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 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.

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.

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 investigate the differences of protein and mRNA expression of matrix metalloproteinase 9 (MMP-9) in osteoclasts (OC) and osteoclast-like cells (OLC) obtained by mechanical and inducement methods. Methods Mechanical separation method was used to separate mature osteoclasts from long bones of SD rats aged one-day;and inducement culture method was applied to induce OLC by using RANKL (100 ng/mL) and M-CSF (100 ng/mL) . The protein expression of MMP-9 was measured by immunocytochemistry and mRNA expression of MMP-9 was assayed by in situ hybridization. Results The integral optical density (IOD) and average optical density (AOD) of positive cells in the visual field were higher in the 12-d group of inducement method as compared with the 3 d-group of mechanical method. Conclusions It is suggested that the protein and mRNA expression of MMP-9 in OLC obtained by 12 d inducement method is much high than in OC obtained by 3 d mechanical method. OLC obtained by inducement method can be applied in the study of osteoporosis.

Objective To explore the application of quantitative DNA analysis in differential diagnosis of benign and malignant breast masses to aid the surgery plan formation.Methods Four hundred and eighty-eight patients with breast mass were enrolled into this study.Tissues of breast mass in patients were gained by fine-needle aspiration puncture.Two sections were made from each sample,one was stained by Papanicolaou for regular cytology analysis and another was stained with Feulgen for quantitative DNA analysis.Pathological results were confirmed in each case after surgery.Results One hundred and sixty-four cases were classified as patients with benign neoplasm,while the other 324 cases were classified as malignant neoplasm,according to the pathological examination results.The sensibility and specificity were 91.4%(296/324) and 92.7%(152/164) for regular cytological method,90.1%(292/324) and 100.0%(164/164) for quantitative DNA analysis method.Meanwhile the positive predictive and negtive value of quantitative DNA analysis was 100.0%(292/292) and 83.7%(164/164),of which regular cytological methods were 96.1%(296/308) and 84.4%(152/180).Conclusion The quantitative DNA analysis might assistant differential diagnosis of benign and malignant breast tumor.

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 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.

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.