Related clinical characteristics of diabetes patients suffering from pancreatic cancer / 重庆医学

Objective To observe and analyze clinical characteristics of diabetes patients suffering from pancreatic cancer . Methods We recruited 107 cases of pancreatic cancer(66 without diabetes and 41 with diabetes) and 100 diabetes patients without pancreatic pancreas as control .Patients′ demographic information ,degree of tumor differentiation ,serum markers etc .were com‐pared in order to find out the relevant clinical features of diabetes patients suffering from pancreatic cancer .Results (1)Patients with pancreatic cancer mostly were middle‐aged males .55 .1% of them suffering from dysglycemia ,18(16 .8% ) and 41(38 .3% ) of whom had impaired fasting glucose and diabetes ,respectively .(2)Compared with their without diabetes counterparts ,pancreatic cancer with diabetes were more prone to be asymptomatic and weight loss(P< 0 .05) .(3)Compared with their without diabetes counterparts ,pancreatic cancer with diabetes had significantly higher levels of fasting blood glucose(FBG) andγ‐glutamyltranspep‐tidase(γ‐GT)(P<0 .05) .(4)When compared with diabetes control ,pancreatic cancer with diabetes were older and shorter duration and lower body mass index(BMI)(P<0 .05) .They were more prone to weight loss(P<0 .05) .Moreover ,serum CA19‐9 and CEA levels in them were significantly higher than those in the diabetes control(P<0 .05) .Conclusion Older age ,shorter duration ,low BMI are all risk factors for diabetes patients to develop pancreatic cancer .Being asymptomatic and weight loss are their clinical characteristics .CA19‐9 and CEA are both sensitive serum markers to detect pancreatic cancer patients with diabetes .

Construction of adenovirus carrying dual-target shRNA for Oct-4 and Survivin and its inhibitory effect on human hepatocellular carcinoma cells / 生物工程学报

The transcriptional factor Oct-4 and Survivin are the key regulatory factors in cancer cell proliferation and mitosis. A dual cancer-specific shRNA adenovirus vector, Ad5-Dual-shRNA, targeting Oct-4 and Survivin genes was constructed by molecular cloning and recombination. After cells were infected with virus, hepatocellular carcinoma cell line EHBH-H1 was used for detecting the expression of Oct-4 and Survivin proteins by Western blotting. The viral cytotoxic effect on cancer cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) reduction assay in vitro, and the inhibition effect on tumor xenografts was observed in nude mice. The results showed that the expression of Oct-4 and Survivin in cancer cell line EHBH-H1 could be silenced markedly by Ad5-Dual-shRNA. In MTT and animal experiments, Ad5-Dual-shRNA also represented much stronger anti-tumor effect on tumor growth than Ad5-Surv-shRNA and Ad5-Oct4-shRNA. From this research we can draw a conclusion that the cancer-specific adenovirus vector expressing dual-shRNA targeting Oct-4 and Survivin genes may provide us a more effective, specific and convenient gene therapy method.

Effects of quercetin on nuclear factor-κB p65 expression in renal ubiquitin-proteasome system of diabetic rats / 中华内科杂志

Objective To investigate the protective effect of quercetin on diabetic nephropathy and to explore its possible mechanism.Methods Type 2 diabetes mellitus rat model was established by feeding high-carbonhydrate-fat diet and injecting with streptozotocin.At 72 hour after injection,blood samples were collected from the tail veins of all rats.Those rats with blood glucose level ≥ 16.7 mmol/L were considered as the diabetes model been successfully established.The model rats were randomly divided into type 2 diabetic group ( group DM,n =9 ) and quercetin group ( group QUE,n =9 ).Other rats were used as normal controls (group NC,n =8).All rats were performed by intragastric administration for 8 weeks.At the end of experiment,the rats were sacrificed and fasting plasma glucose( FPG),fasting insulin( Flns),serum creatinine (SCr),blood urea nitrogen(BUN),TG,TC,LDL-C,24 h urine protein (24 h UP),and kidney index ( KI ) were evaluated.Pathological changes of kidney were observed by periodic acid-silver metheramine( PASM ).The expressions of ubiquitin and NF-κB p65 on glomeruli w ere examined by immunohistochemical method,and its association with the incidence of proteinuria was analyzed.Results In groups DM and QUE,the level of FPG [ ( 25.45 ± 1.23 ) mmol/L and ( 19.99 ± 1.20 ) mmoL/L],FIns [ ( 25.67 ± 2.58 ) mU/L and ( 19.29 ± 1.80 ) mU/L ],SCr[ ( 44.00 ± 2.53 ) μmol/L and ( 34.43 ± 2.23 )μmol/L],BUN[ ( 11.60 ± 0.39 )mmol/L and (8.20 ± 0.37) mmoL/L],TG [ (3.32 ± 0.22 ) mmol/L and (2.43±0.25)mmol/L],TC[(2.95 ±0.21) mmol/L and (2.24 ±0.17)mmol/L],LDL-C[(2.03 ±0.22 ) mmol/L and ( 1.49 ± 0.13 ) mmol/L ],24 h UP [ ( 46.67 ± 2.50 ) mg/24 h and ( 25.57 ± 2.82 )mg/24 h]and KI[ (9.76 ±0.30) × 103 and (8.44 ±0.26) × 103 ] were significantly increased than the indexes of group NC [ (6.56 ± 0.41 ) mmol/L,( 12.63 ± 1.41 ) mU/L,( 22.88 ± 2.36 ) μmol/L,( 5.45 ±0.51 ) mmoL/L,( 1.64 ± 0.1 1 ) mmol/L,( 1.33 ± 0.17 ) mmol/L,(0.46 ± 0.05 ) mmol/L,( 12.38 ±1.19)/24 h and (6.78 ±0.12) × 103].Moreover,the above indexes in group QUE were obviously lower than group DM.There was evidence of pathological changes associated with diabetes,such as focal and segmental sclerosis and thickened basement and mesangial expansion.The expressions of ubiquitin and NF-κB p65 in renal tissues of group DM increased significantly ( P ＜ 0.01 ).The expression of ubiquitin and NF-κB p65 were positively related with the level of 24 h UP ( r =0.893,0.879,P ＜ 0.01 ).Compared with group DM,all above indexes in group QUE were markedly alleviated ( P ＜ 0.01 ).The expression of ubiquitin and NF-κB p65 was reduced but didn't reach level in group NC ( P ＜ 0.01 ).Conclusion The increased expression of NF-κB induced by ubiquitin-proteasome system may participate in the pathogenesis of proteinuria in diabetic nephropathy.Quercetin has renal protective effects partly through reducing NF-κB p65 expression.

