈ column with 50% acetonitrile –0.05mol/L sodium acetate buffer (35:65) as the mobile phase. The flow rate was 1.0 ml/min and detection wavelength at 360 nm. Results The linear range of glycine was 0.006-0.030 mg/ml (r=0.999 6). The average recovery was 100.4%, with RSD 0.44% (n=9). Conclusion This method is simple, accurate and specific. It is suitable for glycine assay in human thrombin.

Objective To investigate the underlying mechanism of ursolic acid in attenuating diabetic mesangial cells injury induced by high glucose (HG).Methods Rat glomerular mesangial cells were cultured in normal glucose,HG,HG with LY294002 and HG with ursolic acid.The cell proliferation and hypertrophy were detected by MTT and the ratio of total protein content to cell number.miRNA-21 was detected by quantitative real-time PCR.The PI3K-Akt-mTOR pathway,autophagy associated protein and collagen I were detected by Western blotting and quantitative realtime PCR.The autophagosomes were observed by electron microscope.Results Compared with normal control group,the cells exposed to HG showed up-regulating miRNA-21 expression(P ＜ 0.01),down-regulating PTEN protein and mRNA expression(P ＜ 0.01),up-regulating p85PI3K,phospho(p)-Akt,p-mTOR,p62/SQSTMI,collagen I expressions and down-regulating LC3II expression(P ＜ 0.01).Ursolic acid and LY294002 inhibited HG-induced mesangial cell hypertrophy and proliferation(P < 0.01),down -regulated the expressions of p85Pl3K,p-Akt,p-mTOR,p62/SQSTMI and collagen I and up-regulated the expression of LC3II(P ＜ 0.01).But LY294002 had no effect on the expression of miRNA-21 and PTEN.Ursolic acid down-regulated miRNA-21 expression(P ＜ 0.01),up-regulated PTEN protein and mRNA expression(P ＜ 0.01).Conclusion Ursolic acid may inhibit high glucose-induced mesangial cell miRNA-21 overexpression,up-regulate PTEN,inhibit the activation of PI3K-Akt-mTOR signaling pathway and the enhanced autophagy to reduce the accumulation of extracellular matrix and ameliorate cell hypertrophy and proliferation.

This article reviewed the measures taken by the hospital against a catastrophic fire hazard and exploration in its response and work organization of nursing care of the wounded.Proposed in this paper are development of such five systems as the pre-plan,exercises,personnel,quality control and incentives,which are expected to improve the nursing capacity of the hospital in emergency rescue,for sustainable development of nursing emergency rescue work.

A new mutant human papiUomavirus type 16 E7 gene, termed HPV16 HBE7, was isolated from cervical carcinoma biopsy samples from patients in an area with high incidence of cervical cancer (Hubei province, China). A previous study showed that the HPVI6 HBE7 protein was primarily cytoplasmic while wild-type HPV16 E7 protein, termed HPV16 WET, was concentrated in the nucleus. With the aim of studying the biological functions of HPV16 HBE7, the transforming potential of HPV16 HBE7 in NIH/3T3 cells was detected through observation of cell morphology, cell proliferation assay and anchorage-independent growth assay. The effect of HPVI6 HBE7 on cell cycle was examined by flow cytometry. Dual-luciferase reporter assay and RT-PCR were used to investigate the influence of HPVI6 HBE7 protein on the expression of regulation factors associated with GI/S checkpoint. The results showed that HPV16 HBE7 protein, as well as HPV16 WE7 protein, held transformation activity. NIH/3T3 cells expressing HPV16 HBE7 could easily transition from G1 phase into S phase and expressed high level of cyclin A and cdc25A. These results indicated HPV16 mutant E7 protein, located in the cytoplasm, induces oncogenic transformation of NIH/3T3 cells via up-regulation of cyclin A and cdc25A.

