Objective To investigate the effect of transforming growth factor-?1 (TGF-?1) on the activity of connective tissue growth factor(CTGF) gene promoter in human renal proximal tubular epithelial cell line HK-2. Methods The regulation fragment of 5' flanking region of human CTGF gene was linked to pGL3-Basic vector. The recombinant plasmid pCTGF-luc was transient transfected to HK-2 cells. The activity of CTGF promoter after treatment of TGF-?1 and mitogen-activated prontein kinases (MAPK) pathway inhibitors was assayed by means of luciferase reporter gene assay system. Results TGF-?1-induced increase of CTGF promoter activity was concentration-dependent, with a plateau at 5 ng/ml by 1.82-fold vs control (P

Objective To study the difference of oxygen free radical metabolism between healthy native Tibetans and mi- grated Hans at different altitude.Methods The activity of Total-antioxidation capability(T-AOC),superoxide dis- mutase(SOD),glutathione peroxidase(GSH-PX)and the content of,reactive oxygen species(ROS),malondialdehyde (MDA)and nitric oxide(NO)in serum in healthy native Tibetans and migrated Hans at different altitude were meas- ured.Results The activity of T-AOC,SOD,GSH-PX and the content of NO were increased in serum in native Tibet- ans group than that in migrated Hans group(P

The aim of this study is to prepare hyaluronic acid (HA) modified core-shell liponanoparticles (pHA-LCS-NPs) as gene delivery system and investigate its gene transfection efficiency in human retinal pigment epithelium (ARPE-19) cells in vitro. The pHA-LCS-NPs was prepared by firstly hydrating dry lipid film with CS-NPs suspension to get LCS-NPs, then modifying the lipid bilayer with HA by amidation reaction between HA and dioleoyl phosphatidylethanolamine (DOPE). Its morphology, particle size and zeta potential were investigated. XTT assay was used to evaluate the cell safety of different vectors in vitro. The gene transfection efficiency of pHA-LCS-NPs modified with different contents of HA was investigated in ARPE-19 cells with green fluorescent protein (pEGFP) as the reporter gene. The results showed that the obtained pHA-LCS-NPs exhibited a clear core-shell structure with the average particles size of (214.9 +/- 7.2) nm and zeta potential of (-35 +/- 3.7) mV. The 24 h cumulative release of gene from pHA-LCS-NPs was less than 30%. After 48 h incubation, gene transfection efficiency of pHA-LCS-NPs/pEGFP was 1.81 times and 3.75 times higher than that of CS-NPs/pEGFP and naked pEGFP, respectively. Also no obvious cytotoxicity was observed on pHA-LCS-NPs. It suggested that the pHA-LCS-NPs might be promising non-viral gene delivery systems with high efficiency and low cytotoxicity.
Cell Survival ; Gene Transfer Techniques ; Genes, Reporter ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; Humans ; Hyaluronic Acid ; chemistry ; pharmacology ; Lipids ; Nanoparticles ; Particle Size ; Phosphatidylethanolamines ; chemistry ; pharmacology ; Retinal Pigment Epithelium ; drug effects ; Transfection

The DNA structures could be altered or even damaged by exogeous or endogenous factors during cell proliferation. Failure of effective and timely repair will lead to cell cycle arrest or apoptosis. By taking the advantage of the quick proliferation of cancer cells, DNA damage induction, cell cycle arrest and apoptosis promotion have become important strategies for ant-cancer chemotherapy. Previous reports showed that an array of natural compounds inhibit cancer cell proliferation by inducing DNA damage, which have therapeutic potentials for anti-cancer drug research and development.
Animals ; Biological Products ; pharmacology ; therapeutic use ; DNA Damage ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Neoplasms ; drug therapy

Objective To explore the imaging features of papillary thyroid microcarcinoma(PTMC) with real time contrast-enhanced ultrasound.Methods One hundredand forty-three cases with 149 thyroid nodules(no diffuse lession) were divided into two groups according to the diameter size(group 1,＜0.5 cm;group 2,0.5-1.0 cm) and examined by contrast-enhanced ultrasound during preoperation.Pathology was followed up as golden diagnosis criteria.Results Seventy-five benign tumors and 74 PTMC were confirmed by pathology.There were significant differences in echoes homogeneity between benign and malignant tumors in group 2(W =1 029.5,Z =-5.524,P =0.000) but no in group 1(W =933.0,Z =-1.738,P =0.082).And nonhomogeneous enhancement were showed in most PTMC in group 2.But most PTMC showed homogeneous enhancement in group 1.Conclusions Contrast-enhanced ultrasound is valuable in diagnosis of PTMC with the diameter size of 0.5-1.0 cm.

ObjectiveTo investigate the choice of graft,and transfusion and immunosuppressant regimen of a RhD negative recipient in liver transplantation.MethodsOne RhD negative patient with hepatocellular carcinoma who received a liver graft from a RhD positive donor was retrospectively studied,and related references were reviewed.During the operation,the patient received five units of RhD negative/O RBC,3000 ml positive/O plasma and 30 units cryoprecipitate.Tacrolimus and prednisone were used to prevent rejection,and prednisone was withdrawn 30d post transplant.Results The patient's liver function recovered smoothly,without any acute rejection or hemolytic reaction.Anti-D antibody was not detected.The patient suffered from cancer recurrence 9 months and died of brain metastasis 13 months after transplantation.ConclusionsA RhD negative recipient can receive a graft from a RhD positive donor in liver transplantation.The selection of RBC and platelet from RhD negative or positive donors should be based on the result of anti-D antibody test.Plasma and cryoprecipitate can be transfused regardless of Rh type.Enhanced immunosuppressant regimen was unnecessary for these patients.

