Dopa-responsive dystonia(DRD) is a clinical syndrome characterized by childhood dystonia and dramatic and sustained response to low-dose levodopa.The deficiency of any link in dopamine synthesis pathway can lead to DRD that is caused by TH gene mutation and is autosomal recessive, which results in the reduction of tyrosine hydroxylase(TH) synthesis and extensive deficiency of dopamine and catecholamine.However, DRD presents atypical clinical manifestations.Mild patients mainly present with dystonia.Some of them might progressively develop into spastic paraplegia and some may have parkinsonian features.Most of them display good response to levodopa.Severe patients present with progressively complex infantile encephalopathy, and badly response to levodopa and remnant intellectual development problems.Most of them manifest with declining homovanillic acid(HVA) in cerebrospinal fluid.However, due to the noncharacteristic clinical course and nonspecific laboratory tests of TH DRD, gene detection still is the only reliable criterion of diagnosis so far.Low-dose levodopa is effective to most mild patients and can improve symptoms to severe patients to some extent.

Objective To investigate the feasibility of laparoscopic radical gastrectomy for gastric cancer.Methods Laparoscopic radical gastrectomy was performed in 8 cases from Febrary 2005 to April 2005,including radical distal gastrectomy in 6 cases,radical total gastrectomy in 1 case,and radical proximal gastrectomy in 1 case.Results All the 8 patients underwent laparoscopic radical gastrectomy smoothly and no conversion to open surgery was required.The operation time was 340?62 minutes in radical distal gastrectomy,was 362 minutes in radical proximal gastrectomy,and 423 minutes in radical total gastrectomy.The intraoperative blood loss was 100～250 ml(mean,140 ml) in radical distal gastrectomy,300 ml in radical total gastrectomy,and 170 ml in radical proximal gastrectomy.No blood transfusion was needed.The number of havested lymph nodes was 18～37(mean,23).No surgical-related complications occurred.The time to first flatus was 38～56 hours(mean,42.4 hours),and the time to liquid diet was 2～5 days(mean,2.5 days).All the 8 cases were followed for 12～14 months postoperatively and no recurrence or metastasis was observed.Conclusions Laparoscopic radical gastrectomy is a feasible technique for patients with gastric cancer at early stage or early progressive stage.

Objective To compare the differences in mucin-histochemistry and etiology of Barrett's esophagus (BE) intestinal metaplasia (IM) , cardiac intestinal metaplasia (CIM) and gastric antrum intestinal metaplasia (GA-IM). Methods Alcian blue /periodic acid-schiff (AB/PAS) and high iron diamine / alcian blue (HID/AB) histochemical methods were used to classify IM in BE, cardia and gastric antrum, and IM were classified into three subtypes: complete small intestinal type (type Ⅰ ) , incomplete small intestinal type (type Ⅱ) and incomplete colonic type (type Ⅲ). Compared the prevalence of different subtypes of IM in above-mentioned sites, and investigated their relationships among the symptoms of gastroesophageal reflux disease (GERD) and Helicobacter pylori (Hp) infection. Results The prevalence of type Ⅲ IM in long-segment BE (LSBE) and short-segment BE (SSBE) is 75. 0% and 63. 3% respectively, it is significantly higher than that in CIM (23. 1% ) and GA-IM ( 17. 7% ) (P

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Objective To express and purify mammalian glycosylation modified trimeric Ebola virus trimeric glycoprotein (EBOV-GP) using novel glycoengineered Pichia pastoris.Methods The EBOV-GP and EBOV-GPΔMLDΔTM genes were cloned into the pPICZ-αA vector,electrochemically converted to glycoengineered Pichia pastoris,and compared with EBOV-GP expressed in HEK-293T cells.Glycosylation was analyzed by PNGaseF and EndoH digestion,while the target protein was purified by affinity chromatography and ion exchange chromatography.N-terminal sequencing was used to determine whether the signal peptide was correctly cleaved during protein translation and gel column analysis was used to find out whether the trimeric structure was formed.Results The results of PNGaseF showed that EBOV-GP expressed by glycoengineered Pichia pastoris and HEK-293T cells had the same relative molecular mass and N-glycosylation degree.EndoH digestion showed that the N-glycosylation modification of EBOV-GPΔMLDΔTM was in a non-high mannose form.N-terminal sequencing showed that the signal peptide of the GP protein itself was correctly excised.Gel column analysis showed that the purified protein was in a trimeric form.Conclusion An EBOV-GP is obtained with complex glycosylation modification based on Glycoengineered Pichia pastoris.

0.05). CONCLUSIONS Levofloxacin is suited to prevent and treat the infection during the perioperation of appendectomy due to acute appendicitis.

