Adolescent ; Adult ; Age Determination by Skeleton ; methods ; Child ; Child, Preschool ; China ; Diagnosis, Computer-Assisted ; methods ; Female ; Humans ; Male

Objective To investigate the effects of simultaneous hepatic artery and portal revaseularization on the expression of TNF-α during bile duet ischemia-reperfusion injury in rats liver transplantation.Methods Male Spragne-Dawley rats were used to establish an autologous orthotopic liver transphmtation model.Model rats were random divided two groups,simultaneous revascularization group(sroup P)and portal vein revascularization group(group N).The animals were separately killed at the 2nd hour,6th hour and 24th hour after reperfusion.Plasma samples were collected for ALT,AST,GGT,AKP,TBiL and DBiL test.Bile duct tissues were collected to detect the histolosical changes,MPO activit,and the expression of TNF-α mRNA.Results The serum levels of GGT in group P was significantly lower than that in group N at the 6th hour and 24th hour after reperfusion(P＜0.05).And the serum levels of AKP,TBiL and DBiL and the morphological scores of bile duct in group P were significantly lower than that in group N at the 24th hour after reperfusion(P＜0.05).The activity of MPO in group N was significantly higher than that in group P at the 6th hour after reperfusion(P＜0.05).Compared with group N,the expression of TNF-αmBNA was reduced significantly in group P at the 2nd hour and 6th hour after reperfusion(P＜0.05).Conclusion Simultaneaus hepatic artery and portal revascularization can alleviate I/R induced bile duct injury of rat liver grafts,and the protective mechanism may be asseciated with inhibition the expression of TNF-α and decrease of neutrophil infiltration.

Objective By using a newly developed animal model of limited macular translocation (LMT) in rabbit eyes, to explore the complications,indications and predictability of LMT. Methods LMT models were performed in 15 rabbit eyes, and were divided into 4.5 mm group and 3.5 mm group by the length of scleral shortening.The effect of retinal translocation (distance and direction) and its complications were analyzed. In a separated group, postoperative corneal astigmatism was studied after release of scleral shortening suture 1 month after LMT. In order to study the impact of neuroretina retinal pigment epithelium (RPE) adhesion on LMT, retinas and choroids in 4 pigmented rabbit eyes were pretreated with photocoagulation. Results Success rate of LMT was 86 7%. In these cases, retinal translocations were achieved [(distance of 610 to 2 690 ?m, [AKx-]?s =(1 395?636) ?m; translocation angle ? of 10 38?, ?s =(22.7?8.3) ?]. The difference in scleral shortening between 4.5 mm and 3.5 mm groups did not significantly influence the final translocation. Major complications of LMT included intraocular bleeding, retinal break, and corneal astigmatism. Release of scleral shortening suture reduced corneal astigmatism and made it more regular, meanwhile, retinal translocation did not regress. The neuroretina RPE adhesion induced by retinal photocoagulation made neuroretinal hydrodissection difficult, and led to retinal break readily. . The difference in scleral shortening between 4.5 mm and 3.5 mm groups did not significantly influence the final translocation. Major complications of LMT included intraocular bleeding, retinal break, and corneal astigmatism. Release of scleral shortening suture reduced corneal astigmatism and made it more regular, meanwhile, retinal translocation did not regress. The neuroretina RPE adhesion induced by retinal photocoagulation made neuroretinal hydrodissection difficult, and led to retinal break readily. Conclusion Sufficient LMT models in rabbit eye were obtained with few postoperative complications. Direction of retinal translocation is an important factor influencing the effect of LMT, except for distance. Release of scleral shortening suture can effectively reduce corneal astigmatism without causing regression of retinal translocation. For the patients with a history of macular or para macular photocoagulation, LMT should be performed carefully.

Purpose To prepare a novel fusion protein of CREKA and tTF as a universal carrier targeting to cancer,and to analyze its activities.Methods CREKA and tTF gene were acquired by PCR,and inserted into plasmid pET22b(+)to construct recombinant plasmid CREKA/tTF/pET22b(+),and the fusion gene was expressed in E.coli BL21.The fusion protein Wag purified through Nickel-affinity chromatography column.After purifying,the fusion protein was refold by subsequent dialysis.The activities of the fusion proteins were measured by coagulation timing and quantitative fluorescence test in vitro.Results The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was obtained.The fusion protein was highly expressed in E.coli BL21.The coagulation of the fusion protein Was determined by the coagulation test.And the capability of the fusion protein effectively binding to clotted plasma proteins is identified in quantitative fluorescence test.Conclusion The recombinant plasmid CREKA/tTF/pET22b(+)with correct sequence was built.The fusion protein CREKA/tTF with both TF and CREKA activity was successfully obtained.

