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.

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.

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 examine the role of c-Src activation in hepatitis B virus X (HBx) protein induced epithelial-mesenchymal transition (EMT) in liver cancer.Methods SMMC-7721 liver cancer cells were transfected with HBx gene to induce EMT and the activated c-Src expression was evaluated by Western blot.Both the morphological changes and the epithelial and mesenchymal markers expression (real-time PCR,western blot and immunocytochemistry) of HBx-transfected SMMC-7721 cell treated by c-Src kinase inhibitor PP2 and negative control PP3 were observed and compared,respectively.Results The activated c-Src expression in HBx gene transfected SMMC-7721 cells was significantly increased compared to that in mock transfected cells,c-Src kinase inhibitor PP2 could enable the HBx-transfected SMMC-7721 cells to transmit from spindle-like shape to original epithelial morphology.Western blot and immunocytochemistry confirmed that the expression of epithelial markers and mesenchymal markers almost returned to the levels of parental cells,indicating the mesenchymal-epithelial transition.Conclusions c-Src activation plays a key role in the process of EMT induced by HBx protein in SMMC-7721 cells.

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 explore the feasibility of electromagnetic navigation for hepatic ducts exploration by choledochoscope based on a three-dimensional (3D) printing model.Methods The retrospective descriptive study was conducted.The clinical data of 1 patient with obstructive jaundice combined with secondary biliary tract dilation who was admitted to the Beijing Tsinghua Changgung Hospital in April 2016 were collected.Digital imaging and standardized format data of enhanced computed tomography (CT) scan were collected,3D reconstruction of liver and bile duct were done under a 3D printing model,and then a hollow model of bile duct was achieved.Choledochoscope with internal electromagnetic probe was inserted into the printed hepatic ducts.Four points of anatomical markers in left and right hepatic ducts and common hepatic duct were chosen as fiducial markers for calibration and registration.Results After registration,the scope can be tracked in main hepatic segmental bile ducts.The locations of choledochoscope matched precisely the navigation results.Conclusion Electromagnetic navigation may bring accurate tracking effectiveness for choledochoscopic examination.