According to the national residency training regulations in the teaching of clinical and scientific research, the department of thoracic surgery of First Affiliated Hospital of Chongqing Medical University has used seminar teaching , and has collected random sampling survey questionnaire of 110 students participating in seminar from September 2014 to March 2016. The investigation shows that through seminar teaching training, doctors will cultivate their clinical and research ability, and enhance their self-confidence. Therefore, the application of seminar teaching in department of thoracic surgery is helpful to improving the quality of clinical teaching and is worthy of further improvement and promotion.

Objective To develop the human colon carcinoma(HT-29) liver metastasis model in nude mice,and to detect the expression of human telomerase reverse transcriptases(hTERT) mRNA in liver of nude mice. Methods Liver metastases were established in 30 Balb/c nude mice by intrasplenic injection of colonic cancer cells(HT-29),and the spleens were resected.After being sacrificed,the tumor growth was observed,and the pathological examinations were performed.The expression of hTERT mRNA in the livers of nude mice was detected by RT-PCR technique. ResultsHuman colon carcinoma(HT-29) liver metastasis model was developed in all the 30 nude mice,with the liver metastasis rate of 100%.The pathological results revealed the occurrence of liver metastases,and the expression of hTERT mRNA was positive in the liver tissues. Conclusion Intrasplenic injection of HT-29 cell is a reliable way for producing colonic cancer liver metastasis model,and the positive expression of hTERT mRNA in liver tissues indicates the significance of hTERT in the early diagnosis and treatment of colon carcinoma liver metastasis.

OBJECTIVE Topreparealipidderivativeofgemcitabine(Gem)anditspolymericmi-celles to overcome the disadvantages of Gem.METHODS N-benzyl-3′-acetyl-gemcitabine(BAG)was synthesized.A BAG-loaded poloxamer polymeric micelle (BAG∶poloxamer 188 =10∶1 ,mol/mol)was prepared using an injection method.The micelles were characterized with a laser particle size and elec-tric charge instru ment and negatively-stained trans mission electron microscopy.Hu man breast cancer cells MCF-7 were cultured with Gem or BAG polymeric micelles of 5,10,20,30,50,70,90 μmol·L-1 for 24,48 and 72 h,respectively.The inhibitory rate of cells was measured with an MTT method.The MCF-7 cytotoxicity of BAG polymeric micelles was investigated.A pharmacodynamic study was per-formed on the mice bearing mouse hepatocellular cancer cells H22.Intravenous (iv)and oral (ig)ad-ministration was used at the dose of Ge m 40 mg·kg -1 or BAG polymeric micelles 62 mg·kg -1 .The mice were administered on the 1 st,4th and 7th day and sacrificed on the 8th day.Tumor inhibitory rates were measured.RESULTS TheBAGstructurewasidentifiedbythinlayerchromatograph,1Hand13C NMR,infrared ray chromatograph and mass spectrum.The appearance of BAG micelles was a slightly blue suspension.The micelles were spheres according to the electron microscopic observation.Their size was 62.82 nm and the zeta potential was -18.8 mV.The half inhibition concentration (IC50)of Gem and BAG polymeric micelles was 40.6 and 90.0 μmol·L-1 ,5.0 and 14.9 μmol·L-1 ,5.0 and 1 3.6 μmol·L-1 at 24,48 and 72 h,respectively according to the MTT results.According to the in vivo results,compared with the tumor model group,Gem (ig),Gem (iv)and BAG polymeric micelles (iv and ig)had significant effect on the tumor weight of H22 cell xenograft mice (P<0.01 ).As for anti-tumor efficiency,BAG polymeric micelles (ig)were better than Gem (ig)(P<0.05);BAG polymeric micelles (iv)were better than BAG polymeric micelles (ig)(P<0.05),and BAG polymeric micelles (iv)were almostequaltoGem(iv).CONCLUSION ThelipidderivativeofGemcanbeloadedinthepoloxamer 1 88 polymeric micelles.BAG polymeric micelles show in vitro MCF-7 cell inhibition and in vivo inhibition of mouse H22 xerografts;iv or ig.BAG polymeric micelles (ig)show better anti-tumor effect than Gem (ig),indicating that BAG polymeric micelles are a promising novel anti-tumor oral preparation.

