Objective To construct the suppression subtracted cDNA library of deltamethrin-resistant Aedes albopictus. Methods Total RNA was extracted from the deltamethrin-resistant (R-lab) and -sensitive (S-lab) isolates, mRNA was obtained after purification. Double stranded cDNAs were synthesized after reverse transcription. Two subtractions were performed by suppression subtractive hybridization with S-lab as tester and R-lab as driver or S-lab as driver and R-lab as tester. Enriched different expressed cDNA was cloned into pMD18-T vector to construct subtractive libraries. Results The subtracted cDNA libraries contained 580 and 477 positive clones respectively. The PCR results of 150 clones picked randomly from each library showed that the positive ratio of constructed cDNA libraries was 93%, with a length of cDNA fragments ranged from 150 bp to 750 bp. Conclusion The suppression subtracted cDNA library of deltamethrin-resistant Ae. albopictus is constructed.

Objective　To study the effect of portal vein blocking on the permeability of the intestinal mucosa in pigs. Methods　Healthy Rongchang pigs were divided into 3 groups: ① sham operation group(SO), ② portal vein clamping for 45 min group (PVC-45'), ③ portal vein clamping for 60 min group (PVC-60'). Urine lactulose/mannitol(L/M) ratio was measured after portal vein blocking. Results　The L/M ratio was increased significantly (P<0.05) in PVC-45' and 60' groups than in SO group, with that of PVC-60' higher than that of PVC-45' group, but not significantly. Conclusion　The increase of intestinal mucosal permeability after portal vein blocking is an early and important index for the damage of the intestinal mucosa barrier.

Myasthenia gravis (MG) is an immune-mediated disorder of the neuromuscular junction that is characterized by fluctuating muscle weakness.Acetylcholine receptor antibody (AchR-Ab) is the first MG specific antibody being elucidated,however,the discovery of many other MG specific or related antibodies enriches its immunopathogenesis greatly.The detection of antibodies is very important for the diagnosis and classification of MG.It can be used to guide the clinical management and evaluate the response to treatment as well.With the development of laboratory medicine,the methods of antibody detection are also being optimized.In this paper,we will review the serum antibodies closely associated with MG and their mechanisms,detection methods and clinical significance.

Objective To study microsatellite instability(MSI) in multiple primary colorectal carcinoma(MPCC) and solitary colorectal tumor(SCT), and to explore the relationship between the expression of mismatch repair(MMR)、p53、Bax、PCNA and MSI. Methods The expression of MMR、p53、Bax、 PCNAwere detected by immunohistochemical staining, and MSI at five microsatellite loci were examined by PCR-SSLP in 51 tumors from 38 MPCC patients and 35 SCT cases. Results The replication errors positive phenotype was observed in 27 of 51(53%) tumor foci from MPCC cases, and in 6 of 35(17%) SCT cases. There was an inverse correlation between replication errors (RER) positive and expression of p53; the PCNA labeling index of RER positive tumors were significantly lower than of RER negative tumors; RER positive related strongly with poor differentiation, the proclivity for proximal colon. Conclusions MSI may play an important role in the development of MPCC and may be used as a tumor marker of MPCC.

Ex-vivo liver resection and liver autotransplantation offers a chance to cure extensive regional liver diseases which are otherwise deemed untreatable in selected cases. A 33-year-old male patient with advanced alveolar echinococcosis (AE) which invaded the right lobes and adjacent retrohepatic inferior vena cava, while the left lobes were free from lesion and had proliferated to over 1200 ml in size except for a 1 cm × 1 cm solid lesion in the segment Ⅳ. No extrahepatic metastases were found in this patient. A lesion with a size of 4300 ml and other small lesions in the right lobes were removed extracorporeally,and the retrohepatic vena cava was repaired, then the remaining AE-free left lobes (more than 700 ml in size) and the retrohepatic inferior vena cava were re-implanted in situ. A temporary end-to-end cavo-caval shunt with interposition of a blood vessel prosthesis and end-to-side portocaval shunt were established to keep the blood flow during the four-hour anhepatic phase. The patient was followed up for six months, and he had no signs of residual liver disease with good hepatic function.

Objective To assess the feasibility,safety and advantages of robotic-assisted precise hepatectomy.Methods Between April and July 2009,13 consecutive patients underwent robotic-assisted hepatectomy for hepatic diseases.The clinical data were analyzed retrospectively.Results All 13 Da Vinci surgical systemassisted precise hepatectomies were successfully performed without conversion to laparotomy.Major hepatectomies were performed in 9 patients,left lateral segmentectomies in 4 patients.All the Da Vinci surgical system-assisted hepatectomies were performed anatomically with hilum dissection.Prior to the parenchymal transaction,vascular control of the portal vessels was carried out whenever possible.The mean operative time was 338 minutes(range,150-720 minutes).The mean blood loss was 208 ml(range,50-800 ml).No patient required blood transfusion,and no mortality,transient bile leakage was observed in patients with hilar cholangiocarcinoma.The mean postoperative stay was 7 days(range,2-13 days).Conclusions These preliminary results show that Da Vinci surgical system-assisted precise hepatectomy is safe and feasible with potential benefits of a minimally invasive approach.Da Vinci surgical system may broaden the indications for laparoscopic hepatactomy,and it enables surgeons to perform precise laparoscopic hepatectomy which required hilum dissection,hepatocaval dissection,endoscopic suturing and microanastamosis.

