Binding Sites ; Hepatitis B virus ; physiology ; Virus Attachment

OBJECTIVETo investigate the role of simultaneous blockade of CD40/CD40L and B7/CD28 pathways in the immune tolerance via co-expression of sCD40LIg and CTLA4Ig mediated by replication-defective adenovirus.

METHODSAd-sCD40LIg-IRES(2)-CTLA4Ig, replication-defective adenovirus co-expressing sCD40LIg and CTLA4Ig, was constructed and identified. The co-expression of sCD40LIg and CTLA4Ig was evaluated with confocal laser scanning microscope and Western blotting. Skin transplantations of C57BL/6 to BALB/c mice were performed. PBS, Ad-Shuttle-CMV and Ad-sCD40LIg-IRES(2)-CTLA4Ig were administered. Skin graft survival was monitored and the mRNA expression of both genes was evaluated in the skin allografts.

RESULTSAd-sCD40LIg-IRES(2)-CTLA4Ig was constructed successfully and identified. The co-expression of sCD40LIg and CTLA4Ig was identified with confocal laser scanning microscopy and Western blotting. Compared to the skin graft mean survival time (MST) of non-treated group ((5.75+/-0.71) d) or Ad-Shuttle-CMV-treated group ((5.50+/-0.53) d), the skin graft MST was dramatically prolonged in the Ad-sCD40LIg-IRES(2)-CTLA4Ig-treated group ((16.38+/-1.19) d, P<0.001). The mRNA expression of both genes was detected.

CONCLUSIONAd-sCD40LIg-IRES(2)-CTLA4Ig, a replication-defective adenovirus carrying genes encoding sCD40LIg and CTLA4Ig, was constructed. Simultaneous blockade of CD40/CD40L and B7/CD28 costimulatory pathway mediated by replication-defective adenovirus significantly prolonged skin allograft survival in mice.

Abatacept ; Adenoviridae ; genetics ; Animals ; Cytopathogenic Effect, Viral ; Graft Survival ; immunology ; Immunoconjugates ; genetics ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction ; Skin Transplantation ; immunology ; methods ; Transfection

OBJECTIVETo study the effect of Poria cocos (Pcs) in preventing acute rejection of rats after renal transplantation and its mechanism.

METHODSRat orthotopic renal transplantation model was performed with Wistar rat as donor and SD rat as donee. All donees were divided into 4 groups, 10 in each group, before transplantation. They were treated respectively with normal saline 5 mL x kg(-1) x d(-1) (A), Pcs 25 mg x kg(-1) x d(-1) (B), Pcs 50 mg x kg(-1) x d(-1) (C) and ciclosporin A (CsA) 5 mg x kg(-1) x d(-1) (D) by intragastric administration. The renal allograft survival time (ST) was recorded, and the serum levels of creatinine (SCr), interleukin-2 (IL-2), gamma-interferon (gamma-IFN), CD4+, CD8+ lymphocytes percentage, CD4+/CD8+ ratio, as well as the pathologic changes were observed one week after transplantation.

RESULTSST of the renal graft in Groups C and D was significantly longer with pathologic change evidently less than those in Groups A and B (P<0.01), and the ST in Group C was shorter that in Group D (P<0.05). Changes of renal function and urine volume were identified to the pathological change of graft, the initiating time of renal dysfunction was later in Groups C and D than that in Groups A and B. Serum levels of IL-2, IFN-gamma and CD4+ percentage in Group C were significantly lower than those in Groups A and B, but higher than those in Group D respectively (P<0.05 or P<0.01), while CD8+ percentage in Group C was significantly lower than that in Group A (P<0.05), but insignificantly different to that in Groups B and D (P>0.05).

CONCLUSIONPcs shows good dosage-dependent effect in suppressing acute rejection of renal transplantation, but the effect is inferior to that of CsA.

Animals ; Cyclosporine ; therapeutic use ; Graft Rejection ; prevention & control ; Kidney Transplantation ; Male ; Materia Medica ; therapeutic use ; Poria ; chemistry ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar

OBJECTIVE: To investigate the meaning of the mutation screening, prevalence, inheritance and the intervention or the prevention for the specific drugs in 10 families with non-syndrome hearing loss in Yunnan Province, China. METHOD: To do a questionnaire about the cases of ten families with non-syndrome hearing loss and to draw a detailed matriarchal family tree detailed. Following that, the A1555G mutation-positive individuals were detected and confirmed using DNA extracting, PCR amplification and sequencing for family volunteer. RESULT: There are 96 members have attended the blood collection in these ten families. Thirty-six of them had the normal hearing and 60 of them had the sensory neural hearing loss. However, 4 out of those had no A1555G point mutation, and 92 had A1555G point mutation (95.8%). While 7 of those were Heterogeneity, the rest were all homogeneous mutation. There were also 73 patients who had amino glycoside antibiotic medication history. However all the rest cases had a history of amino glycoside antibiotic medication were not clear yet. CONCLUSION The proportion of patients with drug-induced deafness is high in Yunnan province and the mutation rate of mitochondrial DNA A1555G is also high. It is worthy to do DNA 12SrRNA A1555G mutation screening for drug intervention and prevention.
Adolescent ; Adult ; Aged ; Child ; China ; epidemiology ; DNA Mutational Analysis ; DNA, Mitochondrial ; genetics ; Deafness ; epidemiology ; genetics ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Point Mutation ; RNA, Ribosomal ; genetics ; Young Adult

OBJECTIVETo evaluate the acceptability of self-management project for patients with hypertension and the short term result, and provide the evidence of preventing and controlling hypertension using chronic disease self-management.

