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 analyze the association of the single nucleotide polymorphism (SNP) of Toll-like receptor 7 (TLR7) and Toll-like receptor 9 (TLR9) with chronic hepatitis C virus (HCV)infection.Methods A total of 150 patients with chronic hepatitis C (CHC) admitted to Renmin Hospital of Wuhan University from January 2011 to May 2012 and 168 healthy controls were enrolled in the study.The genotypes of TLR7 IVS2-151 (rs179009) were detected by Sanger sequencing,and the genotypes of TLR9 T-1486C (rs187084) were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP).SPSS 15.0 was used for statistical analysis,and goodness-of-fit test for HardyWeinberg equilibrium was also performed.Results The frequency of TLR7 IVS2-151G was higher in malepatients with CHC than that in male controls (41.4％ vs.21.6％,x2 =7.250,P =0.007,OR =0.389,95％ CI:0.194-0.781) ; however the female CHC patients had a higher frequency of TLR7 IVS2-151A than the female controls (76.9％ vs.63.1％,x2 =7.202,P =0.007,OR =1.942,95％ CI:1.192-3.164).No significant difference in the distribution of TLR9 T-1486C (rs187084) gene SNP was observed betweenCHC and control groups (P ＞0.05).Conclusion TLR7 IVS2-151 (rs179009) is correlated with HCV infection,which may be involved in the pathogenesis of CHC.

Animals ; Cells, Cultured ; Hepatocytes ; drug effects ; secretion ; Male ; Pancreatic Polypeptide ; pharmacology ; Rats ; Rats, Sprague-Dawley

Objective • To understand the structural changes and related reasons of the average cost of hospitalization of childhood pneumonia in Shanghai, and to propound related policy suggestion. Methods • New grey correlation and degree of structural variation analysis were used to research the average cost of hospitalization of 22 543 children with pneumonia from 2015 to 2018 in a children's specialized hospital in Shanghai. The average cost included 5 categories, i.e. medicine cost, material cost, examination and laboratory cost, labour cost and bed occupation cost. The relational degree and structural changes between each average cost and the average cost of hospitalization were discussed. Results • The new grey correlation analysis showed that the relational degree sort order of each average cost from the highest to the lowest were average examination and laboratory cost (γ3=1.000 0), average medicine cost (γ1=0.862 5), average bed occupation cost (γ5=0.845 1), average labour cost (γ4=0.796 8) and average material cost (γ2=0.786 3). The degree of structure variation analysis showed that the contribution rate of structure variation (CSV) sort order of each average cost in 2015-2018 from the highest to the lowest were average medicine cost (CSV1=36.22%), average bed occupation cost (CSV5=27.65%), average examination and laboratory cost (CSV3=13.91%), average material cost (CSV2=13.78%) and average labour cost (CSV4=8.44%). The average medicine cost and average material cost were negative variation. Conclusion • The average examination and laboratory cost and average medicine cost are the main factors that influenced the average cost of hospitalization of childhood pneumonia. The proportion of the average labour cost is relatively low. It is suggested that the cost structure like raising labour charges and optimizing diagnosis and treatment process should be further adjusted in order to control the growth of medical costs.

Objective To investigate the value of immunophenotypes in pathologic diagnosis of follicular lymphoma (FL) and the differential diagnosis between neoplastic follicle (NF) and reactive hyperplastic follicle (RHF).Methods 50 cases of FL and 10 cases of RHF as control were studied by clinical data, the expressions of CD20, bcl-2, CD3, CD10, bcl-6, CD21 and Ki-67 were detected by EnVision immunohistochemical method for the immunphenotypical pattern of FL and RHF.Results Among 50 FL cases , there were 24 male cases and 26 female cases, with median age of 50 years old (10-80 years old), including 32 cases (64 ％) involved predominantly neck lymph nodes.The histologic grades were 1-2 in 16 cases (32 ％) and ≥3 in 34 cases (68 ％).CD20, CD10 and bcl-6 were positive in NF tissues of FL, with irregular forms of NF and loss of the mantle area without clear demarcation, and infiltrating to NF, irregular shape and no clear perimeter.The bcl-2 was positive in all of grade 1-2 NF, and it was positive in 68 ％ (23/34) cases and was partial or total negative in 32 ％ (11/34) cases in grade≥3 NF.Those were poorly demarcated contours.The Ki-67 proliferative index of NF were ＜30 ％ in grade 1-2, and ＞30 ％ in grade ≥ 3, with the highest to 90 ％.In FL, the CD21 staining result showed follicular dendritic cells (FDC) network were synchronized with NF, and in some cases of grade ≥ 3, the FDC network were ruptured.In RHF, the CD20 was positive for round or oval nodules with clear demarcation.The CD10, bcl-6 and CD21 were positive and bcl-2 was negative in all germinal center without invasive immunophenotype.The Ki-67 index was high in germinal center, and sometimes polarity may be seen.Conclusion The immunophenotypical differences between FL and RHF include invasive characteristics, loss of follicular mantle area and germinal center pattern in the NF, while the normal follicular immunophenotype in the RHF.

Objective Mitotic spindle and midbody are both microtubule-based temporary structures during cell growth and play essential roles in mitosis.The purpose of this study was to establish a mature and efficient method to extract mitotic spindle and midbody.Methods Through the cell cycle synchronization method,mitotic spindle or midbody was made appear inside cells.Low permeability swelling and glycerol gradient centrifugation principles were then used to extract spindle and midbody.Results By Western Blot and immunofluorescence staining,the extracts were identified as mitotic spindle and midbody.Conclusions The successful extraction of mitotic spindle and midbody from synchronized Hela cells will provide foundation for identifying the proteins located in cell during mitosis,and be of great significance to the study of molecular regulation mechanisms of mitosis and tumorigenesis.

