AIM: The present study is to investigate the inhibitory effect of the transfer of replication-defective retroviral recombinant plasmid encoding a wild-type P16 ink4a gene on the formation of restenosis after rabbit carotid arterial injury in vivo.METHODS:A replication-defective retroviral recombinant plasmid encoding wild-type gene P16 ink4a was constructed and the packaged high titer virus stock was obtained. It was transferred into the rabbit carotid arterial wall immediately after injury. The P16 ink4a mRNA expression in the arteries was examined by Northern blot and in situ hybridization. The effect of overexpression of the P16 ink4a gene on arterial intima hyperplasia was determined by pathophysiological method. RESULTS: The exogenous P16 ink4a could be effectively transferred into injured arterial wall by retroviral recombinant plasmid and the gene products could inhibit smooth muscle cells proliferation (11.80?3.54 vs 25.20?5.12,P

Objective To investigate the role of miR-200b on gemcitabine induced epithelialmesenchymal transition (EMT) in pancreatic cancer cell line MiaPaCa-2.Methods Different concentrations of gemcitabine were used to induce MiaPaCa-2,and the concentration of 50％ cell proliferation inhibited (IC50) was applied to obtain drug-resistant MiaPaCa-2 cells.MiR-200b or nonsense small molecular fragments (negative control,NC) was transfected into MiaPaCa-2 cells by liposomes,then gemcitabine of IC50 was used to induce cells to obtain drug-resistant MiaPaCa-2 cells transfected with miR-200b or NC.The morphological characteristics of MiaPaCa-2 cells were observed by inverted microscope.Invasion of cells were detected by transwell chamber.The expression of miR-200b was measured by using real-time PCR.The expressions of Ecadherin,Vimentin,Zebl,Zeb2 proteins were determined by Western blot.Results After gemcitabine treatment,the cells' size gradually diminished,intercellular junctions decreased,pseudopodium increased,which presented the characteristics of mesenchymal morphology.The invaded cell number increased from (26 ± 3) to (85 ± 6),and the expression of Vimentin Zebl,Zeb2 was increased to (1.87 ± 0.17),(2.57 ±0.21),(5.24 ± 0.83) folds of the parent cells.The expression of miR-200b was decreased to (0.36 ± 0.01)folds of the parent cells,and the expression of E-cadherin was decreased to 0.47 ± 0.05 folds of the parent cells,while the invaded cell number of drug-resistant MiaPaCa-2 transfected with miR-200b was decreased to (42 ± 4),and the expression of Zebl,Zeb2 was decreased to (0.36 ± 0.07),(0.08 ± 0.01) folds of drugresistant MiaPaCa-2 transfected with NC.Conclusions The occurrence of EMT is observed in pancreatic cancer cell line MiaPaCa-2 during gemcitabine induction,and miR-200b down-regulation may be a possible mechanism.

ObjectiveTo investigate the methylation of the promoter region in miRNA in pancreatic cancer cell line PANC1 and normal pancreatic tissue,to discover the miRNA with hypermethylation associated with pancreatic cancer.MethodsThe genomic DNA of PANC1 and normal pancreatic tissue was extracted,and fractured by ultrasound.Methylation DNA fragments were obtained by 5-methyl of pyrimidine nucleoside antibodies and immunomagnetic beads.The hypermethylation miRNA differentially expressed between PANC1 and normal pancreatic tissue was selected by using methylation DNA chip.BSP ( bisulfite genomic sequencing PCR) and TA clone sequencing was performed for further validation.The genomic DNA of pancreatic cancer cell lines BXPC3,CFPAC1,PANC1 and SW1990 was extracted.The COBRA (combined bisulfite restriction analysis) was used to validate differentially expressed hypermethylation miRNA.ResultsEight differentially expressed hypermethylation miRNAs were screened from the DNA methylation chips,then five of them were selected for sequencing.The methylation status of miRNA-615,-663,-663b was significantly higher in the PANC1 than in normal tissues (60.6％ vs 7.6％,88.8％ vs 22.2％,94.4％ vs 13.0％ ) ; the methylation status of miRNA-675 was not significantly different between PANC1 and normal pancreatic tissue (76.0％ vs 100％ ).Due to large error in sequencing,miRNA1826 was excluded.The results of COBRA confirmed all the 4 miRNAs were highly methylated in PANC1 ; except for miRNA-675,other 3 miRNAs were highly methylated in BxPC,miRNA-663,miRNA-663b were highly methylated in CFPAC1,while miRNA-615,miRNA-663 were highly methylated in SW1990.ConclusionsHypermethylation miRNAs were differentially expressed between pancreatic cancer cell lines and normal pancreatic tissue,among them,highly methylated miRNA-663 was possibly associated with pancreatic cancer.

