Objective: To establish a method for the determination of norcantharidin in norcantharidin in situ gel .Methods:An optimal HPLC method was set up and an Agilent ZORBAX SB-C18 column (150 mm ×4.6 mm, 5μm) was adopted.The mobile phase was acetonitrile-phosphate buffer solution(1∶9, adjusting pH to 3.1 with phosphorjc acid).The flow rate was 0.8 ml· min-1 and the column temperature was 25℃.The detection wavelength was set at 210 nm and the injection volume was 20μl.Results:Norcanthari-din had a good linear relationship within the range of 0.02-1.00 mg· ml-1 (r=0.999 9).The average recovery was 97.5%and RSD was 0.98%(n=9).Conclusion:The method is accurate, simple and reproducible, which can be used for the determination of nor-cantharidin in norcantharidin in situ gel .

Objective:To investigate the stability of flurbiprofen axetil lipid microspheres injection combined with 0. 9% sodium chloride injection or 5% dextrose injection, and provide theoretical basis for the clinical application. Methods:The content changes of flurbiprofen axetil in the mixture of flurbiprofen axetil lipid microspheres injection and 0. 9% sodium chloride injection or 5% dextrose injection were determined in 5 h at 25℃ away from light, and the changes in the appearance and particle size of flurbiprofen axetil lip-id microspheres were investigated. The changes in the appearance and particle size of flurbiprofen axetil lipid microspheres in the mix-ture of flurbiprofen axetil lipid microspheres injection and 0. 9% sodium chloride injection before and after freezing and thawing were also investigated. Results:The appearance, particle size and content had no significant changes in all mixtures in 5 h at 25 ℃ away from light. The appearance and particle size of flurbiprofen axetil lipid microspheres in the mixture before and after freezing and thawing had no significant changes as well. Conclusion:The mixture of flurbiprofen axetil lipid microspheres injection and 0. 9% sodium chlo-ride injection or 5% dextrose injection is stable in 5 h away from light.

Objective To explore the relevant factors of diabetic retinopathy(DR).Methods 400 patients with type 2 diabetes were selected,and the relevant factors with DR of type 2 diabetes were analyzed.Results The incidence of DR in type 2 diabetes was 49.0％.In type 2 diabetes when diabetes duration＞15 years,the incidence of DR was 84.7％,duration＜5-year incidence rate was 30.0％ respectively,there were significant differences(P＜0.05).Incidence rate in lower blood glucose group and poor glycemic control group were 38.1％ and 68.5％ respectively,and there was statistically difference(P＜0.05),and in normal blood pressure and well control group,the incidence rates were 44.1％ and 41.2％,which were significantly better than 80.0％ of poorly controlled hypertension group.Conclusion Duration of diabetes,glycemic control level,blood pressure,exercise,and regular schedule were related factors of DR.The most important thing of DR treatment and prevention was to control blood glucose.

Objective To investigate the clinical features of aged patients with acute hemorrhage arising from vascular malformation of colon and the efficacy of endoscopic placement of hemoclip in treatment of hemorrhage. Methods Fifty-six patients were diagnosed with acute bleeding arising from vascular malformation of colon by colonoscopy. Patients were assigned to aged group (n = 31,≧60 years of age) and control group (n = 25, <60 years of age). The clinical manifestations and the efficacy of emergent endoscopic treatment with hemoclip application were compared between two groups. Results The complications including hypertension, coronary heart disease and diabetes were higher in aged group [80. 6% (25/31)] than in control group [40. 0% (10/25) ] with significant difference (P<0. 01). The effusive bleeding accounted for 45. 2% (14/31) in aged group and 16. 0% (4/25) in control group (P<0. 05). Endoscopic treatment with hemoclip was performed in 18 and 6 patients in aged group and control group, respectively. Initial hemostasis was achieved in 17 and 6 patients in the aged group and control group, respectively (P<0. 05). Recurrent hemorrhage occurred in 2 patients in the aged group and none in the control group (P>0. 05). The mean number of hemoclips application was 5. 3 ± 2. 4 in aged group and 3. 1 ± 1. 3 in control group (P<0. 05). Conclusions The hemorrhage is more serious in aged patients with acute hemorrhage, because they often complicate with cardiovascular diseases and diabetes. The lesion of hemorrhage can be found and precisely localized using colonoscopy. Promptly endoscopic placement of hemoclip is quite effective in treatment of acute hemorrhage.

