Child ; Cough ; chemically induced ; Disinfectants ; adverse effects ; Humans ; Irritants ; adverse effects ; Male

Objective To investigate the features of immune response of human immunodeficiency virus type-1 (HIV-1) antigen specific T lymphocytes in HIV-1 monoinfected or HIV 1/hepatitis C virus (HCV) coinfected individuals.Methods Twenty-six HIV-1 monoinfected and 23 HIV-1/HCV coinfected individuals were enrolled.Immunomagnetic microbeads were used to isolate T lymphocyte subpopulation CD4+ T cells and CD8+ T cells from human peripheral blood mononuclear cells (PBMC).Frequencies of interferon-γ (IFN-γ) secreting cells of CD4+,CD8+ T lymphocytes and PBMC stimulated by a peptide pool containing 12 overlapping peptides in HIV-1 P24 from 49 patients were assessed by enzyme-linked immunospot (ELISPOT) assay.HIV-1 RNA levels of these patients were also detected by real-time fluorescence quantitative polymerase chain reaction.The data were compared by one-way ANOVA and Mann-Whitney U test,and Spearman test was used for correlation analysis.Results Frequencies of HIV-1 antigen specific CD4+ T lymphocytes [median =25 spot-forming cells (SFC)/106 cells] were significantly lower than those of CD8+T lymphocytes (median=38SFC/106 cells,F=4.592,P=0.037) and PBMC (median=53 SFC/106 cells,F=5.436,P=0.025) in HIV-1 monoinfected group.Frequencies of HIV-1 antigen specific CD4+ T lymphocytes (median=5 SFC/106 cells,Z=-2.432,P=0.015),CD8+T lymphocytes (median=5 SFC/106 cells,Z=-1.996,P=0.046) and PBMC (median=10 SFC/106 cells,Z=-2.306,P=0.021) in HIV-1/HCV coinfected group were significantly lower than those in HIV-1 monoinfected group.Conclusions In HIV-1 infection,antigen specific immune response of CD4+ T cells can be activated,but weaker than that of CD8+ T cells.Co-infection with HCV might down-regulate the responses of HIV-1 antigen specific T lymphocytes in HIV-1 infected individuals.

Objective To investigate the gene expression of vascular endothelial growth factor(VEGF) and ultrastructural changes in cultured endothelial cells from human cerebral microvessels under hypoxic conditions.Methods Human cerebral microvessels were isolated from freshly obtained specimens of normal brain adherent to resected cerebral arteriovenous malformations.The expression of factor Ⅷ-relative antigen(FⅧ-RA) in cultured cells was observed with immunocytochemistry.The level of VEGFmRNA in cells and released VEGF protein in cell supernatant were determined by RT-PCR analysis and ELISA respectively when they were exposed to hypoxic conditions(95% N_2,5% CO_2;two hours,four hours,eight hours) or maintained in basal condition.Ultrastructural changes in cells were also observed by electron microscopy.Results In inverted microscope the cultured cells showed contact inhibition and a rounded cobblestone appearance.More than 90% of them were stained strongly with antibodies against FⅧ-RA.Significant VEGF mRNA and protein accumulated when these cells were exposed to hypoxia for 4 hours.However,their VEGF expression was down-regulated after hypoxia for 8 hours and a number of vesicles and swollen mitochondria were present in the cytoplasm.Conclusion The level of VEGF expression may havesignificant relationship with ultrastructural changes in human cerebral endothelial cells under hypoxic conditions.

Objective To carried out rat as deep inferior epigastic perforator (DIEP) flap necrosis model,to reveal the continuous blood fluid change in process of flap necrosis.Methods Seven SpragueDawley rats of male were used.The designed DIEP flap model was pedicled on the right-sided cranial perforator.Near-infrared fluorescent angiography was performed using SPY imaging system pre-and-aft operation and all angiography videos were compared and analyzed.Results Using SPY imaging system could observe the process of the blood fluid scattered from perforator through choke anastomoses to next vascular territories in living body.The study could clearly observe sequence change of blood fluid pr-and-aft operation,which could give the important information in revealing the reason of necrosis in DIEP flap.Cross-midline traffic vascular sparse and eventually could no longer wear through the contralateral abdominal wall vessel area and open in the region of lateral thoracic vascular occlusion of blood vessels to reach the flap to the far left side,and showed no blood flow in the dark zone.Conclusion The reasons of necrosis of contralateral distal zone of the flap model,are the choke anastomoses across the midline to the contralateral side appeared small,less intensity and the blood supply to contralatral distal zone should through twice choke anastomoses.

