Objective To compare the histological fates and the effects of autologous fat and fascia used as augmentative materials to treat unilateral vocal fold paralysis (UVFP). Methods Vocal fold augmentation with autologous fat and fascia was performed on 2 dogs (group A) to cure glottic insufficiency caused by left vocal fold paralysis. Autologous fat and fascia 0.4 ml were injected respectively into left and right vocal folds of 9 dogs (group B). The dogs of group B were killed after 1,3 and 5 months and the dogs of group A were killed after 8 months. The histology of both substances and the effect of treatment were evaluated. Results Surviving autologous fat grafts was found in the canine vocal fold after 8 months, but the absorption was too rapid to maintain the bulk of grafts, the effect could last no more than 5 months. Autologous fascia also survived after 8 months, Conclusion The results showed that these two augmentative materials were both effective in the treatment of IJVFP. Autologous fat can only be used as short - term augmentative material. However, autologous fascia has a potential use for long - term application to correct glottic insufficiency caused by UVFP and other diseases.

ObjectiveToevaluatelaparoscopiccholecystectomy(LC)versusopen cholecystectomy (OC) in compensated cirrhotics and model for end-stage liver disease (MELD) score and Child-Turcotte-Pugh (CTP)classification in predicting perioperative morbidity.MethodsBetween January 1998 and June 2011,128 cirrhotic patients of symptomatic innocuous gallbladder disease at CPT class A or B liver function were prospectively and randomly divided into LC group (64 patients) and OC group (64 patients ). Data were analyzed by T test, Mann-Whitney U test and Pearson x2 test.ResultsThere was no statistical differences in operation time between the two groups ( t =1.761,P =0.081 ).The intraoperative blood loss ＞ 200 ml occurred in 15 ( 26％ ) LC patients and 35 ( 55％ ) OC patients ( x2 =4.467,P =0.035 ).LC patients had earlier up and about,earlier oral intake,short hospital stay ( t =5.424,t =8.573,t =15.634; P =0.000,respectively) and lower complication rate [ CTP:24％ (14/58) vs 38％ (24/64),x2 =4.582,P =0.032; MELD scores 21％ (12/58) vs 34％ (22/64),x2 =4.238,P =0.040] compared with OC patients.LC patients' VAS- fatigue and VAS-pain scores on first 3 days were lower than OC according to the VAS (visual analogue scale) (Mann-Whitney U =473.0,MannWhitney =6.0,Mann-Whitney =22.5,Mann-Whitney =24.0,Mann-Whitney =46.0; P =0.000,respectively),and the VRS-cosmetic score was higher in LC group than in the OC group according to the VRS ( verbal rating scale) ( Mann-Whitney =145.0,P =0.000).MELD score ＞ 14 predicted higher blood loss and complication rate regardless of LC or OC,while CPT classification did not seem to predict intraoperative bleeding volume and morbidity.ConclusionsLC can be performed safely in cirrhotic patients with CPT class A and B.LC has less,blood loss lower postoperative complication rate,and quicker postoperative recovery.MELD score system is more valuable than CPT classification system in predicting blood loss and postoperative complication rate in cirrhotics undergoing cholecystectomy.

