Objective To explore the best technology to produce recombinant mouse endostatin by thioredoxin fusion expression system.Methods The recombinant plasmid pThioHis-endo was further transformed into different E.coli.,including BL21,JM109,DH5?.After induction with IPTG of different concentration or for different time period,thioredoxin-endo fusion protein was expressed in E.coli.and the product was identified by SDS-PAGE.The recombinant endostatin expression and extraction,wash,degeneration and refolding of inclusion bodies were carried out by the optimized route,and the product was further purified by affinity chromatography through Ni~(+) column and identified by SDS-PAGE.Results No difference of endostatin expression in BL21,JM109,DH5? was found.The optimal concentration of IPTG was 0.9 mmol/L and optimal inductive phase was 5 h.The soluble recombinant endostatin could be obtained by affinity chromatography through Ni~(+) column.Conclusion Soluble endostatin recombinant fusion protein of high purity and yield could be obtained by the optimized technique route.

Objective To discuss the risk factors of free wall rupture (FWR) in acute ST-segment elevation myocardial infarction (STEMI) patients. Methods We retrospectively reviewed all patients (n=1247) with STEMI hospitalized in CCU from January 2005 to July 2010. Results FWR occurred in 29 patients(2.3%). Of these 1247 patients, 128 (10.2%) patients received thrombolytic therapy, 623 (50.0%) patients underwent primary PCI. Compared to No-FWR group, FWR group has signiifcant differences in age (62.4±6.4 y vs. 66.6±8.3 y, P<0.05), hypertenion (29.7%vs. 21.8%, P>0.05), diabetes mellitu (55.2%vs. 23.5%, P=0.022), presence of heart failure on admission (Killip≥Ⅱ) ( 16.4%vs. 34.0%, P<0.05), Peak value of hCRP[20.33 (15.02, 81.25) vs. 43.35 (16.56, 126.78)], no-history of pervious MI (10.3%vs. 18.4%, P=0.018). Multivariate logistic regression showed that age (≥70 years old), Killip≥Ⅱ, hCRP ( > 100 mg/L) and thrombolytic therapy were independent risk factors of FWR. Conclusions STEMI patients with advanced age, Killip≥Ⅱ, hCRP and thrombolytic therapy were more vulnerable of FWR.

Objective: To preliminarily investigate the relationship between the baseline level of serum soluble ST2 (sST2) and 30-day MACE occurrence rate in patients with ST-elevation myocardial infarction (STEMI).
Methods: A total of 121 patients with confirmed diagnosis of STEMI in our hospital from 2015-05-01 to 2015-07-30 were consecutively enrolled. According to baseline sST2 level, the patients were divided into 2 groups:Low sST2 group, the patients with sST2≤56.68 ng/ml, n=61 and High sST2 group, the patients with sST2>56.68 ng/ml, n=60. Clinical condition and 30-day MACE (defined as death and new onset of congestive heart failure) occurrence rate were compared between 2 groups.
Results: ① The systolic blood pressure (SBP), Killip class≥II grade, blood levels of cTNI, NT-proBNP, hs-CRP and LVEF were different between 2 groups, all P<0.05. ② Baseline sST2 level was positively related to heart rate (r=0.271, P=0.003), Hs-CRP (r=0.359, P=0.000), cTNI (r=0.324, P=0.000) and NT-proBNP (r=0.425, P=0.000);negatively related to SBP (r=-0.226, P=0.013) and LVEF (r=-0.406, P=0.000).③30-day MACE occurrence rate was different between 2 groups (8.2%vs 30%, P=0.002). ④ Multivariate Cox regression analysis presented that sST2>56.68 ng/ml was the risk factor for 30-day MACE occurrence (HR=1.152, 95%CI 1.078-1.231, P=0.000).
Conclusion: Increased baseline level of sST2 implied the higher incidence of death and new onset of congestive heart failure in STEMI patients.

AIM: To investigate the changes of cytosolic free calcium concentration ([Ca 2+ ] i) and expression of Bcl-2 in HL-60 cells treated by 6F isolated from Pteris semipinnata L. (PSL), and to discuss the relations between calcium ion and cytotoxicity and DNA fragment induction effects of 6F. METHODS: HL-60 cells were used as in vitro model. [Ca 2+ ] i was measured on fluorescent spectrophotometry using Fura-2/AM as Ca 2+ indicator. Bcl-2 expressing level was measured by flow cytometry. Tetrazolium salt (MTT) and diphenylamine staining methods were applied for cytotoxicity assay and DNA fragmentation detection, respectively. RESULTS: [Ca 2+ ] i increased obviously in a dose and time dependent manner after treated HL-60 cells with 6F. 6F decreased the expressing level of Bcl-2. Adding 2 mmol/L Ca 2+ to the medium, or 1 mmol/L EDTA to chelate Ca 2+ , or 4 ?mol/L calcium ionophore A 23187 to increase the concentration of cytosolic Ca 2+ , the DNA fragment induction by 6F was not affected, whereas the cytotoxicity of 6F was enhanced. 250 ?mol/L Zn 2+ attenuated the DNA fragment induction, and the cytotoxicity of 6F against HL-60 cells was enhanced significantly. CONCLUSION: It was speculated that the decreased expressing of Bcl-2 by compound 6F was related to increased [Ca 2+ ] i in HL-60 cells, and DNA fragment induction was possibly catalyzed by Ca 2+ - independent DNase.