Objective To compare the phenotypes of abnormal CD4+CD25-Foxp3+ T cells with traditional regulatory T cells (CD4+CD25+Foxp3+) in patients with untreated new-onset lupus (UNoL) and investigate their clinical relevance. Methods The expressions of surface markers (CD25, CD127, CCR4, GITR, CTLA-4) and intracellular marker(Foxp3) on the peripheral blood mononuclear cells from twenty-two UNoL patients were analyzed by flow cytometry analysis, and their clinical relevance were assessed. Results There were no significant differences between CD4+CD25-Foxp3+ and CD4+CD25+Foxp3- T cells in the expressions of GITR, CTLA--4 and CCR4 (P>0.05), but they were significantly lower than those of CD4+CD25+ Foxp3+ T cells in UNoL patients (P<0.01). The percentages of CD127low- in CD4+Foxp3+CD25high,CD4+Foxp3+ CD25low and CD4+Foxp3+CD25+ T cells were (93.8±3.5 )%, (93.7±2.3)% and (92.0±2.1)% respectively (P> 0.05), whereas the expressions of Foxp3 on CD4+CD127low- T subpopulations showed significant differences in CD4+CDI27low-CD25high (91.4±2.6)%, CD4+CD127low-CD25low (71.9±3.3)% and CD4+CD127low-CD25- (9.0± 2.2)% T cells(P<0.01 ). The frequency of CD+CCR4+CD25high T cells correlated negatively with SLEDAI (r=-0.695,P<0.001).and it was significantly lower in lupus nephritis patients(1.10±0.17)%compared with SLE patients without nephritis [(1.61±0.23)%,P<0.01]and healthy controls [(1.75±0.10)%,P<0.01], furthermore,the frequency of CD4+CCR4+CD25low-T cells in lupus nephritis was significantly higher than that in healthy controls[(11.5±2.3)%vs (8.0±1.0)%,P<0.01].Conclusion The increased CD4+CD25-Foxp3+ T cells in the Untreated Newonset Lupus(UNoL)patients mimic activated T effector cells.CD4+CD25high-CD127low-T cells can be used to isolate live CD4+CD25highFoxp3+regulatory T cells.CCR4+regulatory T cells may be involved in the pathogenesis of lupus nephritis.

Objective:To evaluate the expression of vascular endothelial growth factor C(VEGF-C),Epstein-Barr virus(EBV) latent membrane protein 1(LMP1),cyclooxygenase-2 (COX-2) in nasopharyngeal carcinoma(NPC).Method:LMP1, COX-2 and VEGF-C were detects by immunohistochemical staining for 57 case NPC tissue.Result:The positives rates of LMP1,COX-2 and VEGF-C detected by immunohistochemical staining were 49.1%(28/57), 75.4%(43/57)and 59.6%(34/57), respectively. The expression of LMP1, COX-2 and VEGF-C were correlated to each1 other in NPC(P<0.05).Conclusion:LMP1 and COX-2 may induce expression of VEGF-C directly or LMP1 induce expression of VEGF-C by induce COX-2 expression, may contribute to lymph metastasis and develop NPC.

Objective:To study the correlation between responses to Hepatitis B and HLA DRB1*07 DRB1*04 DRB1*1001. DQB1*0401 genes in Han population in Ningbo.Methods:A total of 240 Han people living in Ningbo received the routine vaccination of recombinant hepatitis B vaccine.The serum levels of anti-HBs antibody were examined and the subjects were divided into 2 groups according to tile results:negative responses group(n=120.anti-HBs D

