Objective To evaluate the effect of para-esophageal and para-gastric vessels(PEPGV)on endoscopic secondary prophylaxis for varices.Methods The clinical data of patients with cirrhosis-related esophagogastric varices(EGV)who underwent endoscopic variceal ligation and/or obliteration,and had hepatic venous pressure gradient(HVPG)result between January 2020 and December 2020 in Zhongshan Hospital,Fudan University were retrospectively analyzed.Patients were divided into a group without PEPGV and a group with PEPGV based on CT imaging of the portal vein.The main outcome was 2-year re-bleeding.Results A total of 69 patients were included,and 27 of them had PEPGV.There was no statistical difference in baseline characteristics,blood indexes(included hemoglobin level,prothrombin time and albumin level),HVPG,and the secondary prophylactic endoscopic treatment ways between the two groups.A total of 25 patients experienced re-bleeding within 2 years after endoscopic treatment,including 15 in the group with PEPGV and 10 in the group without PEPGV.Kaplan-Meier analysis showed that the cumulative 2-year re-bleeding rate was significantly higher in the group with PEPGV than in the group without PEPGV(60.07％vs 32.79％,P=0.022).Further multivariate Cox analysis showed that PEPGV was an independent predictor of re-bleeding after endoscopic treatment in EGV patients(HR=2.33,95％CI 1.01-5.39,P=0.047).Conclusions The PEPGV is an independent predictor of re-bleeding after endoscopic treatment in EGV patients.It is suggested that when patients with EGV receive endoscopic treatment to prevent re-bleeding,portal vascular CT is suggested to evaluate PEPGV.For patients with giant extraluminal vascular masses,fully evaluating other treatment options such as transjugular intrahepatic portosystemic shunt,or adjusting endoscopic treatment ways is recommended.

Objective:To evaluate the long-term risk of rebleeding in patients with acute esophageal and gastric variceal bleeding and portal vein thrombosis after endoscopic treatment in liver cirrhosis.Methods:From January to December 2022, 57 patients with acute esophageal and gastric variceal bleeding who were treated by endoscopy in the emergency department of the Zhongshan Hospital affiliated to the Fudan University were included in the study. According to the results of portal vein CT angiography (CTA), the patients were divided into thrombosis group and non thrombosis group. We compared the basic information and endoscopic treatment status of two groups of patients. All patients were followed up until 1 year after endoscopic treatment or April 15, 2023, and re bleeding and survival were recorded during the follow-up period. The influencing factors of rebleeding after 1 year of treatment were analyzed.Results:The patient′s age was (55.9±11.4)years old, mainly male [78.95%(45/57)]. The average time from initial bleeding to endoscopic treatment for all patients was (6.6±2.8)days. There was no statistically significant difference between the two groups in terms of age, sex, combined liver malignancy, Child-pugh score, first bleeding form, ascites, and first laboratory examination results (including hemoglobin, platelet, Prothrombin time, creatinine) (all P>0.05). There was no statistically significant difference in the history of endoscopic treatment, bleeding distance from endoscopic treatment, the proportion of patients with esophageal varices and gastric varices, the proportion of patients with esophageal varices treated with ligation, and the proportion of patients with gastric varices treated with tissue glue between the two groups (all P>0.05). A total of 2 patients died after surgery, and 12 patients experienced rebleeding, including 10 in the thrombotic group and 2 in the non thrombotic group. Kaplan Meier analysis showed that the 1-year rebleeding rate in the thrombotic group was significantly higher than that in the non thrombotic group (59.02% vs 24.71%, RR=6.002, 95% CI: 1.06-34.00, P=0.020 8). Cox multivariate regression analysis suggests that the presence of portal vein thrombosis ( HR=7.669, 95% CI: 1.453-40.472, P=0.016) was an independent risk factor for recurrent bleeding after endoscopic treatment of acute esophageal and gastric variceal bleeding for one year. Conclusions:Portal vein thrombosis in liver cirrhosis increases the risk of recurrent bleeding after endoscopic treatment for acute esophageal and gastric variceal bleeding for one year. For patients with acute Upper gastrointestinal bleeding complicated with portal vein thrombosis, regular endoscopic and ultrasonic follow-up, individualized endoscopic sequential and selective anticoagulation therapy should be carried out.

