Objective To analyse the operation technique and therapeutic effect of "inserting" uretero-intestinal anastomosis in orthotopic bladder substitution.Methods Thirty-eight patients undergoing orthotopic bladder substitution operations were followed up,and the way of uretero-intestinal anastomosis in all Datients was the "inserting"uretero-intestinal anastomosis.The therapeutic effect was observed by radiation,cystoscopy,pathologic biopsy and blood test.Results The average follow-up time was(3 1.65±14.14)montll8.and the stricture rate was 4%(3/75),but no vesicoureteric reflux was found.The rate of leakage was 0.Nipples were formed at the site of anastomosis under the view of cystoscope,and among the 7 patients whose nipples were taken to be examined by histology,2 cases were intestinal epithelium which were taken at the base of nipple8.while the others were transitional epithelium which were taken at the top of nipples.The renal function of all patients was normal (Cr 54-135 μmol/L,BUN 3.2-9.4 mmol/L).Conclusion "Inserting"uretem-intestinal anastomosis is an ideal antireflux uretero-intestinal anastomosis method.

Objective To study the enzymatic progress of amyloid precursor protein (APP), and construct the recombinant eukaryotic expression plasmid encoding the double mutations of APP and 2 different types of fluorescent protein. Methods The last 300 bases of APP named as C99 containing APP717 mutation was amplified by polymerase chain reaction (PCR) with the template pcDNA3.0-APP.The cyan and yellow fluorescence sequences named as CFP and YFP respectively were obtained by PCR with the template of digestion of the plasmid pcDNA3.0-CFP-CaM-YFP.The 54 bases in the middle of APP named as 54 bp containingSwedish mutation were synthesized. The 4 fragments, CFP, YFP, C99 and 54 bp were inserted into the vector pcDNA3.0.By genetic engineering the recombinant plasmid pcDNA3.0-CFP-54bp-YFP-C99 and pcDNA3.0-CFP-54bp-YFP were constructed and finally identified by enzyme digestion, PCR and sequencing. Then the 2 constructs were transfected into SH-SY5Y cells respectively. The expression of the fusion gene was examined by multi-photon con-focal microscopy and the fluorescence resonance energy transfer (FRET) signal was collected. The amyloid beta (Aβ)deposition was detected by immunocytochemistry. Results (1) DNA sequencing showed that the sequence of the construct was correct.(2)The cyan and yellow fluorescence could be seen by the multi-photon con-focal fluorescence microscopy. The fusion gene could be correctly expressed.(3)FRET was present in the cells expressing CFP-54bp-YFP,but absent in the cells expressing CFP-54bp-YFP-C99.(4)A13 labeled by YFP was detected by multi-photon con-focal microscope and deposited in the cytoplasm, membrane and in the intercellular space.(5)It was shown that by immunocytochemistry, the CFP-54bp-YFP-C99 expressionproduct could generate Aβ. The Aβ deposition was detected in the cell membrane, cytoplasma and intercellular space. Conclusions (1) The fusion protein could generate Aβ peptide by β and γ proteolytic processing.(2)Aβ may be generated in the conrse of APP transportation from cell plasma to cell membrane.(3)C99 is very important for the correct cleavage of APP. Our test data strongly suggest that C99 may function as the signal-like peptide. It may guide and direct the APP to the right location for the cleavage.(4)Before the Aβ deposition is formed outside of the cell, Aβ intracellular accumulation results in the change of cell shape.

Objective To established a diagnostic methods to idenfity Niemann-Pick disease type C (NPC) in China. Methods Two patients aged 5 and 20 years respectively who presented progressive neurologic regression and splenomegaly were subjected to filipin staining of cultured skin fibroblasts and genetic analyses of NPC1 gene. Results Although there were differences in onset ages and clinical presentations, filipin staining of the cultured skin fibroblasts confirmed the clinical diagnosis of NPC, showing an intense punctate pattern of fluorescence concentrating around the nuclei, consistent with the accumulation of unesterified cholesterol in NPC cells. Genetic sequence analysis further verified the results of filipin staining. The case 1 was compound heterozygous for M1142T(3425T＞C),R1186H(3557T＞C)and case 2 for Q88H(264G＞T),469_470insGT.The latter 2 mutations were novel, and the possibility of polymorphisms was not supposed by analyzing 134 DNA samples obtained form normal controls. Conclusions Filipin stainning of the cultured skin fibroblasts is a reliable method to clinically diagnose NPC with a sensitivity. Genetic diagnose should be performed where genetic analysis is allowed.