Objective To investigate the association between SLC10A1 gene mutations in c.800G>A mutation and c.356 +1098C >T mutation, and the susceptibility to HBV infection by mother-to-child transmission ( MTCT) .Methods Totally 306 individuals born to HBeAg-positive mothers with high load HBV and without receiving nucleotide analogues treatment, including 247 HBV-infected cases and 59 non-HBV-infected ones were enrolled from Southwest Hospital during May 2011 and July 2015.Blood samples were collected from all the subjects, then genomic DNA was extracted and c.800G>A mutation and c.356+1098C>T mutation of SLC10A1 were genotyped .Chi-square test (Pearsonχ2or continuity correctionχ2) was performed to identify the difference in genotypes between two groups.Results Among vaccinated individuals (55 HBV infected and 56 not infected), the frequency of genotype GA of c.800G>A mutation in non-infected ones was 14.3%(8/56), there was a tendency of increasing compared with HBV infected ones (5.5%, 3/55), but the difference was not statistically significant (χ2 =2.424, P =0.119). Similarly, the frequencies of genotypes CC, CT and TT of the c.356+1098C>T mutation in HBV infected ones were 20.0%(11/55), 47.3%(26/55) and 32.7%(18/55), while those in non-infected ones were 12.5% (7/56), 69.6% (39/56) and 17.9% (10/56), and the difference was not of statistical significance (χ2 =5.766, P=0.056).In all subjects (vaccinated and non-vaccinated), the frequency of genotype GA of c.800G>A mutation in non-HBV infected group had an increasing tendency compared with HBV-infected offspring (13.6% vs.6.9%), but the difference was not statistically significant (χ2 =2.010, P=0.156);the frequencies of genotype CC, CT and TT of c.356+1098C>T mutation in HBV infected patients were 20.2%(50/247), 49.8%(123/247) and 30.0%(74/247), while those in non-HBV-infected group were 11.9%(7/59), 69.5%(41/59) and 18.6%(11/59), and the difference was statistically significant (χ2 =7.436, P =0.024 ) .Within the HBV infected group, the frequencies of genotype GA of c.800G>A mutation were 5.5%(3/55) in vaccinated individuals and 7.3%(14/192) in non-vaccinated individuals, and the difference was not of statistical significance (χ2 =0.030, P=0.863);Similarly, the frequencies of genotype CC, CT and TT of c.356 +1098C >T mutation in vaccinated individuals were 20.0%(11/55), 47.3%(26/55) and 32.7%(18/55), while those in non-vaccinated individuals were 20.3%(39/192), 50.5%(97/192) and 29.2%(56/192), and the difference was not of statistical significance (χ2 =0.274, P=0.872).Conclusion c.356+1098C>T mutation in SLC10A1 may be associated with susceptibility to HBV infection of child born in HBeAg positive pregnant women infected with high load HBV.

Objective To study the possibility of the plasma level of heat shock protein 70(HSP70) being used as a bi-ological marker of military stress .Methods Soldiers who returned from a 6-month-navigation were chosen as subjects , the HSP70 level of plasma was measured with the ELISA assay and stress questionnaires and Self -rated Health Measurement Scale (SRHMS) were used to measure the stress level .Results The soldiers′plasma level of HSP70 was 31.40%higher than that of the control .The stress questionnaire indicatesd that the level of thinking and anxiety , negative mood and somat-ic symptoms were higher than normal .The SRHMS indicated that the level of physiological health ,mental health and social health was lower than normal .The plasma level of HSP70 was associated with the level of military stress .Conclusion The plasma level of HSP70 may be used as an important predictor of military stress .It can predict the level of military stress injury.

