purpose To construct the expression vector of Hexastatin gene,to express and to purify the recombinant protein for further activity research.Methods The human Hexastatin gene was isolated by RT-PCR from EC9706 cells total RNA and cloned into pMDl8-T for sequencing.Then the Hexastatin gene was subcloned into pMAL-c4x expression vector and induced to express by IPTG.The recombinant fusion protein was purified with Amylose Resin Heads.Results RT-PCR product was about 687 bp and its sequence was the same as that of Hexastatin reported.The recombinant protein was expressed in E.coli BL21 with high level and the soluble protein accounted for 24.8% of the total bacterial protein,The purification of recombinant protein purified with Amylose Resin Heads reached more than 90%.Conclusion The cloning,expression and purification of human Hexastatin have laid a foundation for its anti-angiogenesis therapy for tumor.

Objective To study the influence of Noradrenergic systems in Lateral Hypothalamus Area (LHA) on small intestine moving. Methods The effect of noradrenalin、noradrenalin+phentolamine、noradrenalin+propolol、phentolamine on small intestines electro-activity of rats was detected by external alimentary canal electrodes and central nervous system stereo-configuration technology. Results After injecting 2 g/L noradrenaline (NE) the cyclic period of MMC extended, the ratio of the active time to the cyclic period and the number of the fast wave within the active time per minute reduced. NE inhibited the electro-activity of small intestine of rats, and the average effective time was (36.86?7.39)min. Hence injecting 5 g/L PE alone to LHA raised the ratio of the MMC active time to the cyclic period. Phentolamine presented a slight excitability on the electro-activity of small intestine of rats. Conclusion The NE in LHA showed the inhibitive myoelectric activity on small intestines, and this effect was introduced through a acceptor,The reacceptor in LHA may function in the inhibition of the electro-activity in small intestines.

Objective To determine the content of trigonelline in Radix Mirabilis.Methods A RP -HPLC method was established.The chromatographic c olumn was Inertsil NH 2 Columns(250?4.6mm,5?m.).The mobile phase was acetonitrile -water(80∶20).The flow rate was 0.8mL?min -1 and the detection wavelength was at 265nm,and the column temperature was 30℃.Results The calibration curve was linear in t he range of 0.1872～0.9360?g .The aver-age recovery for trigonelline was 99.6%and RSD was 1.76%(n=6).Conclusion The method is simple and effec-tive and can be used to determine the c ontent of trigonelline in Radix Mirabilis.

Objective To determine the content of quercetin in Semen Hoveniae.Methods A RP-HPLC method was established.The chromatographic column was Synergi 4u Fusion-RP 80.The mobile phase was acetonitrile-0.5 %phosphoric acid(33 ∶67).The flow rate was 1.0 mL?min-1,the detection wavelength was at 360 nm,and the column temperature was 30 ℃.Results The average recovery of quercetin was 98.6 %,and RSD was 2.39 %(n=6).Conclusion The method is effective,simple and accurate,and can be used to determine the content of quercetin in Semen Hoveniae.

Objective To investigate the angiographic manifestations of renal artery injury caused by percutaneous nephrolithotomy, and to evaluate the therapeutic effect of super-selective renal arterial embolization in treating renal artery injury. Methods A total of 22 patients with persistent or intermittent gross hematuria that occurred after percutaneous nephrolithotomy, who were encountered at authors’ hospital during the period from Jan. 2010 to June 2014, were included in this study. The diagnosis was confirmed by renal angiography in all patients, and super-selective renal arterial embolization with steel micro-coils was carried out in all patients. The patients were followed up for three months. The results were analyzed. Results Of the 22 patients, DSA examination showed that renal artery pseudoaneurysm (RAP) was found in 14 (63.6%), renal arteriovenous fistula (RAVF) in 5 (22.7%) and RAP associated with RAVF in 3 (13.6%). Renal angiography performed after super-selective renal arterial embolization showed that complete obstruction of the bleeding arteries was achieved in all patients, and the active bleeding stopped. Both the technical success rate and the hemostasis rate were 100%. During the follow-up period lasting for three months, no recurrence of hematuria or severe complications occurred. In 20 patients, different degree of embolism syndrome was observed after the treatment. Conclusion Renal artery pseudoaneurysm and renal arteriovenous fistula are the main types of renal artery injury after percutaneous nephrolithotomy. Super-selective renal arterial embolization with micro-coils can be used as the treatment of choice for patients who has failed to respond to conservative therapy.