METHODSVoluntary patients with hypertension were grouped into the control and experimental groups after matching age, sex, education levels and the number of chronic diseases between the two groups. Data collected through questionnaire after intervention were compared with the baseline data through analysis of covariance and chi(2) test.

RESULTSRate of awareness for the diagnostic criteria of hypertension, risk factors and associated diseases increased by 56.4%, 50.4% and 37.6% respectively, and the rate of high salt diet intake, body mass index and the times of visiting doctors decreased by 27.1%, 0.8 kg/m(2) and 2.7 times on average in experimental group, as compared with control group. The health status (energy, fatigue, shortness of breathe, pain) improved (P < 0.05). In the similar comparison, the rate in which the hypertensive patients took antihypertensive medicine increased by 20%, and the scores of cognitive symptoms management and of patient communication with physician increased by 3.9 and 1.1 points respectively. The rate of compliance to medication increased by 21.5% compared with themselves.

CONCLUSIONThe awareness rate of hypertensive patients on knowledge and on unhealthy life style improved after implementing the self-management program. Their emotion, healthy behavior, health status and symptoms related to hypertension improved. Data showed that it was effective for hypertensive patients to use chronic diseases self-management.

Aged ; Awareness ; Body Mass Index ; Female ; Humans ; Hypertension ; therapy ; Life Style ; Male ; Middle Aged

OBJECTIVE: To evaluate the therapeutic effect of the herbal plaster Tangshangxiaobagao on partial thickness burn wound. METHODS: A randomized controlled trial was conducted with two herbal plasters: Tangshang-xiaobagao and Jingwanhong in 57 hospitalized burn patients. Both the effect and safety of two herbal plasters were noted in patients with partial thickness burns. RESTULTS: In superficial second degree burns, the 7 d healing rate of both groups was (61.35+/-36.26)%and (51.21+/-37.24)% and the healing time (10.56+/-3.43)d and (11.98+/-4.13)d P<0.05 respectively. While in deep second degree burns, the 14 d healing rate of both groups was (62.9+/-36.0) % and (53.9+/-32.2) % and the healing time (19.4+/-4.9)d and (21.5+/-5.5)d, respectively. Study group had lower VAS(visual analogue scale)score than control group. No obvious side effects were observed in study group. CONCLUSION: Tangshangxiaobagao is safe and may be an effective adjunct for treatment of partial thickness burn wounds.

OBJECTIVETo study the protective effect of recombinant adenovirus-mediated human cytosolic glutathione peroxidase (hCGPx) gene transfection on vascular endothelial cells ECV304 from oxidative damage.

METHODSpGEM-T Easy Vector containing hCGPx cDNA and recombinant adenovirus shuttle plasmid pACCMV-pLpA were used to construct the shuttle plasmid pACCMV-hCGPx for cotransfection of 293 cells with pJM17, thereby to obtain the recombinant adenovirus AdCMV-hCGPx. Cultured ECV304 cells were transfected with AdCMV-hCGPx for 24, 48 and 72 h, respectively, with the cells transfected with the empty vector serving as control, and hCGPx gene expression was then examined in the transfected cells. The transfected cell viability and apoptotic cell ratio were evaluated after treatment of the cells with H(2)O(2).

RESULTSThe expression ratio of hCGPx gene was significantly higher in the AdCMV-hCGPx-transfected cells than in those with empty vector transfection (P<0.01). The hCGPx gene-transfected cells showed significantly higher viability and significantly lower apoptotic ratio than the control cells following challenge with H(2)O(2)-induced oxidative damage.

CONCLUSIONhCGPx gene transfer mediated by recombinant adenovirus protects the vascular endothelial cells from oxidative damage in vitro, possibly due to the antioxidative and apoptosis-inhibiting effect of hCGPx.

Adenoviridae ; genetics ; Apoptosis ; drug effects ; Cell Line ; Cell Survival ; drug effects ; Cytosol ; enzymology ; Endothelial Cells ; cytology ; drug effects ; metabolism ; Flow Cytometry ; Genetic Vectors ; Glutathione Peroxidase ; biosynthesis ; genetics ; Humans ; Hydrogen Peroxide ; pharmacology ; Oxidative Stress ; Plasmids ; genetics ; Time Factors ; Transfection

OBJECTIVETo study the impacts of kidney transplantation on erectile function and analyse its contributing factors.