Objective To study the pathogenic etiology between nasopharyngeal aspirates (NPA) and bronchoalveolar lavage lfuid (BALF) in children with lower respiratory infection. Methods Multiple pathogen in NPA and BALF from 210 cases with lower respiratory tract infection was detected. Seven common respiratory virus (respiratory syncytial virus, adenovirus, in-lfuenza virus A, inlfuenza virus B, parainlfuenza 1, parainlfuenza 2, parainlfuenza 3) were detected by direct immunolfuorescence assay. MP, CP and HBoV were detected by lfuorescence quantitative PCR.HRV and hMPV were detected by RT-PCR. Aspirates were cultured for bacteria. The results of pathogen detection in secretions of upper and lower respiratory tract were analyzed. Results Total positive detection rate of NPA and BALF in 210 cases was 91.9%(193/210), which is higher than that in NPA 75.2%(158/210) and that in BALF 85.2%(179/210). Bacteria detection rate in NPA was 13.3%(28/210), and 8.6%(18/210) in BALF, without signiifcant difference (P=0.118). Bacteria detection rate in NPA and BALF was of poor consistency (Kappa=0.262). Virus detection rate in NPA was 24.3%, which is higher than that in BALF15.2%. BALF-MP detection rate was 77.6%(163/210), signiifcantly higher than that in NPA 53.3%(112/210). There are 95.5%(107/112) cases with positive results in NPA-MP detec-tioncan also be detected in the BALF-MP. MP copies in BALF were signiifcantly higher than that in NPA (4.28×106 vs. 1.31×105), and its positive rate in NPA was still higher than that in BALF. MP detection rate in NPA in children with clinical course of longer than two weeks was much lower than those with clinical course of two weeks or less. Conclusions The pathogen detection of virus and MP in NPA can be used as a reference for lower respiratory tract infection. The joint detection of NPA and BALF can improve the detection power. The sensitivity of virus detection in NPA is higher than that in BALF. NPA pathogen detection of virus and MP is of great important evidence-based medicine in the diagnosis of lower respiratory infection. MP detection rate and its copies in BALF are signiifcantly higher than that in NPA. BALF detection is the supplement of pathogen diagnosis in severe or refractory lower respiratory infections.

Objective To analyze the correlation between response to hepatitis B virus (HBV) vaccine and cellular immunity and clinical characteristics in children with respiratory infection.Methods Nine hundred and sixty children in Department of Respiratory in Children's Hospital of of Soochow University,who were over 7 months old and had full course of HBV vaccination between January and December 2015 were enrolled in this study.Peripheral blood (1-2 mL) was collected,and antigen-antibody of HBV was detected by using enzyme-linked immunosorbent assay and PCR included HBV surface antigen,hepatitis B antibody,HBV e antigen,HBV e antibody,HBV core antibody,and HBV nucleic acid.According to the results,these children were divided into 4 groups:non response group,low response group,normal response group and high response group according to their responses to HBV vaccine.Cellular immunity was detected by using flow cytometry and patients' clinical data was collected.Results There was no statistical differences of CD3 + CD4 +,which were (3.43 ± 0.28) ％,(3.42 ± 0.30) ％,(3.43 ± 0.36) ％ and (3.52 ± 0.29) ％,among the four groups (F =0.520,P =0.669).CD3 + CD8 + in non response group was (3.18 ±0.28)％,which was significantly higher than that in low response group,normal response group and high response group [(3.08 ± 0.36)％,(3.05 ±0.34)％,(2.93 ±0.30)％],the differences were significant (all P＜0.05);CD4/CD8 in non response group (0.26 ± 0.43) were significantly lower than that in normal response group (0.40 ± 0.50),the differences were significant (P =0.001).There was no significant difference of CD3 +,CD3 + CD8 + and CD4/CD8 among low response group,normal response group and high response group (all P ＞ 0.05).CD3-CD19 + and CD19 + CD23 + level were lowest in non response group [(3.00 ± 0.57) ％,(2.25 ± 0.67) ％] and highest in high response group [(3.33 ± 0.45) ％,(2.57 ± 0.38) ％],the differences were significant (all P ＜ 0.05).Among the 4 groups,children in normal response group had the shortest average hospitalization days [(1.88 ±-0.31) d],which was significantly shorter than that in non response group,low response group and high response group [(1.96 ± 0.39) d,(1.95 ± 0.38) d,(1.96 ±0.15) d],the differences were significant (all P ＜0.05),there was no significantly difference of average hospitalization days among other 3 groups (all P ＞ 0.05).Proportion of severe pneumonia was significantly higher in non response group [6.1％ (22/363cases)] and high response group [13.3％ (2/15 cases)] compared to those in normal response group [2.6％ (7/274cases)],the differences were statistically significant (x2 =4.417,P =0.036;x2 =5.476,P =0.019).The total white blood cell number was lowest in non response group (F =4.695,P =0.003).Platelet number was increased with higher degree of response to HBV (F =6.598,P ＜ 0.001).Conclusions Cellular immunity is lower in respiratory infection children with non response or low response to HBV vaccine.After they have respiratory infection,children with non response to HBV vaccine may have a longer course of disease and worse condition.