A set of primers amplified the VP1 gene of foot-and-mouth disease vims (FMDV) was designed and synthesized. A reverse transcription-polymerase chain reaction (RT-PCR) technique detected the RNA of FMDV was established after selecting the best purification method, reagents and reaction conditions. Samples of fresh milk, lymph node, spinal cord, vesicular skin, milk powder, cotton swab, mouse and meat in daughter-house were detected by RT-PCR, positive rates were41.4% (24/58), 13.33% (2/15), 20% (1/5), 100% (1/1), 100% (1/1), 37.5% (12/32), 100% (2/2) and 10% - 70%, respectively. However, positive rate of cockroach detected by RT-PCR was 0. The results showed that the established FMDV RT-PCR technique provided a more sensitive, specific and reliable method for diagnosis and epizootic study of the foot-and-mouth disease.

Altitude ; Animals ; Arvicolinae ; Gene Expression ; Hypoxia ; Rats ; Tumor Suppressor Protein p53 ; genetics

Objective To analyse the graft impingement after anterior cruciate ligament (ACL) reconstruction based on three-dimensional graft and tunnel images of dual-source computer tomography (DSCT).Methods From November 2012 to November 2014,a consecutive 134 cases of ACL reconstruction with various techniques had their knees scanned by DSCT.Graft and femoral as well as tibial tunnel images were three-dimensionally reconstructed and carefully inspected if there was any impinging on the graft at knee extension,meanwhile the femoral and tibial tunnel location were measured.Of these 134 cases,a serial of 118 patients had been single-bundle reconstructed with same type of graft material.There were 83 males and 35 females aged from 15 to 64 years old (mean,32 years old).Special attention had been paid to examine if there was any graft impinging,and their origin and finally impingement types were identified and classified according to the position and origin of impingement.Bernard and Lorenz method were used to measure the relative locations of femoral and tibial tunnel respectively.Difference was statistically compared between impingement-free and impingement groups.Results Patients were divided into two groups,impingement-free group and impingement group according to graft impinging based on graft visible 3D-CT imaging postoperatively.Of 118 cases,39 (33％) were impingement-free and 79 (67％) existed various types of impingement.Impingement group was further divided into two subgroups according to different impingement location,outlet impingement of intercondylar notch group (77 cases,account for 65％ of total and 97％ of impingement group,respectively) and mid-way impingement of intercondylar notch group (2 cases,account for 2％ of total and 3％ of impingement group,respectively).The outlet impingement group was more further classified into three subtypes,bird beak impingement,10 cases (13％,10/77),tibial plateau impingement,46 cases (60％,46/77) and clamp impingement,21 cases (27％,21/77).Statistic analysis with NAONA revealed that no significant difference between impingement group and impingement-free group regardless of femoral or tibial tunnel placement.Conclusion Three new types of impingements are found based on visible 3D-CT graft imaging after ACL reconstruction which are mid-way impingement of intercondylar notch,tibial plateau impingement and clamp impingement.

Objective:Aldose reductase,involved in the pathogenesis of diabetic complications,was recombinated with an adeno associated virus vector pSNAV2.0,and it was transfected into human embryonic kidney 293(HEK 293)cells.The gene engineering produced AR would be used as a target protein to screen aldose reductase inhibitors.Restriction endonuclease digestion and ligation procedures were performed to construct the AR expression plasmid vector pSNAV-hAR.Methods:After confirmation the recombinant plasmid by PCR,restriction endonuclease digestion,and DNA sequencing,pSNAV-hAR was transfected into HEK293 cells.Western blot and immunofluorescence analysis were performed to detect the expression of AR and its enzyme activity.Results:The results of a series of analysis including AR activity assay,Western blot and immunofluorescence analysis shown the expressed protein mediated by the adeno associated virus vector transfecting HEK 293 cells,was functional AR.The traditional aldose reductase inhibitors,Sobinil and Zopolrestat,were used to test and verify the constructed cell model.Conclusion:The established AR expression model can be used in mechanismresearch of activation of polyol pathway on diabetic complications and screening potential aldose reductase inhibitors.