Objective To investigate the effects of different doses of dexmedetomidine on the minimum alveolar concentration (MAC) of sevoflurane for sedation in patients.Methods ASA physical status Ⅰ or Ⅱ patients of both sexes,aged 18-64 yr,undergoing elective lower abdominal surgery performed under general anesthesia,were randomly divided into 4 groups:control group (group C) and different doses of dexmedetomidine groups (D1,D2 and D3 groups).In D1,D2 and D3 groups,the loading dose of dexmedetomidine 0.4,0.6 and 0.8 μg/ kg was intravenously infused over 15 min,respectively,adverse cardiovascular events were then recorded,followed by infusion at 0.4,0.6 and 0.8 μg· kg-1 · h-1 via a pump,respectively,while in group C,the equal volume of 0.9 ％ normal saline was given instead of dexmedetomidine.Sevoflurane administration was begun after completion of infusion of the loading dose.Up-and-down sequential allocation was used to determine the MAC.The initial end-tidal concentration of sevoflurane was set at 0.8％,0.7％,0.6％ and 0.5％ in C,D1,D2 and D3 groups,respectively,and maintained at this level for 15 min.Each time the concentration of sevoflurane increased/decreased in the next patient depending on whether or not the patients correctly followed the verbal command to open his eyes.The ratio between the two consecutive concentrations was 0.9.The middle point between the positive response and negative response served as a crossover pair.After at least 7 independent crossover pairs were observed in each group,the experiment was stopped.The MAC and 95 ％ confidence interval of sevoflurane were calculated.Results The incidence of adverse cardiovascular events was significantly higher in D3 group than in D1 and D2 groups (P ＜ 0.05).In C,D1,D2 and D3 groups,the MAC (95％ confidence interval) of sevoflurane was 0.68％ (0.64％-0.74％),0.50％ (0.47％-0.52％),0.36％ (0.32％-0.41％) and 0.28％(0.26％-0.31％),respectively.The MAC-awake of sevoflurane was significantly lower in D1-3 groups than in group C,in D2 and D3 groups than in group D1,and in D3 group than in group D2 (P ＜ 0.05).Conclusion Dexmedetomidine 0.6μg/kg can significantly decrease the MAC of sevoflurane for sedation,induces no side effects and is the optimum dose in patients.

Objective To evaluate the effects of different doses of dexmedetomidine on the median effective concentration (EC50) of propofol given by target-controlled infusion (TCI) at loss of consciousness (LOC).Methods Eighty ASA Ⅰ or Ⅱ patients of both sexes,aged 18-64 yr,with body mass index ≤25 kg/m2,scheduled for operations under general anesthesia,were randomly allocated to one of four groups(n=20 each): control group (group C) and dexmedetomidine 0.4 μg/kg group (group D1),dexmedetomidine 0.5 μg/kg group (group D2) and dexmedetomidine 0.6 μg/kg group (group D3).Dexmedetomidine 0.4,0.5 and 0.6 μg/kg were infused intravenously over 10 min in groups D1-3,while the equal volume of normal saline was given instead of dexmedetomidine in group C.Propofol was then given by TCI and the EC50 was determined by up-and-down sequential trial.The target plasma concentration was set at 2.0μg/ml in the first patient in each group.The ratio of the target plasma concentration between the two consecutive patients was 1.1.Loss of response to eyelash stimulation and verbal command (2 times) was considered to be signs of LOC.The EC50 and 95％ confidence interval (CI) of propofol causing LOC were calculated.Complications such as bradycardia,hypotension and respiratory depression were recorded.Results The EC50 (95％ CI) of propofol causing LOC was 2.59 (2.51-2.67),2.09 (2.02-2.16),1.82 (1.70-1.95) and 1.60 (1.49-1.72) μg/ml in groups C and D1.3 respectively.The EC50 of propofol causing LOC was significantly lower in groups D1-3 than in group C.Dexmedetomidine significantly decreased the EC50 of propofol required for causing LOC in a dose-dependent manner in groups D1-3 (P ＜ 0.05).The incidences of bradycardia and hypotension were significantly lower in groups D1.3 than in group C (P ＜ 0.05).Compared with group D1,the incidence of bradycardia was increased in groups D2,3 and the incidence of hypotension was increased in group D3 (P ＜ 0.05),There was no significant difference in the incidences of bradycardia and hypotension between groups D2 and D3 (P ＞ 0.05).No patients developed respiratory depression.Conclusion The optimum dose for dexmedetomidine infused intravenously when combined with propofol given by TCI is 0.4 μg/kg and it can decrease the EC50 of propofol administered by TCI at LOC with no adverse reactions.