METHODSIn order to evaluate the severity of erectile dysfunction (ED), a total of 250 married male kidney transplant recipients (KTR) with functioning graft were assessed with the International Index of Erectile Function (IIEF) questionnaire. Data of clinical characteristics, medical and sexual history and laboratory examination were collected. Univariate and multivariate logistic regression analyses were carried out to determine which have independent impacts on erectile function.

RESULTSThe investigation was accomplished in 84.8% of the KTRs. There was no significant difference in ED incidence before and after renal transplantation (53.8% vs. 44.3%, P > 0.05). According to the IIEF score, erectile function improved in 43.9% of the KTRs, remained unchanged in 42.9%, and deteriorated in 13.2%, as compared with pre-transplantation. Logistic regression analysis showed that significant and independent influencing factors in erectile function were age, hemoglobin level, presence of DM and/or peripheral neuropathy and iterative transplantations, and their relative risks were 3.01, 2.01, 3.15, 3.89 and 2.67, respectively.

CONCLUSIONED is highly prevalent among KTRs and its pathogenesis is multifactorial. Age, presence of DM and/or peripheral neuropathy, hemoglobin level and iterative transplantations were chief contributing factors in erectile function.

Adult ; Diabetes Complications ; Erectile Dysfunction ; epidemiology ; Humans ; Kidney Transplantation ; adverse effects ; Logistic Models ; Male ; Middle Aged ; Risk Factors ; Surveys and Questionnaires

Aminoglycoside antibiotics, due to their strong antibacterial effects and broad antimicrobial spectra, have been very commonly used in clinical practice in the past half century. However, aminoglycoside antibiotics manifest severe ototoxicity and nephrotoxicity, and are one of top factors in hearing loss. In this study, three members of the aminoglycoside antibiotics family, gentamycin, neomycin and streptomycin, were chosen as the representatives to be investigated for their toxicity to the embryonic development and the larva hair cells in zebrafish, and also to their target genes associated with hearing-related genes. The results showed that: (1) the lethal effect of all three drugs demonstrated a significant dependence on concentration, and the severity order of the lethal effect was streptomycin > neomycin > gentamycin; (2) all the three drugs caused the larva trunk bending in resting state at 5 dpf (day past fertilization), probably due to their ototoxicity in the physical imbalance and postural abnormalities; (3) impairment and reducing of the hair cells were observed in all three cases of drug treatment; (4) four genes, eya1, val, otx2 and dlx6a, which play an important role in the development of hearing organs, showed differential and significant decrease of gene expression in a drug concentration-dependent manner. This study for the first time reports the relevance between the expression of hearing genes and the three ototoxic antibiotics and also proved the feasibility of establishing a simple, accurate, intuitive and fast model with zebrafish for the detection of drug ototoxicity.
Aminoglycosides ; toxicity ; Animals ; Anti-Bacterial Agents ; toxicity ; Embryonic Development ; drug effects ; Gene Expression Regulation ; Gentamicins ; toxicity ; Hair Cells, Auditory ; cytology ; drug effects ; Hearing Disorders ; chemically induced ; genetics ; metabolism ; Homeodomain Proteins ; metabolism ; Intracellular Signaling Peptides and Proteins ; metabolism ; Larva ; drug effects ; Lateral Line System ; drug effects ; MafB Transcription Factor ; metabolism ; Models, Animal ; Neomycin ; toxicity ; Nerve Tissue Proteins ; metabolism ; Nuclear Proteins ; metabolism ; Otx Transcription Factors ; metabolism ; Protein Synthesis Inhibitors ; toxicity ; Protein Tyrosine Phosphatases ; metabolism ; Streptomycin ; toxicity ; Zebrafish ; embryology ; Zebrafish Proteins ; metabolism

OBJECTIVETo establish a stable method for obtaining large quantity of highly purified immature dendritic cells (imDCs) in vitro, and identify the morphology, function and surface markers of the cells.

METHODSCD117(+) hemopoietic stem cells (HSCs) were isolated and purified from the bone marrow of healthy C57 mice by magnetic affinity cell sorting. After cell expansion by treatment with stem cell factor (SCF) and interleukin-3 (IL-3), the HSCs were induced for directional differentiation into imDCs by treatment with GM-CSF, IL-4 and IL-10. The imDCs obtained were identified by morphological and functional observation under inverted microscope, scanning electron microscope and transmission electron microscope, followed by detection of the expressions of the surface markers using flow cytometry.

RESULTSAfter 3, 5 and 7 days of culture in the presence of SCF+IL-3, the cells were expanded by 10.34-/+1.43, 22.65-/+2.71 and 54.39-/+3.08 folds, respectively. The HSCs were successfully induced to differentiate into imDCs with phagocytotic activity. The dendrites of the imDCs were short small, and appearing spinous. The expressions of surface markers were detected from the cells showing the phenotype of CD11c(+), I-A/I-E(low), CD40(-), CD80(-), CD86(-).

CONCLUSIONThe method described allows steadily acquisition of large quanty of highly purified imDCs and of their effective identification in vitro.

Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Dendritic Cells ; cytology ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Microscopy, Electron, Transmission ; Proto-Oncogene Proteins c-kit