BACKGROUND:In recent years,many studies have confirmed that assembloids can make up for the shortcomings of organoids,which cannot fully reproduce the interaction between cell and cell and between cell and matrix.Since the assembloids construction methods are in the early stage of development,there is no unified standard. OBJECTIVE:To review the current construction methods,applications,advantages,and disadvantages of assembloids,guide the development and improvement of vitro cell models. METHODS:PubMed,CNKI,and WanFang databases were searched with English search terms"assembloids,organoids,tumor microenvironment,organoids AND assemble,organoids AND microenvironment"and Chinese search terms"assembloids,organoids,tumor microenvironment,organoid reorganization,multicellular model".Totally 94 articles were screened out for review after excluding irrelevant articles and deduplication. RESULTS AND CONCLUSION:(1)According to the different sources of cells,the construction of assembloids can be divided into three methods:self-assembly,direct-assembly,and mixed-assembly.According to the differences of cell culture methods,it can be divided into suspension culture method,matrix culture method,organ chip culture method,and 3D bio-printing.(2)The process of self-assembly covers early stages of cell and tissue development,so it has broad prospects in the fields of organ development and developmental disorders.The function of differentiated mature cells is relatively perfect,and the assembloids directly assembled by them have more potential in the study of functional disorders and cell-damaging diseases.Self-assembly may be better in organ transplantation,and direct-assembly will be more suitable for the repair of tissue damage.Mixed-assembly combines the advantages of the former two and is mostly used to explore the physiological and pathological mechanisms of cells in the microenvironment,as well as drug screening.(3)Although different assembloids have their own advantages,they all face the problem of imperfect vasculature system,then,each method has its own limitations,for example,the degree of cell differentiation in self-assembly assembloids may still be different from that in vivo,and the fixed cell types in direct-assembly models cannot simulate complex microenvironments in vivo.These are urgent problems to be solved.(4)In the future,with the continuous improvement of assembloids culture technology,scientists can assemble biomimetic organoids with more complex tissues in vitro,providing infinitely realistic models for the study of physiological and pathological processes of human tissue and organ.

OBJECTIVETo summarize the treatment status of gastric gastrointestinal stromal tumor (GIST) in Shandong province,by analyzing the clinicopathological features and prognostic factors.

METHODSClinicopathological and follow-up data of 1 165 patients with gastric GIST between January 2000 and December 2013 from 23 tertiary referral hospitals in Shandong Province were collected to establish a database. The risk stratification of all cases was performed according to the National Institutes of Health(NIH) criteria proposed in 2008. Kaplan-Meier method was used to calculate the survival rate. Log-rank test and Cox regression model were used for univariate and multivariate prognostic analyses.

RESULTSAmong 1 165 cases of gastric GIST, 557 were male and 608 were female. The median age of onset was 60 (range 15-89) years. Primary tumors were located in the gastric fundus and cardia in 623 cases(53.5%), gastric body in 346 cases(29.7%), gastric antrum in 196 cases(16.8%). All the cases underwent resection of tumors, including endoscopic resection (n=106), local resection (n=589), subtotal gastrectomy(n=399), and total gastrectomy(n=72). Based on the NIH risk stratification, there were 256 cases (22.0%) at very low risk, 435 (37.3%) at low risk, 251 cases (21.5%) at intermediate risk, and 223 cases (19.1%) at high risk. A total of 1 116 cases(95.8%) were followed up and the median follow-up period was 40 (range, 1-60) months. During the period, 337 patients relapsed and the median time to recurrence was 34 (range 1-60) months. The 1-, 3-, and 5-year survival rates were 98.6%, 86.1% and 73.4%, respectively. The 5-year survival rates of patients at very low, low, intermediate, and high risk were 93.1%, 85.8%, 63.0% and 42.3% respectively, with a statistically significant difference (P=0.000). Multivariate analysis showed that primary tumor site (RR=0.580, 95%CI:0.402-0.835), tumor size (RR=0.450, 95%CI:0.266-0.760), intraoperative tumor rupture(RR=0.557, 95%CI:0.336-0.924), risk classification (RR=0.309, 95%CI:0.164-0.580) and the use of imatinib after surgery (RR=1.993, 95%CI:1.350-2.922) were independent prognostic factors.

CONCLUSIONSThe choice of surgical procedure for gastric GIST patients should be based on tumor size. All the routine procedures including endoscopic resection, local excision, subtotal gastrectomy and total gastrectomy can obtain satisfactory curative outcomes. NIH classification has a high value for the prediction of prognosis. Primary tumor site, tumor size, intraoperative tumor rupture, risk stratification and postoperative use of imatinib are independent prognostic factors in gastric GIST patients.

