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.

Nutritional risk screening is the priority for nutrition support after stroke.The advance in nutritional risk screening after stroke was reviewed in order to provide assistant information for post-stroke nutrition support and nursing care.

Objective To observe the effects of MSH2 gene re expression on estrogen-induced apoptosis of colon cancer cells LOVO,and to explore its mechanisms.Methods According to different plasmid and whether with estradiol intervention,colon cancer LOVO cells were divided into empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,estrogen receptor (ER) β with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group,and received corresponding treatment.The expression of MSH2,ERβ protein and apoptosis related caspase 3 protein were detected by Western blotting.Cell viability was measured by cell counting kit-8.Cell DNA fragments of each group were isolated with apoptosis DNA fragments isolation kit.And the DNA ladder was observed.The rate of apoptosis was detected by flow cytometer.Single factor variance analysis was performed for comparison among multiple groups,and t test was used for comparison between the two groups.Results After transfection,the expression of the MSH2 and ERβ at protein level in LOVO cells significantly increased and neither of their expression was effected by estradiol.The expression levels of caspase 3 cleavaged active fragments of ERβ with estradiol group and ERβ with MSH2 and ethanol group were higher than other groups,and there was no significant difference between these two groups.The LOVO cell viability of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 1.72 ±0.25,1.74 ± 0.31,1.77 ± 0.35,1.74±0.33,1.70±0.34,1.02±0.48,1.71±0.31 and 1.07±0.18,respectively,and the differences between the groups were statistically significant (F=3.791,P＜0.05).Among them,the LOVO cell viability of ERβ with estradiol group was lower than that of ERβ with ethanol group,accordingly,that of ERβ with MSH2 and estradiol group was lower than that of ERβ with MSH2 and ethanol group,that of ERβ with estradiol group was lower than that of empty plasmid with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t=3.158,3.075,3.648,3.253,all P＜0.05).DNA ladder formed from DNA fragments of apoptosis cells was seen in ERβ with estradiol group and ERβ with MSH2 and estradiol group.The apoptosis rate of empty plasmid with ethanol group,empty plasmid with estradiol group,MSH2 with ethanol group,MSH2 with estradiol group,ERβ with ethanol group,ERβ with estradiol group,ERβ with MSH2 and ethanol group and ERβ with MSH2 and estradiol group was 7.86±0.19,7.87±0.39,8.39±1.02,9.05±1.54,7.54±0.99,19.77±2.35,7.76±1.32 and 19.30±1.75,respectively,and the differences between groups were statistically significant (F=45.436,P＜0.05).Among them,the apoptosis rate of ERβ with ethanol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and ethanol group was lower than that of ERβ with MSH2 and estradiol group,that of empty plasmid with estradiol group was lower than that of ERβ with estradiol group,that of ERβ with MSH2 and estradiol group was lower than that of MSH2 with estradiol group,and the differences were statistically significant (t =8.260,9.133,8.596,7.617,all P＜ 0.05).Conclusions Estrogen may promote colon cancer cell apoptosis through ERβ pathway.The process of apoptosis maybe related with caspase protein,MSH2 may not be involved in the regulation of this signal pathway.

Objective To explore the therapeutic value of modified endoscopic submucosal dissection with ligation (ESD-L) for small tumors originated from gastric muscularis propria.Methods Total of 60 patients with tumors originated from gastric muscularis propria,which was confirmed by endoscopic ultrasonography and smaller than 12 mm and were recruited to the present study.The conventional ESD technique was used to dissect the tumor to the depth of muscularis propria.Then the bottom of the tumor and the beneath muscularis propria were ligated fully with a nylon loop.Further dissection was applied till the whole tumor was isolated.Results All 60 lesions were dissected completely with perforation occurred in 10 cases,which were managed successfully with metal hemoclip.Pathologic diagnosis was obtained in all lesions,and no recurrence was found during the follow-up.Conclusion Modified technique of ESD-L is effective for complete resection of small tumors originated from gastric muscularis propria,which can also decrease the risk of stomach perforation.

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.

OBJECTIVETo establish and experimental verification the mathematical model of the balance groups that is the steady-state of traditional Chinese medicine in extraction.

METHODUsing the entropy and genetic principles of statistics, and taking the coefficient of variation of GC fingerprint which is the naphtha of the Houttuynia cordata between strains in the same GAP place as a pivot to establish and verify the mathematical model was established of the balance groups that is the steady-state of traditional Chinese medicine in extraction.

