Objective To investigate the effects of caveolin-1 overexpressing on the growth of cervical squamous cell cancer Hela cell line. Methods Eukaryotic expression vector of human caveolin-1 gene was introduced into Hela cells by Lipofectamine. The clones stably overexpressed caveolin-1 were identiffed by RT-PCR, immunofluorescence cell staining techniques and Westernblotting. Cells proliferation viabihty was tested by MTT assay, and flow cytometry was used to assay the cell cycle and apoptosis, and the relative phosphorylation level of extracellular regulated protein kinases (Erk1/2) were detected by Westernblotting. Results The clones stably overexpressed caveolin-1 were obtained. Compared with the parental Hela cells, the tranfected cells exhibited a slower rate of growth. FAGS analysis results revealed that overexpression of caveolin-1 resulted in the cell cycle arrest in the G0/G1 [ ( 68. 04 ± 2. 57 ) % vs ( 53.41 ±1.01)%] phase and increased the apoptotic cell fraction[ (19. 18 ±2.20)% vs (5.63 ±0.55)%, P <0. 05 ]. Western blotting results showed that overexpression of caveolin-1 reduced the phosphorylation of Erk1/2(0.28 ±0.05 vs 0.81 ±0.07, P <0.05). Conclusions Overexpression of caveolin-1 suppressed the growth of Hela cells and induced apoptosis, down-regulation of Erk1/2 phosphorylation might be involved in its mechanism.

This study was performed to investigate the chemical constituents in the twigs and leaves of Harrisonia perforate. Six compounds were isolated from the 95% EtOH extract of the twigs and leaves of Harrisonia perforate by silica gel, ODS, Sephadex LH-20 column chromatographies and preparative HPLC. On the basis of chemical properties and spectra data, these compounds were identified as harriperfin E (1), kihadanin A (2), kihadanin B (3), 6α-acetoxyobacunol acetate (4), gardaubryone C (5), and β-sitosterol methyl ether (6), respectively. Compound 1 is a new chromone, and compounds 2-6 are isolated from this plant for the first time.
Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Phytochemicals ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Simaroubaceae ; chemistry

Objective To investigate the diagnosis and treatment of pulmonary tuberculosis in renal transplantation recipients.Method The clinical data of 8 renal transplantation recipients suffering from pulmonary tuberculosis infection were retrospectively analyzed.Result Fever,cough and expectoration were the most common symptoms,however,lacking typicality.Images of chest Xray and CT scan were various and couldn't verify TB infection from pneumonia.Seven of 8 cases were diagnosed through invasive methods,either bronchofibroscope or fiberthoracoscopy.Immunosuppressants were decreased in all cases.Three-drug regimens,including isoniazide,rifampicin and ethambutol or pyrazinarnide,were administrated as anti-tuberculosis chemotherapy.All the cases were cured,without episodes like acute rejection and liver function impairment.Conclusion Routine examinations are not sufficient to diagnose pulmonary tuberculosis in kidney transplantation recipients.While,invasive methods like bronchofibroscope and fiberthoracoscope are helpful.When diagnosed,patients should receive normative anti-tuberculosis treatment and immunosuppressive agents adjustment,which can benefit the prognosis of pulmonary tuberculosis in renal transplantation recipients.

Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.

OBJECTIVETo investigate the tissue toxicity and degradation of ZK60 magnesium alloy with micro-arc oxidation coatings (MAO-ZK60), in order to discuss the possibility of its potential application for orthopedic implantation.

METHODSEighteen Sprague-Dawley rats were randomized equally to three groups of A, B, C. MAO-ZK60 sticks were implanted in the femoral condyles of rats in group A (experimental group). Sticks of ZK60 magnesium alloy without any surface treatment (ZK60) were implanted in the femoral condyles of rats in group B (control group). The poly L-lactic acid (PLLA) sticks were implanted in the femoral condyles of rats in group C (control group). The changes of blood bio-chemical indexes of different groups were observed and compared. All the rats were sacrificed at 12 weeks and histological observation of liver and kidney were carried out to evaluate the hepatic and renal toxicity. Micro-CT was used to evaluate the degradation of the implants and to observe the bone-implant interface. GEHC MicroView software was operated to calculate the volume variation of magnesium alloy.

RESULTSThere was no apparent biochemical index change with time in each group, and there was no significant change among each group. No significant pathology change of liver and kidney was detected among three groups. By using a micro-CT, a gap was found on the bone-implant interface at 4 weeks after implantation in group A, which decreased gradually at 8 weeks after implantation and continued to decrease at 12 weeks after implantation. A good combination between bone and implant formed at 12 weeks after implantation. Group A has less change of volume with time than group B (P < 0.05).

CONCLUSIONZK60 magnesium alloy with micro-arc oxidation coatings is safe in vivo. It has higher corrosion resistance than ZK60 magnesium alloy without any surface treatment.

Alloys ; chemistry ; metabolism ; toxicity ; Animals ; Magnesium ; chemistry ; Male ; Rats ; Rats, Sprague-Dawley

Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.

OBJECTIVETo investigate the polymorphism of human cytomegalovirus (HCMV) UL150 open reading frame (ORF) in low-passaged clinical isolates, and to study the relationship between the polymorphism and different pathogenesis of congenital HCMV infection.

METHODSPCR was performed to amplify the entire HCMV UL150 ORF region of 29 clinical isolates, which had been proven containing detectable HCMV-DNA using fluorescence quantitative PCR. PCR amplification products were sequenced directly, and the data were analyzed.

