This study examined the implication of EMT induced by TGF-beta1 in pancreatic cancer invasion. TGF-beta1 expression was determined in 29 cases of human pancreatic carcinoma (PC) by immunohistochemistry and the results were compared with those of pathological examination. Moreover, the effects of TGF-beta1 on the phenotype and invasion of pancreatic cancer cell line Panc-1 were also investigated. TGF-beta1 was detected in 12 cases (41.4%) of PC. Significant correlation was found between the expression of TGF-beta1 and lymph node involvement (P=0.047) and the depth of invasion (P=0.035). TGF-beta1 obviously promoted EMT of Panc-1 cell lines and their invasion ability was substantially enhanced. TGF-beta1 may promote the malignancy of pancreatic cancer by triggering EMT.

Genes, Reporter ; Genetic Vectors ; Luciferases ; genetics ; Transcription, Genetic ; bcl-2-Associated X Protein ; genetics

Objective: To investigate the inhibitory effect of survivin antisense oligonucleotide (ASODN) on the expression of survivin gene and proliferation of human hepatocellular carcinoma cell line SMMC-7721. Methods: The 20 mer antisense oligonucleotide (ASODN) targeted to the promoter region of survivin mRNA was designed and synthesized. The expression of survivin gene in hepatocellular carcinoma cell line SMMC-7721 was blocked by means of ASODN transfection mediated by DOTAP liposomal reagent. The changes of survivin protein and mRNA expression after transfection were as-sessd by Western blot and in situ hybridization, respectively. The apoptotic rate was detected by flow cytometer. The changes of cell adherent rate, cell growth activity, and the inhibitory rate of cell growth were also studied. Results: The expression of survivin protein and mRNA were decreased markedly after survivin ASODN transfection. Meanwhile, the cell adherent rate also decreased markedly while the apoptotic rate increased markedly. Conclusions: Transfection of ASODN targeted to the promotor region of survivin mRNA by DOTAP liposomal transfection reagent could down-regulated the expression of survivin protein and mRNA significantly in 7721 cell line and inhibit the proliferation of cancer cells. Survivin could be an important target in the therapy of hepatocellular carcinoma.

OBJECTIVE:To presume the structures and determine the contents of two metabolites of omeprazole in human urine after extraction and purification.METHODS:24 healthy volunteers were assigned to receive single oral dose of 40mg omepazole capsules,whose urinary samples collected within 12 hours after administration were extracted and concentrated with diethyl ether,and separated by HPLC.The relative purified metabolites were detected by mass spectrum,the structures of which were presumed and the contents were computed.RESULTS: The isolated 2 metabolites of omeprazole were presumed to be pyridine 5'— or 3'— methyl oxidation-generated hydroxy sulfone metabolite and 5'—methy hydroxylation thioether metabolite in pyridine ring,and the contents of which were 96.54% and 97.26%,respectively.CONCLUSION:The metabolites of omeprazole isolated from urinary samples by the method mentioned above were of high purity.

AIM:To investigate the impact of various levels of glucose on endothelial progenitor cells (EPCs) proliferation, senescence, and nitric oxide (NO) secretion,and the effect of insulin under high glucose conditions.METHODS: Mononuclear cells were collected from rat bone marrow by density gradient centrifugation, cultured with medium 199, and identified to be EPCs at 7th day by flk-1 and AC133 double staining. EPCss were harvested and incubated with glucose (5, 10, 20, 40 mmol/L) or insulin (0.1, 1, 10, 100 nmol/L) under high glucose conditions for 24 h or 7 days. Proliferative capacity, senescence level and NO secretion (after 24 h of incubation) were subsequently determined.RESULTS: High glucose (40 mmol/L) markedly inhibited EPCs proliferation, accelerated EPCs senescence, and decreased NO production (all P