Objective To discuss the safety and efficacy of straight light beam greenlight photoselective vaporesection of the prostate (PVRP) and photoselective vaporization of the prostate (PVP) in treatment of benign prostatic hyperplasia. Methods 113 cases of BPH were randomly divided into two groups, 62 cases in PVRP group and 51cases in PVP group. Clinical data was collected and compared between the two groups, including pre-operation and six month after operation international prostate symptom score (IPSS), quality of life (QOL), urine flow rate (Qmax), postvoid residual urine (PVR), as well as operational time, operative bleeding volume, bladder irrigation time, indwelling catheter time after operation and complications after operation. Results All the patients were operated successfully without serious complications. There was significant difference in operative time [(49.4 ± 18.9) min vs (75.1 ± 20.7) min (P < 0.05)] between the two groups. There were no significant difference in blood loss and bladder washing time after operation. The Qmax after 6 months of surgery, PVR, IPSS, QOL of the two groups had significantly improved compared with preoperative (P < 0.05), while the difference between the two groups had no significance in statistics (P > 0.05). Conclusions Treatment of straight light beam greenlight PVRP and PVP are safe and effective for BPH. Straight light beam greenlight PVRP has the advantages of shorter operation time.

Objective To construct and express human papillomavirus type 11(HPV11) E7 gene with recombinant adenovirus vectors. Methods HPV11 E7 gene was amplified by PCR and directionally cloned into vector pENTR-TOPO to form TOPO-E7 plasmid. E7 gene was transferred into the pAD/CMV/V5-DESTTM gateway vector by LR recombination reaction with pAD/CMV/V5-DESTTM gateway vectors and TOPO-E7 plasmid. The recombination vector was digested by Pac I enzyme and transfected into 293A cell by Lipofectamine method to obtain recombinant adenovirus vectors pAD-E7. Expression of E7 on HaCaT cells infected with pAD-E7 vectors was analyzed by confocal microscopy. Results The recombinant plasmid TOPO-E7 was identified and confirmed with enzyme digestion and sequencing. Recombinant adenovirus vectors pAD-E7 were generated efficiently with a titer of 1.4 ? 107 pfu/mL in transfected 293A cells. E7 protein could be identified in HaCaT cells with confocal microscope 48 h after infected with recombinant adenovirus vector. Conclusions The results indicate efficient expression of HPV11 E7 gene in eukaryotic cells by recombinant adenovirus mediated transfer, which facilitates further research of its function.

Objective:To study the formula of triptolide (TRI) self-microemulsifying drug delivery system (SMEDDS) and evaluate the pharmaceutical properties.Methods:The formula and preparation process of triptolide self-microemulsion were screened by the solubility test and pseudo-ternary phase diagram.With the average particle size and self-microemulsifying time as the indices,the further formula optimization of triptolide self-microemulsion was carried out.The pharmaceutical properties of triptolide self-microemulsion were evaluated.Results:The optimal formula of TRI SMEDDS was as follows:the amount of medium chain triglycerides (MCT) was 20%,that of polyoxyethylene castor oil (EL-35) was 40%,and that of polyethylene glycol 400 (PEG-400) was 40% in the oil phase.The average particle size was 43.48 nm,and the time of self-microemulsification was less than 30 s.Conclusion:The average particle size and the appearance of triptolide self-microemulsion are accordance with the requirements of pharmaceutics.Triptolide self-microemulsion has good sustained-release effect,which lays foundation for the further study on pharmacodynamics.

Objective To evaluate the plasma D-dimer level in the patients of type 2 diabetes mellitus with hypertension and investigate their correlation.Methods Eighty-five subjects were divided into three groups according to clinical manifestation:control group:20 subjects ; type 2 diabetes mellitus group:21 subjects; type 2 diabetes mellitus combined with hypertension group:44 subjects.The level of plasma D-dimer was measured and the difference was compared between groups.The results were showed as mean ± sd,and the difference was compared using ANOVA Test ( SPSS13.0).Results The plasma D-dimer concentrations in normal control group was ( 102.15 ± 32.48 ) μg/L,in single type 2 diabetes mellitus was ( 148.62 ± 80.99 ) μg/L,while plasma concentrations in type 2 diabetes mellitus combined with hypertension was ( 206.28 ± 92.99 ) μg/L.plasma D-dimer concentration was higher in single type 2 diabetes mellitus than that in normal control cases( P ＜0.05) ;And plasma D-dimer concentration was also found increased in type 2 diabetes mellitus combined with hypertension when compared with control group (P ＜ 0.01 ) ;And there was also significant difference on plasma D-dimer concentration between single type 2 diabetes mellitus and type 2 diabetes mellitus combined with hypertension cases ( P ＜ 0.05 ).Conclusion The plasma levels of D-dimer was increased obviously in single type 2 diabetes mellitus and type 2 diabetes mellitus combined with hypertension,it may be related to the imbalance of coagulation and fibrinolytic system.Monitoring of plasma D-dimer concentration in type 2 diabetes and patients with hypertension may have important clinical implications for the prevention of thrombotic diseases.