Objective To investigate the clinical value of serum lipoprotein ( a ) concentration in evaluation of plaques characteristics for patients with coronary atherosclerotic heart diseases ( CAHD ) . Methods Using case-control method, Patients with suspicious CAHD, received coronary computed tomography angiography in the Shanghai East Hospital during October 2013 to June 2015 were enrolled.According to the results of coronary artery CTA, the patients were divided into two groups : the CAHD group (352 cases) and control group(438 cases) , the particle concentrations and mass concentration of lipoprotein(a), triglyceride, total cholesterol, HDL-C, LDL-C, glucose, HBA1c and hs-CRP and other tests were measured, the patients of CAHD group were divided into three subgroups by characteristics of coronary artery plaques including soft plaque (176 cases), calcified plaque (90 cases) and mixed plaque (86 cases), analysis were made with all these data.Using T test or variance analysis to compare the means between or among groups, the risk for CAHD was analyzed by logistic regression, the relationship between LP (a) -P and LP( a) -M were explored by linearly egression analysis, Conformance test were analyzed using kappa test.Results Compared with control group, the mean results of the CAHD group are significantly higher than that of control group, including LP (a) -P 18.5(8.3 -43.0))nmol/L vs.13.6 (7.6-32.4)nmol/L( t =-2.110), LP(a)-M 183(71 -361)mg/L vs.126(67 -293)mg/L(t =-2.063), age (62 ±9)years vs.(52 ±9)years(t=-7.691), hs-CRP 0.86(0.44-1.97) )mg/L vs.0.70(0.38-1.64)mg/L(t=-2.236), glucose (6.1 ±2.29 )mmol/L vs.(5.36 ±1.32 )mmol/L(t=-4.914), BA1c (6.13 ±0.98) % vs.(5.81 ±0.58) %(t=-4.842), APO(B) (1.09 ±0.33) g/L vs.(1.03 ±0.29) g/L( t=-2.407), all of the P values <0.05;The relative risk(RR)of age, glucose, LP( a)-P and LP ( a)-M are 1.067, 2.377, 1.384 and 1.342 respectively; Among the three types of plaques groups,the mean differences of age, TC, HDL-C, LDL-C and LP ( a)-P are statistically significant ( F=6.276,3.060,3.127,4.723,2.878;all of the P<0.05);The median of LP ( a)-P in the soft plaque group 20.3(8.3-48.2)nmol/L is higher than that of the mixed plaque group 15.7(7.3-26.0)nmol/L(P<0.05 ) and calcified plaque group 15.6 ( 8.1 -23.1 ) nmol/L ( P <0.05 ).The linearly regression equation of LP ( a) -M and LP( a)-P is Y=6.646X, r=0.939; Consistency test indicate the two methods are not consistent when used for grouping ( Kappa value is 0.557 ).Conclusions Serum concentration of lipoprotein(a) is an independent risk factor of CAHD, and the particle concentration of LP(a) is closely related to the characteristics of the plaques, especially to the soft plaque.

Objective To discuss the effect of QGY/CDDP hepatoma lines multi-drug resistance (MDR) reversed by gene transfection of mdr1,mrp-ASODN+ultrasound contrast agent+ultrasound.Methods The QGY/CDDP cells were transfected by mdr1,mrp ASODN+ultrasound contrast agent+ultrasound irradiation respectively and detected of the various indicators:cell adhesion rate,cell sensitivity to cell drug-resistance,the mRNA expression of mdr1 and mrp gene,the expression of P-gp and MRP protein. Results After mdr1-ASODN transfection,the drug sensitivity and expression of P-gp,MRP protein of QGY/CDDP cells were smaller changes(P＞0.05),and the rate of cells adherent and expression of resistance gene mRNA were obvious changes(P＜0.05).After mrp-ASODN transfection,the cells adherent rate,the drug sensitivity,the expression of resistance gene mRNA and P-gp,MRP protein were obvious changes respectively(P＜0.05),the experiment group(group 2')had bigger effects(P＜0.05).Conclusions mdr1. mrp-ASODN+ultrasound contrast agent+ultrasound irradiation could safely partly reverse MDR of hepatoma cells,which is a potential new approach for gene therapy.