RESULTA mathematical model that is suitable for the balance groups of the steady-state of traditional Chinese medicine and preparation in extraction, and the balance groups which is 29 683 strains (approximately 118.7 kg) were gained with the same origin of H. cordata as the model drug.

CONCLUSIONUnder the GAP of quality control model, controlling the stability of the quality through further using the Hardy-Weinberg balance groups of the H. cordata between strains, the new theory and experiment foundation is established for the steady-state of traditional Chinese medicine in extraction and quality control.

Chemical Fractionation ; methods ; Chromatography, Gas ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; standards ; Houttuynia ; chemistry ; genetics ; Models, Statistical ; Quality Control

Objective: To assess the efficacy and safety of the endoscopic anti-reflux mucosectomy for gastroesophageal reflux disease. Methods: Data of 18 patients with gastroesophageal reflux disease who underwent endoscopic anti-reflux mucosectomy at the First Affiliated Hospital of ZhengZhou University from December 2015 to July 2018 were retrospectively studied. The therapeutic effects (improvement of heartburn and reflux symptoms, 24 h esophageal pH monitoring) and complications were analyzed. Results: Anti-reflux mucosectomy was performed successfully in all patients with successful rate of 100%. ESD was performed in 8 cases and EMR in 10 cases.24 h esophageal pH monitoring results showed that the Demeester score, the time percentage of pH < 4, total reflux events and reflux times of pH < 4 with time longer than 5 minutes after treatment were significantly lower than those before treatment (20.16±9.12 VS 74.16±20.03, (2.70±0.88)% VS (6.42±1.37)%, 43.78±19.68 VS 156.56±41.22, 2.89±1.68 VS 9.89±2.95, all P<0.05). No bleeding, perforation or infection was observed after the procedure. During a follow-up period of 3-34 months, symptom relief rate was 89% (16/18). A tightened cardiac was noted in 18 cases and recovery of mucosal damage was found in 16 patients. Conclusion Anti-reflux mucosectomy is safe, effective and easy to operate for gastroesophageal reflux disease.

Objective To summarize the preliminary experience of endovascular recanalization in treating chronic symptomatic internal carotid artery occlusion.Methods Four patients with chronic symptomatic internal carotid artery occlusion,admitted to and underwent endovascular recanalization in our hospital from August 2013 to August 2014,were chosen in our study; their clinical data were retrospectively analyzed.Results Four patients were successfully opened the internal carotid arteries;cerebral CT angiography showed that all arteries were unobstructed.One appeared intra-operative iatrogenic internal carotid artery cavernous sinus fistula,and successful occlusion of the fistula with internal carotid artery patency was achieved after one week.Three months after the operation,two showed unobstructed internal carotid artery by DSA and the other stated no abnormalities during the telephone follow up.Conclusion Endovascular recanalization is a safe and effective treatment method for chronic symptomatic internal carotid artery occlusion.

Data of 55 cases of gastric neuroendocrine neoplasms (G-NENS) with diameter ≤12 mm in the First Affiliated Hospital of Zhengzhou University from August 2014 to August 2019 were retrospectively analyzed. According to the methods of endoscopic resection, the patients were divided into two groups: the endoscopic mucosal resection with a cap (EMR-C) group (35 cases) and the endoscopic submucosal dissection (ESD) group (20 cases). The results showed that the success rates of operation, the whole resection rates and the complete resection rates were all 100.0% in the two groups. Compared with the ESD group, the EMR-C group had a shorter median operation time (12.00 min VS 28.35 min, P<0.001), less mean hospitalization costs (21 165.19 yuan VS 28 400.35 yuan, P=0.004), and a similar overall incidence of complications [2.86% (1/35) VS 0, P=1.000]. By March 2020, the recurrence rate of EMR-C group and ESD group were 28.6% (10/35) and 15.0% (3/20), respectively, without significant difference ( P=0.418). It is suggested that for G-NENS with diameter ≤12 mm, without muscular invasion, lymph node metastasis or distant metastasis, EMR-C and ESD are both safe and effective, but EMR-C has more advantages in terms of operation time and hospitalization costs.

