Objective To explore the feasibility of recombinant adeno-associated virus (rAAV) as a vector for the gene therapy of liver cancer. Methods The rAAV/enhance green fluorescein protein (EGFP) recombinant was prepared by the routine method of two plasmids cotransfection.Results The experiment showed that one 10cm plate could produce 107-108 infection unit recombinant by the method of two plasmids cotransfection, and the transduction of HepG2 cell was increased with the increase of infection dosage of rAAV. About 100 multiplicity of infection (MOI) AAV vector could make all the tumor cell light. Conclusion Liver cancer cell can be efficiently transduced by rAAV, and AAV vector may be a valuable vector for the gene therapy of liver cancer.
目的:探讨经脐入路行腹腔镜胆囊切除术的可行性。方法:对6例患者采用仅在脐部切开一个切口进行腹腔镜胆囊切除术。结果:6例患者手术均获成功,无中转常规腹腔镜手术或开腹手术。手术时间80～130min,无出血、胆管损伤等并发症发生。术后1d出院,术后1月门诊随访,患者恢复顺利,除脐部外,腹壁无手术瘢痕。结论:经脐入路腹腔镜胆囊切除术技术上是可行的,但难度较大,在开展手术初期应慎重选择病例。
Objective To investigate the feasibility of laparoscopic cholecystectomy through the transumbilical approach. MethodsThe clinical data of 18 patients underwent endoscopic cholecystectomy through only one transumbilical incision at West China Hospital were retrospectively analyzed. Results All of the operations were successfully completed without conversion to routine laparoscopic surgery or open surgery. The operation time was 40-130 （58±10） min. There was no intraoperative complication. The patients did well postoperatively and were discharged 1 day after operation. There was no postoperative complications and without visible abdominal scar on 1 month follow-up. Conclusions Laparoscopic cholecystectomy through the transumbilical approach is technically feasible and safe. But this technique is difficult, the patients should be selected carefully.
Objective To evaluate the suitability of the biodegradable microsphere encapsulation of adenovirus as a targeting vector for gene therapy of hepatocellular carcinoma. Methods Encapsulate the recombinant adenovirus in PLG 〔poly (lactic/glycolic)〕 copolymer by the solution evaporation method, the release test and the bioactivity of viruses incorporated in vitro were studied. Results More than 19.3% of adenovirus was encapsulated in PLG microspheres. The release test shows that the adenovirus was released for more than 200 h, 50% were shed within the first 100 h, and their activity was retained. Conclusion Recombinant adenovirus can be formulated in a polymer preparation of PLG with retention of bioactivity. It may be a valuable vector for the gene therapy of liver cancer.
Objective To investigate the reversal of the multidrug resistant gene mdr1 in vivo by antisense oligodeoxynucleotide （ASODN） on the basis of study in vitro. Methods The cultured drug-resistant human hepatocellular carcinoma cells were injected under the skin of axilla to establish the tumor model of nude mice. mdr1 ASODN accompanied by Lipofectamine were injected locally and ADM was injected intraperitoneally. Control 1 and control 2 were locally injected by Lipofectamine and normal saline separately, and ADM was also injected intraperitoneally. Results As time went on the tumor size increased and from the 5th day on alterations were marked, tumor size in different time phase showed marked difference to the prior time phase with significant difference (P＜0.05）. Tumor size in group ASODN was marked smaller than that of other 3 groups after the 5th day (P＜0.05)，while tumor size of group control 1,2 and group SODN in different phase showed no significant difference (Pgt;0.05). The results suggested that SODN and Lipofectamine showed no marked effect on tumor growth of nude mice and ASODN had marked inhibition effect on tumor growth. Conclusion mdr1 ASODN can also reverse multidrug resistance of drug-resistant human hepatocellular carcinoma cells in vivo. After the treatment the tumor’s growth in nude mice will slow down in a range of time.
ObjectiveTo investigate target gene therapy for hepatocellular carcinoma (HCC). MethodsHerpes simplex virus thymidine kinase (HSVTK) gene was inserted into the gene of AFP enhancer/ALB promoter with adenoassociated virus (AAV) plasmid (WAV2) as a carrier, and a hybrid plasmid pWAV2/AFPALB/HYTK was constructed. Besides, plasmid pEGFP1/AFPALB was also constructed. Two kinds of plasmids were transferred into AFP positive cells HepG2 and AFP negative cells 7721, SPC and 7901.ResultsIt was found that enhance green fluorescence protein could only be seen in AFP positive cells HepG2. 710 bp DNA was amplified only in AFP positive HepG2 cells.ConclusionPlasmid pWAV2/AFPALB/HYTK for HCC demonstrates specificity in vitro.
ObjectiveTo summarize the research progress of tissue engineering technology to promote bone tissue revascularization in osteonecrosis of the femoral head (ONFH).MethodsThe relevant domestic and foreign literature in recent years was extensively reviewed. The mechanism of femoral head vascularization and the application progress of tissue engineering technology in the promotion of ONFH bone tissue revascularization were summarized.ResultsRebuilding or improving the blood supply of the femoral head is the key to the treatment of ONFH. Tissue engineering is a hot spot in current research. It mainly focuses on the three elements of seed cells, scaffold materials, and angiogenic growth factors, combined with three-dimensional printing technology and drug delivery systems to promote the revascularization of the femoral bone tissue.ConclusionThe strategy of revascularization of the femoral head can improve the local blood supply and delay or even reverse the progression of ONFH disease.