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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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 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.

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 develop an interleukin-4(IL-4) antagonist named M5-IgG1Fc protein constructed by genetic engineering of antibody Fc fragment-cytokine mutein fusion protein which has a long half-life time in plasma.M5-IgG1 Fc protein binds to IL-4 receptor but cannot activate downstream signalling pathway , which provides a basis for drug develop-ment for allergic diseases .Methods The synthesized interleukin-4 mutant gene ( named M5 ) was cloned into the expres-sion vector pBV220 and transformed into E.coli DH5α.Chimeric gene M5-IgG1Fc obtained by overlap extension (SOE) method was transformed into glycoengineered Pichia pastoris GJK01 through expression vector pPICZαA .Then M5-IgGFc fusion protein was obtained by protein purification after being induced by methanol in 72 hours.The anti-IL-4 biologicial ac-tivity assay of M5 and M5-IgG1 Fc was performed with CTLL-2/IL-4R cells and detected with MTT colormetry .Finally,the half-life time of M5 and M5-IgG1 Fc protein in mice was compared by detecting the remaining amount in plasma with ELISA kit.Results The M5 protein expressed in E.coli and M5-IgG1 Fc fusion protein expressed in P.pastoris GJK01 both had IL-4 antagonistic bioactivity .The EC50 of both, which inhibited 5.6 ×10 -2 nmol/ml of IL-4, were 0.31 ±0.05 and 0.77 ± 0.03 nmol/ml,respectively.The maximum of M5 in plasma at 0.5 h was 5.8 ×10 -2 nmol/ml but the remaining amount was 2.8%of the maximum at 2 h.M5 protein could not be detected after administration at 8 h because of the detection line . The maximum of M5-IgG1 Fc fusion protein was 4.7 ×10 -2 nmol/ml,while fusion protein M5-IgG1 Fc decreased to 4.3%of its maximum at 120 h and could not be detected at 168 h.Conclusion M5 protein has IL-4 antagonistic bioactivity .M5-IgG1 Fc fusion protein expressed in glycoengineered P.pastoris GJK01 has IL-4 antagonistic bioactivity and long retention time in mice,which can be potentially used for treatment of allergic diseases .

Background and purpose: Cyclin-dependent kinase 4 (CDK4) is a kind of protein kinases regulating the cell cycle progression, which has been reported to be overexpressed in endometrial carcinoma tissues. But the role of CDK4 in endometrial carcinogenesis and relative mechanisms has not been identiifed yet. In this study, we used a small interfering RNA targeting CDK4, and explored the effects of CDK4 on endometrial cancer cells HEC-1B biological function and relative mechanisms.Methods:The chemically synthesized small interfering RNA targeting CDK4 (si-CDK4) was transiently transfected into HEC-1B cells;the quantitative real time-PCR assays and Western blot assays were performed to explore the mRNA and protein expression levels of CDK4 and its downstream genes, Rb and p-Rb, in HEC-1B cells upon transfection;Moreover, the CCK-8, lfow cytometry (FCM) and invasion assays were performed to indentify the effects of si-CDK4 on the proliferation, cell cycle distribution, apoptosis and invasion abilities of HEC-1B cells, respectively. Results:The results showed that the mRNA and protein expressions of CDK4 were suppressed in HEC-1B cells upon transfection with si-CDK4 (P<0.01);Suppression of CDK4 inhibited cell proliferation and invasion of HEC-1B cells;the number of cells migrating through the transwell membrane in si-CDK4 group was 117±21, which was much fewer than the cells in si-control (269±39) and untreated groups (262±35) (P<0.01);the early apoptosis rate of cells treated with si-CDK4 [(21.7±3.5)%] was much higher than the untreated [(12.4±2.1)%] and si-control groups [(11.8±1.9)%] (P<0.01);moreover, suppression of CDK4 increased cells in G1 phase (P<0.01) and correspondingly decreased cells in S phase (P<0.01);further Western blot results showed that suppression of CDK4 down-regulated the expression of p-Rb in cells, but did not inlfuence the expression of total Rb. Conclusion:CDK4-siRNA speciifcally and efifciently blocks the constitutively activated CDK4 in human endometrial cancer cells HEC-1B, resulting in tumor suppression.

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.