Objective:To explore the biological effects of the targeted nanocomposite on breast cancer 4T1 cells with hepatitis B virus-like particles (HBc-RGD-VLPs) as a carrier for the delivery of anti-cancer drugs, and to provide a new theoretical basis for reducing the toxicity of doxorubicin (DOX) anti-tumor drugs and changing the path of administration.Methods:The hepatitis B pseudoviral particles prepared in the early stage of this laboratory enveloped DOX to form a target nanocomposite HBc-RGD-VLPs/DOX. The homogeneity and morphology of particles were detected by transmission electron microscopy and granular size analyzer, and applied to 4T1 cells for in vitro bioactivity exploration.Results:The structure of the target nanocomposite HBc-RGD-VLPs/DOX was detected by means of a transmission electron microscope, in a homogenous form, and the particle size distribution was 30-35 nm. In vitro cell experiments showed that the safety of target vector HBc-RGD-VLPs was better, the cell survival rate was more than 80%, and the HBc-RGD-VLPs/DOX after encapsulation had a significant inhibitory effect on the growth of 4T1 cells, and the effective inhibitory concentration (IC50) for half of 4T1 tumor cells was 1.445 g/ml. Fluorescence microscopy showed that HBc-RGD-VLP/DOX can be specifically targeted to tumor cells relative to the isolated DOX.Conclusions:The safety of target vector HBc-RGD-VLPs is better, HBc-RGD-VLPs/DOX showed good proliferation inhibitory effect and certain tumor-targeting effect on tumor cells.

Objective To study the method of dose reconstruction in human body under the photon external radiation accident condition,and to verify the accuracy of the method for the local dose distribution.Methods Based on the open source Monte Carlo tool kit Geant 4 and using the human voxel phantom recommended by ICRP Publication 103,the dose reconstruction method under the condition of external radiation accident was studied to evaluate the average absorbed dose,organ absorbed dose and local dose distribution.To validate the code,several irradiation experiments were implemented in some standard radiation fields by putting TLDs in the tissue equivalent physical phantom ART.A voxel phantom was used to reconstruct the radiation doses,which was created based on the CT scan image of the ART phantom with resolution of 1.57 mm× 1.57 mm× 10.00 mm.The result of experiment were compared with those of dose reconstruction simulation.Results The relative uncertainty of the measured values was 10.9％.The relative uncertainty of the dose reconstruction simulation values was 7.10％ at the non-tissueinterface area and 16.6％ at the tissue-interface area.For 451 measuring points,the average of the simulated value divided by the measured value was 0.972,with the standard deviation of 0.083 8.In the range of 0.95-1.05,0.90-1.10 and 0.80-1.20,and the proportions were 49.2％,79.4％ and 96.4％,respectively.Conclusions The method of Monte Carlo dose reconstruction based on human voxel phantom meets the accuracy requirement of actual uses both at the whole body or organ level and at the local dose distribution level.It can be used as a powerful tool for dose assessment of the exposed people in an external radiation accidents and provide support for diagnosis and treatment.

