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Objective: To construct the recombinant lentiviral vector containing cardiac adriamycin responsive protein (CARP) gene and small-hairpin RNA (shRNA) targeting CARP gene and to pack the lentivirus, and to lay the foundation for further study on the function and mechanism of CARP in adriamycin (ADR) induced cardiomyopathy. Methods: After the rat CARP gene was amplified by PCR and shRNAs targeting CARP were designed and synthesized, they were inserted into the shuttle plasmids GV-358 or GV-248. respectively. After confirmed by sequencing, the recombinant shuttle vectors containing CARP gene or shRNA and auxiliary packaging plasmid Helper 1 0 and Helper 2 0 were co transfected into the IIEK293T cells for virus packaging and amplification; the viral titer was detected by end point dilution. The IIEK293T cells were infected with the recombinant lentiviruses. and the green fluoresence intensity was observed by fluorescence mircroscope. The H9C2 cells were infected with the recombinant lentivirues and divided into control group. CARP overexpression group and shRNA group; Western blotting method was used to detect the CARP expression levels in the cells in various groups. Results: The DNA sequencing results showed that the sequences of CARP overexpression and shRNA vectors were in accordance with the designed sequences. The expression of green fluorescence protein was seen under fluorescence microscope after transfection of the vectors of the IIEK293T cells. After infection of the H9C2 cells, the expression level of CARP protein in CARP overexpression group was 5. 3 times higher than that in control group ( P=0. 01); while it was down-regulated by 53% in CARP shRNA group compared with control group ( P= 0 02). Conclusion: The lentivirus expression vectors carrying CARP or shRNA targeting CARP are successfully constructed and the lentiviruses obtained could significantly interfere the expression of CARP in the II9C2 cells.
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Objective@#To study serum zinc level in pregnancy and umbilical cord blood and their association with newborn birth weight.@*Methods@#Pregnant women accepting obstetric examination in Ma'anshan Maternal and Child Care Center were recruited from May 2013 to September 2014. The follow up was conducted during their first, second and third trimesters of pregnancy and the self-designed questionnaire was used to collect information of social and demographic characteristics. Blood samples in the first, second pregnancy period and umbilical cord blood samples were collected and serum concentrations of zinc were assayed. 3 239 mother-infant entered the final analysis. We divided serum zinc level into low (<P25), medium (P25-P75) and high (>P75) groups according to their exposure concentrations at each trimesters. Non-conditional multivariate logistic regression model was conducted to evaluate the association between serum zinc level in first, second trimesters of pregnancy and umbilical cord blood with small for gestational age (SGA) and large for gestational age (LGA).@*Results@#Serum zinc level in P50 (P25-P75) during the first, second trimesters and cord blood were 1 016.18 (907.09-1 145.60), 813.36 (732.47-897.89) and 903.44 (808.71-1 015.64) μg/L, respectively. The prevalence of zinc deficiency during the first, second trimesters and cord blood were 1.5% (44/2 957), 15.9% (492/3 087) and 6.5% (176/2 707), respectively. The prevalence of total SGA and LGA were 9.7% (313/3 239) and 16.5% (536/3 239), respectively. Compared to high-level serum zinc group, the risk of SGA (OR (95%CI) in low-level serum zinc group during first trimesters was 1.51 (1.05-2.19)). Serum zinc level among second pregnancy period and umbilical cord blood had no statistically significant effect on SGA and LGA (both P values >0.05).@*Conclusion@#Zinc nutritional status of pregnant women in Ma'anshan city was at a good level. The low serum zinc level in first trimester increased the risk of SGA.
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Objective To investigate the relationship of thallium exposure and outcomes of births.Methods A total of 3 236 mothers who had visited in Ma'anshan Maternal and Child Health-Care Hospital between May 2013 and September 2014 were included in this study and their thallium concentrations measured from samples of maternal and umbilical cord blood by inductively coupled plasma mass spectrometry.The results were correlated and evaluated with birth outcomes of the infants,using the multiple linear regression method.Results The median (P25-P75) of thallium levels in first trimester,second trimester and umbilical cord blood were 61.7 (50.8-77.0),60.3 (50.8-75.2) and 38.5 (33.6-44.1) ng/L,respectively.After adjustment for potential confounders,the thallium levels showed an inversely significant association with birth head circumference (unstandardized β coefficient=-0.41,95％ CI:-0.76--0.06) in thc first trimester blood,and associated with reduced birth length (unstandardizedβ coefficient=-0.65,95％ CI:-1.25--0.05) in umbilical cord blood.However,there appeared no significantly associations with birth weight,length and head circumference (P＞0.05) in second trimester.On stratification by sex,in girls but not in boys,the thallium levels were adversely associated with birth head circumference (unstandardized β coefficient=-0.53,95％ CI:-1.05--0.01) in the first trimester and were associated with decreased birth weight (unstandardized β coefficient =-277.08,95％CI:-485.13--69.03) and length (unstandardized β coefficient=-1.39,95％CI:-2.26--0.53) in umbilical cord blood thallium.Conclusions Thallium exposure appeared a gender difference in newborn birth outcomes.In the first trimester,it was negatively associated with the birth head circumference,in the umbilical cord blood,and reduced birth weight and length in girls.