OBJECTIVE:To screen formulation of Insulin cream and to investigate the effects of insulin cream on wound healing in rabbits.METHODS:The best formulation of insulin cream was selected from 8 kinds of formulations with appearance and stability as index.Rabbit trauma model were induced and divided into matrix group,blank group,Insulin cream groups(high-dose,medium-dose and low-dose).Wound healing rate and healing time were compared among those groups.RESULTS:The fifth formulation possessed good appearance and stability,which included albolene,cetanol,glycerin monostearate,glycerol,etc.Compared with matrix group,the wound healing rates were obviously increased at different time points in Insulin cream groups(P

Objective To evaluate the efficacy of mitomycin (MMC) intravesical chemotherapy for superficial bladder carcinoma by in vitro chemosensitivity using histoculture drug response assay (HDRA).Methods Forty-one cases of superficial bladder transitional cell carcinoma(TCC) were obtained,including 30 males and 11 females with mean age of 55 years.Of them,10 cases were Ta and 31 were T1 according to TNM stage system (UICC 2002) while 9 cases were G1,22 were G2 and 10 were G3 (WHO1973).All cases had no chemotherapy history before operation and were operated to retain bladder.Tumor specimens were cultured by three-dimensional histoculture.HDRA with im-proved MTT assay was utilized for chemosenstivity test of MMC with 1 g/L concentration and 2 hours exposure.Growth inhibition rate (GI) exceeding 70% was defined as high-sensitive while less than 50% GI was defined as insensitive,others were moderate-sensitive.All cases were performed standard intravesical chemotherapy with MMC 40 mg plus 40 mt saline.Every case was followed up every 3 months.The patients were followed up for 5 years or untill recurrence.Results The MMC chem-osensitivity was different among 41 patients.Thirteen cases were insensitive including 1 of Ta,12 of T1 (P=0.009) and 1 of G1,7 of G2,5 of G3(P=0.101).Total recurrence rate was 39%(16/41) af-ter 3 to 5 years follow-up.There were 1 of Ta,15 of T1 (P=0.059) and 10 of G2 6 of G3 (P=0.016).Insensitive group recurrence rate was 77% (10/13) while sensitive group was 21% (6/28,P= 0.004).Patients in sensitive group showed a longer median time(49.2 months) than patients in insen-sitive group (16.5 months,P<0.001) according to Kaplan-Meier analysis with Log-rank test.The MMC chemosensitivity was independent prognostic factor examed by Cox regression analysis (P= 0.008).There was a 78% correlation rate of chemosensitivity by HDRA to clinical effect of MMC in-travesical chemotherapy.Conclusion HDRA could evaluate MMC intravesical chemotherapy for su-perficial bladder TCC,improve therapeutic effect and lower tumor recurrence rate.

Objective To evaluate the internal reliability of Psychosocial Function Evaluation for mental disabled people in day care unit. Methods 16 participants were evaluated twice by the same rater (social worker) in the unit at the Permanent stage and the Steering stage. Results Cronbach's α was greater than 0.7 in the both evaluation. The Spearman correlation coefficient between each dimension and total score were between 0.502 and 0.869. Conclusion The internal reliability of Psychosocial Function Evaluation is satisfactory for mental disabilities.

Cysticercosis is caused by the metacestode form of Taenia solium-Cysticercus cellulosae and it causes great economic losses and threatens the people's health. There are some problems on how to control cysticercosis, in order to resolve the key problem that the native antigen to diagnose and prevent cysticercosis is very limited and is not satisfied, Pichia pastoris Expression System was used to express recombinant P2 protein. The interested P2 gene was got by digesting the pGEM - P2 vector using restriction endonuclease, then it was inserted into the secretory pPIC9K Pichia pastoris expression vector and transformed into E. coli. Positive recombinant plasmids were selected sequenced and named pPIC9K-P2 and it was linearized by Sal I and Bgl II, then the linear DNA transfored into Pichia pastoris GS115 by electroporation. The recombinant expression vector pPIC9K - P2 integrated into GS115 via homologous recombination between the transforming DNA and regions of homology within the genome. The transformants were screened for multicopy recombinants using G418 and were distinguished for Mut phenotypes by MD and MM. Two different phenotypes were generated-HIS+ MUT+ (Methanol utilization plus) and HIS+ MUT(S) (Methanol utilization slow). PCR analysis of the multicopy recombinants indicated that the P2 gene was integrated within the genome of pichia Pastoris. The multicopy recombinants were named GS115/pPIC9K - P2HIS+ MUT+ and GS115/pPIC9K-P2HIS+ MUT(S), both HIS+ MUT+ and HIS+ MUT(S) were induced with methanol. The results of SDS-PAGE and Western blot demonstrated that the culture supernatant of the induced Pichia pastoris contained P2 protein which was accumulated up to 33 % of total proteins in the culture supernant and its molecular weight is 12.6kD. The results of the clinical study indicated that the expression P2 protein could be used to diagnose human cysticercosis and swine cysticercosis as diagnosis antigen.
Animals ; Blotting, Western ; Cysticercosis ; diagnosis ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Genome, Fungal ; genetics ; Helminth Proteins ; genetics ; metabolism ; Humans ; Phosphoproteins ; genetics ; metabolism ; Pichia ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; metabolism ; Swine ; Taenia ; metabolism