Objective To evaluate the correlating clinical factors of coronary artery calcification score(CACS).Methods 141 patients suspected of coronary artery disease were included.They underwent multi-slice row computed tomography,pulse wave velocity ( PWV ),UCG and blood biochemistry within a period of 3 months.The subjects were divided into three groups according to CAC score:A(CACS =0-10),B ( CACS = 11-400),C ( CACS > 400).Results CACS was significantly associated with age,history of hypertension and diabetes mellitus.It was also associated with the presence of mitral annular calcification and aortic valve calcification,low ankel brachial pressure index(ABI) and high mean artery pressure(MAP) as well as high values of brachial ankel PWV (baPWV) and Upstroke time (UT).Muhifactorial logistic regression analysis showed that the presence of aortic valve calcification and mitral annular calcification,the history of diabetes mellitus and high value of UT were independently correlated with severe coronary artery calcification.Conclusions Aortic valve calcification,mitral annular calcification,history of diabetes mellitus,high value of UT were independently correlated with severe coronary artery calcification.Measurement of PWV and UCG should be performed before muhi-slicerow computed tomography,because the assessment of coronary artery lumen narrowing with multi-slice row computed tomography can not be carried out accurately in the presence of severe coronary artery calcification.

OBJECTIVETo obtain the information on the glucuronidation of Ginkgo flavonoid and the interaction profile of Ginkgo flavones with other drugs in vitro.

METHODSGinkgo flavonoids (quercetin, isorhamnetin and keampferol) and other drugs were co-incubated with rat hepatic microsome at 25 degree; the residual concentrations of flavonoids were determined by HPLC. The enzymatic parameters of quercetin, isorhamnetin and keampferol metabolism were assessed. The interactions between flavonoids and these drugs on glucuronidation were observed.

RESULTThe K(m) values were ( 24+/-0.05), (148+/-0.09) and (110+/-0.03) micromol/L and the V(max) values were (60+/-0.21), (48+/-0.02) and (34+/-0.02) micromol x g(-1) x min(-1) for quercetin, isorhamnetin and kaempferol, respectively. The IC(50) of nifedipine propafenone ipriflavone and diphenytriazol on flavonoids metabolism were 54 - 70, 69 - 122, 85 - 98 and 210 - 362 micromol, respectively. The inhibition constants (Ki) of diphenytriazol propafenone and ipriflavone on quercetin, isorhamnetin and keampferol metabolism were (57.6, 50.5, 33.1) (33.6, 59.5, 45.2) and(13.7,24.0,15.7) microg/ml respectively. The ratio [I]/[Ki] of the plasma concentration and inhibition constant for propafenone was 0.002 - 0.003.

CONCLUSIONThe metabolic level of quercetin is the strongest among three Ginkgo flavonoids. Nifedipine propafenone and ipriflavone inhibit the metabolism of quercetin, isorhamnetin and keampferol at different levels. Because of the interaction between Ginkgo flavonoids with nifedipine, caution must be taken when two drugs are used together clinically.

Animals ; Drug Interactions ; Flavonoids ; metabolism ; Ginkgo biloba ; Glucuronides ; metabolism ; Microsomes, Liver ; metabolism ; Rats

Objective To compare the clinical effects of extensive or limited arthroscopic debridement in varying degrees of knees osteo-arthritis. Methods 32 cases with knees osteoarthritis who underwent extensive debridement under arthroscopy were selected as the observa-tion group, and 30 cases with knees osteoarthritis who underwent limited debridement were selected as the control group. The excellent rate of joint function, Lysholm and VAS scores were compared between the two groups. Results The excellent rate of joint function were 90. 0%and 91. 4% in observation group and control group respectively, without significant difference (P>0. 05). Lysholm scores of patients with grade Ⅱ osteoarthritis was much higher in observation group than that in control group after operation (P<0. 05). There was no significant difference in Lysholm scores between gradeⅠand gradeⅢosteoarthritis (P>0. 05). No statistically significant difference of VAS scores on 1 year after operation were found (P>0. 05). Conclusion Extensive debridement and limited debridement under arthroscopy have similar effects for knees osteoarthritis. Extensive arthroscopic debridement is an ideal treatment method for Ⅱ grade of knees osteoarthritis.

Objective To investigate the effect of injectable Xuebijing on the proliferation of mouse brain microvascular endothelial cell in vitro.Methods Cultured mouse brain microvascular endothelM cell line bEnd. 3 was treated by injectable Xuebijing of different concentrations,0 (control),5,25 and 50 mg/ml.The regulatory. effect of Xuebijing on the proliferation of cell line bEnd.3 was observed and studied by means of MTT method and cell cycle analyzed with flow cytometry.Results Compared to the control group,MTT and proliferation index (PI) of 5 and 25 mg/ml groups were significantly increased at 12 and 24 h,and PI,but not MTT,of these 2 groups was decreased remarkably at 48 h.Meanwhile,50 mg/ml group showed significantly decreased MTT at 24 and 48 h,and PI of this group was increased obviously at 12 and 24 h,but decreased significantly at 48 h. Conclusions Injectable Xuebijing at certain concentrations might promote the proliferation of cultured mouse brain microvascular endothelial,cells within specific time frame.