Objective To establish a novel flow cytometric crossmatch assay allowing detection of complement-fixing donor specific anti-HLA IgG alloantibodies (Flow cytometry complement-dependent crossmatch,Flow-CDC). Methods One hundred pretransplant crossmatchings were performed using Flow-CDC and NIH-CDC between 62 patients awaiting renal transplantation and 33 donor cells.These crossmatchings were divided into two groups according to PRA.Group 1 consisted of 25 sera with negative PRA,and group 2 consisted of 75 sera with positive PRA.All of the sera were pretreated with DTT to inactivate IgM. Lymphocytes were isolated from peripheral blood (or,in a few instances,from the spleen) of the cadaveric donors. The correlation between different techniques for detection of donor specific anti-HLA antibodies was evaluated.The effect of both methods on clinical transplantation outcome was observed. Results In group 1,NIH-CDC and Flow-CDC were negative for all 25 sera.In group 2,24 (32.0%) had a positive NIH-CDC,31 (41.3%) had a positive Flow-CDC.There was a significant difference between two methods (?2=5.14, P= 0.016 ).Overall concordance between both tests was 93% with 69 concordant negatives and 24 concordant positives. The correlation coefficient (r) was 0.80.In group 2,5 patients received transplantation. One of them with negative NIH-CDC and positive Flow-CDC suffered from acute rejection after transplantation and lost the graft,and the other patients with negative NIH-CDC and Flow-CDC had good outcome. Conclusions Flow-CDC can detect specifically complement-fixing IgG alloantibodies against donor HLA and is more sensitive than NIH-CDC.Additionally,the computer printouts represent a permanent record of the crossmatch for retrospective review.Flow-CDC may become the standard crossmatch method as a possible alternative to conventional NIH-CDC testing.

Objective To study the effect of DNA methylation regulation on the toxic effect of paraquat on the sensitized V79 cells t pretreated with 5-aza-2'-deoxycytidine.Methods V79 cells were treated by 5-aza-2'-deoxycytidine (5-Aza-2'-dc) for 12h,which is a DNA methylation inhibitor,and then treated with paraquat for 12h.The morphological changes of V79 cells were observed by microscopy and the cell viability was determined by MTT assay and trypan blue staining method.Results Microscopic examination showed that the combination of 5-Aza-2'-dc and paraquat had stronger effect in inhibiting the growth of V79 cells(the cells became smaller and poorer adhensive ability) than single 5-Aza-2'-dc or paraquat.MTT assay showed that cell viability in the combination group (54.47 ± 3.04) ％ was significantly lower than the 5-Aza-2'-dc group (95.52 ± 0.90) ％ and paraquat group (89.68 ± 4.26) ％ (P＜0.05).Trypan blue staining assay showed that the death rate of ceils in the combination group (53.58 ± 1.57) ％ was significantly higher than the 5-Aza-2'-dc group (7.44 ± 2.31) ％ and paraquat group (12.90 ± 1.21) ％ (P＜0.05) Conclusion 5-Aza-2'-dc promotes V79 cells damage caused by paraquat.

Objective To observe the effect of electroacupuncture(EA) in combination with small dose of clozapine on clinical symptoms of refractory schizophrenia patients so as to evaluate its validity and security.Methods Eighty schizophrenia patients were randomized into medication group and EA+medication group(n=40/group).Patients of medication group were treated with conventional oral administration of clozapine(50-100 mg/d to 200-500 mg/d,for 8 weeks) and those of EA+ medication group treated by EA of Baihui(GV 20) and bilateral Taiyang(EX-HN 5) and oral administration of clozapine(50 mg/d to 100-150 mg/d).EA was given to the patients,3 times a week for 8 weeks.Scores of the Positive and Negative Symptom Scale(PANSS) and the Untoward Effect Symptom Scale(TESS) were used to assess the therapeutic effect.Results Following treatment,PANSS scores for positive symptoms and the common psychiatric symptoms of the medication and EA+medication groups all decreased evidently from the 2rd week on(P0.05).But TESS score of EA+medication group was evidently lower than that of medication group(P

OBJECTIVETo explore the scavenging action of tenuigenin (TEN) on intracerebral amyloid β protein (Aβ) aggregation and the abnormal phosphorylated tau protein and its mechanism in Alzheimer's disease (AD) rats' brain.