RESULTSTotally 25 among 29 isolates were amplified, and 18 isolates were sequenced successfully. HCMV UL150 ORF sequences derived from congenitally infected infants were high variability. The UL150 ORF in all 18 clinical isolates shifted backward by 8 nucleotides leading to frame-shift, and contained a single nucleotide deletion at nucleotide position 226 compared with that of Toledo strain. The nucleotide diversity was 0.1% to 6.8% and the amino acid diversity was 0.2% to 19.2% related to Toledo strain. However, the nucleotide diversity was 0.1% to 6.4% and amino acid diversity was 0.2% to 8.3% by compared with Merlin strain. Compared with Toledo, 4 new cysteine residues and 13 additional posttranslational modification sites were observed in UL150 putative proteins of clinical isolates. Moreover, the UL150 putative protein contained an additional transmembrane helix at position of 4-17 amino acid related to Toledo.

CONCLUSIONHCMV UL150 ORF and deduced amino acid sequences of clinical strains are hypervariability. No obvious linkage between the polymorphism and different pathogenesis of congenital HCMV infection is found.

Amino Acid Sequence ; Base Sequence ; Cytomegalovirus ; genetics ; isolation & purification ; Cytomegalovirus Infections ; congenital ; virology ; DNA, Viral ; genetics ; Genes, Viral ; Genetic Variation ; Humans ; Infant ; Infant, Newborn ; Molecular Sequence Data ; Open Reading Frames ; Polymorphism, Single Nucleotide ; Sequence Homology, Amino Acid ; Viral Proteins ; chemistry ; genetics

The genus Nodulisporium, is known to produce secondary metabolites with structural diversity. A new alkaloid, 2-hy- droxy-1,1-dimethyl-1,2,3,9-tetrahydro-4H-carbazol-4-one(1), was isolated from the extract of a fungal strain Nodulisporium sp. fermented with rice, together with three known phenols, tyrosol(2), hydroxytyrosol(3), and hydroxytyrosol acetate(4). Their structures were identified by detailed spectroscopic analyses.
Alkaloids ; chemistry ; isolation & purification ; Xylariales ; chemistry

OBJECTIVETo investigate the variability of human cytomegalovirus (HCMV) UL138 open reading frame (ORF) in clinical strains.

METHODSHCMV UL138 ORF was amplified by polymerase chain reaction (PCR) and PCR amplification products were sequenced directly, and the data were analyzed in 19 clinical strains.

RESULTSUL138 ORF in all 30 clinical strains was amplified successfully. Compared with that of Toledo strain, the nucleotide and amino acid sequence identities of UL138 ORF in all strains were 97.41% to 99.41% and 98.24% to 99.42%, respectively. All of the nucleotide mutations were substitutions. The spatial structure and post-translational modification sites of UL138 encoded proteins were conserved. The result of phylogenetic tree showed that HCMV UL138 sequence variations were not definitely related with different clinical symptoms.

CONCLUSIONHCMV UL138 ORF in clinical strains is high conservation, which might be helpful for UL138 encoded protein to play a role in latent infection of HCMV.

Amino Acid Sequence ; Cytomegalovirus ; classification ; genetics ; Cytomegalovirus Infections ; congenital ; genetics ; Humans ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Protein Structure, Secondary ; Sequence Alignment ; Viral Proteins ; chemistry ; genetics

OBJECTIVETo investigate the effect of Rhizoma Curcumae (RC), arsenite trioxide (As2O3) on proliferation ana signal transduction molecule in lens epithelial cell (LEC), in order to provide experiment evidence for prevention and treatment of after cataract.

METHODProliferation of cultured bovine LEC were induced by induced by recombinant human basic fibroblast growth factor (rhbFGF); Inhibitory rates of LEC proliferation induced by RC, As2O3 were detected by methyl thiazolyl tetrazolium (MTT); Inhibitory effects of expression of proliferating cell nuclear antigen (PCNA) induced by RC, As2O3 in LEC were assayed via flow cytometer (FCM); Concentrations of LEC calcium ([Ca2+]i) were determined by spectrofluoremeter, intracellular concentrations of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) of LEC were measured by radioimmunoassay.

RESULTInhibitory rates of RC, As2O3 on LEC proliferation induced by rhbFGF increased significantly, showing dose-dependent (P < 0.01). PCNA expression of LEC proliferation induced by rhbFGF were down regulated obviously by RC, As2O3, showing dose-dependent (P < 0.01). Concentrations of [Ca2+]and cAMP increased and cGMP decreased significantly in LEC of proliferation inhibited by RC, As2O3 (P < 0.01).

CONCLUSIONRC, As2O3 can inhibit LEC proliferation obviously. Signal transductions of [Ca2+]i, cAMP, cGMP may be the important molecular mechanism. There are broad prospect for RC, As2O3 on prevention and treatment of after cataract.

Animals ; Arsenicals ; pharmacology ; Calcium ; metabolism ; Cattle ; Cell Proliferation ; drug effects ; Cells, Cultured ; Curcuma ; chemistry ; Cyclic AMP ; metabolism ; Cyclic GMP ; metabolism ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Fibroblast Growth Factor 2 ; genetics ; pharmacology ; Flow Cytometry ; Growth Inhibitors ; pharmacology ; Lens, Crystalline ; cytology ; drug effects ; metabolism ; Oxides ; pharmacology ; Proliferating Cell Nuclear Antigen ; metabolism ; Radioimmunoassay ; Recombinant Proteins ; pharmacology ; Rhizome ; chemistry ; Signal Transduction ; drug effects