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

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.

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 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

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

BACKGROUNDGlobally, 180 million people suffer from diabetes mellitus. Islet transplantation is believed to be an almost ideal therapy for insulin-dependent patients. How to maintain the viability and the function of isolated human islets is a challenge in clinical practice. Sertoli cells are considered 'nurse cells' in the seminiferous tubules and have been used in cell graft protocols for neurodegenerative diseases and diabetes in many studies. Many researchers have used immature murine testes as the primarily source of Sertoli cells in islet transplantation because they are easily purified. Mature human Sertoli cells have been seldom investigated. In the present study, we developed a method for the isolation and culture of Sertoli cells derived from adult human testes, and investigated their effects on the function of allogeneic islets when they were cultured together in vitro.

METHODSAdult Sertoli cells were prepared successfully by two-step enzyme digestion with trypsin, collagenase and hyaluronidase. They were identified by morphological characteristics and their activity was determined by MTT colorimetry over a 28-day culture time in vitro. A glucose-stimulated insulin secretion test was performed to detect the effects of Sertoli cells on allogeneic islets' function when they were co-cultured for 21 days in vitro.

RESULTSIn cultured cells, mature human Sertoli cells accounted for more than 90% of total cells. The activity of Sertoli cells reached 95% and they remained highly cytoactive for a long time in vitro (P > 0.05). Compared with the islets cultured alone, the co-cultured islets with allogeneic Sertoli cells maintained higher sensitivity to glucose stimulation for the duration of the experiment (P < 0.01).

CONCLUSIONSA method of isolation and culture of Sertoli cells from adult testes has been established. Sertoli cells could enhance allogeneic islets' function when they were co-cultured in vitro. They could be a helper cell in islet transplantation.

Adult ; Cell Separation ; methods ; Cell Survival ; Cells, Cultured ; Coculture Techniques ; Humans ; Islets of Langerhans ; physiology ; Islets of Langerhans Transplantation ; Male ; Sertoli Cells ; cytology ; physiology

OBJECTIVETo investigate the status of pubertal development in children born with assisted reproductive technology (ART).

METHODSA retrospective analysis was performed on the pubertal development data of children born with ART in Peking University Third Hospital from 1994 to 2003 (ART group). The data in the cross-sectional study "Reports on the Physical Fitness and Health Research of Chinese School Students in 2010" were used as a control. The age at menarche and the age at spermarche were compared between the two groups. The status of pubertal development in the overweight and obese children in the ART group was evaluated to investigate the correlation between pubertal development and body mass index (BMI).

RESULTSA total of 200 children born with ART were enrolled in this study, and 72 of them (41 males and 31 females) completed the survey (response rate=36.0%). In the ART group, the mean age at spermarche and the mean age at menarche were 13.9 years (95%CI: 13.7-14.3 years) and 12.2 years (95%CI: 11.8-12.6 years), respectively. There were no significant differences in the age at spermarche and the age at menarche between the ART and control groups (P>0.05). In the ART group, there were no significant differences in the age at spermarche and the age at menarche between the overweight and obese children and the normal weight children (P>0.05). There were also no significant differences in overweight rate and obesity rate between the children in the ART group and the adolescents in Beijing (P>0.05). In the ART group, there was no significant correlation between the age at spermarche or menarche and BMI (P>0.05).

CONCLUSIONSNo delayed or precocious puberty is observed in children born with ART. This is consistent with the normal control data. And there is no significant correlation between pubertal development and BMI in children born with ART.

Adolescent ; Body Mass Index ; Child ; Child Development ; Cross-Sectional Studies ; Female ; Humans ; Male ; Menarche ; Obesity ; physiopathology ; Overweight ; physiopathology ; Puberty ; physiology ; Reproductive Techniques, Assisted ; Retrospective Studies