An important index determination for clinical diagnosis of renal function is to assay the creatinine concentration in serum. In the analytical process applied with coupled-enzyme, the quality control of sarcosine oxidase (SOX) as a key enzyme is the first problem to be solved. In order to establish an efficient and laboratory-scale production of SOX, the recombinant sarcosine oxidase (r-SOX) gene was a high-level expression in E. coli induced with lactose on a large-scale fermentation in 300 L fermenter. The results suggested that the biomass concentration reached OD600 of 22 and the expression of recombinant sarcosine oxidase in E. coli accounted for about 25% of total soluble protein in culture after fermentation. The cell-free extract obtained from high pressure homogenizer was processed by selective thermal denaturation and then purified with Ni-Sepharose FF chromatography. The sarcosine oxidase with 97% purity, 25 U/mg specific activity and 92.4% activity recovery was obtained. The molecular weight with single peptide chain of 53 kD and 55 kD of recombinant sarcosine oxidase was assessed by SDS-PAGE in presence or absence of 2-mercaptoehanol and Sephacryl S-200 chromatography. This sarcosine oxidase was found to be a conjugated protein, yellow enzyme, which combined with FAD as prosthetic group by covalent linkage. The contaminant of catalase was not detected in the sample pool of this enzyme. In addition, a further test to the thermal stability of sarcosine oxidase was done. According to the above results, the development and utilization of this enzyme has been set up on a reliable foundation.
Escherichia coli ; Fermentation ; Recombinant Proteins ; biosynthesis ; Sarcosine Oxidase ; biosynthesis

Objective To explore the serum level of Glucagon like peptide-1 in late-onset Alzheimer′s patients and its clinical significance.Methods Case control study.Collecting cerebral vascular disease fifty-five cases, diagnosed with late-onset Alzheimer′s disease sixty-one cases, type 2 diabetes mellitus fifty-one cases , type 2 diabetic patients combined with late-onset Alzheimer′s disease thirty-seven patients from the Shanghai East Hospital and partly Pudong area elderdly hospital during October 2013 to March 2014, and forty healthy persons as normal control from physical examination center of Shanghai East Hospital during September 2013 to February 2014.Measuring the concentrations of GLP-1,β-amyloid, Tau protein and other routinely used clinical tests in the serum of patients from the normal controls , cerebrovascular disease , late-onset Alzheimer′s disease, type 2 diabetes and type 2 diabetes mellitus combined with late-onset Alzheimer′s disease by ELISA method developed in our laboratory.The blood samples were also collected at three fixed time including fasting time ,1 hour after taking glucose , 2 hour after taking glucose, the concentrations of GLP-1 were determined in the LOAD group , T2DM group and the T2DM combined with LOAD group and normal control group.The concentrations of serum GLP-1 among groups were compared with single factor analysis of variance , and the concentrations of serum GLP-1 between the two groups were compared using LSD-t test.Analysing the correlation between GLP-1 and other indicators with Pearson analysis.Results The fasting GLP-1 levels of LOAD group were ( 123.4 ±20.8 ) nmol/L, and they were highest between the normal control group (78.6 ±6.0) nmol/L and the cerebral blood vessel disease group(89.0 ±8.7)nmol/L (F values were 3.46 and 1.98, P<0.05).The fasting GLP-1 levels of T2DM combined with LOAD group (157.9 ±28.6) nmol/L were higher than the LOAD group (123.4 ± 20.8) nmol/L (t =1.63,P <0.05), but there were no difference of the fasting GLP-1 levels between T2DM combined with LOAD group (157.9 ±28.6) nmol/L and T2DM group(153.8 ±18.0)nmol/L(t=0.96,P>0.05).Deficient secretion of GLP-1 after taking glucose 1 hour in most of the patients of T2DM combined with LOAD group (99.1 ±14.2) nmol/L, LOAD group(73.9 ±6.6 ) nmol/L and T2DM group (96.3 ±7.0 ) nmol/L could be concluded .The GLP-1 levels of T2DM combined with LOAD group after taking sugar 2 hour were (115.4 ±18.6)nmol/L ,and were higher than that of normal levels (63.3 ±6.2) nmol/L after taking sugar 2 hour(t=4.49,P<0.05).There were no difference between the GLP-1 levels of the LOAD group (73.6 ±5.8 )nmol/L and the GLP-1 levels of the normal group(63.3 ±6.2)nmol/L after taking sugar 2 hour (t=0.94,P>0.05).Pearson correlation analysis showed that the relationship of the levels of GLP-1 with Aβ( 1-42 ) and the levels of GLP-1 after taking glucose 1 h and 2 h were positively relative, and its coefficients of correlation were 0.401,0.436,0.722.Conclusions LOAD and T2MD are similar, and they have GLP-1 secretion shortage phenomenon after taking glucose , so monitoring dynamic change of GLP-1 after taking glucose may contribute to the auxiliary diagnosis of LOAD.

