Objective To analyze behavior of having multiple sexual partners among outside school adolescents and the impact factors in one county.Method Participatory method was adopted in the survey,trainees of an occupational training center were trained to investigate their peers with anonymous questionnaires.Results The subjects who had more than 3 sexual partners accounted for 38.3%,and the factors related to multiple sexual partners were complicated.The most im- portant protective factor was to raise level of HIV/AIDS related knowledge (OR=0.85);the key risk factors were: promiscuous behaviors (OR=4.91) and prostitution(OR=3.37) among their friends.Conclusion For reducing behav- ior of having multiple sexual partners among outside school adolescents,it is essential to promote HIV/AIDS related health education and to enhance their ability to respond to pressures from their bad peers.

Adult ; Bone Neoplasms ; diagnosis ; surgery ; Humans ; Ilium ; Magnetic Resonance Imaging ; Male ; Osteoma, Osteoid ; diagnosis ; surgery ; Tomography, X-Ray Computed

Diagnosis, Differential ; Hemangiopericytoma ; metabolism ; pathology ; Hemangiosarcoma ; metabolism ; pathology ; Histiocytoma, Malignant Fibrous ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Leiomyosarcoma ; metabolism ; pathology ; Nerve Sheath Neoplasms ; metabolism ; pathology ; Neuroectodermal Tumors, Primitive ; metabolism ; pathology ; Rhabdoid Tumor ; metabolism ; pathology ; Rhabdomyosarcoma ; metabolism ; pathology ; Soft Tissue Neoplasms ; metabolism ; pathology ; Urinary Bladder ; metabolism ; pathology

OBJECTIVEModeling HAdV-3 infect Hep-2 cells in vitro. The effect of realgar nanoparticles on the expression of HAdV-3 is detected by using fluorescent quantitative PCR.

METHODSThe experiment is divided into four groups: Hep-2 cells control group, HAdV-3 virus control group, realgar nanoparticle group and ribavirin group. In order to detect HAdV-3 viral load, add realgar nanoparticles and ribavirin in vitro and remain that vitro for 24 hours when HAdV-3 has infected Hep-2 cells, extract total DNA of Hep-2 cells infected by HAdV-3, and establish Real-time PCR reaction system of every experimental groups.

RESULTThe Hep-2 cells group has no amplification curve, the Ct value is greater than 35, which illustrate HAdV-3 pathogen detection is negative. However, realgar nanoparticles group, ribavirin group and the HAdV-3 group have amplification curve, the Ct values are 29.30 +/- 0.08, 33.05 +/- 1.29, 26.01 +/- 0.25 respectively, which illustrate HAdV-3 pathogen detection is positive. The viral copy amount of the adenovirus group(66 699 932 +/- 23.85) is more than that of realgar nanoparticles group (912 435.44 +/- 16.57), and much greater than that of ribavirin group (459 124.84 +/- 12.82) (P < 0.05).

CONCLUSIONThe model of Hep-2 cell infected by HAdV-3 is reliable. The method of quantitative PCR is sensitive and specific. Realgar nanoparticles have a certain inhibition role for adenovirus nucleic acid replication.

Adenoviridae Infections ; virology ; Adenoviruses, Human ; drug effects ; genetics ; physiology ; Arsenicals ; chemistry ; pharmacology ; Gene Expression Regulation, Viral ; drug effects ; Hep G2 Cells ; Humans ; Nanoparticles ; chemistry ; Sulfides ; chemistry ; pharmacology ; Virus Replication ; drug effects