OBJECTIVE: To explore the clinical effects and short-term complications of using free jejunum graft (FJG) to reconstruct the defects by resections of pharyngeal, laryngeal or cervical esophageal cancers. METHOD: Fifty-eight cases of pharyngeal, laryngeal or cervical esophageal cancers were reconstructed with FJG. All cases were analyzed retrospectively. RESULT: The success rate of FJG transplantations was 91.4% (53/58). The incidence of post-operative short-term complication was 43.1% (25/58), which was not related to age or BMI. The most common complication was anastomotic leakage (18.9%), which was not related to per-operative radiation therapy. However, BMI > 25 cases had significantly higher incidence of anastomotic leakage than BMI ≤ 25 cases (P = 0.009). The second and third most common complications were respiratory system complications (10. 3%) and FJG necrosis (8. 6%). Para-operative death rate was 3.4% (2/58). Two-year overall survival rates of hypopharyngeal cancer and cervical esophageal cancer were 49% and 67% respectively. The group with no short-term complications had a slightly better survival rate than the group with short-term complications from the Kaplan-Meier curve, but there was no significant difference (P = 0.103). CONCLUSION FJG is ideal to reconstruct cervical digestive tract circumferential defects with a high success rate and a low mortality. However, the post-operative complication rate is high. Intensive observation, early detection and timely treatment of complications are crucial.
Esophageal Neoplasms ; surgery ; Humans ; Hypopharyngeal Neoplasms ; surgery ; Jejunum ; transplantation ; Laryngeal Neoplasms ; surgery ; Pharyngeal Neoplasms ; surgery ; Postoperative Complications ; Postoperative Period ; Retrospective Studies ; Survival Rate

Objective The anti-tumor effect by sequential treatment with Newcastle disease virus (NDV) strain 7793 and 5-FU in liver-metastases mice model was evaluated and immune-active response stimulated by sequential therapy was investigated. Methods Liver metastasis mice model was established by intra-peritoneal injection. The model mice were randomly divided into 3 groups, being given PBS (0.1 mL/d,10 d), NDV7793 [512 HU/(kg·d),5 d] and NDV7793[512 HU/(kg·d),5 d] + fluorouracil [5-FU,10 mg/(kg·d),5 d]. The effect on survival time,body weight,liver weight change and the formation of liver metastasis in tumor-bearing mice model were detected after different treatments in evaluating the regression of mice liver metastasis by sequential therapy. The detection of thymus index and IFN-γ concentrations in liver tissue of tumor-bearing mice model may indicate the stimulation of immune-active response by sequential therapy. Results The mean survival time of tumor-bearing mice treated with NDV7793 and 5-Fu sequentially was 32 d , which was significantly higher than those of tumor-bearing mice treated with NDV7793(30 d) or PBS injections (17 d), respectively (P< 0.05); The metastatic foci of tumor-bearing mice treated with NDV and 5-FU sequentially (30.60 ± 9.32) which was significantly less than those of tumor-bearing mice treated with PBS injection (273.30 ± 30.73), (P <0.05), seem quite similar to those treated with NDV (24.83 ± 6.90),(P > 0.05), and the liver weight was lighter than PBS (P < 0.05); Compared with NDV treatment, the decreased thymus index and increased amount of the effector IFN γ were observed in tumor-bearing mice treated with NDV 7793 and 5-FU sequentially (P <0.05). Conclusions The sequential therapy with Newcastle disease virus 7793 strain and 5-FU was observed to co-exert a significant suppressive effect in liver metastases of colon cancer cells in tumor-bearing mice model. Compared with NDV treatment , the survival time of mice model and the induction of antitumor effector molecules were significantly improved after sequential therapy.

ObjectiveTo optimize human Hexastatin gene,to express,purify protein and conduct activity experimental research,and to provide a theoretical basis for further study of Hexastatin.MethodsHuman Hexastatin gene was optimized and synthesized.It was connected to the pET28a expression vector,induced to express by isopropyl β-D-1-thiogalactopyranoside(IPTG),and optimized induction conditions.After the ultrasonication of bacterial cells and inclusion bodies,the recombinant fusion protein was purified with Ni-NTA chromatographic column,analyzed and identified by SDS-PAGE and Western Blot,and conduct activity experimental research in vitro by MTT.ResultsConstructed production was pET28a-Hexastatin expression plasmid.The human Hexastatin protein was expressed in E.coli BL21 the high level and accounted for 45.1％ of the total bacterial protein.The purification of recombinant protein purified with Ni-NTA chromatographic column reached 90％,and the concentration was 80 μg/ml.Human Hexastatin protein can restrain the growth of C6,MCF-7 and human vascular endothelial cell (HMEC) cells,and inhibition ratio reach to 72.9％±3.6％,48.8％±2.9％,52.7％±2.5％,respectively through MTT test.ConclusionThe optimized human Hexastatin protein was expressed successfully,which confirmed the inhibition to tumour cells.It is a new way for anti-angiogenesis therapy of tumour.