Objective To explore the operation points of the steps offront-typehypospadias operation in oral mucosa urethroplasty and scrubbed shaped urethra with meat membrane covering.Methods After correction of chordee of penis of 40 patients with Front -type hypospadias, oral mucosa was transplanted and fixed on albuginea surface at the one-third of ventral penile for all the patients to increase the width of the urethra and form the urethra with the selected appropriate size ureter. The skin of dorsal penile was transferred to ventral penile. After clearing the pedicled skin flap, the subcutaneous layer of meat was kept down, and stamped wholly on forming place of urethral reel (including both sides inferior of cut-off cavernous body of glans penis),forming the glans again.Results There was no ankylo-urethria among the 40 front-type hypospadias operation, ureteroscopy examination after two months of the operation showed that all the transplanted oral mucosa survived, and the stamped subcutaneous layer of meat located at both sides inferior of cut-off cavernous body of glans penis adhered with satisfaction,no glans incision dehiscence,there occurred 2 cases of urinary fistula which had been cured by neoplasty,there was 1 case of transferred flap necrosis which had been cured after dressing change.40 patients were satisfied with penis appearance after operation.Conclusion Following up the operation points of “front-type”hyospadias operation,the success rate of operation can be improverd obviously,the plastic effect is good,and the complications after operation can be reduced.stamped wholly on formed urethra.There is a small probability of incidence of urethral stenosis and urinary fistula after operation.

Objective To investigate the clinical feature , pathologic characteristics and prognosis of urothelial carcinoma of bladder with squamous differentiation . Methods From Jan.2010 to Jun.2013, the pathological and clinical data of 96 cases of urothelial carcinoma of bladder with or without squamous dif-ferentiation were compared .Of the group with squamous differentiation , there were 39 males and 9 females with a median age of 70 (29 to 87) years.44 cases presented with painless gross hematuria .4 cases presen-ted with finding of bladder tumors in annual physical examination .TURBT, partial cystectomy and radical cystectomy were performed in 25, 8 and 13 cases, respectively.In addition, one case was only underwent bi-lateral ureteral skin gastrostomy .The last one only performed cystoscopy .In accordance with sex , age, path-ological stage and classification and surgical approach , 48 controls were selected .For the other group , there were 40 males and 8 females with a median age of 68 (39 to 86) years.45 cases presented with painless gross hematuria.3 cases presented with finding bladder tumors by annual physical examination .TURBT, par-tial cystectomy and radical cystectomy were performed in 28, 7 and 13 cases, respectively.All patients with retaining bladder had postoperative intravesical instillation for one year .Some patients with or without bladder performed 3-6 cycles chemotherapy with the GC protocol . Results In squamous differentiation group , there were 1 (2.1%) pTa, 25 (52.1%) pT1, 17 (35.4%) pT2, 4 (8.3%) pT3 and 1 (2.1%) pT4 tumors. Whereas, 1 (2.1%) pTa, 28 (58.3%) pT1, 16 (33.3%) pT2, 2 (4.2%) pT3 and 1 (2.1%)pT4 tumors were selected in the control group .There were 2 (4.2%) cases with low grade and 46 (95.8%) cases with high grade carcinomain in both groups .Patients were followed up with a mean duration of 16 and 12 months in squamous differentiation and control group , respectively .In squamous differentiation group , eight recur-rences were recorded with a mean follow-up of 12 months.Of the 3 died patients, only one died from bladder cancer.In control group, seven recurrences were recorded with a mean follow-up of 22 months, and no pa-tient died.For patients with TURBT, 3 year recurrence rate of patients with squamous differentiation was 49.5%, while the control was 34.8%.The difference was statistically significant (P<0.05). Conclusions Urothelial carcinoma of bladder with squamous differentiation is at a high level of malignant and recurrence . The rate of myometrial invasion with squamous differentiation is higher than pure urothelial bladder cancer . Patients with squamous differentiation should be closely followed up .