ObjectiveTo prepare high specific monoclonal antibodies(mAbs) against BP26 of Brucella(B.)melitensis.Methods A recombinant plasmid pET-28a-BP26 was constructed and transformed into competent Escherichia coli BL21 (DE3),and then the bacteria were induced by 1 mmol/L isopropylthio-β-D-galactoside (IPTG).After induction,the recombinant BP26 protein (rBP26) was purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PGAE) and nickel ion affinity chromatography(Ni-NTA).Mice were inoculated with rBP26 antigens for three times at 2-week intervals.The first subcutaneous injection contained 100 μg rBP26 with 0.1 ml complete Freund adjuvant.The second subcutaneous injection was 50 μg rBP26 with 0.1 ml incomplete Freund adjuvant.The antibody titers to rBP26 were determined 2 weeks after each reimmunization.Three days before cell fusion,the mice with the highest titer were intraperitoneally injected with 50 μg rBP26 in 0.1 ml PBS.Pre- and post-immunization sera were collected and used as negative or positive controls for screening mAbs.Mice with the highest titer were sacrificed and spleen cells were isolated.The spleen cells of rBP26 immunized mice were fused with SP2/0 myeloma cells in a ratio of 5 ∶ 1 by polyethylene glycol(PEG) 1450.Antibody-producing hybridomas were primarily screened by an indirect enzyme-linked immunosorbnent assay(ELISA) with rBP26.Reactive hybridomas were subcloned for 3 times,then the strains of hybridoma cells secreting antibodies against BP26 were obtained.Supernatant of cloned hybridoma cultures was collected for mAb analyses.These mAbs were named by the hybridoma clone number and tested their reactivity to membrane proteins extracted(NMP) from B.melitensis vaccine strain(M5-90) by Western blotting and Dot-ELISA.mAbs isotyping and kappa(κ) or lambda(λ) light chain was identified by Mouse Monoclonal Antibody Isotyping Kit.Results A total of two mAbs reactive to rBP26 of B.melitensis were selected from antibody screening hybridomas by indirect-ELISA.The two mAbs were named 3C3 and 5A5,and identified as IgG1 (κ) and IgG2(κ),respectively.They could react with NMP from M5-90.Conclusions Results of identification show that two mAbs against rBP26 can be produced.The two mAbs can recognize natural BP26 protein,giving the experimental materials for further research on identification of its epitopes.

The complex level of constructing biopharmaceutics classification system of Chinese materia medica CMMBCS) was the study of traditional Chinese compound, on the premise of insisting that the multicomponent simultaneous determination, when carrying out the study of intestinal permeability, the primary task was to define the source of the components that was absorbed through the intestinal wall, namely, which medicinal material the components belonged to in traditional Chinese compound. The technology of chemical fingerprint and in vitro everted gut sac model were used in this research to make multicomponent an intuitive source attribution which permeated the intestine in the classic formula Gegen Qinlian decoction, and to lay the foundation for the further qualitative and quantitative research of intestinal permeability.
Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Intestines ; metabolism ; Male ; Permeability ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

Objective The objective of this study was to investigate a risk of prostate cancer(PCa) at a repeat biopsy in patients with chronic prostate inflammation and widespread high grade prostatic intra epithelial neoplasia(wHGPIN).Methods From July 2006 to December 2014,172 cases of prostate biopsy were collected.All of them were diagnosed as HGPIN for the first biopsy,punctured by transrectal ultrasound for 12 points.After the first puncture for 6 months,patients were punctured for rebiopsy.Multi-focal wHGPIN was defined as a high -grade prostate intraepithelial neoplasia with 2 or more cores detection in a prostate biopsy.Isolated HGPIN was defined as a high-grade prostate intraepithelial neoplasia with only one core detection in a prostate biopsy.Results Seventy-two patients with HGPIN were isolated from primary HGPIN,102 patients with isolated HGPIN,17 patients with chronic prostatitis,70 with multifocal HGPIN and 54 with chronic prostatitis.Forth-eight of 172 patients initial diagnosis of HGPIN was diagnosed as PCa at rebiopsy.The detection rate of wHGPIN was 52.86％ (37/70)and isolated HGPIN for 10.88％ (11/102).They showed a statistically difference between two groups(P ＜0.001).The detection rate of PCa in HGPIN patients with chronic prostatitis was higher than that in patients without chronic prostatitis(P =0.011).Chronic prostatitis and multifocal wHGPIN were a risk factor for prostate cancer independent by rebiopsy,confirmed by the logistic regression model.Conclusion Rebiopsy is a high risk factor of prostate adenocarcinoma for patients with chronic prostatitis and multifocal HGPIN initially diagnosed by the first biopsy.Therefore,these patients are recommended under ultrasound induced by rectal prostate rebiopsy.

