Objective To explore the influence of blocking B7/CD28 co-stimulatory pathway by RNA interference on the rejection response in mice heterogeneous heart transplantation and its mecha- nism.Methods siRNA of which sequence specified to CD80 and CD86mRNA was synthesized in vitro respectively and transfected into donor derived myeloid dendritic cells (DCs).The expression levels of CD80 and CD86 mRNA and surface antigen CD80,CD86 were assayed by semi-quantitative reverse transcription polymerase chain reaction and flow cytometry before and after CD80siRNA and CD86siRNA transfection.Seven days prior to heterogeneous heart transplantation in mice,DCs modi- fied by siRNA were transfused into recipients intraveneously (DC interference group).At the same time,group of allograft transplantation,cyclosporine A (CsA)-treated (CsA injected subcutaneously postoperatively,5 mg.kg~(-1)?d~(-1)) group,group of isograft transplantation,and non-interference DC group (transfusion of non-interfered DCs pre-transplanting) were assigned.The graft survivals were individually recorded and the graft rejection grading was pathologically evaluated.Interleukin 2 (IL- 2),interferon?(IFN-?),and IL-10 mRNA expression levels in grafts tissue were determined.Results After siRNA transfected into DC,the expression levels of CD80 and CD86 mRNA were down- regulated significantly and the the antigen CD80~+ and CD86~+ reduced from 84%,67% to 35% and 30% respectively.As compared with groups of allograft and non-interference DC,survival of the grafts was significantly longer in DC interference group (P＜0.01),pathological grade of rejection significantly lower (P＜0.01),IL-2 and IFN-?mRNA expression levels lower,and IL-10 mRNA ex- pression levels higher in grafts tissue (P＜0.01).Conclusion Knocking down the molecule B7 expres- sion level in donor-derived myeloid DCs through RNAi,which could block B7/CD28 co-stimulatory pathway,could exhibit inhibitive effect on rejection response in mice heart transplantation.The mechanism might be due to induction of T lymphocyte anergy and Th cell differentiation deviating to T_H2.

Objective To evaluate the effect of exercise on the relationship between body mass index (BMI) and health-related quality of life (HRQOL).Methods A total of 1 114 participants were recruited from the Physical Examination Center of two hospitals.SF-36 was used to measure HRQOL,and the classification of sports level was done according to the subjective evaluation of participants.Results The total score of HRQOL,physical component summary and mental component summary were 76.26± 16.10,75.97 ± 16.67 and 76.55± 18.33 respectively.The results of correlation analysis were as following:the positive correlation between BMI and general health (r=0.109),vitality (r=0.116),social functioning (r=0.092),role emotional (r=0.122),mental health (r =0.145) were significant (P＜ 0.01).The negative correlation between BMI and PF(r=-0.07,P=0.019)was significant.The mediation effects of exercise in was relationship between BMI and general health,vitality,mental health were significant,and the mediating effect were 5.84％,9.14％ and 6.58％.Conclusion This study demonstrates that mediation effects of the exercise are significant in some dimensions of HRQOL,but the direct influence of BMI on the HRQOL plays a leading role.And BMI is a protective factor under certain conditions.

ObjectiveTo evaluate the value of endoscopy in the diagnosis of biliary tract roundworm disease.Methods32 cases of biliary tract roundworm diseases were diagnosed and treated with fibre (electron) stomach duodenum endoscopy and fibre biliary tract endoscopy under the direct-view.Results32 cases were all clearly diagnosed and the worms were taken out with direct-view endoscopy or direct cholangiography with no complications.ConclusionEndoscopy is effective and accurate for the biliary tract roundworm disease and has the advantage of rapidly dispelling the sufferings of the patients.

Objective To investigate the characteristics of sagittal anatomic structure of the upper airway in patients with ankylosing spondylitis using three-dimensional reconstruction based on computed tomography (CT).Methods Thirty-one male patients with ankylosing spondylitis,aged 20-60 yr (AS group),and 41 common patients (male) without difficult airways,aged 20-60 yr (control group),who underwent spiral CT scan of the head and neck using Helical CT from January 2007 to February 2011 in our hospital,were enrolled in the study.Reconstructed images of the upper airway were obtained using AW4.4 workstation and six distances (D1-D6) and four angles (α-δ) were recorded and analyzed:(1)D1,the arc distance between the upper central incisor and root of epiglottis; D2,the distance between the upper central incisor and root of epiglottis; D3 and D4,the lengths of maxilla and mandible ; D5,the distance between the root of epiglottis and midpoint of glottis; D6,the distance between the end of mandible and midpoint of glottis; (2) angle α,the angle of line D2 and D5; angle β,the angle of line D2 and the lower edge of the upper central incisor to the midpoint of glottis; angle γ,the angle of line D4 and D6; angle δ,the angle of the point of the lower edge of the upper central incisor to the trailing edge of the hard palate and then to the root of epiglottis.Results Compared with control group,no significant change was found in D1,D2,D3,D4 and D5 (P ＞ 0.05),and D6,angle α and angle δ were significantly increased,whereas angle β and angle γ were decreased in AS group (P ＜ 0.05).Conclusion The anatomic structure of the upper airway has the characteristics of specific changes and a laryngoscope blade with a large degree of curvature may be helpful for successful tracheal intubation in patients with ankylosing spondylitis.