Objective To evaluate the short and long term efficacy of self-expandable metallic stent implantation through bronchoscope in the management of the patients with malignant airway stenosis.Methods Seventy-four hospitalized patients with malignant airway stenosis during a 10-year period were selected as our subjects from Dec.2002 to Nov.2012 in the Respiratory Department of the Central Hospital of Wuhan.All these patients underwent bronchoscope for nitinol self-expandable metallic stent implantation.Clinical improvement,6min walking distance (6MWT),mMRC index,arterial blood gas,as well as early and late postoperative complications were investigated.Results A total of 77 stents were implanted,including 34 covered metallic stents and 43 uncovered metallic stents.All patients showed the obvious relief of dyspnea after the operation.6MWT at pre-operation was (172 ± 45)m and (288 ± 36)m at 28 week post-operation.Arterial blood gas was (61.34 ±6.44)mmHg at pre-opemtion and (72.12 ±6.87) mmHg at 28 week post-operation.mMRC was 3.56 ± 0.44 at pre-operation and 1.96 ± 0.37 at 28 week post-operation.All these differences were significant(P ＜ 0.01).The operation succeeded in releasing bucking of patients with esophago-tracheal fistula.Symptoms such as fever,cough,pharyngodynia,foreign body sensation and hemoptysis were observed,and all the above side effect could be solved while no serious complications arose.Conclusion Bronchoscopic insertion of metallic airway stents for malignant airway stenosis is a safe and effective procedure that could obviously relieve respiratory distress and improve the quality of life.

Objective According to the fine artery anatomy of the lower lip, we developed an innovative partial-thickness myocutaneous flap based on the vascular network of the submucosal and subcutaneous layers of the lower lip.We attempted to treat the secondary bilateral cleft lip deformities using this innovative cross-lip flap.Methods From July 2009 to June 2015, this new technique was used in 98 patients with secondary bilateral cleft lip deformities.The central defects usually occurred in these patients.The defects were reconstructed partially or completely according to the severities of the defects.The operative procedures were as follows: The split flap was elevated from the posterior portion of the oris orbicularis muscle after the inferior labial arteries were divided.Then the partial-thickness flap was rotated 180 degrees horizontally and inverted 180 degrees upward to the upper lip defect.Results All 98 musculomucosal pedicle flaps were viable.The upper lips were reconstructed according to the severities.Conclusions This myocutaneous cross-lip flap with musculomucosal-pedicle has a more reliable blood supply, better flexibility and plasticity.It is an effective method for aesthetic reconstruction for the secondary bilateral cleft lip deformities.

Objective:To investigate the expression of MEK/ERK signaling pathways in renal cell car-cinoma with bone metastasis,and to analyze the differences of expressions of VEGFR-2,MEK,ERK on the primary and metastasis tissue and its mechanism.Methods:The tissue samples were obtained from 7 renal cell carcinoma patients kindly provided by Department of Urology,Peking University People’s Hos-pital from January 1,2009 to January 1,2010.The expression of MEK/ERK signaling pathways was de-tected in the 7 renal cell carcinoma patients`primary and matched metastatic tissues with ICH,The anti-body concentrations were 1 ∶200,1 ∶25,and 1 ∶250,respectively.The mutation of the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the se-cond exon of the MEK1 gene were detected with PCR.Results:The expression intensities of VEGFR-2, MEK,and ERK were measured by H-score [intensity (1,2,3,or 4)multiplied by the distribution (%)].VEGFR-2,MEK,and ERK expressions were divided into 3 groups according to the positive dis-tribution of the tumor cells:1,0 -5%;2,6% -50%;and 3,>50%,To assess intratumor heteroge-neity,three distinct microscopic fields (×200)from each specimen were used to evaluate the expres-sions,Subsequently,the scores were averaged to obtain a single concatenated score for each tissue. VEGFR-2,MEK,and ERK expressions were assessed by 2 independent pathologists who were blinded to the clinicopathological data.The data were expressed as the mean value of the triplicate experiments.The expressions of MEK,and ERK were higher in the metastatic tissues than in the matched RCC tissues (6.10 ±4.10 vs.1.33 ±0.51,P =0.015;9.10 ±2.24 vs.4.43 ±2.84,P =0.021 )while the ex-pression of VEGFR-2 was not different between the primary and metastatic tissues (P =0.901).No mu-tation was detected on the twentieth exon of the PDGFRA gene,the second exon of the K-ras gene,the fifteenth exon of the Brafgene and the second exon of the MEK1 gene.Conclusion:MEK/ERK signa-ling pathways may play an important role in the metastasis and the resistance of sunitinib in RCC patients with bone metastasis.