Objective To investigate the changes in serum level of microparticles (EMPs) in mice with ventilator-induced lung injury (VILI), and explore its significance in VILI. Methods Forty-eight grade SPF male C57BL/6J mice were randomly divided into two groups, with 24 mice in each group: the mice in mechanical ventilation (MV) group were given high tidal volume (VT 30 mL/kg) MV for 4 hours after tracheal intubation, and those in spontaneous breathing group were spontaneously breathed for 4 hours. The apical blood of 12 mice in each group were collected, and serum levels of interleukins (IL-1β, IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme linked immunosorbent assay (ELISA), and serum EMPs levels were determined by flow cytometer. The correlations between EMPs and IL-1β, IL-6, and TNF-α were analyzed by linear regression analysis. The lung tissues of other 12 mice in each group were harvested, and wet/dry weight (W/D) ratio was assessed. After hematoxylin-eosin (HE) staining, the morphological changes in lung tissue were observed under light microscope. After double staining of uranium acetate and lead citrate, the ultrastructural changes in lung tissue were observed with electron microscope. Results Compared with spontaneous breathing group, the levels of lung W/D ratio in MV group was significantly increased (5.47±0.14 vs. 4.34±0.11), the levels of IL-1β, IL-6, TNF-α and EMPs were also significantly increased [IL-1β (ng/L): 42.4±4.4 vs. 7.7±3.6, IL-6 (ng/L): 1 239.5±66.3 vs. 21.7±4.6, TNF-α (ng/L):237.6±25.8 vs. 37.1±19.1, EMPs (cells/μL): 28.6±1.8 vs. 5.9±1.8, all P < 0.01]. It was shown by correlation analysis that EMPs were positively related with IL-1β, IL-6, and TNF-α (r value was 0.968, 0.932, 0.945, respectively, all P = 0.000). It was shown by fitting linear regression analysis that when EMPs increased by 1 cell/μL, IL-1β increased by 2.4 ng/L [95% confidence interval (95%CI) = 1.9-2.8, P < 0.001], IL-6 increased by 34.5 ng/L (95%CI = 25.1-44.0, P < 0.001), and TNF-α increased by 13.6 ng/L (95%CI = 10.3-16.9,P < 0.001). It was shown by light microscope that the structure of lung tissue and alveolar of mice in spontaneous breathing group appeared normal, while the shrinks of alveolar and disappearance of alveolar architecture were found in MV group. It was shown by electron microscopy that alveolar wall edema and thickening and broken alveolar septa were found in MV group, by contrast, the structure of alveolar was normal in spontaneous breathing group. Conclusion 30 mL/kg VT ventilation for 4 hours could induce VILI with increase in EMPs, suggesting EMPs closely related to VILI, and EMPs level may be putative biomarker of VILI.

Objective To investigate the correlation between the level change of serum homocysteine (HCY) and vascular endothelial growth factor (VEGF) with the degree of the coronary artery stenosis in the patients with acute coronary syndrome (ACS) .Methods A total of 157 ACS patients were divided into the ST‐elevation myocardial infarction (STEMI) group ,non‐ST el‐evation myocardial infarction (NSTEMI) group and unstable angina pectoris(UA) group based on the symptoms ,cardiac enzymes level and electrocardiogram changes .The cases were induded into the mild ,moderate and severe stenosis lesion groups according to the coronary arteriography examination;meanwhile the enzyme‐linked immunosorbent assay (ELISA) was used to detect the change of the serum VEGF concentration .The HCY level was determined by enzymatic cycling methods with the biochemical analyzer (BXC800 ,Beckman ,USA) .The differences in the concentrations of VEGF and HCY compared among different groups .Results The VEGF level had statistical differences among the mild ,moderate and sever stenosis groups(F=39 .9 ,P=0 .00) ,and between the UA group with the NSTEMI group and STEMI group(F=123 .3 ,P=0 .00) .The HCY level had statistically significant differ‐ence between the severe stenosis group with the mild and moderate stenosis groups (F=39 .7 ,P=0 .00);the HCY level had statis‐tically significant difference among the UA group ,NSTEMI group and STEMI group(F=102 .65 ,P=0 .00) .The VEGF and HCY levels in the mild stenosis group ,different degrees of coronary stenosis groups and different clinical diagnosis groups were positively correlated with the Gensini scores(r=0 .723 ,0 .716) .Conclusion The serum VEGF and HCY levels are correlated with the degree of the coronary artery lesion and myocardial necrosis in ACS patients ,furthermore are related with the Gensini scores .