Objective To investigate the effects of ultrasound irradiating hydroxycamptothecin (HCPT)-loaded microbubbles on apoptosis in human hepatoma SMMC-7721 cells, and to clarify its probable mechanism. Methods The SMMC-7721 cells were randomly divided into six groups as follows: control group, blank liposome microbubbles plus ultrasound, HCPT-loaded microbubbles, HCPT, HCPT plus ultrasound,and HCPT-loaded microbubbles plus ultrasound. Apoptosis was ascertained by Annexin V-Fire/ PI and TUNEL methods. The expression of Bcl-2 and Bax protein was detected by immunocytochemistry staining technique. Results In the group of HCPT-loaded microbubbles plus ultrasound,the apoptosis rate at early term was found to reach (12.18 ± 1.38)% by Annexin V-Fitc/PI assay, brown-colored positive apoptotic cells were observed and the apoptosis rate at advanced term was found to reach (34.25 ± 1.83)% with TUNEL method. The expression of Bcl-2 was found to decrease,the expression of Bax increase,and the ratio of Bcl-2/bax significantly decrease by immunocytochemistry staining technique in the same group. Differences with other groups were significant. Conclusions The method of ultrasound irradiating HCPT-loaded microbubbles can notability induce human hepatoma SMMC-7721 cells apoptosis. The decrease in the ratio of Bcl-2 to Bax might be responsible for cells apoptosis.

Objective To explore the genetic prenatal diagnosis method for acbendroplasia (ACH).Methods During May to November 2007, three ACH pedigrees were diagnosed at the Prenatal Diagnosis Center, Department of Obstetrics and Gynecology, Affiliated Drum Tower Hospital of Medical College, Nanjing University. In family 1, there was a 6-month-old male ACH infant. In family 2, the expectant mother, with 18 weeks of pregnancy, was an ACH patient. Amniocentesis was performed for prenatal diagnosis. The fetus of family 3 was diagnosed as ACH by ultrasound examination on the 39th week of gestation. Umbilical cord blood of this fetus was collected for examination. Totally, three methods, restriction enzyme (Sfc Ⅰ and Msp Ⅰ ) digestion analysis, denaturing high performance liquid chromatography (DHPLC) and sequencing analysis were performed simultaneously to detect the pathogenic mutation of flbroblastic growth factor receptor 3 (FGFR3) for the three ACH families. Results ( 1 ) The DHPLC detection: heteroduplex was detected in the patient of family 1 ; beth the patient and the fetus of family 2 showed heteroduplex results; the result of the fetus of family 3 was also heteroduplea. (2) The enzyme digestion analysis for the PCR products of 10 exon of FGFR3: after Sfc Ⅰ digestion, the PCR products of patients and the fetus of family 1 and 2 showed not only the band of 247 bp, but also bands of 162 bp and 85 bp. But their PCR products could not be digested by Msp Ⅰ , and it only showed the band of 247 bp. For the fetus of family 3, the PCR products could not be digested by Sfc Ⅰ , while after digestion by Msp Ⅰ , bands of 162 bp and 85 bp were shown up. The PCR products of the normal control could be digested by neither Sfc Ⅰ nor Msp Ⅰ. (3) The sequencing results: the heterozygote mutation of 1138 C→A was confirmed in the patient of family 1. The pregnant woman and her fetus in family 2 showed the same result. The heterozygote mutation of C→C was confirmed in the fetus of family 3. The site of 1138 was G homozygote in the normal control The three detection results of the fetus in family 2 were the same as that of the mother, which means that the fetus inherited the same pathogenic mutation from his or her mother. Conclusions Both DHPLC and restriction enzyme digestion analysis could detect the mutation of FGFR3 gene, but DHPLC is more rapid, convenient and sensitive. So DHPLC can be applied to genetic diagnosis and prenatal diagnosis for ACH patients.

Objective To study the nerve protective mechanism of puerarin on ischemic brain injured and observe the expression change of TNF-α cytokines in ischemic brain area. Methods 90 healthy male SD(Spraque-Dawley)rats were used to built the animal model of brain medium sized artery focal cerebral ischemia reperfusion with the patching methods, and the rats were randomly divided into three groups: Sham operation group(n=30),operation group(n=30),operation with puerarin interfered group(n=30).There were five different observing time in every group(6h,12h,24h,48h,72h after operation).The operation with puerarin group rats were treated with puerarin solution, the sham operation and operation group rats were injected the same volume Sodium Chloride. To observe the expression of TNF-α cytokines in different time, immunohistochemistry staining in ischemic region was used. Results Compared with the sham operation group. the expression of TNF-α cytokine in cerebral isehemic reperfusion brain tissue in the operation and the operation with puerarin interfered groups was higher(P＜0.01).Compared with the operation group, the expression of TNF-αcytokine was significantly decreased in different time after cerebral ischemic-reperfusion in the operation with puerarin interfered group(P＜0.01 or P＜0.05).Conclusions The puerarin can restrain the expression of the TNF-α cytokine.

