Objective:To investigate the inhibitory effect of antisense human telomerase RNA(hTR) gene on implanted hepatocellular carcinoma in nude mice.Methods: HepG2 cells were subcutaneously inoculated into BALB/c nude mice at the axilla to establish implanted hepatocellular carcinoma model.The retrovirus plasmid containing antisense telomerse RNA(PLXSN-hTR-BamHⅠ) was injected into the tumor(0.2 ml every time,5 times).Retrovirus plasmid containing sense telomerase RNA(PLXSN-hTR-EcoRⅠ) and normal saline were inoculated as control groups.Tumor volume was determined and the inhibitory rate was calculated.Tumor necrosis was observed by histological analysis and cell apoptosis was analyzed by terminal transferase dUTP nick end labeling(TUNEL).Results: Tumor growth in antisense hTR group was significantly inhibited compared with the two control groups.The tumor inhibitory rate(26.78%) of antisense hTR group was significantly higher than that of sense hTR group(1.93%,P

BACKGROUND: Some studies have showed that after indirect co-culture, bone marrow mesenchymal stem cells can differentiate into myocardial cells and hepatocypte-like cells. OBJECTIVE: To investigate the possibility of human umbilical cord blood mesenchymal stem cells (UCB-MSCs) to differentiate into hepatocytes by co-culture with human hepatocyte line LO2 in vitro.METHODS: Full-term umbilical cord blood samples were obtained sterilely. The UCB-MSCs were isolated by density gradient centrifugation and directly adherence growth, then passaged with trypsin digestion at 80% cell fusion. By utilizing cell culture plate insets with microporous membrane combined with 6-well plate, the LO2-/UCB-MSCs co-culture system was established. UCB-MSCs were plated into the wells of 6-well plate at a density of 1×10~7/L. LO2 cells were plated into the cell culture plate insert at a density of 1×10~5/L. UCB-MSCs were plated in both layers in the control group. Surface markers of adhered cells were detected by flow cytometry. Morphological changes of UCB-MSCs were observed by inverted phase contrast microscopy. mRNA expression was detected by reverse transcriptase PCR. RESULTS AND CONCLUSION: HUCB MSCs expressed CD44 and CD29 strongly, but CD34 and CD45 were expressed negatively. After 5 days, fusiform-shaped cells were reduced in the co-culture group; while, the time passing by, cells shaped irregular round or polygonal were increased, which were similar to hepatocytes. At 4 weeks after culture, UCB-MSCs were still fusiform-shaped in the control group. At day 5 after culture, alpha fetoprotein mRNA expressed positively, but other expressed negatively in the co-culture group; at day 14 after culture, cytokeratin-19 mRNA and albumin mRNA expressions were observed; moreover, with the time passing by, the expression of albumin mRNA was increased, but the expression of alpha fetoprotein-19 mRNA was decreased. Antigenic expressions in the control group were negative. This suggested that UCB-MSCs could differentiate into hepatocypte-like cells by co-culture with human hepatocyte line LO2 in vitro.

Objective To investigate the therapeutic efficacy and safety of endoscopic resection of giant gastric stromal tumors without explicit evidence of metastases.Methods A total of 12 giant gastric stromal tumors with no evidence of metastases diagnosed by endoscopic ultrasound (EUS) and computed tomography (CT) scan were managed by endoscopic resection.Operation time,blood loss and the incidence rate of perforation were recorded respectively.The diagnoses of tissue specimens were made by pathological examination and immunohistochemistry.In order to assess local recurrence and distant metastases,endoscopy and endoscopic ultrasound follow-up examinations were performed routinely at 2,6 and 12 months,and the whole abdominal CT scan was also performed at 12 months after operation.Results Endoscopic resections were successfully performed in 10 of 12 cases (83.3％),among which,6 underwent endoscopic submucosal excavation (ESE) without unexpected perforation and 4 endoscopic full-thickness resection (EFR)with intentional perforation.The rate of intentional perforation was 33.3％ (4/12),and all the perforations could be sealed by endoscopic methods.The blood losses were all more than 100 ml,which could be controlled by argon plasma coagulation,electrocoagulation or hemostatic clips.In the 10 encapsulated tumors,8 could be smoothly removed from esophagus,whose long diameter of the minimum cross section was less than 3.5 cm,however,2 tumors whose diameters were larger than 3.5 cm were taken out after segmentation.In the 10 tissue samples,9 were confirmed as low risk GIST,1 larger than 5 cm was pathologically confirmed as high risk GIST.During 1-year follow-up,no local recurrence or peritoneal metastasis was found.2 tumors,larger than 5.0 cm,could not be removed by endoscopic methods due to uncontrolled bleeding.The rate of uncontrolled bleeding was 16.7％ (2/12).The patients were transferred to surgery,and pathologically confirmed as having high risk GIST.Conclusion For low-risk giant gastric stromal tumors whose diameters were less than 5cm without evidence of metastases,endoscopic resection is considered as a safe and effective procedure.Tumors with long diameter of the minimum cross section less than 3.5 cm are more suitable for endoscopic resection,which can be smoothly taken out through cardia.However,for high-risk GIST larger than 5.0 cm,the rate of uncontrolled bleeding is high,so endoscopic resection should be adopted with discretion.

