OBJECTIVETo screen the specific antibody binding peptides of tuberculosis from phage-displayed random phage display (Ph.D.)-7 peptide libraries with purified IgG from tuberculosis serum, and to provide the basis for the development of serological detection reagent of tuberculosis.

METHODSPurified IgG of tuberculosis serum was used as solid ligand to screen the binding peptide from the Ph.D.-7 peptide library, according to the biopanning process of absorption, elution and amplification. Purified IgG of health people serum was used as the molecule of counter selection during the second and third selection. Phages were enriched after 3 rounds of screening, then 20 single phages separately eluted by IgG of tuberculosis and health people were randomly selected on each direction of the determination plates and amplified. The single chains DNA were extracted as template for sequencing. The combination abilities of selected clones to IgG of tuberculosis and health people were tested by indirect enzyme linked immunosorbent assay (ELISA), and the positive clone was identified. Serum samples, from 47 patients with tuberculosis and 37 healthy people vaccinated with BCG, were verified by positive phage clones using phage-ELISA. The verifying results of 24 serum samples used for panning were separately analyzed statistically.

RESULTSAfter 3 rounds of panning, remarkable enrichment of phages that could specifically bind with target molecules were observed. Single phages were randomly selected for sequencing analysis and 12 sequences were obtained. 12 phage clones with different sequences were amplified and detected with indirect ELISA and single phage H12 showed higher affinity with IgG of tuberculosis (S/N ≥ 2.1) and was identified as the positive clone. It was found that, in indirect ELISA with single Phage H12, the A(450) value of tuberculosis patients (0.105 ± 0.010) was significantly higher than that of healthy individuals (0.070 ± 0.005), and the t value was 2.912 (P < 0.0001). The A(450) value of 12 serum samples of tuberculosis patients and 12 samples of health individuals used for panning were 0.144 ± 0.016 and 0.052 ± 0.004, and the t value was 5.69 (P < 0.0001).

CONCLUSIONBy using phage-displayed random peptide libraries, we obtained the specific antibody binding peptides of tuberculosis, which showed specific binding activity with IgG of tuberculosis. It can provide a basis for the establishment of a new serological detection method of tuberculosis with peptide.

Adult ; Case-Control Studies ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Molecular Sequence Data ; Mycobacterium tuberculosis ; immunology ; Peptide Library ; Peptides ; immunology ; Tuberculosis ; diagnosis ; immunology ; microbiology ; Young Adult

Microbes play important roles in human health and disease. The interaction between microbes and hosts is a reciprocal relationship, which remains largely under-explored. Current com-putational resources lack manually and consistently curated data to connect metagenomic data to pathogenic microbes, microbial core genes, and disease phenotypes. We developed the MicroPhenoDB database by manually curating and consistently integrating microbe-disease associ-ation data. MicroPhenoDB provides 5677 non-redundant associations between 1781 microbes and 542 human disease phenotypes across more than 22 human body sites. MicroPhenoDB also pro-vides 696,934 relationships between 27,277 unique clade-specific core genes and 685 microbes. Dis-ease phenotypes are classified and described using the Experimental Factor Ontology (EFO). A refined score model was developed to prioritize the associations based on evidential metrics. The sequence search option in MicroPhenoDB enables rapid identification of existing pathogenic microbes in samples without running the usual metagenomic data processing and assembly. Micro-PhenoDB offers data browsing, searching, and visualization through user-friendly web interfaces and web service application programming interfaces. MicroPhenoDB is the first database platform to detail the relationships between pathogenic microbes, core genes, and disease phenotypes. It will accelerate metagenomic data analysis and assist studies in decoding microbes related to human dis-eases. MicroPhenoDB is available through http://www.liwzlab.cn/microphenodb and http://lilab2. sysu.edu.cn/microphenodb.

OBJECTIVETo explore the risk factors of children with high dmft.

METHODSIn suburban of Guangzhou, oral health of 401 3 - 4-year-old children were examined and structured questionnaire were completed by their parents. 120 children with highest number of dmft (dmft > or = 5) and 118 caries-free children were chosen for case-control analysis.

RESULTSThe results of logistic regression analysis showed that the factors associated with high dmft were developmental defect of enamel, visible plaque index, frequency of toothbrushing, frequency of sugar consumption, and income.

CONCLUSIONSAdvocating brushing teeth at least twice daily, controlling the frequency of sugar consuming, reducing the developmental defect of enamel and paying more attention to the oral health of lower income population may effectively reduce dental caries of the children.

