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Objective To analyze the clinical characteristics and survival prognosis of patients with AIDS-related malignant tumor. Methods We retrospectively analyzed the data of 354 patients with AIDS-related malignant tumor. Univariate analysis was conducted by Log rank test and multivariate analysis was conducted by Cox proportional risk regression model. Results The average age of the patients was 54.10±12.96 years old. The ratio of male to female patients was 2.1:1. The number of patients with AIDS complicated with lymphoma was the most, accounting for 28.25%. The 1-, 3- and 5-year survival rates were 78.48%, 62.13% and 55.31%, respectively. Univariate analysis showed that there were statistical differences in prognosis of patients with different types of malignant tumor, age, gender, medical insurance type, number of admissions after diagnosis of AIDS, average length of stay, radiotherapy or not, leaving hospital according to medical advice. Multivariate analysis showed that gender, number of admissions after diagnosis of AIDS, average length of stay, proportion of out-of-pocket and leaving hospital according to medical advice were independent risk factors affecting the survival and prognosis of patients. Conclusion AIDS is easily complicated with lymphoma, lung cancer and cervical cancer. The patients received insufficient anti-tumor courses in hospital.
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Objective To investigate the associations of peripheral CD4+ CD25 + CD127low/-regulatory T cells (Treg) with HBV viral load and liver pathology in hepatitis B virus (HBV) carriers.Methods Forty six chronic HBV carriers admitted in the first hospital of Jiaxing during October 2012 and February 2014,and 23 healthy subjects (controls) were enrolled in the study.CD4+ CD25 + CD127low/-Treg in peripheral blood of the two groups were detected by flow cytometry.Ultrasound-guided liver biopsies were performed in chronic HBV carriers and HBV DNA load was determined by real-time PCR method.Independent samples t test was used for the comparison between groups,and Spearman rank correlation or Pearson linear correlation analyses were performed.Results The frequency of the peripheral CD4+ CD25+CD127low/-Treg in 46 HBV carriers was (5.11 ±1.47)％,which was significantly higher than that in healthy controls [(3.46 ± 1.23) ％,t =4.629,P ＜ 0.01].The HBV DNA load in HBV carriers was (6.21 ±1.98)lg copies/mL,which was positively correlated with the CD4+ CD25+ CD127low/-Treg level (r =0.405,P ＜ 0.01).Among 46 HBV carriers,21 (45.65％) were of inflammation grade 2 or above,and 16 (34.78％) were of fibrosis stage 2 or above.Peripheral CD4+ CD25+ CD127low/-Treg level was negatively correlated with inflammation and fibrosis in HBV carriers (r =-0.343 and-0.452,P ＜ 0.05).Conclusion CD4 + CD25 + CD127low/-Treg may be associated with the chronicity of HBV infection and the degree of liver damage.
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Objective To investigate the effect of methylation of the APC gene on expression and the correlation with clinical data in pancreatic cancer.Methods Sixty postoperative tissue samples with pancreatic cancer were collected in the First Affiliated Hospital of Zhengzhou University from August 2010 to January 2011,20 benign pancreatic disease tissues were collected as control groups.APC promoter methylation and gene expression levels were detected by Methylation Specific PCR (MSP),Real Time PCR (RT-PCR) and Western blot in 60 pancreatic carcinoma,42 metastasis and 20 benign pancreatic disease tissues,then analyze the relation between methylation of the APC gene and the clinical data.Results APC promoter methlation was observed 48.53％,46.67％ and 1.16％ in pancreatic carcinoma,metastasis and benign pancreatic disease tissue,respectively.Methylation of APC in pancreatic carcinoma and metastasis increased significantly compared with control tissues (x2 =12.903,14.402; P ＜ 0.05).There were no statistically significant differences of APC expression in these tissues (P ＞ 0.05).There was a significant correlation between methylation of APC and clinicopathological stage (x2 =6.801,P ＜ 0.05),but no correlation with gender,age,tumor size,histological grade and metastasis (x2 =0.727,1.311,0.372,0.148,0.017 ; P ＞ 0.05).Conclusion The methylation of APC gene is closely related with pancreatic carcinoma inogenesis and the clinicopathological stage,but do not effect the expression of APC in tissues.
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Objective To further research the biological functions of PES1,the ovarian SKOV3 cell line with inducible stable PES1 expression is established by using Tet-on system.Methods PES1 was cloned into pTRE-Tight vector via PCR and its expression was identified. After transfected the regulating plasmid pTet-on,SKOV3 cells were screened with G418 and re-transfected pTRE-Tight-PES1.The positive cell clones were screened out with hygromycin and were induced by doxycycline (Dox) to definite the best induction concentration.Growth velocity of SKOV3 cells stably expressing PES1 induced by Dox was detected with viola crystallina.Results The SKOV3 cells with inducible PES1 expression were screened out after the cells were transfected pTRE-Tight-PES1 constructed.Dox could dose-dependently induce the PES1 expression with the concentration under 2 mg/L,and 2 mg/L of Dox induced the highest PES1 expression.Growth velocity of SKOV3 cells transfected pTRE-Tight has no significant difference between the SKOV3 cells transfected nothing induced with Dox.However,the SKOV3 cells transfected pTRE-Tight-PES1 grew faster than the cells transfected pTRE-Tight or without transfection in the fourth day (P =0.001 ).Conclusion The inducible stable PES1 expression SKOV3 cells are successfully established and could be used to be an effective cell model to research the biological functions of PES1.The expression of PES1 could promote the growth of SKOV3 cells.
