[Cloning, expression and immunogenicity analysis of cathepsin L-like protease of Fasciola hepatica].

OBJECTIVE: To clone and express the cathepsin L-like protease gene of Fasciola hepatica (FhCL) and investigate the immunogenicity of the recombinant FhCL protein. METHODS: Specific primers were designed according to the reported FhCL gene in GenBank. Using total RNA from adult worms of F. hepatica, FhCL gene was amplified by RT-PCR. The PCR product was cloned into pMD18-T vector and then subcloned into pET30a(+) vector. The recombinant plasmid was transformed into E. coli BL21 (DE3) and followed by expression of the protein induced by IPTG. The expression situation of recombinant FhCL was analyzed by SDS-PAGE. Its immunoresponse to the sera of infected goat and the antisera of SD rats against FhCL was examined by Western blotting analysis. RESULTS: PCR and double enzyme digestion showed that the FhCL gene fragment was about 1,000 bp in length. The constructed recombinant plasmid pET30a (+)-FhCL was identified by sequencing. The recombinant protein (Mr 42,000) was expressed in the form of inclusion body. The protein was recognized respectively by the sera of infected goat and the sera from rat immunized with FhCL. CONCLUSION: The recombinant plasmid pET30a(+)-FhCL has been constructed, which shows high antigenicity.

Modified laparoscopic cholecystectomy technique for treatment of situs inversus totalis: A case report.

Laparoscopic cholecystectomy is a broadly used technique for gallbladder treatment. However, situs inversus, a rare anomaly, is reportedly difficult to treat by conventional laparoscopic cholecystectomy. A 36-year-old woman with chronic cholecystitis and multiple gallstones was found to have dextrocardia on a chest X-ray. Magnetic resonance imaging demonstrated situs inversus, cholecystitis, and cholelithiasis. We successfully performed laparoscopic cholecystectomy using our modified technique, which mainly involved a left-handed operation and adjustment of the port positions. This case will be very instructive for right-handed surgeons in the management of cholelithiasis by laparoscopic cholecystectomy in patients with situs inversus.

[Combined medial and lateral malleolus operative treatment for severe fracture of ankle].

OBJECTIVE: To explore the clinical effects of combined medial and lateral malleolus in treating severe fracture of ankle. METHODS: From May 2007 to January 2011,89 patients with ankle fracture who underment surgical treatment and had complete follow-up, the data were retrospectively analyzed. In this study, there were 66 males and 23 females with an average age of 30.9 years (ranged, 26 to 62 years). Cause of injury included road accidents in 52 cases,sprains from fall down in 17, construction injury in 16 and other injuries in 4. According to type of Denis-Weber, type A were in 13 cases, type B in 24, type C in 52. Operative technique was as follows. Chosen two-incision of combined medial and lateral malleolus, and reduced lateral malleolus fracture firstly, fixed it with 1/3 tubular armor plate or rebuilt-titanium plate. To chose one or two hollow screws according to the size of fracture block. If fracture of fibula was syntripsis,it can be multi-segmental fixed. If fracture line of posterior malleolus was no more than 1/3 articular surface of ankle or the bone displacement was not serious, it can be disregarded. And if the lesion level was over 1/3, or shift position more than 3 mm,it can be fixed with lag screw after reduction. Either front-to-back or back-to-front fixed direction was OK. At the third day after operation, the patients were instructed to exercise with affected limb, weight training after 6 weeks. RESULTS: Eighty-nine patients were followed up from 6 to 24 months with an average of 16 months. Healing time of the fracture was 10 to 12 months. According to standard of the Baird-Jackson scoring system, 68 cases got excellent results, 18 good and 3 fair. CONCLUSION: Surgical treatment is quite effective for severe fracture of ankle. It should be chosen correctly according to types of the fracture and degree of ligament injury. At the same time, damage degree of the ligament around the ankle should be considered and properly treated.