Aged ; Female ; Humans ; Male ; Middle Aged ; Silicosis

Objective To construct the eukaryotic expression vector of human PRKCB1 containing enhanced green fluorescence protein gene and transfer into human umbilical vein endothelial cells(HUVECs),then identify activity of expression protein.Methods The pReceiver-M29-PRKCB1 eukaryotic expression plasmid was constructed by frame amplified from pMD18-T-PRKCB1 plasmid.Then the recombinant plasmids were identified by enzyme analysis and DNA sequencing.According to optimized conditions,the eukaryotic expression plasmids were transfered into HUVECs and observed under fluorescence microscope.After that,transfection efficiency was calculated under random vision.The plasma membrane/cytosol ratio of fluorescence was calculated under confocal microscope.The translocation was identified.Results The gene sequence was completely consistent with that reported in GenBank.The enhanced green fluorescence protein was observed in HUVECs after 48 hours.Transfection efficiency was 18.6%?1.6%.The translocation was observed.Conclusion The eukaryotic expression plasmid is successfullyconstructed and transfered into HUVECs,the translocation was identified.It is a potential tool for screening HUVECs stably expressing human protein kinase C ?2 and isolating protein complex.

Objective To study the Ile796Val polymorphism in human SCAP gene in Hubei area, and analyze its correlation with coronary heart disease (CHP) and hypertension and the relationship between polymorphism and lipid metabolism. Methods Using PCR RFLP, we detected genotypes of Ile796Val polymorphism in human SCAP gene. Results The allele A frequencies of Ile796Val in human SCAP gene for controls, CHD patients and the hypertension patients were 0.32, 0.45 and 0.42 respectively. The allele G frequencies were 0 68, 0.55 and 0.58 respectively. There were significant differences in frequencies of genotype and alleles between controls and hypertension group. And there was significant difference in the level of TC, LDL C and ApoB. In CHD group, there was significant difference in the TC level between different genotypes. In hypertension patients, although a difference was noted in genotype, there was no significant difference in allele frequencies and lipid level exceps a significant difference in the levels of TC, LDL C and ApoB in hypertension patients. Conclusion Ile796Val polymorphism in human SCAP gene may be a great agent to cause disorder in the lipid level of blood and lipid metabolism of tissue. It is of great significance in disorder in lipid metabolism of inter cellular and genetic investigation of hyperlipidemia.

【Objective】To establish a simple,rapid and practical method for the in-vitro isolation,purification and proliferation of rat marrow mesenchymal stem cells(MSCs).【Methods】Rat MSCs were isolated and purified by differential adhesion method.The effects of conditioned medium on the morphologic characteristics and growth of rat MSCs were observed.【Results】Rat MSCs were effectively purified by differential adhesion method,and grew well with good proliferation and normal morphology in conditioned medium.【Conclusion】Differential adhesion method and conditioned medium are effective for the purification and culture of rat MSCs.

The MPN method was used to enumerate ammonia-oxidizing bacteria (AOB) in water and sediments of several shallow lakes. The suitable incubation time, medium types and substrate (ammonium sulphate) concentrations were studied. The results showed that, MPN values increased with the incubation time, reaching a stable maximum at some time stages, which was 40 days in all the samples for MSF medium. Among the three media used (XZ-AOB、MSF、SW), MSF give the highest MPN value. In addition, am- monium sulphate concentration in medium was an important factor affecting MPN estimation of AOB. Compared to AOB in lake sediments, AOB in lake water was more sensitive to ammonium sulphate concentration.