Objective: To analyze the characteristics of a norovirus outbreak in a kindergarten, and factors affecting the detoxification time, so as to provide a scientific basis for targeted control measures. Methods: On February 24,2023, the basic personal information, clinical manifestations，morbidity and treatment of 16 kindergarten cases with a norovirus outbreak in Wuhan were collected. Anal swabs were collected every 7 d after the outbreak to detect norovirus. Chisquare test was used for comparison of intergroup rates. The comparison of detoxification time between different groups was conducted by Logrank test, and the influencing factors of detoxification time in cases were analyzed by Cox multiple regression analysis. Results: From February 19-28, 2023, a total of 18 cases were reported and 16 of them participated in the detoxication time monitoring. In the first, second, third and fourth sampling after the outbreak, the positive rates were 60.00%, 100.00%, 75.00% and 0, respectively. The reverse transcriptionpolymerase chain reaction cycle threshold (Ct values) were (25.83±5.74, 28.83±5.55, 36.13±4.30), and undetected, respectively. The median time of detoxification was 19.42 d with 95%CI=(18.21-20.64)d. The results of Cox regression showed that the detoxication time was shorter in the treatment group than in the nontreatment group[HR(95%CI)=5.09(1.39-18.58), P<0.05]. Conclusion Children infected with norovirus has a long duration of detoxification,and case management, which could be shortened by drugs, and disinfection should be strengthened after the case returned to school.

Objective:To investigate the risk factors of lymph node metastasis and the clinical significance of deep submucosal invasion in patients with T1 stage colorectal cancer.Methods:From January 30, 2010 to December 31, 2019, at Shandong Provincial Hospital Affiliated to Shandong First Medical University, among patients with T1 stage colorectal cancer, 41 patients underwent radical surgery for colorectal cancer (surgery group) and 23 patients received endoscopic submucosal dissection (ESD) (ESD group) were enrolled. The tumor gross type, maximum diameter, histologically poorly differentiated components, degree of invasion (the type of mucosal muscle destruction, the width and depth of invasion), the budding grade of tumor, and whether with vascular tumor thrombus were recorded. The additional treatment and prognosis of patients were collected by telephone follow-up. The risk factors of lymph node metastasis in stage T1 colorectal cancer, the correlation between the complete muscularis mucosa destruction and the width and depth of invasion in the ESD group, and the effects of additional treatment after operation on the prognosis of patients were analyzed. Independent sample t test and chi-square test were used for statistical analysis. Results:The rate of lymph node metastasis in patients with poorly differentiated components or vascular tumor thrombus was higher than that in patients without poorly differentiated components or vascular tumor thrombus (3/6 vs. 12.1%, 7/58; 3/4 vs. 11.7%, 7/60), and the differences were statistically significant ( χ2=5.934 and 11.409, both P<0.05). All patients in the surgery group had complete muscularis mucosa destruction. In ESD group, the width of tumor invasion was ≥ 2 mm in 16 cases, including complete destruction of muscularis mucosa in 15 cases and partial destruction in one case; the width of tumor invasion was <2 mm in seven cases, including complete destruction of muscularis mucoa in two cases and partial destruction in five cases; the depth of infiltration was ≥ 2 000 μm in 14 cases, including complete destruction of muscularis mucosa in 13 cases and partial destruction in one case; the depth of infiltration was <2 000 μm in nine cases, including complete destruction of muscularis mucosa in four cases and partial destruction in five cases. The complete muscularis mucosa destruction was related with tumor of invasion width ≥ 2 mm and invasion depth ≥ 2 000 μm (15/16 vs.2/7, 13/14 vs. 4/7), and the differences were statistically significant ( χ2=10.729, 6.659, both P<0.05). Among the 64 patients with T1 stage colorectal cancer in this study, six cases (9.4%) had poor prognosis; five cases (7.8%) died, and three of them (4.7%) were tumor-related deaths. Adjuvant therapy was added in 10 cases in surgery group and 10 cases in ESD group, and there were no poor prognosis in those patients. There were no significant difference in the incidences of poor prognosis of patients without additional treatment and patients with additional treatment of the two groups (9.7% (3/31) vs. 0 (0/10) and 23.1% (3/13) vs. 0 (0/10)) (both P>0.05). Conclusion:When T1 stage colorectal cancer with tumor submucosal invasion, clinicians should comprehensively evaluate the prognostic risk based on various pathological characteristics such as the degree of tumor differentiation, vascular tumor thrombus and mucosal muscle destruction.