Differentiation of bovine male germ-line stem cells in vitro / 生物工程学报

Male germ-line stem cells (mGSCs) have the capability of self-renewal and latent capability of differentiation. mGSCs is the unique diploid immortal cell which can transfer genetic information to filial generation. The combination of transgenic technology and mGSCs heterotransplanting will supply new opportunities and paths to cloning animal, transgenic animal and gene therapy of some human hereditary disease. We studied the isolation and cultivation of mGSCs that were isolated and purified from 5-6 month old bovine fetal testis, new born bovine testis by adopting mixed enzymes digestion and different attaching velocities methods. The results showed that Sertoli cells were indispensable to mGSC's proliferation and differentiation in vitro. The Sertoli cells in logarithmic phase have a significant effect on mGSC's attaching, proliferation and differentiation. Co-culture with Sertoli cells, mGSCs differentiated to long sperm after 16 days. A preliminary system for mGSC's inducing differentiation was established.

Cotransfection of glial cell-derived neurotrophic factor and endothelin receptor type B gene into mouse neural stem cells / 中国组织工程研究

BACKGROUND: Deletion of glial cell-derived neurotrophic factor and endothelin receptor type B gene will induce abnormal development of enteric nervous system. Neural stem cell transplantation can repair nervous system from anatomy and function,and be considered as a vector of gene transfection.OBJECTIVE: To transfect recombinant adenovirus carrying glial cell-derived neurotrophic factor and endothelin receptor type B gene into mouse neural stem cells, and to observe expression of target gene.DESIGN: A cell-gene study.MATERIALS: New-born Kunming mice were provided by the Animal Center of Tongji Medical College, Huazhong University of Science and Technology, China. jetPEI reagent was purchased from PolyPlus Co, France. The pAdTrack-CMV-GE with green fluorescent protein (GFP) was gifted by Doctor Sun Nianfeng and Zhang Jinghui in our laboratory.METHODS: Neonatal mouse brain tissues were sterilely obtained to prepare monoplast suspension. Adenovirus expressing glial cell-derived neurotrophic factor and endothelin receptor type B gene with GFP was dissolved in NaCI to prepare JetPEI/DNA complex. Subcultured neural stem cells in DMEM/F12 were regulated to 5×10~8/L, and 400 μL cell suspension and 100 μL JetPEI/DNA complex were seeded on a 24-well plate at 37 ℃ in 5% CO_2 incubator. Neural stem cells were harvested at 24, 48 and 72 hours following transfection.MAIN OUTCOME MEASURES: The efficiency of transfection was detected using fluorescence microscope and flow cytometry.Target gene expression in neural stem cells was determined using RT-PCR.RESULTS: Bright green fluorescence of the transfected cells could be observed under fluorescence microscope after 24 hours of transfection. The positive rate of GFP was 15.36%, 24.67%, 25.73% at 24, 48 and 72 hours following transfection respectively.Neural stem cells expressed glial cell-derived neurotrophic factor and endothelin receptor type B gene at various time points.Strap brightness was low at 24 hours, and exogenous gene expression was great at 72 hours.CONCLUSION: The target genes were successfully transfected into neural stem cells by using jetPEI reagent. Moreover, glial cell-derived neurotrophic factor and endothelin receptor type B gene effectively transcribed and expressed in target cells.