Objective To establish a method for quality control of xinshuaining mixture. Methods The herbal contents including red ginseng,milk veteh,root of red rooted salvia were identified by thin layer chromatography ( TLC) and the content of ginsenoside Rb1 in xinshuaining mixture was determined by high performance liquid chromatography ( HPLC ) . Results The herbs can be overtly identified by TLC. Ginsenoside Rb1 had a linear relationship in the range of 1. 095-5. 475μg, and the average recovery was 97. 3%(RSD 1. 98%). Conclusion The method is simple and rapid,and can be used for the quality control of xinshuaining mixture.

Objective To study the effects of two kinds of vertical partial laryngectomy on laryngeal function,postoperative extubation rate and survival rate of patients with glottic laryngeal carcinoma.Methods From January 2013 to January 2014,one hundred and fifty patients with glottic carcinoma who treated in Huzhou Gospel Hospital were selected.The patients were divided into observation group and control group by random number table method,with 75 cases in each group.The control group used the traditional vertical partial laryngectomy,the observation group was treated with modified vertical partial laryngectomy.The postoperative survival rate,extubation rate,extubation time and complications were observed in the two groups.Results The extubation rate was 100.00% (75/75) in the observation group and 97.33% (73/75) in the control group,the difference was statistically significant between the two groups(χ2 =2.027,P>0.05).The extubation time in the observation group was (11.85 ± 0.49)d,which in the control group was (14.55 ± 0.56) d,the difference was statistically significant between the two groups( t=31.424, P<0.05).There were 67 cases(89.33% ) with grade 0 and 8 cases(10.67% ) with grade 1 in the observation group.The swallowing function of the observation group was stronger than that of the control group( Z=5.238,P<0.05).The 1-year and 2-year survival rates of the two groups were similar(all P>0.05).The 3-year survival rate of the observation group was 97.33% (73/75),which of the control group was only 88.00% (66/75),the differ-ence was statistically significant between the two groups (χ2 =4.807,P<0.05).There was no pharyngeal fistula in both two groups. There were 5 incision infections in the control group, and 1 incision infection in the observation group.There was no statistically significant difference in the incidence rate of complications between the two groups (χ2 =2.778,P>0.05).Conclusion Improved vertical hemilaryngectomy therapy for patients with glottic laryngeal carcinoma can effectively shorten the time of extubation,better preserve swallowing function,improve the long-term survival rate,and the extubation rate and complication rate are similar with the traditional vertical partial laryngectomy.

Objective To analyze the clinical characteristics,diagnosis,treatment and prognostic factors of primary hepatic neuroendocrine tumor (PHNET).Methods To analyze the clinical data of 21 patients with PHNET who were admitted to the First Affiliated Hospital of Zhengzhou University from January 2012 to January 2019.There were 11 males and 10 females,with ages which ranged from 36 to 75 years.Log-rank test was used for single-factor analysis of the relationship between clinical and pathological indicators and prognosis.Cox proportional risk model was used for multi-factor analysis.Results Among the 21 PHNET patients,10 presented with central and upper abdominal pain,3 abdominal pain accompanied by nausea and vomiting,5 abdominal distension,and 3 were asymptomatic.Carbohydrate antigen (CA) 19-9 was elevated in 6 patients and CA125 in 7 patients.Abdominal enhanced CT showed solid liver space occupying lesions.Four patients had solitary and 17 had multiple lesions.The mean diameter was 58 mm.Fourteen patients had lymph node metastasis.Five patients underwent radical hepatectomy,2 transcatheter hepatic arterial chemoembolization + chemotherapy,12 chemotherapy,and 2 supportive treatment.Factors which were associated with prognosis of PHNET patients were surgery,tumor grading,cytokeratin positivity and lymph node metastasis.Multivariate Cox regression analysis showed that inoperability (HR =8.99,95％ CI:1.13-71.80) was an independent risk factor of prognosis of PHNET patients.The prognosis in patients who underwent surgical resection was better.Conclusion Patients with PHNET had no specific clinical manifestations.Surgical resection gave the best results in treatment.As surgical resection affected prognosis,it should be carried out if technically feasible.