Hypothalamic-pituitary-adrenal(HPA)axis-related factors in patients with type 2 diabetes were studied according to different levels of HbA1C.It showed that HPA axis was normal in HbA1C≤ 7% group[ACTH (18.03±8.39)ng/L,blood cortisol(49.22±8.68)μg/L],hyperactive in 7%11% group with weak feedback regulation[ACTH(26.08±15.41)ng/L,blood cortisol(55.64±24.27)μg/L].These results suggest that HPA axis-related factors in type 2 diabetic patients are different with different grades of glucose metabolic turbulence.

Objective To evaluate left ventricular function in patients with hypertensive hypertrophic cardiomyopathy(HHC)using real-time 3-dimensional echocardiography(RT-3DE).Methods Thirty patients with HHC and 32 control subjects were studied.Full-volume RT-3DE data from apical window were acquired,and regional volumetric time curves of 17 segments were obtained by fast 3-dimensional border detection software.Several left ventricular function parameters were calculated semiautomatically,including global left ventricular end-diastolic volume(EDV),end-systolic volume(ESV),left ventricular ejection fraction(LVEF),the ratio of ESV/EDV of 17 segments,the standard deviation(SD)and difference(Dif)(adjusted by the R-R interval) of time to minimum systolic volume(Tmsv)in 16 segments(Tmsv16-SD and Tmsv16-Dif).Results EDV and ESV were significantly larger in patients with HHC than that in control subjects[(88±29)ml vs (72±15) ml,t=-2.680,P=0.008;(28±10)ml vs (22±6 )ml,t=-2.613,P=0.01].HHC had a higher ratio of ESV/EDV at interventricular septum(IVS)compared with control group[mid-segments of anterior IVS:(40.51±20.28)% vs (26.43±10.10)%,t=-3.378,P=0.002;mid-segments of posterior IVS:(41.44±23.55)% vs (24.46±8.12)%,t=-3.688,P=0.001;apical segments of IVS:(30.96±21.31)% vs (19.53±7.33)%,t=-2.745,P=0.01].In patients with HHC,Tmsv16-SD and Tmsv16-Dif were significantly longer[(2.48±1.38)% vs (1.16±0.26)%,t=-5.117,P＜0.001;(7.67±5.07)% vs (3.95±1.48)%,t=-3.865,P＜0.001].And the prevalence of left ventricular dyssynchrony was higher than that in control subjects(43% vs 3%).Conclusions HHC patients may have regional left ventricular systolic dysfunction before global changes,and have a higher prevalence of left ventricular dyssynchrony.RT-3DE is a useful imaging modality for assessing left ventricular systolic function.

OBJECTIVETo investigate the root canal curvature in Chinese mandibular permanent incisors.

METHODSTwo hundred and ninety nine Chinese mandibular permanent incisors were included in this study. The root canals were inserted with stainless steel 15# K files, and then taken radiography from the facial and proximal view by X-ray paralleling technique. Canal curvatures were evaluated by measuring the angle, the radius of the curvatures, and the length of the curved part of the canal. Based on the curved angle and the shape of curved canals, the root canals were classified.

RESULTSThe curved angle, radius, and the curved canal length was 5.28 degrees +/- 5.52 degrees, 16.19 mm +/- 12.38 mm, 4.10 mm+/-2.01 mm, respectively in the facial view; 9.99 degrees +/- 5.84 degrees, 18.86 mm +/- 9.71 mm, 3.27 mm +/- 2.39 mm, respectively in the proximal view. The prevalence of straight, light, moderate, and serve curved root canals in mandibular permanent incisors was 15.7%, 66.9%, 16.7%, and 0.7%, respectively (P<0.05) based on the curved angle. The prevalence of straight, L shape, and S shape curved root canals was 7.7%, 50.5%, and 41.8%, respectively (P < 0.05).

CONCLUSIONMost of the root canals in mandibular permanent incisors are curve.