Child, Preschool ; Dental Caries ; epidemiology ; etiology ; Health Behavior ; Humans ; Logistic Models ; Oral Health ; Oral Hygiene ; Risk Factors ; Surveys and Questionnaires ; Tooth, Deciduous

Objective To observe the efficacy and safety on the efficacy of HBeAg-positive chronic Hepatitis B patients treated with adefovir dipivoxil for 4 years.Methods Ninety-five patients with HBeAg-positive chronic hepatitis B were treated with adefovir dipivoxil 10 mg per day orally.The patients were observed before and after treatment for their serum levels of ALT and HBV DNA,the new increasing rates of serum ALT normalization,HBV DNA clearances,HBeAg loss,HBeAg seroconversion and adverse drug events.Results At 4 years on study,the rates of ALT normalization,HBV DNA clearances,HBeAg loss,HBeAg seroconversion and HBV DNA rebound were 89.5％,63.2％,47.4％,41.1％ and 8.0％,respectively.No drug related to renal function impairment was found during the treatment,eight patients had adverse drug events but all were mild.Conclusion Adefovir dipivoxil could effectively inhibit HBV replication,normalize ALT and enhance transformation from HBeAg to HBeAb for cases with naive and treated-first patients.The efficacy were increased with prolongation of the treatment period.It is safe and has a good tolerance.

ObjectiveThis study aims to investigate the role of suPAR in the pathogenesis of podocyte injury in FSGS. Methods① The sera of primary FSGS patients （17 cases） were collected. Healthy volunteers （10 cases） and patients with other types of primary nephrotic syndrome （10 cases） were set as normal and disease controls. SuPAR levels were detected by ELISA； ② Podocytes were stimulated by suPAR in vitro， and cells were collected to analyze apoptosis by flow cytometry and for RNAseq analysis； ③ Differentially expressed genes （DEGs） were screened from RNAseq data. Both up-regulated and down-regulated genes were analyzed by KEGG and GO enrichment analysis. Heat map was used to show expression of genes related to podocyte focal adhesion， slit diaphragm and actin dynamics and endocytosis. Differentially expressed genes were verified by qPCR. Results① The level of suPAR in FSGS patients was significantly increased， and that in other nephrotic syndrome（NS） patients was also significantly increased； ② suPAR stimulation significantly altered the transcriptome pattern of human podocytes. A total of 272 up-regulated genes and 288 down-regulated genes were screened； ③ KEGG and GO enrichment analysis of up-regulated and down-regulated genes showed that Focal adhesion and DNA replication and DNA repair related pathways were significantly down-regulated； ④ suPAR did not increase podocyte apoptosis. ConclusionThe level of suPAR is significantly increased in patients with primary FSGS. SuPAR may promote podocyte injury by interfering with genomic homeostasis and disrupting focal adhesion， slit diaphragm， actin dynamics and endocytosis-related functional molecules of podocytes.

OBJECTIVETo study the effects of cluster of differentiation 40 ligand immunoglobulin (CD40LIg) gene-modified bone marrow mesenchymal stem cells (MSCs) on liver graft rejection in rats.

METHODSThe orthotopic liver transplantation models were established with DA rats as the donors and Lewis rats as the recipient. MSCs infected with the recombinant adenoviruses containing CD40LIg gene were infused into the liver graft after transplantation. The liver function, survival of the recipient rats and the morphological changes of the liver grafts were observed after the transplantation. The serum levels of the cytokines interferon-γ (INF-γ) and interleukin-2 (IL-2) in the recipient rats were quantified by ELISA.

RESULTSThe survival of the recipient rats receiving transplantation of genetically modified MSCs (group D) was significantly prolonged compared with that of the control group (group A), MSCs group (group B) and gene transfection group (group C); the survival of groups B and C were significantly longer than that of group A (F=7.615, P<0.05). The level of serum alanine aminotransferase, total bilirubin, IL-2 and INF-γ were significantly higher in group A than in the other 3 groups (F=8.738, P<0.05). HE staining of the liver grafts showed severe acute rejection in group A, mild acute graft rejection in groups B and group C, but no rejection in group D.

CONCLUSIONCD40LIg gene-modified MSCs can prolong the survival of the recipient rats and suppress graft rejection following liver transplantation.

Adenoviridae ; genetics ; metabolism ; Animals ; Bone Marrow Cells ; cytology ; metabolism ; Graft Rejection ; prevention & control ; Liver Transplantation ; adverse effects ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Rats ; Rats, Inbred Dahl ; Rats, Inbred Lew ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Transfection

OBJECTIVETo explore the method of clinical application and the efficacy of free fibula osteomyocutaneous flap in one-stage reconstruction of transmidline bilateral mandibular defect caused by giant neoplasms.

METHODSFrom july 2000 to october 2002, transmidline bilateral mandibular defects caused by ameloblastoma (4 cases) and gingival carcinoma (2 cases), according to the character of defects, were reconstructed with free fibula osteomyocutaneous flaps. Peroneal artery and vein were used as vascular pedicle, the fibula was reshaped, and micro-titanium plates were used in rigid fixation between fibula and residue of bilateral mandible. Microvascular anastomoses were carried out between peroneal artery/vein and small artery/vein in neck.

RESULTSSix free fibular osteomyocutaneous flaps survived well. Follow up duration ranged from 6 months to 2 years, the lower face appearance recovered well, occlusion relationship were normal, all patients were satisfactory with appearance and chewing function after repair of removable denture.

CONCLUSIONFree fibular osteomyocutaneous flap is a favorable material in the reconstruction of transmidline bilateral mandibular giant defect. The blood supplement of fibula is offered both by segmentral periosteum and nutrient artery from bone marrow, It is greatly benefit to reshaping as arched mandible.