Objective:To compare the clinical efficacy of endoscopic resection and laparoscopic surgery in the treatment of gastric gastrointestinal stromal tumor (GIST) with a maximum diameter of 2 to 5 cm, and to analyze the influence of factors such as tumor surface, growth pattern and lesion origin on the choice of resection method, so as to provide a safer and more effective treatment for patients with gastric GIST.Methods:From January 2012 to November 2019, at the First Affiliated Hospital of Zhengzhou University, the clinical data of 301 patients with gastric GIST who underwent endoscopic resection (137 cases in the endoscopic resection group) or laparoscopic surgery (164 cases in the laparoscopic surgery group) were retrospectively analyzed, including age, gender, whether there was depression on the tumor surface (the local subsidence depth of the mucosa on the tumor surface was >5 mm), whether the tumor surface was irregular (non-hemispherical or non-elliptical tumor surface), whether there was combined ulcer, location, shape, origin of the lesion, growth pattern (intralumina growth or combined intraluminal and extraluminal growth), risk classification (very low risk, low risk, medium risk, high risk), whether the tumor was en bloc resection, operation time, whether bleeding or not, fasting time, indwelling time of gastric tube, time of hospitalization, time of postoperative hospital stay, postoperative complications and follow-up. Independent sample t test, chi-square test or Fisher′s exact test and Wilcoxon rank sum test were used for statistical analysis. Results:Among the 137 patients with gastric GIST in the endoscopic resection group, 85 cases (62.0%) underwent endoscopic submucosal dissection, 9 cases (6.6%) underwent endoscopic submucosal excavation, 42 cases (30.7%) underwent endoscopic full-thickness resection, and 1 case (0.7%) underwent submucosal tunnel endoscopic resection. There were no significant differences in gender, age, lesion location, tumor size, and risk classification between the endoscopic resection group and the laparoscopic surgery group (all P>0.05). The tumor surface was depressed, with ulcer or irregular in 1, 49, 26, and 2 cases of patients with gastric GIST of very low risk, low risk, medium risk and high risk, respectively. There was statistically significant difference in the proportion of depression, irregularity and ulcer on the tumor surface at different risk levels ( Z=-2.55, P=0.011). The complete tumor resection rate of the endoscopic resection group was lower than that of the laparoscopic surgery group (86.1%, 118/137 vs. 100.0%, 164/164), and the difference was statistically significant ( χ2=24.28, P<0.001). However the operation time, fasting time, the indwelling time of gastric tube, time of hospitalization, and the time of postoperative hospital stay of the endoscopic resection group were shorter than those of the laparoscopic surgery group, and the total hospitalization cost was lower than that of the laparoscopic surgery group (90.0 min (62.5 min, 150.0 min) vs. 119.5 min, (80.0 min, 154.2 min); 3 d (3 d, 4 d) vs. 5 d (4 d, 7 d); 3 d (2 d, 4 d) vs. 4 d (2 d, 6 d); 11 d (10 d, 14 d) vs. 16 d (12 d, 20 d); 7 d (6 d, 9 d) vs. 9 d (7 d, 11 d); (38 211.6±10 221.0) yuan vs. (59 926.1±17 786.1) yuan), and the differences were statistically significant ( Z=-2.46, -7.12, -4.44, -6.89 and -5.92, t=-13.24; all P<0.05). The incidence of postoperative abdominal pain and other severe postoperative complications (including shock, respiratory failure, pulmonary embolism, gastroparesis, etc.) of the endoscopic resection group were all lower than those of the laparoscopic surgery group (16.8%, 23/137 vs. 27.4%, 45/164; 0.7%, 1/137 vs. 4.9%, 8/164), and the differences were statistically significant ( χ2=4.84, Fisher′s exact test, P=0.028 and 0.043). There were no significant differences in the incidence of intraoperative bleeding, postoperative bleeding, fever and perforation between the two groups (all P>0.05). The incidence of operation-related complications of lesions with intraluminal growth and originating from muscularis propria in the endoscopic resection group were lower than those of the laparoscopic surgery group (19.5%, 25/128 vs. 32.6%, 45/138; 12.6%, 12/95 vs. 31.4%, 37/118), and the differences were statistically significant ( χ2=5.86 and 10.42, P=0.016 and 0.001). There was no significant difference in the postoperative tumor recurrent rate between the endoscopic resection group and the laparoscopic surgery group (0, 0/137 vs. 2.4%, 4/164; Fisher’s exact test, P=0.129). Conclusions:Endoscopic treatment is safe and effective for gastric GIST with a maximum diameter of 2 to 5 cm, which is superior to laparoscopic surgery. However, laparoscopic surgery is recommended for tumor with depressed, ulcerative, or irregular surface and combined intraluminal and extraluminal growth.