Objective To explore the characteristic of memory impairment and its relationship with Nmethyl-D-aspartate receptor 2B (NR2B) expression in alcohol dependence patients,in order to provide an unprecedented view of alcohol-associated memory impairment therapy.Methods Participants (n=67) included 35 alcohol dependence patients and 32 matched healthy controls.Wechsler memory scale (WMS) was used to access the memory.The expression levels of NR2B were detected with quantitative reverse transcription-polymerase chain reaction (qRT-PCR).Results Compared with the memory quotient(MQ) of controls(69.45±8.96),that of alcohol dependence patients(50.59±8.64) significantly decreased (t=-6.08,P＜0.01).Compared with the NR2B expression level of controls (1.00-0.00),that of alcohol dependence patients (3.52 ± 1.17) significantly increased (t =9.67,P＜0.01).MQ was negatively correlated with the levels of NR2B expression (r=-0.44,P＜0.05).Conclusion Alcohol dependence patients suffer memory impairment detected by WMS,and modulate NR2B expression may improve the memory.

Objective In order to understand which kind of function genes play an important role for es-timating wound age, the variation of difference genes’ mRNA expression were compared after injury. Methods T he mRNA expression levels of seven candidate genes (ICAM-1, NF-κB, MX2, MT1, MT2, sTnI, and Cox6c) were analyzed in contused rat skeletal muscle at different time points using real-time fluorescent quantitative PC R (R T-qPC R ). T he rawC t values were normalized relative to that of RPL32 mRNA , and converted to standard C t values. A t each time point after injury, the standard deviations (SD ) of the standard C t values were calculated by SPSS. Results T he expression trends of the seven genes were all found to be related to wound age, but there were lower variation coefficients and greater reliability of sTnI and Cox6c when compared with other genes. Conclusion T he genes encoding struc-tural proteins or proteins that performbasic functions can be suitable for wound age estimation.

Objective T o investigate the relationship betw een the expression of secreted frizzled-related protein 5 (SFR P5) m RNA and the tim e interval after skeletal m uscle injury in rats by real-tim e PC R . Methods A total of ninety SD rats w ere random ly divided into the contusion groups at different tim es including 4 h, 8 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, 48 h after contusion, incision groups at different tim es including 4h and 8h after incision and the control group. T he sam ples w ere taken from the contused zone at different tim e points. T he total RNA w as isolated from the sam ples and re-versely transcribed to analyze the expression levels of SFRP5 m RNA . Results C om pared to the control group, the expression of SFRP5 m RNA in contusion groups w ere dow n-regulated w ithin 48 h after con-tusion and reached the low est level at 20 h, and the expression of SFRP5 m RNA gradually increased from 20 h to 48 h after contusion. T he expression of SFRP5 m RNA in the incised groups w ere signifi-cantly low er than that of the contusion groups at 4 h after injury. A t the tim e of 8 h, the expression levels betw een the contusion and incision groups show ed no statistically significant difference. Conclusion It is suggested that SFRP5 m RNA analysis m ay show regular expression and can be a m arker for estim ation of skeletal m uscle injury age.

BACKGROUND:It is important to produce and save a large amount of high-activity feeder cel s for the culture of human embryonic stem cel s.
OBJECTIVE:To establish the optimal method for isolation and culture of Kunming mouse embryonic fibroblasts, and to evaluate the feasibility of preparing feeder layers for culture of human embryonic stem cel s.
METHODS:Embryonic fibroblasts were isolated and cultured by different concentrations of trypsin from Kunming mouse fetuses in vitro. The biological characteristics and growth rule of mouse embryonic fibroblasts were investigated, and then the feeder layers for human embryonic stem cel s culture were produced. The growth of human embryonic stem cel s on the prepared feeder layer was tested.
RESULTS AND CONCLUSION:The optimal fetal age for preparing Kunming mouse embryonic fibroblast feeder layer was 13.5 days. Kunming mouse embryonic fibroblasts at different concentrations grew wel with high purity and active proliferation by trypsin digestion method. There was no significant difference in the survival rate of cel s after cryopreservation for 2 weeks, 1 month, 3 months and 6 months. The cel s were proliferative from the second to fourth passage and declined sharply after the fifth passage. Human embryonic stem cel s which grew on Kunming mouse embryonic fibroblasts feeder layers were stil to remain the typical undifferentiated morphology and were strongly positive for alkaline phosphatase and periodic acid-Schiff after long-term subculture. The
mouse embryonic fibroblasts can be used as the stable and high-quality feeder cel s for human embryonic stem cel s.