OBJECTIVE To investigate the prognostic value of serum creatinine-cystatin C ratio(CCR)for nasopharyngeal carcinoma(NPC).METHODS NPC patients who underwent consultation at the Army 72nd Group Military Hospital from January 2007 to December 2015 were retrospectively analyzed and divided into high-expression and low-expression groups according to whether the CCR was＜0.81(median),and the predictive variables of the high-expression and low-expression groups were compared.Kaplan-Meier survival analysis and Cox regression were used to analyze the effect of CCR on overall survival(OS),and clinical prediction models and column line diagrams were constructed and evaluated based on relevant predictive factors.RESULTS A total of 516 NPC patients were included.The percentage of elderly patients in the high-expression group(22.5%)was significantly lower than that in the low-expression group(32.6%),and the percentage of highly-differentiated patients(22.5%)was significantly higher(14.7%),and the percentage of patients with TNM stage Ⅰ and Ⅱ(40.7%)was significantly higher than that in the low-expression group(29.8%),and the differences of the above indexes were all statistically significant(P Kaplan-Meier survival analysis suggested that low-expression CCR was significantly associated with poorer OS,and multifactorial Cox regression analysis showed that age,CCR,pathologic grade,time from diagnosis to treatment,and TNM stage were significantly associated with OS(P＜0.05).The clinical prediction model constructed on the basis of the above predictive factors had higher AUC values than the traditional TNM staging model,and its column line graph predicted OS with good agreement with the actual.CONCLUSION CCR can be considered as a potentially useful prognostic factor for NPC patients.

Objective To explore the value of non-contrast CT findings of acute ischemic stroke(AIS)for predicting early prognosis after mechanical thrombectomy.Methods Data of 161 AIS patients from clinical center 1 who underwent mechanical thrombectomy were retrospectively analyzed.The patients were divided into training set(n=113)and internal test set(n=48)at the ratio of 7∶3,while 79 AIS patients who underwent mechanical thrombectomy from clinical center 2 were retrospectively enrolled as external test set.According to the National Institutes of Health stroke scale(NIHSS)scores 7 days after thrombectomy,patients'prognosis were classified as good(＜15 points)or poor(≥15 points).Pre-treatment non-contrast CT images of patients were reviewed,and CT findings were comparatively analyzed.Independent predictors of patients'early prognosis after mechanical thrombectomy were obtained with sequential univariate and multivariate logistic regressions,and a predicting model was established and visualized as a nomogram.The receiver operating characteristic curve was drawn,and the distinction was assessed with the area under the curve(AUC),then calibration was assessed with Hosmer-Lemeshow goodness of fit test,and the net benefit was evaluated with decision curve analysis(DCA).Results Alberta stroke program early CT score(ASPECTS),hyperdense middle cerebral artery sign(HMCAS)and basal ganglia calcification were all independent predictors of early prognosis of AIS after mechanical thrombectomy(all P＜0.05).The predictive model was established combining the above 3 variables and then visualized as a nomogram to predict prognosis of AIS after mechanical thrombectomy,with AUC of 0.776 in internal test set(χ2=6.052,P=0.417)and 0.800 in external test set(χ2=2.269,P=0.811).DCA showed that the nomogram might provide clinical net benefit within certain threshold probability ranges.Conclusion ASPECTS,HMCAS and basal ganglia calcification were all independent predictors of early prognosis of AIS after mechanical thrombectomy.The nomogram originated from predicting model combining the three could be used to somewhat accurately predict poor early prognosis after mechanical thrombectomy.