Objective:To investigate the potential pro-apoptotic activity of arsenic trioxide (ATO) in human leukemia HL-60 cells,as well as the potential mechanism with focus on mitochondrial pathway.Methods:After treatment with different concentrations of ATO (1 μg/mL,5 μg/mL or 10 μg/mL) for 24 h,apoptotic cell death was detected by flow cytometry,oxidative stress was determined by measuring MDA and GSH levels,the expression of apoptotic factors was detected by western blot,and mitochondrial membrane potential (MMP) was determined by immunofluorescence staining.Results:ATO at the concentrations of 5 μg/mL or 10 μg/mL induces apoptotic cell death and increases oxidative stress in human leukemia HL-60 cells.ATO significantly increases the expression of pro-apoptotic factors (Bax and Caspase-3),whereas decreases the expression of anti-apoptotic factor Bcl-2.Compared with the control group,ATO treatment significantly decreases the MMP level in HL-60 ceils.Conclusions:Arsenic trioxide induces apoptotic cell death through mitochondrial pathway in human leukemia HL-60 cells.

Objective: To clarify the research status of economic burden of stomach cancer in China from 1996 to 2015. Methods: Based on three electronic literature databases (China Knowledge Resource Integrated Database, Wanfang Database and PubMed), a total of 2 873, 1 244 and 84 articles published during 1996 to 2015 were found, respectively, using keywords of"cancer","neoplasms","malignant tumor","tumor","economic burden","health expenditure","cost","cost of illness", and"China". According to the inclusion and exclusion criteria, 30 literatures were included in the final analysis. Then the basic information and study subjects, indicators and main results of economic burden were abstracted and analyzed. All the expenditure data were discounted to the values in 2013 by using China's percapita consumer price index. Results: Totally, 30 articles were included, covering 14 provinces and of which 16 were published during 2011-2015. One article was based on population-level and the remaining studies were all based on individual-level. The number of individual-level articles that reported direct medical, non-medical and indirectly economic burden was 29, 1 and 2, respectively. The main indicators of direct medical expenditure were expenditure per patient (22), per clinical visit (9) and per diem (11), respectively. The median expenditure per patient was 7 387-28 743 RMB (CNY), with average annual growth rate (AAGR) of 1.7% (1996-2013). The median expenditure per clinical visit was 18 504-41 871 RMB (2003-2013), with AAGR of 5.5%. The median expenditure per diem was 313-1 445 RMB (1996-2012), with AAGR of 3.7%. Difference was found among provinces. Conclusions The evidence for economic burden of stomach cancer was still limited over the past two decades and mainly focused on individual and regional levels. An increase and differences in provinces were observed in direct medical expenditure. Evaluation on direct non-medical and indirect medical expenditure needs to be addressed.

OBJECTIVE:To provide reference for imipenem-resistant Pseudomonas aeruginosa infection control. METHODS:114 patients infected with imipenem-resistant P. aeruginosa were selected from 3 tertiary hospitals in Zhoushan during Feb. 2013 to Feb. 2014. 114 strains of P. aeruginosa were isolated from clinical specimens,and drug resistance characteristics and carbapene-mase-producing gene diversity were analyzed. 101 inpatients with imipenem-resistant P. aeruginosa infection were included in con-trol group;univariate and multivariate Logistic regression analysis were adopted to explore the risk factors of imipenem-resistant P. aeruginosa infection. RESULTS:114 strains were sensitive to polymyxin B,and had different levels of resistance to other 9 kinds of antibiotics. Carbapenemase-producing gene were mainly IMP and VIM type gene. Long-term hospitalization,mechanical ventila-tion,used imipenem and early combined use of antibiotics were risk factors of imipenem-resistant P. aeruginosa infection. CON-CLUSIONS:In Zhoushan area,imipenem-resistant P. aeruginosa shows serious drug resistance. To avoid long-term hospitalization and early combined use of antibiotics can reduce imipenem-resistant P. aeruginosa infection.

Objective To investigat the changes of hemodynamics and arterial blood gas during the establishment of ECMO model and provide an ideal platform for the study of the mechanism of DCD protection using ECMO, a porcine model of controlled donors of cardiac death (CDCD) with extracorporeal support was established in the paper. Methods Eight domestic crossbred pigs were anes-thetized, atracurium (1mg/kg) and heparin (150~200 U/kg) was administered and the ventilator was discontinued while the animal was under deep anesthesia to kill the animal.Meanwhile, blood was drained from the left jugular vein catheter into the ECMO circuit and re-turned into the left femoral artery to establish the porcine model of CDCD with extracorporeal support . The average blood press and arterial blood gas were ministered at normal, cardiac arrest and ECMO supporting 1 h, 3 h and 5 h. Results The model of CDCD with extra-corporeal support was established successfully.Two animals restored the beating of the heart and the high doses of potassium chloride were given untill cardiac arrest during extracorporeal support.Blood gases throughout the apneic time showed a severe hypoxemia and acidosis, pH and PaO2 reduced to 7.09 ±0.24 and 5.3 ±0.93mmHg respective-ly, which can be improved during extracorporeal support.MAP and PaO2 were maintained over 300mmHg and 65mmHg respectively. Conclusion A new porcine model of controlled donors of cardiacdeath with extracorporeal support has been successfully established and showed to be feasible for related pathophysiological research work.