Objective To evaluate the effect of Da Vinci surgical system for the treatment of hepatopancreatobiliary diseases.Methods The clinical data of 29 patients with hepatopancreatobiliary diseases who had undergone operations with Da Vinci surgical system from March to November 2009 at the General Hospital of PLA were retrospectively analyzed.Results The operations were successfully done on 28 patients,except 1 patient was converted to open pancreaticoenterostomy.The total operation time was(339±149)minutes,and the time for operations done with Da Vinci surgical system was(285±117)minutes.The postoperative bowl movement recovery time was(33±21)hours,and the length of postoperative hospital stay was(8±6)days.No blood transfusion was needed.Three patients had postoperative complications and were cured by conservative treatment.Conclusion Laparoscopic operations for hepatopancreatobiliary diseases can be applied with the help of the threedimensional imaging system and flexible surgical instruments of the Da Vinci surgical system,and its superiority is more obvious when applied for intractable hepatopancreatobiliary diseases.

Objective To evaluate Western blot analysis in diagnosing limb-girdle muscular dystrophy type 2A (LGMD2A). Methods The clinical records including their pathological and biochemical results of 4 patients with LGMD type 2 were reviewed. Histochemical and immunohistochemical staining were performed on muscle biopsy specimens from the four patients. The expressions of dysferlin and calpain-3 in muscles were analyzed by Western biol. Results All 4 LGMD patients shared some common clinical features, such as dorsal muscular atrophy of lower limbs and remarkably elevated CK. The immunohistochemical results showed partial or complete deficiency of dysferlin staining in all 4 LGMD patients. However, Western blot revealed that the calpain-3 protein in the muscle of patient 1 was completely absent, who was later diagnosed with LGMD2A. The other 3 patients had complete dysferlin deficiency with reduced calpain-3 expression and they were confirmed to be LGMD2B. Conclusions Western blot analysis of calpain-3 and dysfcrlin can be used to differentiate LGMD2A which shows absence of calpain-3 from other LGMD types which show dysferlin deficiency. Western blot is an invaluable method in clinical diagnosis of LGMD2A.

Objective To investigate the removal effect of immunoadsorption (IA) on associated antibodies and the efficacy in late-onset myasthenia gravis (MG). Methods A total of 25 late-onset MG patients were randomly selected to enroll in this study. IA therapy was given to 10 patients (IA group), while immunoglobin (0.4 g·kg-1·d-1) was administrated to the other 15 patients for 5 days(Ig group). The titers of Titin antibody (Titin-ab), acetylcholine receptor antibody (AchR-ab) and presynaptic membrane antibody (PrsmR-ab) were detected before and after the treatment. Quantitive MG (QMG) score was assessed before and immediately after the entire course of treatment. The clinical efficacy, the duration of respiratory support and in-hospital were compared between two groups. The correlation between three antibodies and QMG score was also analyzed. Results Compared with that before treatment, the Titin-ab PIN values, the AchR-ab PIN values, and the PrsmR-ab P/N values of IA group were all decreased significantly after treatment (P<0.05, respectively). The P/N value of Titin-ab in IA group was decreased by 54.7%～3.5%, which was significantly higher than that in Ig group(19.9%±3.1%) (P<0.01). QMG score reduced by 42.4%± 4.2% and 23.8%±3.7% in IA group and Ig group respectively (P<0.01, respectively). Symptoms were effectively ameliorated by both treatments, but the effective power of IA group was higher than that of Ig group (70% vs 40%, P<0.05). Remission time of IA group was significantly shorter than that of Ig group [(5.38±0.42) d vs (8.4±1.54) d, P=0.008), so was the duration of in-hospital [(13.50±0.50) d vs (16.50±0.50) d, P<0.05). The number of respiratory support in IA group was less than that in Ig group (1/10 vs 6/15, P<0.05). By the Pearson correlation analysis, the decrease of Titin-ab showed a better longitudinal correlation with the decrease of QMG score than the other two antibodies (r=0.6315, P<0.01). Conclusion IA can rapidly and effectively clear the pathogenic antibodies of late-onset MG patients and its short-term clinical efficacy is better than immunoglobin.

By means of joint research of new advanced technology project,joint development and application of appropriate health technology project,comprehensive prevention and control of chronic diseases between municipal hospitals and community clinics project and research supporting platform project,Shanghai Hospital Development Center has effectively improved the clinical science and technology innovation capability, promoted the development of medical disciplines and talents and improved the discipline influence of such hospitals.