Adult ; Bone Transplantation ; Female ; Fibula ; surgery ; Follow-Up Studies ; Humans ; Male ; Mandible ; surgery ; Mandibular Neoplasms ; surgery ; Osteotomy ; Reconstructive Surgical Procedures ; methods ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo study the specific cellular immunoresponse of peripheral blood lymphocytes in the chronic hepatitis B patients treated with different doses of recombinant hepatitis B vaccine.

METHODSSeventy-two chronic hepatitis B patients who did not use any anti-HBV drugs within 6 months were randomized into 3 groups (90 micrograms, 60 micrograms, and placebo) in a ratio of 1:1:1. The patients in different groups were treated with different doses of recombinant hepatitis B vaccine in combination with IFN alpha 1b 50 micrograms with 3 times a week for 24 weeks. All patients were followed up for 24 weeks (W24). HBV DNA, HBeAg and liver functions were detected at different time points, and the number of cells that secrete IFN-gamma were detected by ELISPOT.

RESULTSThere were no significant difference in ELISPOT positive ratio among the 3 groups on baseline detection. At W24, 12 cases, 12 cases, and 7 cases showed ELISPOT positive in the group of 90 micrograms, 60 micrograms, and placebo. The proportion of patients who were ELISPOT positive was higher in the groups treated with recombinant hepatitis B vaccine (including the dose of 90 micrograms and 60 micrograms) than that in the placebo group (P=0.0446). HBV DNA turned negative in 6/24 of the patients treated with recombinant hepatitis B vaccine (at both the doses of 90 micrograms and 60 micrograms), and HBeAg/Anti-HBe seroconversion or HBeAg became negative in 7/24 of them. In the placebo group, none of the patients showed undetectable HBV DNA, HBeAg/Anti-HBe seroconversion or HBeAg disappearance. At the 24W of follow up, in the patients who were ELISPOT positive, HBV DNA became undetectable in 4 of the patients treated with recombinant hepatitis B vaccine (at doses of 90 micrograms and 60 micrograms), and HBeAg/Anti-HBe seroconversion or HBeAg disappearance were found in 9 of the cases. In the placebo group, none of the cases showed undetectable HBV DNA, and only 1 case had HBeAg/Anti-HBe seroconversion.

CONCLUSIONThe recombinant hepatitis B vaccine may increase the function of specific T lymphocytes in patients with chronic hepatitis B. There were no significant differences between the patients treated with the dose of 90 micrograms and 60 micrograms hepatitis B vaccine.

Adult ; DNA, Viral ; blood ; Female ; Hepatitis B Vaccines ; immunology ; Hepatitis B, Chronic ; immunology ; Humans ; Interferon-gamma ; biosynthesis ; Male ; Recombinant Proteins ; immunology ; T-Lymphocytes ; immunology ; Vaccines, Synthetic ; immunology

OBJECTIVETo assess the accuracy and reliability of the nest-PCR-sequence specific primer(SSP) method in HLA-A site genotyping of single blastomeres retrieved from human pre-implantation embryos.

METHODSBy nest PCR on HLA-A exon 2, the success rate of first-round amplification was estimated for single blastomeres. Based on the first-round amplification, the HLA-A genotype of every single blastomeres was analyzed by commercially available PCR-SSP kits.

RESULTSThe amplification of HLA-A exon 2 were performed to 120 blasotmeres retrieved from in vitro fertilization(IVF) surplus embryos donated by 10 couples. The average success rate of family 1-5 and 6-10 was 78.2%(43/55) and 93.8%(61/65), respectively. And 86.7%(104/120) in total. Eighty blastomeres were further tested by nest-PCR-SSP, among which 11 blastomeres failed to HLA-A exon 2 amplification and then failed to genotyping while the other 69 blastomeres succeed in HLA-A exon 2 amplification and succeed in genotyping. Except for 6 blastomeres that were uncertain for allele lost because of parents' homozygosity, the left 63 blastomeres had accurate HLA genotyping. Among these 63 blastomeres, 59 blastomeres had genotypes confirmed from their parents(93.6%), 3 blastomeres lost one of parents' alleles(4.8%), and only one blastomere had two more than parents' alleles(1.6%).

CONCLUSIONThe above research results indicated that based on the successful first round amplification of single blastomeres, nest-PCR-SSP strategy offers a convenient and reliable option for HLA genotyping on single blastomeres, which is a key process in pre-selecting HLA-identical sibling for allogeneic cord blood cell transplantation.

Base Sequence ; Blastomeres ; metabolism ; DNA ; analysis ; DNA Fingerprinting ; methods ; DNA Mutational Analysis ; Female ; HLA Antigens ; analysis ; HLA-A Antigens ; analysis ; genetics ; Histocompatibility Antigens Class I ; analysis ; genetics ; Humans ; Male ; Polymerase Chain Reaction ; methods ; Single Person

Male ; Humans ; Urinary Bladder/diagnostic imaging* ; Prostate/diagnostic imaging* ; Multiparametric Magnetic Resonance Imaging ; Cystitis/diagnostic imaging* ; Hemorrhage/etiology* ; Urinary Bladder Neoplasms/complications*