Objective To evaluate the effect of extracorporeal membranous oxygenation (ECMO) on the small bowel preservation in a porcine model of controlled donors after cardiac death (CDCD).Method The domestic crossbred donor pigs were anesthetized and ventilated with 100％ oxyger.An intravenous cannula was placed through iliac arteries and jugular vein,and connected to ECMO system.The ECMO was performed to infuse abdominal organs when cardiac death was declared.Pathology,electron microscope,energy metabolism and cell apoptosis level of intestinal mucosa were evaluated before operation,in cardiac death and at the end of extracorporeal support,respectively.Result The normothermic extracorporeal support could quickly restore graft blood supply and oxygenation.One-h extracorporeal support could improve the energy status in intestine of donation after cardiac death (DCD).The histologic damage and apoptosis of 1-h extracorporeal support had no significant difference with those before operation and in cardiac death (P＞0.05).With the extension of extracorporeal support,the intestinal mucosa damage degree was gradually increased,and the content of adenosine triphosphate in intestinal mucosa reduced gradually.Conclusion The normothermic extracorporeal support for 1 h in DCD is beneficial for improving the viability of small bowel.However,the integrity of intestinal mucosa is destroyed gradually as extracorporeal supporting time over,which may be initiated by the activation of intestinal epithelial apoptosis.

OBJECTIVELuteolin, a flavone, has considerable neuroprotective effects by its anti-oxidative mechanism. However, it is still unclear whether luteolin can protect neurons against oxygen-glucose deprivation/reperfusion (OGD/R) induced injury.

METHODAfter 2 hours oxygen-glucose deprivation and 24 hours reperfusion treatment in primary cultured hippocampal neurons, the neuron viability, survival rate and apoptosis rate were evaluated by MTT assay, lactate dehydrogenase (LDH) leakage assay and Hoechst staining, respectively. The activity of Na+/K+ -ATPase was examined in cultured neurons or in the hippocampus of SD rats treated by 10 minutes global cerebral ischemia and followed 24 hours reperfusion.

RESULTTreatment by OGD/R markedly reduced neuronal viability, increased LDH leakage rate and increased apoptosis rate. Application of luteolin (10-100 micromol x L(-1)) during OGD inhibited OGD/R induced neuron injury and apoptosis in a dose-dependent manner. Compared to the control group or OGP/R-treated neurons, the activity of Na+/K+ -ATPase was significantly suppressed in global ischemia/reperfusion group or OGD/R-treated neurons. Application of luteolin during ischemia or OGD preserved the Na+/K+ -ATPase activity. Furthermore, inhibition of Na+/K+ -ATPase with ouabain attenuated the protective effect afforded by luteolin.

CONCLUSIONThe data provide the evidence that luteolin has neuroprotective effect against OGD/R induced injury and the protective effect may be associated with its ability to improve Na+/K+ -ATPase activity after OGD/R.

Animals ; Antioxidants ; pharmacology ; Apoptosis ; drug effects ; Cell Hypoxia ; Glucose ; metabolism ; Luteolin ; pharmacology ; Neurons ; drug effects ; enzymology ; metabolism ; pathology ; Neuroprotective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; enzymology ; metabolism ; pathology ; prevention & control ; Sodium-Potassium-Exchanging ATPase ; metabolism

Objective To evaluate the immediate impact of radiofrequencycatheter ablation on left atrial (LA)volumes and function by velocity vector imaging(ⅤⅥ)and compare the LA function with and without atrial fibrillation after the operation.Methods Ten consecutive patients with paroxysmal AF were studied at baseline and within 3 days after ablation.Ten consecutive patients with persistent AF were studied within 3 days after ablation,in sinus rhythm.Ten patients with normal ventricular function were included in the study.ⅤⅥ was performed to assess LA sizes and strain,strain rate,velocity of the septum,lateral wall and the atrial roof from the apical four-chamber view.Results In patients with paroxysmal AF,global and regioal LA function was not significantly impaired after the ablation procedure.Subgroup analysis demonstrated that there were no significant difference in LA function betwwen patients with paroxysmal AF and control subjects,but the indexed LAVmax was significantly larger in all AF patients compared with control subjects.The global function of LA,including LAEF,LAaEF and LA expansion index significantly decreased in persistent AF patients.By contrast,the LA septal strain,velocity and lateral strain were lower than controls.Conclusions Radiofrequency catheter ablation has no influence on LA function assessed by ⅤⅥ within three days after the operation for patients with paroxysmaI AF.The function of persistent AF patients is absolutely lower than control subjects.