OBJECTIVE： To study the pharmacokinetics of domestic carvedilol and relative bioavailability of carvedilol capsule in Chinese volunteer.METHODS： Eight volunteers orally took a single dose of 30mg test preparation and 25mg control preparation in a random crossover and self-control method.Samples were determined by RP-HPLC fluorescent method.RESULTS： Profiles of carvedilol in vivo could be described as open two-compartment model.The main pharmacokinetics parameters of test and control preparations were as follows： Cmax（98.89± 27.60） ng/ml、 （70.06± 27.29） ng/ml， Tmax（0.4 849± 0.2 635） h、 （0.6 037± 0.1 707） h， CL（0.1 621± 0.08 057） （mg· h） /（ng· ml） 、 （0.1 796± 0.09 198） （mg· h） /（ng· ml） ， V/F(c)（0.2 127± 0.1 260） mg/(ng· ml)、 （0.2 777± 0.1 860） mg/（ng· ml） ， T1/2β （2.011± 1.709） h、 （1.959± 1.156） h， AUC（233.1± 97.12） ng/（ml· h） 、 （168.0± 70.61） ng/（ml· h） ； Mean relative bioavailability in man was (111.3± 15.18)％ .CONCLUSION： The results can be used for design of therapeutic scheme.

ObjectiveTo investigate the changes in liver function and the efficacy of either hand-assisted laparoscopic surgery (HALS) or open splenectomy (OS) in combination with pericardial devascularization in the treatment of portal hypertension. MethodsThe clinical data of 94 patients who received splenectomy combined with pericardial devascularization to treat portal hypertension due to cirrhosis from Jan 2002 to May 2008 were analyzed retrospectively. 56 patients received OS and 38patients HALS. The operating time, intraoperative blood loss, postoperative complications, liver dysfunction and mortality were analyzed according to the Child's grading. ResultsThere was no difference in the operating time between HALS and OS (P＞0. 05). The intraoperative blood loss and postoperative complications were 5.6% and 10.8%, respectively (P＜0. 05). There was no significant difference in the serum ALT between HALS and OS, but there was a significant difference in the ALB (P＜0. 05). The AST also had a significant difference on postoperative day 5 (P＜0. 05). The serum ALT and AST were elevated after HALS, but there was a significant difference only for AST (P＜0.05). The serum ALT and AST in OS were significantly higher after than before operation (P＜0. 05). The serum ALB in OS was significantly lower after operation (P＜0.05), but it was significantly lower only on postoperative days 1 and 3 (P＜0.05) in HALS. ConclusionsCompared with OS, HALS combined with pericardial devascularization caused less damage to the intestinal tract and the liver function. It is a feasible and safe operation and it had fewer postoperative complications.

Objective To prepare recombinant human prothrombin-2 expressed in Pichia pastoris, and assay the enzymatic and clotting activities of prothrombin-2 activated by prothrombin activator ecarin.Methods Human prothrombin-2 gene and Echis carinatus ecarin gene were synthesized separately on the basis of the cDNA sequences published in GenBank.The gene of prothrombin-2 was cloned into the expression vector pPICZαA.The expression vector pPICZαA/prothrombin-2 was transformed into glycoengineered P.pastoris, and then prothrombin-2 engineered P.pastoris was screened.The expression products were induced by methanol, purified by two-step chromatography and identified by diges-tion by PNGase F and analysis of pepetide fingerprint.The ecarin gene was cloned into the expression vector pcDNA3.1. The expression vector pcDNA3.1/Ecarin was transformed into HEK 293T cells and the culture supernatant of HEK 293T/Ecarin was collected.The reaction product of HEK 293T/Ecarin cell culture supernatant and purified prothrombin-2 was analyzed by S-2238,which was the chromogenic substrate for thrombin.Fibrinogen was used to measure blood clotting time. Results The purified protein of P.pastoris expressed prothrombin-2 culture supernatant was 37 ×103 .The relative molecular mass(Mr) of the purified protein was reduced to 35 ×103, which was consistent with the theoretical Mr of prothrombin-2 molecular weight.The purified protein was proved to be prothrombin-2 by peptide fingerprint identification. The purified prothrombin-2 processed by HEK 293T/Ecarin culture supernatant could hydrolyze S-2238 to produce yellow pNA, and D405 of pNA increased with the volume of the processed prothrombin-2 that could promote the plasma coagulation.The blood clotting time was close to that of the thrombin kit.Conclusion Prothrombin-2 is prepared by P.pastoris and activated toα-thrombin by ecarin.This technique may replace the method of extraction of prothrombin from plasma and can be used for the treatment of war wounds or for future clinical research.

Objective To explore the clinical value of autologous fat injection with platelet rich plasma in facial rejuvenation.Methods 35 female beauty seekers were collected in this study.They were divided into two groups:sample autologous fat group (FAT group,n=20) and combined autologous fat and platelet-rich plasma group (PRP group,n =15).The average level of injection was 34 ml.The fat was collected at the abdominal or thigh sites using 18 G needle connecting with negative pressure suction-pump.The fat was centrifugated 2 min to get rid of the lower layer of water and the upper layer of oil.Weigao PRP extract kit was used to obtain the PRP.The ratio of PRP to fat was 1:5.The injection technique was according to Colemen method.The survival rate,satisfaction and complication were compared between the 2 groups.Results There were no infection,hematoma,fat liquefaction necrosis occurred in the 2 groups.Compared with the FAT group,the skin texture and facial contour of the PRP group was improved.Conclusions The platelet rich plasma can improve the survival rate of fat and has a stable effect,which is one of the safe and ideal methods of facial rejuvenation.