To evaluate the ability of microbial mix-culture fermenting syngas into ethanol, we studied the microbial mix-cultures A-fm 4, G-fm 4, Lp-fm 4 and B-fm 4 obtained by enrichment and compared with Clostridium autoethanogenum DSM10061 with 10% and 25% inoculation size. The results show that, with 10% inoculation size, the ethanol production of A-fm 4, G-fm 4, Lp-fm 4, B-fm 4 and C. autoethanogenum were 349.15, 232.16, 104.25, 79.90 and 26.99 mg/L respectively. With 25% inoculation size, the ethanol production were 485.81, 472.73, 348.58, 272.52 and 242.15 mg/L respectively. Higher inoculation size will increase the production of ethanol. The tested mix-culture exhibited a significant yield advantage compared with the maximum production of C. autoethanogenum reported in the literature (259.64 mg/L). This research provided a practical method to improve ethanol production from syngas.
Bacteria ; classification ; metabolism ; Clostridium acetobutylicum ; metabolism ; Ethanol ; isolation & purification ; metabolism ; Fermentation ; Gases ; metabolism ; Hydrogen ; metabolism

Objective:To explore the relationship between microsatellite instability (MSI) and Ki-67 expression level and the clinicopathological features of colorectal cancer, and investigate their impact for prognosis, so as to provide reference for prognostic judgment of colorectal cancer.Methods:The data of 183 patients who underwent radical colorectal cancer surgery and were diagnosed pathologically in the Department of General Surgery, the First Affiliated Hospital of Xinxiang Medical University from January 2017 to December 2019 were retrospectively analysed, including 101 males (55.2%)and 82 females(44.8%), ranged from 20 to 86 years and the mean age was(60.27±13.13)years. According to the results of mismatch repair protein immunohistochemical staining, the patients were divided into MSI-H group ( n=32) and MSI-L/MSS group ( n=151). According to the results of Ki-67 antigen immunohistochemical staining, the patients were divided into low Ki-67 expression group (<82.5%, n=136) and high Ki-67 expression group (≥82.5%, n=47) , among which 62 cases (78.5%) with low Ki-67 expression and 17 cases (21.5%) with high Ki-67 expression were in patients with Ⅲ+ Ⅳ stage colorectal cancer. The data of clinicopathological features, disease-free survival, and overall survival were collected and analyzed. The cotegorical variables were presented as n(%), and the comparisons between groups were performed using Chi-square test or Fisher′s exact test. The multivariate Logistic regression model was used to estimate the correlation between microsatellite instability and Ki-67 expression level and clinicopathologic characteristics of colorectal cancer. Kaplan-Meier survival curve and COX proportional hazards regression model were used to analyze the correlation between microsatellite instability and Ki-67 expression level and disease-free survival and overall survival. Results:Single factor analysis showed that the differences in gender ( χ2=4.37, P=0.037), tumor site ( χ2=26.40, P<0.001), tumor maximum diameter ( χ2=11.12, P=0.001) and nerve invasion ( χ2=5.53, P=0.019) between MSI-H group and MSI-L/MSS group were statistically significant. Multivariate Logistic regression model analysis showed that only gender ( OR=3.013, 95% CI: 1.183-7.672, P=0.021), tumor location ( OR=0.167, 95% CI: 0.067-0.419, P<0.001) and nerve invasion ( OR=0.202, 95% CI: 0.042-0.968, P=0.045) were independently correlative factors for MSI status. In Ⅲ+ Ⅳ stage colorectal cancers, the difference in tumor site between low Ki-67 expression group and high Ki-67 expression group was statistically significant( χ2=3.91, P=0.048). Multivariate Cox proportional hazards regression model analysis revealed that high Ki-67 expression ( HR=0.301, 95% CI: 0.118-0.768, P=0.012; HR=0.275, 95% CI: 0.083-0.912, P=0.035) and MSI-H ( HR=0.072, 95% CI: 0.010-0.525, P=0.009; HR=0.122, 95% CI: 0.017-0.900, P=0.039) were independently protective factors for disease-free survival and overall survival. Conclusions:MSI-H colorectal cancer is common in males, right-sided colonic cancers and non-neuroinvasive patients. In stage Ⅲ+ Ⅳ colorectal cancer, the expression level of Ki-67 in right-sided colonic cancer was lower than in left-sided colorectal cancer. Patients with MSI-H and high Ki-67 expressive colorectal cancer had longer disease-free survival, longer overall survival and better prognosis.