Objective To evaluate the influential factors of iron acquisition during Francisella tularensis LVS infection of mouse macrophages.Methods F.tularensis LVS expressing green fluorescent protein was used to infect murine macrophage J774A.1 cells.Transferrin receptor 1(Tfr1)was detected with mono-antibody and visualized with a goat-anti mouse IgG conjugated to Alexa 594.The expression profile of 5 iron metabolism related genes of J774A.1 murine macrophages uninfected or infected with F.tularensis LVS was determined with real-time PCR.Immunoblot analysis was used to compare the Tfr1 expression of live Francisella infected macrophage with dead bacteria.Tfr1 knock-off in J774A.1 cells was performed with siRNA.The transfected cells were infected with Francisella for immunoblotting and microscopy and infection assay.Results It was revealed that the live vaccine strain of F.tularensis induced the expression of Tfr1 in host macrophages.Gene expression analysis indicated that F.tularensis LVS drove an active iron acquisition program with induction of Tfr1 and iron regulatory proteins(Irp1 and Irp2).It was shown by Western-blotting that the siRNA-Tfrc-1 could knock off about 75% of Tfr1 in J774A.1 cells.It was determined by infection assay that,Tfr1 was knocked off,the bacteria number at 1h infection with Francisella was not different from that of control(F=1.06,P=0.326 5),while it was decreased significantly after 24h of infection(F=24.12,P=0.000 6).Conclusions It is demonstrated that upregulation of the Tfr1 may be mediated by post-transcriptional regulation during early infection,but sustained later through increased expression of Irp 1 and Irp 2.Increased expression of Tfr1 expands the intracellular iron pool through transferrin-mediated delivery and may thus be readily available for uptaking by Francisella.Knocking off the expression of Tfr1 does not affect bacterial invasion.Francisella,however,may fail to proliferate in macrophages in which the expression of transferrin receptor has been suppressed.

Background Scarring of the filtering bleb is a main cause of filtering surgical failure in glaucoma.It has been reposed that tetrandrine could suppress the proliferation of cultured human fibroblast of Tenons capsule in vitro and thus has the potential effect to prevent scarring after the filtering surgery. Objective Present study was to investigate the anti-cicatricial effect of tetrandrine drug delivery system(Tet DDS)during filtration surgery. Methods Filtration surgery was performed in bilateral eyes of 18 New Zealand white rabbits.The Tet DDS with 0.3 mg Tet,0.2 mg Tet or free-Tet were implanted subcunjunctially during the surgery.The filtering blebs were scored in 1 day,4,7,10,14 days after referring to the corneal thickness and bleb range under the slit-lamp biomicroscopy.The morphology of filtering bleb was assessed by in vivo confocal microscopy in 7 and 14 days after operation.The filtering bleb specimen was prepared in 7 and 14 days for the histopathological examination. Results The filtering bleb scores in Tet DDS implantation groups were significantly higher than those in free-Tet DDS group from 4 days through 14 days after trabeculectomy(P＜0.01),and the scores showed a considerably increase in 0.3 mg Tet DDS group compared with 0.2 mg Tet DDS group from 7 days through 14 days after trabeculectomy(P＜0.05).The filtering blebs of Tet DDS implantation groups were found with distinct subepithelial cystic spaces under the light microscopy and in vivo confocal microscopy on the 7th day and 14th day after surgery.Compared with free-Tet DDS group,the numbers of subepithelial mierocysts were much more(P＜0.01)and the area of microcysts was larger(P＜0.01)in Tet DDS group.The filtering tissue presented with more subepithelial microcysts and larger microcysts range in 0.3 mg Tet DDS group than 0.2 mg Tet DDS group in 7 and 14 days after operation(P＜0.05).The inflammatory cell infiltration wag milder in 0.3 mg Tet DDS group in comparison with 0.2 mg Tet DDS group and free-Ted DDS group.Conclusion Tet DDS has strong inhibitory effects on inflammatory cells activity and fibroblagt activity the early stage after filtering surgery and therefore improve the surgery success rate.