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ObjectiveTo evaluate the effect of shenfu injection on airway resistance and oxygenation during one-lung ventilation (OLV) in patients undergoing lobectomy.MethodsSixty ASA Ⅱ patients of both sexes,aged 50-80,BMI 20-29 kg/m2,scheduled for lobectomy under thoracic epidural block combined with general anesthesia,were randomly divided into 2 groups( n =30 each):normal saline control group (group C) and shenfu injection group (group S).In group S,shenfu injection was infused intravenously at 4.5 ml·kg-1 ·h-1 for 20 min before anesthesia induction.In group C,equal volume of normal saline was infused instead of shenfu injection.Airway peak pressure was recorded and arterial blood samples were taken for determination of PaO2 before OLV,at 30,60 min of OLV and the end of surgery.Oxygenation index was calculated.ResultsCompared with group C,airway peak pressure was significantly decreased and oxygenation index increased at 30,60 min of OLV in group S ( P ＜ 0.05).ConclusionShenfu injection can decrease airway resistance and increase oxygenation during OLV in patients undergoing lobectomy,indicating that shenfu injection has a lung-protective effect.
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Objective To construct estrogen receptor α (ERα)trans-activation system. Methods The full length ERα and its different function regions [ ( transcriptional activation function 1 ( AF1 ), DNA inding domain ( DBD), and transcriptional activation function 2 ( AF2 ) ] were amplified from pcDNA3 -ERα by PCR and cloned into the pGAL vector. The expressions of the recombinant plasmids constructed were detected via immunoblotting. The 293T cells transfected with recombinant plasmids of full length ERα, its different function regions and empty vector were divided into 5 groups; each group was divided into 2 parts which were treated with or without estrogen (E2). The transcriptional activity of each group was detected in 293T cells after the recombinant plasmid was co-transfected with 0. 2 μg of estrogen receptor element luciferase(ERE-LUC) and 0. 1 μg of plasmid expressing β-galactosidase and treated with or without 10 nmol/L E2 for 24 hours. Results The full length ERα and its different function regions were expressed in the 293T cells. Compared with the empty pGAL vector, the transcription activities of full length ERα, AF1, AF2 and DBD recombinant plasmids were raised about 20. 44±1.01, 2. 09±0. 11, 8. 09±0. 30 and 1.05±0. 09 fold, respectively, with the induction of E2 after transfection in the 293T colls. Conclusion The trans-activation system of ERα has been successfully established.
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In order to investigate if there exists interaction between mitogen-activated protein kinase (MAPK) and signal transducer and activator of transcription 3 (STAT3) protein, and how the interaction regulates tumor necrosis factor-? (TNF-?) transcription activity, the human p38 and extracellular-signal regulated protein kinase 2 (ERK2) genes were amplified from human flag-p38 and flag-ERK2 by polymerase chain reaction (PCR) and cloned into pcDNA3-HA. Protein expression of the plasmids was examined by Western blotting. Co-immunoprecipitation was used to identify if there exists interaction between MAPK and STAT3 proteins. If the interaction was approved to be true, report gene system was applied to find how the interaction affect transcriptional expression of TNF-?. After STAT3 pathway was inhibited by RNA interfering, the action on TNF-? activity was determined. The results of DNA sequencing and enzyme digestion showed that the cloned p38 and ERK2 genes were correct, to be 1 080 bp or so. p38 and ERK2 proteins were expressed in 293T cell to be approximately 40 ku. Co-immunoprecipitation data showed that p38 and ERK2 proteins integrated with STAT3 protein in vivo. TNF-? reporter gene activity results found that protein complex of p38-STAT3 and ERK2-STAT3 coordinately increased TNF-? activity. After STAT3 was interfered, the TNF-? activity markedly decreased. These data indicated that there exists interaction between p38 and STAT3 protein, ERK2 and STAT3 protein. The complex of the proteins can coordinately regulate TNF-? expression. After interfereing STAT3 pathway, the coordinated action on TNF-? transcription activity might be obviously reduced.
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Under the patterns of net-teaching,the construction of net-teaching database is an important tache to develop teaching favourably and improve the quality and efficiency of teaching quickly.This paper introduces the process of constructing a net-teaching database for medical haematology,the course learning of clinical hematology for the undergraduates and graduate students and the applications of the clinician in learning.
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Objective To quantify the mRNA of 5-lipoxygenase (5-LO) in the rat peritoneal macrophages cultured in vitro by capillary electrophoresis (CE). Methods The rat peritoneal macrophages were isolated and cultured for indicated time. The mRNA of 5-LO was detected by RT-PCR, and the products of RT-PCR were quantified by CE. Results The DNA fragments in the 100 bp DNA marker and the products of the RT-PCR were separated successfully by CE with the sieving buffer containing 1.8% hydroxypropylmethylcellulose (HPMC). It proved that there were no significant changes on the expressions of 5-LO mRNA when the cells were cultured for 72 h quantified by CE. The mRNA of 5-LO significantly decreased by almost 80% by CE with the cells cultured for 120 h in vitro.Conclusions The products of RT-PCR could be separated and quantified by CE directly.The 5-LO mRNA could express normally in the rat peritoneal macrophages for 72 h in vitro.