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Objective To investigate the relationship of thallium exposure and outcomes of births.Methods A total of 3 236 mothers who had visited in Ma'anshan Maternal and Child Health-Care Hospital between May 2013 and September 2014 were included in this study and their thallium concentrations measured from samples of maternal and umbilical cord blood by inductively coupled plasma mass spectrometry.The results were correlated and evaluated with birth outcomes of the infants,using the multiple linear regression method.Results The median (P25-P75) of thallium levels in first trimester,second trimester and umbilical cord blood were 61.7 (50.8-77.0),60.3 (50.8-75.2) and 38.5 (33.6-44.1) ng/L,respectively.After adjustment for potential confounders,the thallium levels showed an inversely significant association with birth head circumference (unstandardized β coefficient=-0.41,95％ CI:-0.76--0.06) in thc first trimester blood,and associated with reduced birth length (unstandardizedβ coefficient=-0.65,95％ CI:-1.25--0.05) in umbilical cord blood.However,there appeared no significantly associations with birth weight,length and head circumference (P＞0.05) in second trimester.On stratification by sex,in girls but not in boys,the thallium levels were adversely associated with birth head circumference (unstandardized β coefficient=-0.53,95％ CI:-1.05--0.01) in the first trimester and were associated with decreased birth weight (unstandardized β coefficient =-277.08,95％CI:-485.13--69.03) and length (unstandardized β coefficient=-1.39,95％CI:-2.26--0.53) in umbilical cord blood thallium.Conclusions Thallium exposure appeared a gender difference in newborn birth outcomes.In the first trimester,it was negatively associated with the birth head circumference,in the umbilical cord blood,and reduced birth weight and length in girls.
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Objective To explore the effect of the Internet technology′s application on the hand hygiene compliance of medical staff. Methods Establishing Internet+hand hygiene management model to implement Internet based hand hygiene supervisory and reminding, Internet based hand hygiene publicity and education, and Internet based hand hygiene feedback and communication. This model was applied in Guangdong Provincial Dermatology Hospital to intervene the hand hygiene related activities. Before and after the intervention, the hand hygiene compliance of medical staff in the trial hospital was observed with WHO hand hygiene monitoring table by directly manual observation. Thereafter, the changes of hand hygiene compliance before and after the intervention were compared based on the statistical analysis of test results. Results With the application of Internet+hand hygiene management model, the hand hygiene compliance of medical staff increased from 41.75% (324/776) before the intervention to 72.14% (554/768) after the intervention, and the difference was statistically significant (χ2=145.264, P<0.01). The hand hygiene compliance of both doctors and nurses was significantly increased (doctors: χ2=94.784, P<0.01;nurses:χ2=58.798, P<0.01). In all the five indications, i.e., before patient care, before an aseptic task, after exposure to blood/body fluids, after patient care, and after environmental contact, the hand hygiene compliance was 35.42%(68/192), 63.33%(38/60), 84.38%(54/64), 44.83%(130/290), and 20.00%(34/170) respectively before the intervention;it was 65.38%(102/156), 83.33%(120/144), 95.45%(105/110), 68.52%(148/216), and 55.63%(79/142) after the intervention; and the difference before and after the intervention was statistically significant (χ2=6.305-42.529;P<0.01 or 0.05). Most of the medical staff of the trial department supported this model. Conclusions Internet+ hand hygiene management model could significantly improve the hand hygiene compliance of medical staff and were very popular. And, the model is worth of application in more hospitals.