AIM To investigate the effects of tranfection of IL 2R antisense RNA expression plasmids on mouse spleen cells' proliferation in vitro and its possible mechanism. METHODS Spleen cells were transfected with IL 2R antisense RNA eukaryotic expression plasmids using adhesion assisted lipofection method, and then the spleen cells were stimulated by mitogen. Cells' proliferation was tested by tetrazolium salt (MTT) method. IL 2R mRNA and protein expression level were measured by slot blot hybridization assay and flow cytometry method respectively. RESULTS The proliferation of spleen cells was inhibited obviously after transfecting with recombinant plasmids. The inhibitory rate of pcAnti mIL 2R?? and pciAnti mIL 2R?? transfected group was higher than that of pcAnti mIL 2R? and pcAnti mIL 2R? transfected group; the inhibitory rate of pcAnti mIL 2R? tranfected group was higher than that of pcAnti mIL 2R? tranfected group. No inhibitory effect on the growth of NIH3T3 cells was observed when they were transfected with recombinant plasmids. IL 2R mRNA and protein expression level were decreased in spleen cells after transfection of recombinant plasmids. CONCLUSION IL 2R antisense RNA can efficiently inhibit the proliferation of mouse spleen cells in vitro. IL 2R?? chimeric antisense RNA showed higher inhibitory rate than ? or ? antisense RNA. IL 2R? antisense RNA was more effective than ? antisense RNA. It can be concluded preliminarily that the inhibitory effect of IL 2R antisense RNA was exclusively on the growth of cells functionally expressing IL 2R. The inhibitory effect on the spleen cells proliferation was likely due to the blocking of IL 2R expression by antisense RNA.

Objective To summarize the diagnosis and surgical treatment of mesenteric cyst in children.Methods The clinical records of mesenteric cyst cases from January 2011 to December 2015 were reviewed retrospectively.The diagnosis and treatment options were analyzed, and the prognosis of laparoscopic surgery and laparotomy was compared.Results The main clinical symptoms included abdominal mass, abdominal pain, and abdominal distension.Abdominal ultrasound and/or CT scan were the diagnostic tools in all cases.Traditional laparotomy was performed in 14 cases, while laparoscopy in 7 cases (1 case switched to laparotomy).2 cases had emergency surgery due to acute abdomen, laparotomy and laparoscopy in each case.Simple cyst resections were completed in 14 cases, of which 2 cases with a small amount of residual in the mesenteric root.Intestinal resection and anastomosis were required in other 7 cases.The average time of hospital stay for laparotomy group was 12 days, and 10.14 days for laparoscopy group.There was no significant difference.All patients were discharged without postoperative complications.With 1-4 years follow-up, there was no recurrence.Conclusion The operation for mesenteric cysts depends on the relationship between the cyst and the adjacent bowel or organs, and the overall outcome is favorable.The selective use of laparoscopy will benefit more children.

Acute gastrointestinal dysfunction is a common and important complication of sepsis. As no exiting formal definition and classification of gastrointestinal dysfunction, most of the treatment strategies for gastrointestinal dysfunction are not based on clinical evidence, but on their own clinical experience. Experts of traditional Chinese medicine, integrated traditional Chinese and Western medicine and Western medicine from various disciplines in Shanghai are organized by the Shanghai Society of Integrated Traditional Chinese and Western Medicine and the Emergency Department Branch of Shanghai Physicians Association. After repeated discussion, literature search and formulation of the outline, we developed consensus on gastrointestinal dysfunction secondary to sepsis with integrating Traditional Chinese Medicine and Western medicine by consulting extensively on clinical experts in the fields of emergency medicine, gastroenterology, general surgery, infectious medicine and traditional Chinese medicine, and holding several expert forums and consultation meetings. This clinical expert consensus focused on acute gastrointestinal injury (AGI) classification and inducer of sepsis. In this consensus, the common symptoms, diagnosis, classifications, treatment strategies and suggestions of acute gastrointestinal injury or dysfunction secondary to sepsis were explored from the aspect of both Traditional Chinese Medicine and Western medicine.