Objective To rapidly identify and assign the chemical components in Sugemule-4 decoction by using UPLC Orbitrap Exploris MS/MS,and then conduct network pharmacological analysis,and clarify the anti-insomnia effect of Sugmule-4 decoction.Methods ACQUITY UPLC BEH C18(2.1 mm×100 mm,1.7 μm)Chromatographic column,with 0.1％formic acid aqueous solution(A)-0.1％formic acid acetonitrile solution(B)as mobile phase gradient elution.Flow rate:0.5 mL·min-1,injection volume:5 μL.Under the control of the control software(Xcalibur,Thermo Fisher Scientific),the primary and secondary mass spectrometry data are collected based on the FullScan-ddMS2 function.Combined with the self-built database and online database,the chemical components in Sugemule-4 decoction are identified by accurate relative molecular weight,fragment ion information,pyrolysis pattern and comparison of control samples.The identified chemical components were analyzed by network pharmacology,and the targets of drug and insomnia were obtained from TCMSP,SwissTargetPrediction,GeneCards,OMIM,TTD,DrugBank and DisGeNET databases respectively,and the intersection target was obtained;Protein interaction analysis was performed through STRING database.Use Metascape platform to analyze GO function and KEGG pathway,and predict the mechanism of Sugmule-4 decoction in treating insomnia.To determine the anti-insomnia effect of Sugmule-4 decoction,SD rats were divided into normal control group(K),model group(M),Sugmule-4 decoction group(S)and Anshenbunao liquid group(A).After chronic unpredictable mild stress and PCPA were used for modeling,the rats were given corresponding drugs by gavage for 7 days,then the open field text was conducted to observe the general state and behavioral changes,and detection of AChE,5-HT,and GABA levels in serum and brain tissue of rats in each group using ELISA method.Results 136 components were identified from Sugemule-4 decoction,including 37 terpenoids,34 alkaloids,26 flavonoids,20 phenylpropanoids,14 phenolic acids and 5 amino acids.The results of network pharmacology analysis show that the mechanism of Sugemule-4 decoction in treating insomnia is mainly achieved by its components such as Romucosine D,Nornantenine,Nuciferine,and(R)-ar Turbone acting on targets such as ACHE,ADORA2A,and CHRM1 through signal pathways such as Neuroactive ligand-receptor interaction,synaptic signaling,and trans synaptic signaling.This is a multi component-multi target-multi signal pathway joint implementation process.The results of pharmacodynamic experiment showed that Sugmule-4 decoction and Anshenbunao liquid could revise the general state and behavioral changes of insomnia model rats,increase the content of AChE,5-HT and GABA in serum(P<0.01).Conclusion This study comprehensively reflects the chemical components of Sugmule-4 decoction,and preliminarily summarizes the mass spectrum decomposition characteristics of various chemical components.The network pharmacology and pharmacodynamics experiments show that Sugmule-4 decoction has significant anti-insomnia effect,which provides a reference basis for the quality control and mechanism study of Sugmule-4 decoction.

Objective:To compare the clinical efficacy of anti-reflux mucosectomy (ARMS) and endoscopic cardial constriction ligation (ECCL) on treatment of gastroesophageal reflux disease.Methods:A retrospective study was conducted on the data of 48 consecutive patients with gastroesophageal reflux disease, who underwent ARMS or ECCL at the First Affiliated Hospital of Zhengzhou University from December 2015 to August 2018. Twenty cases were in the ARMS group and 28 cases in the ECCL group. The short-term and long-term efficacies were compared between the two groups.Results:The success rate of operation was 100.0% in the both groups. The operation time of the ECCL group was significantly shorter than that of the ARMS group (8.43±1.59 min VS 34.05±12.35 min, t=-9.227, P<0.001). After 2 months follow-up, the symptom improvement rate of the ECCL group and the ARMS group was 89.3% (25/28) and 60.0% (12/20), respectively ( χ2=4.128, P=0.042). The GERD Q score of the ECCL group was significantly lower than that of the ARMS group (6.24 ±1.22 VS 7.35±1.79, t=-2.400, P=0.023). One year after operation, there were no significant differences in the symptom improvement rate, GERD Q score, DeMeester score and the time percentage of pH<4 between the two groups ( P>0.05). Conclusion:The long-term clinical effect of ARMS and ECCL is similar, but the short-term clinical effect of ECCL is superior to ARMS.