Objective:To investigate whether chemokine like factor (CKLF)-like myelin and lympho-cyte and related proteins for vesicle trafficking and membrane link (MARVEL)transmembrane domain-containing protein 2 (CMTM2)is involved in spermatogenesis in varicocele induced sub-fertility rats and to discuss the possible mechanisms.Methods:Forty male SD rats (body weight:220 -330 g,age:6 -7 weeks)were randomly divided into 4 groups:varicocele for 4 weeks,varicocele for 12 weeks,sham operation for 4 weeks and sham operation for 12 weeks,with 10 rats in each group.These rats were intro-duced by partially ligating left kidney veins for the experimental groups,and the sham surgery groups as controls were executed with exactly the same surgery as in the experimental groups except for the ligation. The rats in control and experimental groups for 4 and 12 weeks were killed after laparotomy at the end of 4 and 12 weeks,respectively,the left testes and epididymis were taken out for counting the sperm,ob-serving the seminiferous tubule change and immunochemistry for CMTM2.The changes included sperm density and motility,the outer diameter and inner diameter change and the changes of epithelium and the CMTM2 expression in immunochemistry.Results:Compared with the control groups,the sperm density [(63.9 ±7.1)×106 /mL vs.(74.3 ±5.0)×106 /mL]and motility[(58.7% ±7.9%)vs.(66.1% ± 4.3%)]were reduced slightly in group of varicoele for 4 weeks,respectively (t =1.432,1.563;P =0.076,0.059,respectively ).Varicocele significantly caused a decrease in sperm concentration [(40.5 ±7.2)×106 /mL vs.(71.1 ±4.5)×106 /mL]and motility [(35.2% ±8.5%)vs.(63.4% ± 4.1%)]at 12 weeks,compared with the related sham groups (t =3.754,3.933;P =0.004,0.002, respectively).Additionally,testis CMTM2 exhibited the same disparity,that is,the CMTM2 protein ex-pression in varicocele group was significantly reduced,with the ratio of sham group to varicocele group at the end of 12 weeks 2.3 ±0.4 (t =1.978;P =0.039).In the evaluation of seminiferous tubules diame-ter,the external [(198.2 ±10.2)μm vs.(255.8 ±12.7)μm,t =2.125,P =0.003]and epithelium diameter [(54.1 ±1.5)μm vs.(75.5 ±4.1)μm,t =2.246,P =0.021]were decreased compared with the sham-related groups and previous varicocele groups.In all the varicocele groups,all types of sperm motility decreased compared with the related sham-operated group (P <0.05).Conclusion:This study suggests varicocele has a detrimental effect on CMTM2 levels and decreases spermatogonia cell number,seminiferous tubules diameter,and sperm indices.CMTM2 is associated with sperm changes in rats with varicocele,and further studies are needed to study the mechanism.

Objective: To explore the value of dual energy CT lung perfusion imaging (DEPI) for diagnosing pulmonary embolism (PE) in comparison with CT pulmonary angiography (CTPA). Methods: There were 49 patients with suspected PE received DEPI scanning and 19 with CTPA conifrmed diagnosis were enrolled in this study. CTPA image was obtained by 80 kv data, and DEPI image was obtained by PBV software. The location, type of PE in CTPA image, and the location, shape of perfusion defect in DEPI were observed and compared by segment basis. The correlation and agreement of CTPA and DEPI for diagnosing PE were calculated and the un-agreement was analyzed. Results: A total of 380 segments were included for analysis. CTPA detected 162 segments of PE and DEPI detected 155 segments of perfusion defect or reduction, partial PE were mainly presented by perfusion defects as speckles, patches or without perfusion defect, and complete PE were mainly showed segmental or sub-segmental perfusion defects. CTPA and DEPI were correlated for PE diagnosis (χ2=305.5,P=0.000), the diagnostic agreement was 83.42% and KAPPA value was 0.659. Conclusion: The perfusion defect in DEPI is related to the degree and type of PE presented in CTPA, their combination is helpful for diagnosing PE.