Objective To detect the expression of RXR? gene protein in gastric cancer, superficial gastritis, normal gastric mucosa, in order to explore the relationship between the development of gastric cancer, combined with clinical and pathological analysis of its clinical significance. Methods Using immunohistochemical detection RXR? protein in gastric cancer, superficial gastritis, gastric mucosa in the normal expression. Results RXR? protein mainly located in the nucleus of cells, there are a small number of cytoplasmic staining, a brown or yellow granular, in gastric cancer, superficial gastritis, normal gastric mucosa, RXR? protein expression rates were 25.0%, 81.1%, 96.1%; group and the normal mucosa and superficial gastritis, gastric cancer group RXR? expression rate has dropped, there were significant differences (P0.05). The lymph node metastasis and tumor pathology and clinical phases closely related (P

Objective To construct the endothelial cell model with overexpressed human protein kinase C ?2(PKC?2) after high glucose inducement in order to study the function of human PKC?2. Methods The PRKCB1 gene was amplified from pMD18-T-PRKCB1 plasmid and then directly cloned into shuttle plasmid pDC315 to construct shuttle plasmid. Then the recombinant shuttle plasmid and adenovirus genomic plasmid pBHGlox△E1,3Cre were cotransfected into 293 cells to construct recombinant adenovirus Ad-PRKCB1. The virus titer was calculated by TCID50. The Ad-PRKCB1 was verified by immunocytochemistry and RT-PCR. Ad-PRKCB1 was transfected into human umbilical vein endothelial cells (HUVECs) followed by the treatment of high glucose (25 mmol/L) for 96 h, and then the cells were observed by laser scanning confocal microscopy (LSCM). Results Restrictive endonuclease digestion and PCR result showed that the target gene was correctly cloned into shuttle plasmid. Cytopathic effect (CPE) was observed under inverted microscope through homologous recombination. The virus titer was 7.9?109 IU/ml. Immunocytochemical staining and RT-PCR indicated the expression of human PKC?2 in the transfected HUVECs. The translocation was observed under LSCM. Conclusion A recombinant adenovirus vector with human PKC?2 and the endothelial cell model with human PKC?2 overexpression by high glucose are constructed successfully.

Homeostasis between the host and viruses is naturally maintained.On the one hand,the immune system activates the immune re-sponse to kill or eliminate viruses;on the other hand,the immune system controls the immune response to maintain immune homeostasis. The cause of persistent infections with hepatitis viruses such as HBV and HCV is that viral molecules damage the immune system of the host and their variants escape immune clearance.Long-term coexistence of the host and viruses is the process involving various immune cells and molecules and is the result of homeostasis maintenance in antiviral immune response.The immune homeostasis maintained during persis-tent infections with hepatitis viruses is analyzed by the cellular and molecular mechanisms.

Objective To study the effect of climate change on physiological indexes of elderly people.Methods From January,2006 to January,2007,30 elderly couples were selected and blood pressure,pulse,body weight were investigated.The weather data was collected.Results The lowest value of body weight appeared in February in normotensive subjects,in September in hypertension subjects.The lowest value of body temperature in two groups appeared in February and March.The lowest value of pulse appeared in March and April in normotensive subjects,in September in hypertension subjects.The curve of blood pressure change(SBP and DBP) presented"V"in two groups.The lowest value appeared in summer(July-September).The highest value appeared in winter(December-February).The blood pressure in winter was higher than that in summer(P

Objective Micronucleus test(MNT) was used to evaluate the single and combined genotoxicity of dichlorvos(DDVP),dimethoate(DM) and malathion(Mal) in HepG2 cell line,and 2?2 factorial design was adopted to elucidate the combined genotoxic effects.Methods In cytotoxicity test,HepG2 cells were exposed to the three OPPs(DDCP,DM,Mal) for 4 h respectively,and the doses at which cell viability above 80% were selected for the MNT,DDVP:3.125-25 ?g/ml,DM:25-200 ?g/ml,Mal:50400 ?g/ml,the micronucleated cell(MNC) rates and the replicative index(RI) were calculated.The combined genotoxicity of them was investigated with their doses as follows:low dose(DDVP:3.125 ?g/ml,DM:25 ?g/ml,Mal:50 ?g/ml);high dose(DDVP:12.5 ?g/ml,DM:100 ?g/ml,Mal:200 ?g/ml).Results In MNT,after treatment of HepG2 cells with DDVP,DM or Mal alone for 4 h,the MNC rates were increased in a dose-response manner(DDVP:r=0.955,P