Objective To analyze negative lymph nodes of 34 non-small cell lung cancer(NCLC) patients with total correction by means of fluorescent quantitation PCR and immunohistcchemistry,and to form molecular bi- ology staging.Methods Clinical data and tissue samples of 193 lymph nodes were collected from 34 patients under- going resection for non-small cell lung cancer.Using fluorescent quantitation reverse transcription-polymerase chain reaction(RT-PCR) and immunohistochemistry method,lymph nodes were examined for CEA gene mRNA,P53 and CK to form molecular biology staging.All the patients were followed-up for an average of forty months.Results The CEAmRNA was identified in 21.7% (42/193) lymph nodes negative patients from 17 patients(17/34,50%); TMN staging was up-regulated in 8 patients;positive lymph nodes were increased in 9 patients.P53 and AE1/AE3 were identified 9.8%(19/193) from 11 patients,18.6 % (36/193)from 15 patients,separately;TMN staging was up-regulated in 2 patients of P53 examination and 5 patients of AE1/AE3 analysis;positive lymph nodes were in- creased in in 7 patients of P53 examination and 11 patients of AE1/AE3 analysis.There was obvious statistical sig- nificance in them,but the molecular biology staging based on the three markers was not an independent factor on re- currence and metasis of lung cancer.Conclusion CEAmRNA.P53 and AE1/AE3 analysis could find lung cancer micrometasis more sensitively to form molecular biology staging which was relative to the prognosis,but not an inde- pendent prognostic indicator.It might be good to the therapy strategy after operation.

Objective To establish the HepG2 cell lines which can stably express and replicate hepatitis 13 virus (HBV).Methods One point two X unit length of HBV genome was cloned intn SalⅠsite of the eukaryotic expression vector pREP10 to construct the recombinant plasmid pREP-HBV. Human hepatoblastoma cell HepG2 was transfected with pREP-HBV by Lipofectamine 2000 and seh,cted by bygromycin at the concentration of 250?g/mL.HBsAg and HBeAg were monitored by enzyme linked immunosorbent assay (ELISA)kits.H13V particles presemed in supernatant were ex- amincd by electronic microscopy.DNA isolated from intracellular HBV core particles was analyzed by Southbern blot using HBV-specific probe.Results The recombinant vector pREP HBV containing 1.2?unit length of HBV DNA was constructed successfully.After transfection of pREP-HBV to HepG2 cells and consistently cultured in hygromycin selective medium.5 drug-resistant cell lines, RHBV1-RHBV5.were established,and all of them could stably express HBsAgand HBeAg.South ern blot analysis revealed that HBV could replicate in all cell lines,as confirmed by the presence of replicateintermediatc DNA in intracellular HBV core particles.Clustered 42 nm Dane particles as well as 22-26 nm spherical H13sAg particles in condensed cuhure supernatant were visualized by elec tronic microsopic analysis.Conclusion HepG2 ceil lines in which HBV can replicate and express specific antigens are successfully established.Up to now,the cells have been passaged every three days for 50 times.

Objective To investigate the morbidity and characteristic of nosocomial infections in end stage renal disease(ESRD) patients undergoing blood purification.Methods Medical records of ESRD patients undergoing blood purification from 2004 to 2005 were enrolled in this retrospective study of hospitalized cases.The clinical data of nosocomial infections in hemodialysis (HD)and peritoneal dialysis(PD)patients were analyzed separately.Results Nosocomial infection was identified in 76 of the 400 enrolled patients.In HD patients,pulmonary infection was the most common nosocomial infection(53.3%),most due to gram-negative microorganisms,followed by bloodstream(16.7%)and urinary tract infection(15%),most of both were due to gram-positive bacteria.Pulmonary infection was usually complicated.Bloodstream infection was associated with the duration of placement of central vein catheters.Asymptomatic bacteriuria accounted for most of the urinary tract infection.In PD patients, most infections were pulmonary infection(65.5%),mainly caused by fungal pathogens,followed by peritonitis(20.7%), mainly due to gram-positive bacteria.Certain proportion of infections in both groups was caused by multiple microorganisms or identified in multiple sites.Conclusions The prevalence of nosocomial infection is high in hospitalized patients undergoing blood purification.The infection is complicated in terms of pathogen and clinical picture.Pulmonary infection is the most common in- fection.The prevalence of fungal infection is increasing.Effective prevention and therapeutic measures should be applied more vigorously in ESRD patients.

?ig-h_3 was first identified as a transforming growth factor-beta1-inducible gene in human lung adenocarcinoma cell line. It encodes for a secreted extracellular matrix (ECM) protein, which is thought to act on cell attachment and ECM composition. Previous study showed that ?ig-h_3 were highly expressed in human hepatoma cell lines and lowly expressed in human normal hepatic cells. The present study aimed to transfect ?ig-h_3 into 7721 cells to investigate its effect on secretion of MMPs in the transfected human hepatoma cells. Full-length ?ig-h_3 gene,cloned by reverse transcription polymerase chain reaction (RT-PCR) was inserted into the eukaryotic expression vector pEGFP-C_2. The recombinant plasmid was transfected into 7721 cells with Lipofectamine2000 and Gelatin-Zymography were adopted to detect the production of MMPs in the transfected cells. Results showed that ?ig-h_3/pEGFP-C_2 recombinant expression plasmid was successfully constructed and achieved high transfection efficiency. MMPs expression of the transfected cells was promoted significantly. These results suggest that overexpression of ?ig-h_3 promoted the production of MMPs, indicating that ?ig-h_3 may play roles in the invasive and metastatic processes of hepatoma.