Objective: To understand the distribution of tobacco retailer within 100 meters outside middle schools in Wuhan City and its impact on smoking behavior of middle school students, so as to provide basis and feasible suggestions for the development of tobacco control policy for adolescents. Methods: From February to May 2023, a multi stage stratified cluster random sampling method was used to select 20 middle schools from 4 districts in Wuhan City. To investigate the distribution of tobacco retailer within 100 metres outside the school and the sale of tobacco to minors. A total of 4 882 students were surveyed using the core questions of the 2021 Chinese Adolescent Tobacco Prevalence Questionnaire. Fisher exact probability test, Chi square test and Chi square trend test were used for statistical analysis. Results: Nearly 70.00% of middle schools had tobacco retailer within 100 metres, with an average of (1.10±0.97) per middle school. The awareness rate (100.00%) and labeling rate (87.50%) of licensed tobacco retailer were higher than those of non licensed tobacco retailer (33.33%, 16.67%) ( P <0.05). The rates of tried smoking, current smoking and buying cigarettes within 30 days were 7.13%, 1.99% and 2.54%, respectively. The rates of students who tried smoking ( 8.58 %), current smoking (2.29%) and buying cigarettes within 30 days (2.85%) in schools with tobacco retailer within 100 metres were higher than those in schools without tobacco retailer (3.79%, 1.28%, 1.83%)( χ 2=35.80, 5.37, 4.37 , P <0.05). And as the grade increased, the rates of tried smoking, current smoking and buying cigarettes among middle school students all showed an upward trend ( χ 2 trend =66.20, 36.10, 16.17, P <0.05). Conclusions Middle school students in Wuhan City have high tobacco availability. The findings suggest that school ban should be extended from 50 meters to 100 meters, and the regulatory authorities must strictly prohibit selling tobacco products to minors at tobacco retailer.

Objective To evaluate the efficacy and safety of X-ray guided endoscopic gastrojejunostomy using stent in treatment of malignant gastric outlet obstruction ( GOO ) . Methods Six hospitalized patients with malignant GOO underwent X-ray guided endoscopic gastrojejunostomy using stent in the department of gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University between March 2017 and June 2017. The technical success rate, clinical success rate, procedure time, adverse events and follow-up were recorded and analyzed in this retrospective study. Results The stent was successfully placed in the 6 patients with 100％ ( 6/6) technical success rate. The mean procedure time was 91. 7±51. 8 min. After the procedure, all patients were fed liquid or semi-liquid diet, and the GOO score system was increased from 0-1 before operation to 2-3 after operation. The clinical success rate was 100％(6/6). Peritonitis was observed in 2 patients during operation, and resolved by abdominal drainage. Gastrointestinal bleeding occurred in 1 patient after operation, which was resolved with conservative treatment. During a mean follow-up period of 78. 6 days (range 32-100 days), there was no recurrence of obstruction symptoms except that 1 patient died because of tumor progress 60 days after procedure. Conclusion The X-ray guided endoscopic gastrojejunostomy using stent is feasible and safe to treat malignant GOO with a reliable short-term efficacy.

Model informed precision dosing (MIPD) is a new concept to guide precision dosing for individual patient by modeling and simulation based on the available information about the individual patient, medications and the disease. Compared to the empirical dosing, MIPD could improve the efficacy, safety, economics and adherence of the pharmacotherapy according to the individual's pathophysiology, genotyping and disease progression. This consensus report provides a brief account of the concept, methodology and implementation of MIPD as well as clinical decision supporting systems for MIPD. The status and future advancing of MIPD was also discussed to facilitate the appropriate application and development of MIPD in China.

With the increasing cost of drug development and clinical trials, it is of great value to make full use of all kinds of data to improve the efficiency of drug development and to provide valid information for medication guidelines. Model-based meta-analysis (MBMA) combines mathematical models with meta-analysis to integrate information from multiple sources (preclinical and clinical data, etc.) and multiple dimensions (targets/mechanisms, pharmacokinetics/pharmacodynamics, diseases/indications, populations, regimens, biomarkers/efficacy/safety, etc.), which not only provides decision-making for all key points of drug development, but also provides effective information for rational drug use and cost-effectiveness analysis. The classical meta-analysis requires high homogeneity of the data, while MBMA can combine and analyze the heterogeneous data of different doses, different time courses, and different populations through modeling, so as to quantify the dose-effect relationship, time-effect relationship, and the relevant impact factors, and thus the efficacy or safety features at the level of dose, time and covariable that have not been involved in previous studies. Although the modeling and simulation methods of MBMA are similar to population pharmacokinetics/pharmacodynamics (Pop PK/PD), compared with Pop PK/PD, the advantage of MBMA is that it can make full use of literature data, which not only improves the strength of evidence, but also can answer the questions that have not been proved or can not be answered by a single study. At present, MBMA has become one of the important methods in the strategy of model-informed drug development (MIDD). This paper will focus on the application value, data analysis plan, data acquisition and processing, data analysis and reporting of MBMA, in order to provide reference for the application of MBMA in drug development and clinical practice.