Culture and identification of monoclonal neural stem cells derived from cerebral cortex / 华中科技大学学报（医学）（英德文版）

To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to detect the specific marker of neuroepithelial stem cells (Nestin) of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution.

Culture and Identification of Monoclonal Neural Stem Cells Derived from Cerebral Cortex / 华中科技大学学报（医学）（英德文版）

To isolate and culture the purified monoclonal neural stem cells from the cerebral cortex of new born mice, new-born mice cerebral cortex was isolated and dissociated to single-cell suspension by mechanical trituration. The dissociated single cells were cultured in serum-free medium. After the formation of neurospheres, single-cell clone culture was performed by limiting dilution and the proliferated single-cell clones were harvested for subculture. Immunocytochemistry was used to detect the specific marker of neuroepithelial stem cells (Nestin) of the primary and monoclonal neurospheres. In the differentiated cells we detected the specific antigen of NF-200 and GFAP. Our results showed that the primary neurospheres expressed Nestin antigen positively. By limiting dilution, we cultured the cell lines from single-cell clone and the monoclonal neurospheres expressed Nestin and had capabilities of self-renewal, proliferation and the potentiality of differentiation into neurons and glial cells. It is concluded that monoclonal neural stem cells which have the ability of proliferation and multi-directional differentiation can be isolated and cultured from the cerebral cortex of new-born mice by limiting dilution.

CT quantitative study of coal miner's pneumoconiosis / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To study the value of CT quantitativeness in the diagnosis of coal miner's pneumoconiosis.</p><p><b>METHODS</b>104 cases were examined by HRCT scan at top of aortic arc, carina of trachea, 3 cm below the bifurcation of bronchi, among them there were 87 patients with different stages of coal miner's pneumoconiosis, 17 cases of normal males as the control group. All images were determined by CT density histogram at specific region (- 1,024-0 HU). Calculated the percentage of each pixel included a varying number of CT value, and the ratio of density values in the specific region.</p><p><b>RESULTS</b>The ratio of density values in the region of -983 (-) -778 HU was 87.31% in normal control group, and 80.51%, 75.27% and 72.99% respectively in the I, II, III stages of coal miner's pneumoconiosis. There were statistically significant differences among the groups (P < 0.01).</p><p><b>CONCLUSION</b>CT quantitative histogram information was able to observe the fibrosis and its degree of coal miner's pneumoconiosis. It has a good diagnostic value for its reliability and objectiveness.</p>

Comparative Study on the Vitro Antibacterial Activity of Domestic and Imported Teicoplanin against 224 Strains of Enterococci / 中国药房

OBJECTIVE:To study the vitro antibacterial activity of domestic teicoplanin against224strains of enterococ-ci.METHODS:Determination of MIC of domestic teicoplanin against169strains of entercococcus faecalis and51strains of E.faecium was tested by agar dilution method,and its antibiotic effect was compared with that of the imported teicoplanin and other antibiotics.RESULTS:The MIC 50 of domestic teicoplanin against169strains of E.faecalis and51strains of E.faecium were0.125,0.25?g/ml respectively,MIC 90 were2,1?g/ml respectively;The MIC 50 of imported teicoplanin against E.faecalis and E.faecium were0.25,0.25?g/ml respectively,MIC 90 were1,0.5?g/ml respectively.CONCLUSION:Two kinds of te-icoplanin have strong antibacterial activity against224strains of enterococci;the sensitivity of224strains of enterococci to both kinds was100%.

Practical Value and Imaging Expression of Target Reconstruction Technique of HRCT in the Diagnosis of Pneumoconiosis / 实用放射学杂志

Objective:To study the practical value and imaging expression of target reconstruction technique of HRCT in the diagnosis of pneumoconiosis.Methods:Fifty-seven cases with different level type of pneumoconiosis were first examined by routine CT scan,followed by HRCT examination at top of aortic arch,carina of trachea,3 cm below bifurcation of bronchi,and 2 cm above thd right diaphragm.Target reconstruction have been finished using the raw data stored.Results:Target reconstruction photoimages were cleared to display the morphology of delicate lesions of pneumoconiosis:(1)Nodular shape(100%).(2)Interlobular septal thickening(98.24%).(3)Lobular structure abnormality(98.24%).(4)Subpleural nodes (84.21%).(5)Centrilobular emphysema(63.15%).(6)Subpleural line(64.91%).(7)Bronchiectasis(63.15%).(8)Ground-glass opacity(35.08%).(9)Honeycombing(15.78%),and to display earlier period change details.Conclusion:HRCT target reconstruction technique may make lesion dispaly more detail than comventional and high-resblution CT.It is reliable to display the delicate lesions of pneumoconiosis and provide more diagnostic information.