METHODSAβ1-40 was injected into the right CA1 region hippocampus to establish the AD model. Successfully modeled rats were divided into the model group, the low, middle, high TEN group. Rats were administered with TEN (18.5, 37.0, 74.0 mg/kg) by gastrogavage. Besides, a sham-operation group was set up. Expression levels of Aβ1-40 and Tau p-Ser262 were detected by immunohistochemistry. Expression levels of ubiquitin (Ub) and Ub-protein ligase E3 were measured by Western blotting.The content of 26S proteasome was detected by ELISA.

RESULTSImmunohistochemical results showed that the number of Aβ and Tau p-Ser262 positively reacted neurons significantly increased in model group, when compared with the sham-operation group (P < 0.01). Results of Western blot showed expression levels of ubiquitinated protein were up-regulated and those of Ub-protein ligase E3 were down-regulated in the model group (P < 0.01). ELISA results showed that the content of 26S proteasome significantly decreased in AD rats' brain (P < 0.01). Compared with the model group, expression levels of Aβ1-40, Tau p-Ser262, and Ub significantly decreased; expression levels of Ub-protein ligase E3 apparently increased; the content of 26S proteasome significantly increased in each TEN treatment group (P < 0.05, P < 0.01). Best effect was shown in 37.0 mg/kg and 74.0 mg/kg TEN groups.

CONCLUSIONSUb proteasome pathway (UPP) participated in the occurrence of AD. TEN could obviously reduce intracere- bral Aβ1-40 accumulation and abnormal tau phosphorylation.

Alzheimer Disease ; metabolism ; Amyloid beta-Peptides ; Animals ; Drugs, Chinese Herbal ; pharmacology ; Hippocampus ; metabolism ; Neurons ; metabolism ; Phosphorylation ; Proteasome Endopeptidase Complex ; metabolism ; Rats ; Ubiquitin-Protein Ligases ; metabolism ; Ubiquitins

Objective To study effect of excessive iodine intake on sodium-iodide symporter(NIS)mRNA and protein expression of breast in lactating rats.Methods60 Wistar rats,having been weaned for one month,were randomly divided into three groups according to their body weights,I.e,①normal iodine(NI,30 rats);②ten fold high iodine(10 HI,15 rats);③one hundred fold high iodine(100 HI,15 rats).Eating food containing iodine of 300μg/L and drinking water of iodine at 5,1845,20 295μg/L,respectively.After fed for 3 months,the rats mated and had offspring,and urine and milk iodine of lactating rats were determined by As-Ce-catalytic spectrophotometric method.Their marmnary glands were sampled at lactation day 10.Then NIS mRNA expression by RT-PCR was determined and NIS protein by immunohistochemistry(SABC)was observed.Results The urine iodine of 10 HI group(3597.5μg/L)and 100HI group(25 404.3μg/L)increased obviously compared with that of NI group(344.7μg/L).The milk iodine of 10HI group(27.1×103μg/L)and 100HI group(191.0×1μg/L)was higher than that of NI group(6.0×103μg/L),but the increased fold of milk iodine was not paralleled with that of urine iodine.Difference of NIS mRNA expression was significant(F=24.19,P＜0.01)among the groups,and the NIS mRNA expression in 10HI(1.250±0.034)and 100HI(1.272±0.039)group were less than that in NI (1.532±0.044)group(P＜0.01).The breast NIS mRNA expression in lactating rats(1.532±0.044)was significantly higher than that in unlactating rats(0.879±0.018,P＜0.01).With the increasing iodine uptake,NIS protein expression decreased.Conclusions The NIS mRNA and protein in rat breasts is down-regulated by excessive iodine intake.So increasing extent of milk iodine concentration is inhibited,which is important to prevent off-spring from getting excessive iodine intake from parental generation.

[Introduction] To estimate the prevalence ratios, using a log-binomial model with or without continuous covariates. Prevalence ratios for individuals' attitude towards smoking-ban legislation associated with smoking status, estimated by using a log-binomial model were compared with odds ratios estimated by logistic regression model. In the log-binomial modeling, maximum likelihood method was used when there were no continuous covariates and COPY approach was used if the model did not converge, for example due to the existence of continuous covariates. We examined the association between individuals' attitude towards smoking-ban legislation and smoking status in men and women. Prevalence ratio and odds ratio estimation provided similar results for the association in women since smoking was not common. In men however, the odds ratio estimates were markedly larger than the prevalence ratios due to a higher prevalence of outcome. The log-binomial model did not converge when age was included as a continuous covariate and COPY method was used to deal with the situation. All analysis was performed by SAS. Prevalence ratio seemed to better measure the association than odds ratio when prevalence is high. SAS programs were provided to calculate the prevalence ratios with or without continuous covariates in the log-binomial regression analysis.