Objective To prepare the human dual-specificity protein phosphatase18 (Dusp18) monoclonal antibodies (McAb) with high titer and specificity and identify its characterization, which is based on further studying Dusp18 function. Methods BALB/c mice were immunized with purified recombinant Dusp18 protein. Cell fusion was performed between mouse splenic cells and myeloma cells (SP2/0), and then the hybridoma cell lines secreting McAb against Dusp18 antigen were screened and cloned. The ascites were prepared and purified with Protein G affinity chromatography. The titer and subtypes of McAb against Dusp18 were identified and measured by ELISA and Western blotting analysis. Results Two hybridoma cell lines, F003 and F004, that stably secreted McAb against Dusp18 were successfully obtained, which belong to the subtypes of IgG1 and k light chain. The antibody titers in culture supematant were 1:5120 and1:10 240, and those in the ascites fluid were 1:25 600 and 1:51 200 respectively. Western blotting analysis and immunohistochemistry showed that the two antibodies can specifically bind with Dusp18 derived from human eucaryotic cells or tissue. Conclusion Two McAb against Dusp18 have been successfully prepared which can be used for further studying the biological properties of Dusp18 and reveal its relationship with tumorigenesis and development.

BACKGROUND:There are several reports about the application of fresh bone marrow aspirate being injected directly to repair partial ligament injury, but the application about fresh bone marrow aspirate directly being planted on scaffolds to build tissue-engineered ligament is rarely mentioned.
OBJECTIVE:To evaluate the feasibility of applying fresh bone marrow aspirate planted directly on scaffolds to construct tissue-engineered ligament
METHODS:We constructed fibroin fiber/smal intestinal submucosa composite scaffold, then planting fresh bone marrow directly to built bone marrow seeding group and planting seed cel s (bone marrow mesenchymal stem cel s) on the scaffold to built cel seeding group. The control group had no treatment. After that, we detected the density of cel adhesion, cel proliferation ability and extracel ular matrix secretion. Then, the composite in the bone marrow seeding group was implanted into the broken anterior cruciate ligament in rabbits, and material biocompatibility in vivo was evaluated after 12 weeks.
RESULTS AND CONCLUSION:After 4 hours of incubation, bone marrow seeding group was significantly higher than the cel seeding group in cel adhesion density and proliferation rate (P<0.05). Bone marrow seeding group and cel seeding group showed higher type I, III col agen secretion compared with the control group (P<0.05), but the col agen secretion of bone marrow seeding group and cel seeding group showed no significant difference. Composite cel scaffold implantation in vivo did not cause fatal immune rejection and severe inflammatory reaction, and no significant ligament regeneration and vascularization occurred. These findings indicate that fresh bone marrow aspirate can be seeded directly on scaffolds to construct tissue-engineered ligament, and the short-term biocompatibility in vivo is good.

Objective To investigate the risk factors of articular cartilage damage secondary to anterior cruciate ligament (ACL) rupture.Methods Clinical data of 490 patients sustaining ACL rupture from July 2008 to July 2014 were collected for a retrospective cohort study.The factors relating to the incidence and degree of the secondary articular carticular damage were analyzed,including gender,age,weight,blood group,places of residence,causes of injury,complication of meniscus tear,damage part and duration of disease.Results Factors associated with the incidence of secondary articular cartilage damage were blood type O (OR =0.605,95％ CI 0.381-0.960,P ＜ 0.05) and complication of meniscus tear(OR =2.241,95％ CI 1.378-3.643,P ＜0.01).Factors associated with the degree of articular cartilage damage were duration of disease (rs =0.168 7,P ＜0.01),damage part (Hc =53.237,P＜0.01),and complication of meniscus tear (x2 =16.944,P＜0.01).Conclusions Meniscus tear is the moderate risk factor for the incidence of secondary articular cartilage damage while blood type O has weak protective effect.Damage part,course of disease,and meniscus tear are factors associated with the degree of secondary articular cartilage damage.