BACKGROUND:Although there is a certain progress in the preparation of tissue-engineered bone tissue using a variety of materials, some deficiencies have appeared such as mismatching between scaffold degradation rate and new bone formation rate, slow tissue growth, toxic metabolites.
OBJECTIVE:To build a new type of inducible bone repair composite scaffold with bionic bone structurematerials and to evaluate its physicochemical and biological properties.
METHODS: Icarin encapsulated by chitosan was used to prepare drug-loaded microspheres, and the drug release rate of the microspheres was detected. Chitosan microspheres were mixed with colagen to build the core part of scaffold materials. Hydroxyapatite (HA), polycaprolactone (PCL) and colagen were mixed in hexafluoride isopropanol (HFIP) to prepare the HA/PCL/colagen outer part of composite scaffold material at the rate of 0:3:3, 1:3:3, 2:3:3, 3:3:3. Each proportional electrospinning was used for one layer, and finaly the 4-layer outer tube of the scaffold was produced. The tube core and outer tube were crosslinked by 1% genipin. Universal material testing machine, surface contact angle meter, infrared spectroscopy, scanning electron microscope, water absorption, permeability, porosity,in vitro degradation tests for cross-linked and uncross-linked were used to observe the structure and characteristics of tubular materials. Bone marrow mesenchymal stem cels were seeded on the surface of cross-linked and uncross-linked bone repair materials to evaluate the biocompatibility of the scaffolds. Cross-linked and uncross-linked bone repair materials were implanted subcutaneously into Wistar rats to evaluate the histocompatibility of the scaffolds.
RESULTS AND CONCLUSION:The drug in the scaffold had a suitable release; the bone scaffold material had good uniformity, and cross-linked scaffolds materials had better mechanical properties, water absorption and permeability than the uncross-linked(P < 0.05). The degradation rate of the cross-linked group was significantly lower than that of the uncross-linked group (P< 0.05). Hematoxylin-eosin staining showed that the bone marrow mesenchymal stem cels could adhere wel to the cross-linked and uncross-linked materials. No inflammatory reactions occurred after subcutaneous implantation of cross-linked and uncross-linked materials. These findings indicate that the cross-linked scaffold for inducible bone tissue engineering has good biocompatibility and mechanical properties.

Objective To investigate the effect of highly active antiretroviral therapy (HAART) on human immunodeficiency virus type-1 ( HIV-1 ) antigen specific cytotoxic T lymphocyte (CTL) immune responses. Methods Peripheral blood mononuclear cells (PBMCs) were collected from 38 HIV-1 infected individuals receiving HAART ( HAART group) and 31 HIV-1 infected individuals not receiving HAART (non-HAART group), and stimulated with a peptide pool containing 12 overlapping peptides in HIV-1 P24;then the frequency of interferon γ ( IFNγ ) secreting cells were assessed by enzyme-linked immunospot (ELISPOT) method. Difference in HIV-1 antigen specific CTL immune response between non-HAART group and HAART group was analyzed by χ2 and Mann-Whitney U tests. Results Positive response rate of HIV-1 antigen specific CTL immune responses in HAART group ( 65.8%, 24/38 ) was higher than that of non-HAART group (32.3%, 10/31, χ2 = 6. 522, P ＜ 0.05 ). For HIV-1 infected individuals with blood CD4 +T cells ＞ 350/μL, the frequency of HIV-1 antigen specific CTL responses in HAART group was higher than that in non-HAART group (Z = -2. 819, P ＜0.05 ). In the HAART group, those receiving HAART more than 12 months were of higher frequency of HIV-1 antigen specific CTL responses ( Z =-2. 195, P ＜ 0. 05 ). Conclusion HAART especially long-term treatment may enhance HIV-1 specific CTL responses in HIV-1 infected individuals.

Objective:To investigate frequencies of microsatellite instability(MSI)and loss of heterozygosity(LOH)in renal ceil carcinoma(RCC),and to discuss the relationship of clinicopathological characteristics of RCC with MSI and LOH. Methods:Twelve microsatellite markers located at chromosomes 3p,9p and 14q were selected to investigate microsatellite alterations(MSI and LOH)in 31 RCC specimens and their paired metastasis specimens by polymerase chain reaction- polyacrylamide gel elect rophoresis-ethylene dibromide(PCR-PAGE-EB)staining and sequencing.Results:The frequency of MSI could reached 61.3% and that of LOH could reach 54.8%.The highest frequency of MSI was at locus of D9S168(32.3%);the highest frequency of LOH was at locus of D3S1289(21.4%).No correlation was found between MSI or LOH and the patients' age,sex,pathology type and metastastis,except that MSI was correlated with TNM stage of RCC(P