Objective To transfect a recombinant pcDNA3.1-DCN into HepG2 cells and detect the expressions of decorin(DCN) and p21WAF1/CIP1 so as to investigate the mechanism of tumor suppression of DCN.Methods HepG2 cells were divided into pcDNA3.1-dec-HepG2 group(transfected group) and pcDNA3.1-HepG2 group(control group).After the recombinant pcDNA3.1-DCN and pcDNA3.1 were transfected into HepG2 hepatoma cells by liposome,the stably transfected cell lines were established using G418 screening.RT-PCR method was used to detect the expressions of DCN and p21WAF1/ CIP1 mRNA;Western blotting method was used to detect the expressions of DCN and p21WAF1/CIP1 protein;immunohistochemistry was performed to detect the expression of DCN protein.Results Compared with control group,the expressions of DCN and p21WAF1/CIP1 mRNA were significantly increased in transfected group by RT-PCR(P

Objective To construct the vector for ?-galactosidase (?-gal) gene expression regulation with tetracycline-regulated gene expression system,and to control ?-galactosidase gene expression in hepG2 cells with the vector.So that offer a experimental base for regulating gene expression for liver cancer gene therpy with this system.Methods The primers for explanding two tetracycline operator (TetO2) gene according to the gene nucleotide sequences of TetO2 gene and pcDNA3.1 vector were designed.TetO2 gene was amplified by PCR with pcDNA3.1 plasmid as template.The TetO2 PCR products were cloned into pcDNA3.1 and the vector was named as pcDNA3.1-TetO2.The pcDNA3.1-Teto2 with TetO2 PCR products sequence was analyzed by ABI3130 sequencing analysis.Then ?-gal gene was cloned into pcDNA3.1-TetO2,the vector was named as pcDNA3.1-TetO2-?-gal.The pcDNA6/TR plasmid vector with tetracycline repressor(TR) was transfected into HepG2 cells by Lipotap,and the stable transfection cells were screened by blasticidin.TR gene expression was detected by RT-PCR in HepG-2 cells with and without pcDNA6/TR plasmid transfection.The pcDNA3.1-TetO2-?-gal plasmid vector was transfected into HepG2 cells with and without pcDNA6/TR plasmid transfection,and 3 to 4 d later, these transfected gene cells were treated with doxycline(4 mg?L-1).After 48 h,?-gal gene expression was detected with ?-gal cell staining.Results The pcDNA3.1-TetO2 sequencing analysis results showed that a cassette was made for a cytomegalovirus-type 2 tetracycline operator (TetO2)-TetO2 promoter in pcDNA3.1-Teto2.The double digestion reslut of pcDNA3.1-TetO2-?-gal plasmid vector demonstrated that ?-gal gene was successfully cloned into pcDNA3.1-TetO2 vector .The RT-PCR result of TR gene showed TR gene could be expressed in HepG2 cells with pcDNA6/TR plasmid transfection and TR gene didn’t express in HepG2 cells without pcDNA6/TR plasmid transfection.?-gal gene expressed in HepG2 cells without pcDNA6/TR plasmid transfection,but didn’t express in HepG2 cells with pcDNA6/TR plasmid transfection.However,?-gal gene expression could be induced with doxycline in HepG2 cells with pcDNA6/TR plasmid transfection.Conclusion The tetracycline-regulated gene expression system vector for regulating ?-gal gene expression is successfully constructed and the vector system can control ?-gal gene expression in HepG2 cells.

Objective To investigate the effect of continuous health education on the control of glucose and the changes of behavior. Methods One hundred and twenty-six newly diagnosed type 2 diabetic patients were randomly divided into control group and observation group,each group included 63 patients. The control group just received health education in hospitalization period,and the observation group received continuous health education both in and after hospitalization. The control of glucose and the changes of behavior were evaluated 3 months and 1 year later. Results After 3 months and 1 year treatment,the FPG,2hPG and HbA1c were significantly decreased compared to pre-treatment both in control group and observation group(P<0. 05). The FPG,2hPG and HbA1c were significantly decreased in the ob-servation group compared to the control groups(P<0. 05). The behaviors contained that mastery of diabetic knowledge,control of food in-take,persist in physical exercise, rational administration, self-monitoring of blood glucose and regular reexamination were significantly im-proved both in control group and observation group compared to pre-treatment (P<0. 05),and the behaviors were significantly improved in the observation group compared to the control group (P<0. 05). Conclusion Compared to control group,observation group had more ad-vantages in the control of glucose and the changes of behavior in newly diagnosed type 2 diabetic patients.

Objective To investigate the significance of serum interleukin-1β,interleukin-2,interleukin-6,TNF-α,erythrocyte sedimentation rate(ESR),c-reactive protein(CRP),IgA,IgG and IgM in patients with ankylosing spondylitis(AS),and to analyze their relationship with the Bath ankylosing spondylitis disease activity index (BASDAI). Methods Participants of this study included 45 AS patients(patient group)and 30 healthy subjects (control group).The patient group was further divided into an active subgroup and an inactive subgroup based on the disease activity assessed by using BASDAI.Then serum levels of IL-1β,IL-2,IL-6,TNF-α,ESR,CRP,IgA,IgC and IgM were tested in all the subjects,and the values were compared between different groups.In addition,the relationship between various parameters and those with BASDAI were also evaluated. Results The serum levels of ESR,CRP,IgA,lgM,IgG,IL-1β,IL-2,IL-6 and TNF-α in patients with AS were significantly higher than those in the healthy controls(P<0.05).The serum level of CRP,IL-2 and IL-6 were significantly different between the active and inactive subgroups(P<0.05).There was a positive correlation between CRP and IL-6,IL-6 and IL-2,as well as IgA and IgG levels.The IL-2,IL-6 and CRP levels were positive correlated with BASDAI.Conclusion It was suggested that the serum levels of IL-2,IL-6 and CRP can be used to assess the disease activity in patients with AS.

The quality control process throughout the Ebola virus nucleic acid detection in Sierra Leone-China Friendship Biological Safety Laboratory (SLE-CHN Biosafety Lab) was described in detail, in order to comprehensively display the scientific, rigorous, accurate and efficient practice in detection of Ebola virus of first batch detection team in SLE-CHN Biosafety Lab. Firstly, the key points of laboratory quality control system was described, including the managements and organizing, quality control documents and information management, instrument, reagents and supplies, assessment, facilities design and space allocation, laboratory maintenance and biosecurity. Secondly, the application of quality control methods in the whole process of the Ebola virus detection, including before the test, during the test and after the test, was analyzed. The excellent and professional laboratory staffs, the implementation of humanized management are the cornerstone of the success; High-level biological safety protection is the premise for effective quality control and completion of Ebola virus detection tasks. And professional logistics is prerequisite for launching the laboratory diagnosis of Ebola virus. The establishment and running of SLE-CHN Biosafety Lab has landmark significance for the friendship between Sierra Leone and China, and the lab becomes the most important base for Ebola virus laboratory testing in Sierra Leone.
China ; Ebolavirus ; classification ; genetics ; isolation & purification ; Hemorrhagic Fever, Ebola ; diagnosis ; virology ; Humans ; Laboratories ; manpower ; standards ; Laboratory Infection ; Quality Control ; RNA, Viral ; genetics ; Sierra Leone

Obesity has become one of the most serious issues threatening the health of humankind, and we conducted this study to examine whether and how celastrol protects against obesity.

DNA ; isolation & purification ; Formaldehyde ; Genes, erbB-1 ; genetics ; Genetic Techniques ; Humans ; Mutation ; Paraffin ; Tissue Fixation