Objective:To observe the clinical effect of hormone replacement therapy(HRT) on patients with cervical squamous cell carcinoma after operation and its effect on sex hormone, blood fat, bone content and tumor markers, and to evaluate the clinical significance of HRT therapy.Methods:From January 2016 to January 2019, a total of 100 patients with cervical squamous cell carcinoma admitted to Zhuhai Maternal and Child Health Hospital were analyzed retrospectively.After surgical treatment, 50 patients were willing to receive hormone replacement therapy as the experimental group and 50 patients received routine treatment as the control group, Kupperma score was used to evaluate the severity of perimenopausal symptoms, and the serum levels of sex hormones, blood fat, bone content and tumor markers in the two groups before and after treatment were detected and the occurrence of adverse reactions were evaluated.Results:Before treatment, there was no significant difference in kupperma score, estradiol, follicle-stimulating hormone, luteinizing hormone level, lipid level, bone content and tumor marker level between the two groups(all P>0.05). After HRT treatment, kupperma score in the experimental group was significantly lower than that before treatment((11.47±5.12) vs.(20.46±7.52), t=7.262). In the detection of sex hormones, the levels of estradiol in the experimental group was significantly increased((39.26±7.43) ng/L vs.(13.78±7.52) ng/L, t=12.345), and the levels of follicle stimulating hormone((34.25±7.61) U/L vs.(62.18±19.12) U/L, t=8.245)and luteinizing hormone((20.31±6.25) U/L vs.(35.08±6.27) U/L, t=5.452, P=0.004) in the experimental group were significantly decreased.In the detection of blood lipid level, bone content and tumor markers, high-density lipoprotein((1.62±0.33) mmol/L vs(1.34±0.26) mmol/L, t=4.592, P=0.008) and alkaline phosphatase levels((66.21±25.75) U/L vs.(46.88±9.06) U/L, t=5.912, P=0.001)was significantly increased, the low density lipoprotein((2.78±0.43) mmol/L vs.(2.87±0.78) mmol/L, t=4.265, P=0.012)and total cholestenone((4.02±0.45) mmol/L vs.(4.23±0.91) mmol/L, t=5.761, P=0.002) levels were significantly decreased in the experimental group, with statistically significant differences(all P<0.05). There was no significant difference in the level of serum SCC antigen between the experimental group and the control group before and after treatment ( P> 0.05); there was no significant difference in the above indicators between the experimental group and the control group before and after treatment ( P> 0.05); there was no significant increase in adverse reactions such as breast swelling and pain (3 cases), body mass increase (2 cases), bleeding (1 case), body pain (4 cases), vomiting (4 cases), etc. between the experimental group and the control group (2, 1, 2, 3, 1 case) There was no statistical significance ( P> 0.05). Conclusion:HRT treatment of cervical squamous cell carcinoma patients after surgery can significantly improve the peri menstrual syndrome caused by low estrogen level, and did not significantly increase the risk of recurrence of cervical cancer patients and adverse reactions.

Objective To summarize the etiology,diagnosis and treatment of three cases of duodenal perforation. Methods The data of 3 cases of children with duodenal perforation in our hospital from September 14,2016 to June 20,2017 were retrospectively analyzed. The causes,clinical features and treatment of children's duodenal perforation were summarized. Results A total of 2 males and 1 females aged 2 years,3 years and 5 years were included in the 3 cases. All 3 cases had fever history of upper respiratory tract infection before onset. Acute abdominal pain occurred after oral administration of ibuprofen several times,and all the abdominal plain films prompted pneumoperitoneum. Three cases of perforation sites were duodenal anterior wall,diameter were 0. 5-1 cm. Two cases of small amount of pneumoperitoneum were explored and repaired the duodenal perforation by 3D laparoscopic,1 case underwent laparotomy to repair the duodenal perforation due to a large number of liquid pneumoperitoneum and severe shock. Repair of 3 cases were covered with omentum. Three cases were all cured without anastomotic fistula, ulcer, adhesive intestinal obstruction or other complications. Followed-up in the department of gastroenterology,3 cases had no Helico-bacter pylori infection. Conclusion Repeatedly oral administration of ibuprofen can cause duodenal perfora-tion in children in the short term. Once the digestive tract perforation confirmed,emergency surgical explora-tion is needed. Laparoscopic repair of duodenum perforation is safe and effective and may have a faster recov-ery. We can choose exploratory laparotomy if conditions are not allowed.

In this paper, the domestic and international demand and development trend of clinical diagnostic radionuclides are analyzed, and the medium and high-energy cyclotrons, adequate and systematic facilities, and preparation techniques required for the production of medical radionuclides based on solid targets are introduced. This paper focuses on the research and development carried out by some important medical institutions and scientific research institutes in China over the years in the aspects of medium and high-energy cyclotrons, beam transmission lines, high-power irradiation target stations and new medical isotope production processes etc. It also looks forward to some new directions for the development of medical radionuclides in China during the 14th Five-Year Plan period.