Glucose-6-phosphate dehydrogenase (G6PD) derives from the expression of the house-keeping gene G6PD. Recent studies have indicated that G6PD is related to tumor genesis, growth, clinical phenotype, therapy, and prognosis. To elucidate the relationship between G6PD and cancer, three siRNA sequences and one negative control sequence were designed based on the 3' noncoding region of the human G6PD gene. Two complementary single-strand DNA (sense and antisense) were designed and synthesized based on siRNA sequences. The DNA fragments were annealed and ligated to the GFP expression vector pRNAT-U6.2/Lenti. One siRNA with higher interference efficiency than the other two was found after siRNA plasmid transfecting human skin A375 melanoma cells. After lentivirus particle packaging and virus production, the A375 cells were infected, and the single cell clone was acquired and cultured to establish the stable cell strain. Western blotting showed that the endogenous G6PD in the stable A375 cell strain was 0.257 ? 0.074, which was 11.17% of G6PD expression (2.301 ? 0.286) in wild type A375 cells. The final siRNA interference efficiency in this stable cell strain was 88.83%. The G6PD activity of A375-G6PD?驻 was 21.53% of A375-WT. Further study showed that A375-G6PD△ doubling generation time prolonged, and its proliferation was greatly inhibited and the cloning efficiency lowered 25%(P

Objective To investigate the protective effect of IL-37 on hepatocyte injury against hypoxia/reoxygenation (H/R) by promoting polarization of M2-type macrophages and its molecular mechanisms.Methods Real-time fluorescent quantitative PCR (qRT-PCR) and Western blot were used to detect the levels of IL-37 mRNA and protein in human monocyte-macrophage THP-1 cells with different polarizations.The lentivirus with IL-37 gene was infected into THP-1 cells.The levels of CD206,CD86,ARG1 and iNOS mRNA was detected by qRT-PCR.The levels of CD163 and CD86 protein was detected by flow cytometric analysis.THP-1 cells and L02 cells were co-cultured by Transwell and treated with H/R.The survival rate and apoptotic rate of L02 cells were detected.The levels of alanine transaminase (ALT) and aspartate aminotransferase (AST) in culture medium were measured.The levels of STAT6 and its phosphorylation in THP-1 cells were detected by Western blot.Results The levels of IL-37 mRNA and protein were up-regulated in M2-type macrophages.IL-37 promoted the polarization of M2-type macropahges.M2-type macrophages induced by IL-37 were cocultured with L02 cells,the survival rate was significantly increased by H/R treatment (P =0.015),while the apoptotic rate,ALT level and AST level were significantly decreased (P＜0.001).The level of phosphorylated STAT6 in THP-1 cells overexpressing IL-37 was up-regulated (P ＜ 0.01).Conclusions IL-37 can induce polarization of M2-type macrophages and protect hepatocyte injury against H/R.Its mechanism may be related to STAT6 signal pathways.

Objective To analyze the high risk factors of blood infection in Chinese citizens' organ donation,provide the basic evidence for early protection,increase the success rate of donor distribution,and expand the Chinese organ donation pool.Methods A retrospective study was performed on 70 cases of donation recruited during October 2014 to January 2016.The incidence of blood infection in these donors was analyzed.The univariate analysis and multivariate logistic regression analysis were used to find out the high risk factors influencing the donor blood infection.Finally,the donor blood infection assessment model and the receiver operating characteristic (ROC) curve were established to assess the sensitivity and specificity.Results The overall infection rate was 64.3％ (45/70).The pulmonary,blood,and urinary tract infection rate was 42.9％,31.4％ and 1.4％ respectively.The total length of hospital stay (＞10 days) (P =0.017),oxygenation index (＜ 233.5 ± 107.0) (P =0.046),aspartate aminotransferase (＞196.9 ± 329.1 U/L) (P =0.044),and valley alanine aminotransferase (＞95.0 ± 78.1 U/L) (P =0.026) were four risk factors for predicting the donor blood infection.The multivariate logistic regression analysis revealed that the total length of stay ＞10 days along with the donors' oxygenation index (＜233.5 ± 107.0) was independent risk factor for predicting the blood infection.The donor blood infection model was:0.193 + 1.753 hospital stay (＞10 days)-0.007 oxygenation index.The sensitivity and specificity were 0.682 and 0.75 (P ＜0.001) respectively.Conclusion For a long-term stay in ICU,the rate of blood infection for donors was much higher,at this time,the most effective antibiotics should be chosen.Besides,improving donor oxygenation index and liver function can reduce the incidence of infection.