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Objective:To construct lentiviral vector which can overexpression miR-137 and produce lentivirus by lentivirus packaging system, and to explore its infection efficiency and expression in HEK293T cells.Methods: miR-137 sequence was chemically synthesized and cloned into lentiviral vector GV209, and the recombinant plasmid containing human miR-137 was obtained and identified.Then miR-137 recombinant plasmid together with two helper plasmids were transfected into HEK293T cells using Lipofectamine 2000.After the HEK293T cells were infected in multiplicity of infection(MOI) 40 for 48 h, the expression of green fluorescent protein (GFP) was observed by fluorescence microscope and the expression level of miR-137 was detected by fluorescence quantitative PCR.Results:The sequencing results showed that the inserted gene sequence was completely consistent with the published human miR-137 gene sequence in GenBank.The GFP was observed in the HEK293T cells infected with miR-137 overexpression lentivirus under fluorescence microscope.The fluorescence quantitative PCR results showed that the expression level of miR-137 in the cells infected with overexpression lentivirus was 12.74 times higher than that in the control cells.Conclusion:The lentivirus containing miR-137 gene is successful packaged, and it could efficiently infect the HEK293T cells.
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Purpose To explore the immunohistochemical features of neuroendocrine markers, CKpan and TTF-1 and their relationship to TNM stage and prognosis in small cell lung cancer (SCLC). Methods The expression of NSE, CgA, Syn, CD56, TTF-1 and CK-pan in SCLC tissue specimens were detected using immunohistochemical EnVision indirect method. Clinical data and TNM stage of 90 patients were collected and the overall survival ( OS) was followed up by telephone. Results Of 90 cases of SCLC, the vast majority were occured in the elderly men. The ratio of man to woman was 5 to 1. The median age was 64 years old. The stageⅠ+Ⅱ was 21 cases, 30 in stageⅢand 39 cases in stage IV. The positive rate of immunohistochemical staining of neuroendocrine markers for NSE, CgA, Syn and CD56 were 83. 3%, 70%, 65. 5% and 86%, respectively. The positive rate of CKpan, TTF-1 were 92. 2% and 81. 1%. Kaplan-Meier analysis indicated that the expression of TTF-1 and NSE were significantly correlated with the TNM stage and over-all survival of patients with SCLC (P<0. 05). The median OS was 8 months in positive expression of TTF-1, which was higher than those in negative expression of TTF1 (5. 5 months)(P=0. 000). The median OS was 7 months in NSE positive expression which was lower than those in negative expression of NSE (11 months)(P=0. 009). The median OS of stageⅠ+Ⅱ,ⅢandⅣwere 16 months, 9 months and 4 months with significant difference ( P=0. 000 ) . Cox multivariate analysis indicated the TTF-1 expression and TNM were independent prognostic factors for the OS of the SCLC patients. Conclusion Most of SCLC has neuroendocrine differentiation, expression CKpan and TTF-1. The expression of TTF-1 may be negative correlation but NSE positive correlation with the prognosis of SCLC patients. And the TTF-1 expression and TNM may be independent prognostic factors for the OS of the SCLC patients.
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Objectiv e:To investigate the effect of different culture conditions on the differentiation of Treg and Th17 to lay a foundation for exploring the methods to reverse the immune tolerance induced by tumor microenvironment.Methods:The IL-6 gene was cloned and stablely transferred into the tumor cell line expressing TGF-β.The conditioned mediums ( CM) were prepared by collecting the culture supernatants of tumor cell lines with or without IL-6 expression and used in the in vitro culture of peripheral blood mononuclear cells ( PBMC ) .The changes of Treg and Th17 in PBMC treated with different CM were detected with flow cytometry ( FCM) .Results:The expression of TGF-βin BEL-7402 was higher than that in HepG2.Thus the BEL-7402 was selected for preparation of cell line stablely transfected with IL -6 gene.ELISA detection confirmed the effective expression of IL -6 by the identified cell lines.It was showed that the Treg increased in PBMC treated with culture supernatants of tumor cells .However,the presence of IL-6 reversed the increase of Treg and promoted the differentiation of Th 17.Conclusion: The culture supernatants of tumor cells increases the proportion of Treg.However,the presence of IL-6 in this CM can reverse the increase of Treg and raise the proportion of Th 17.
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Objective To investigate the influence of telephone follow-ups on the compliance of syphilis patients.Methods The 120 patients diagnosed with syphilis during February 2010 to December 2011 were randomly divided into experiment group and control group with 60 cases in each group.The control group was given disease-related education during outpatient visits,while the experimental group received nursing intervention by phone calls.The two groups were compared in respect of curative compliance. Result The curative compliance of the experiment group after follow-ups was significantly better than that of the control group (P<0.01).Conclusion The telephone follow-ups can significantly improve the curative compliance of syphilis patients,which can significantly prevent the spread of syphilis.