Objective To study the role of ubiquitin-conjugating enzyme 9(UBC9)in the pyroptosis of homocysteine-induced macrophages mediated by small ubiquitin-like modifier(SUMO)modification.Methods First,the effects of homocysteine at different concentrations(0 μmol/L,50 μ.mol/L,100 μmol/L,150 μmol/L and 200 μmol/L)on the viability and pyrodeath of mouse macrophages(RAW264.7)were detected by CCK-8 and Western blot.Western blot was used to detect the expression levels of UBC9,SUMO-1,and the inflammatory cytokine IL-1β in different groups of cells.qRT-PCR was used to detect the mRNA expression of UBC9 before and after RNA interference and the expression of UBC9,pyrogen-related protein,and SUMO-1 after RNA interference.Results After stimulation with 100 μmol/L homocysteine,the effect of macrophage activity was minimal,and NLRP3 and Caspase-1 were the proteins with the most obvious increase in expression(P＜0.05).Compared with the Control group,the Hcy group's expression of IL-1β and SUMO-1 was increased(P＜0.01).Compared with the Control group,the Hcy group's UBC9 protein and mRNA levels were increased(P＜0.05).The expression of NLRP3,Caspase-1,IL-1β,UBC9,and SUMO-1 was decreased in the si-UBC9+Hcy group compared with the si-NC+Hcy group(P＜0.01).Conclusions Homocysteine induces pyroptosis in macrophages,and its mechanism of action is related to the up-regulation of UBC9 to induce SUMO modification.

Objective:To explore the sex-based heterogeneity in demographic and pathological trends of lung cancer during the past 30 years.Methods:Patients with primary lung cancer who received surgical treatment in the Department of thoracic surgery, Shanghai Pulmonary Hospital Tongji University from 1989 to 2018 were retrospectively analyzed. The differences between male and female patients in age, smoking history, pathological stage and type were compared. Mann- Kendall trend test was performed for trend analysis. Results:A total of 58 433 patients were included in this study, encompassing 30 729(52.6%) men and 27 , 704(47.4%) women. Compared with male patients, female patients were younger(56.0 years old vs. 59.7 years old), and had a higher proportion of non-smokers(98.3% vs. 52.3%), stage Ⅰ lung cancers(60.6% vs. 49.3%), and adenocarcinoma(93.7% vs. 56.1%, all P-values <0.001). Trend analyses revealed that the proportion of female patients increased year by year, and surpassed males in 2015, with the current ratio of male to female being 1∶1.5. After 2013, the age of onset in females was getting younger, and the average age decreased from 58.7 years old to 54.7 years old( P=0.02). The decrease in the proportion of smoking patients was mainly reflected by male patients(from 68.5% to 31.1%, P<0.01). Stage Ⅰ lung cancers in male and females outnumbered advanced stage in 2012 and 2010, respectively, with a much higher proportion in female patients. Among male patients, adenocarcinoma has replaced squamous cell carcinoma as the most common pathological type since 2012, while in female patients adenocarcinoma remained the most common pathological type of lung cancer, and its proportion continued to increase reaching over 98%. Conclusion:A dramatic change in gender distribution was noticed during the past 30 years. Female patients became the primary population in surgically-treated lung cancers, with a trend of getting younger. The proportion of smokers and squamous cell carcinoma decreased significantly in male patients, and adenocarcinoma has become the most common pathological type of lung cancer. The proportion of stage Ⅰ lung cancers was on a dramatic rise, with the popularization of CT screening for lung cancer.

Objective: Taxifolin is a natural flavonoid compound that can be isolated from onions, grapes, oranges and grapefruit. It also acts as a medicine food homology with extraordinary antioxidant and anti-inflammatory activity. This study aims to explain the protective effects and potential mechanisms of taxifolin against inflammatory reaction. Methods: Levels of interleukin (IL)-6, IL-1β and intracellular reactive oxygen species (ROS) were assessed in different time after the treatment of taxifolin in RAW264.7 cells induced by lipopolysaccharide (LPS). Subsequently, the mRNA and protein levels of inducible nitric oxide synthase (iNOS), vascular endothelial growth factor (VEGF), cyclooxygenase (COX)-2, tumor necrosis factor (TNF)-α and the phosphorylation expression levels of the MAPK signal pathway were also evaluated. A silico analysis was used to explain the binding situation for the investigation of taxifolin and MAPK signal pathway. And then MAPK inhibitors were used to reveal the expression level of iNOS, VEGF, COX-2 and TNF-α in RAW264.7 cells. Results: It was demonstrated that cell inflammatory damage induced by LPS was significantly alleviated after the treatment of taxifolin. Then, the mRNA and protein levels of iNOS, VEGF, COX-2 and TNF-α were reduced and the phosphorylation expression levels of the MAPK signal pathway were down-regulated remarkably as well. In silico analysis, taxifolin could form a relatively stable combination with MAPK signal pathway. MAPK inhibitors showed increasing or decreasing effect in the mRNA levels of iNOS, VEGF, COX-2 and TNF-α, which suggesting that taxifolin down-regulated iNOS, VEGF, COX-2 and TNF-α expressions were not entirely through the MAPK pathway. Conclusion: This finding demonstrated that taxifolin improved the inflammatory responses that partly involved in the phosphorylation expression level of MAPK signal pathway in RAW264.7 cells exposed to acute stress.

Objective:To explore the clinical characteristics and prognosis of metastatic sites symptom as the first manifestation in esophageal carcinoma patients with stage T 1 and T 2, and to provide a reference for clinical practice. Methods:The clinical data of 50 esophageal carcinoma patients with stage T 1 and T 2 who had lymph node or distant metastasis as the first symptom in Anyang Tumor Hospital of Henan Province from November 2007 to December 2019 were retrospectively analyzed. Survival analysis was performed by using Kaplan-Meier method. Univariate analysis was performed by using log-rank test. Results:Among 50 patients with esophageal carcinoma, lymph node metastases as the first symptom were found in 42 cases and distant organ metastases as the first symptom were found in 8 cases. The 1-, 3-, 5-year overall survival rates of patients with stage Ⅰ-Ⅱ and stage Ⅲ-Ⅳ were 58.7%, 49.0%, 16.3% and 56.1%, 12.2%, 0, respectively, and there was no statistically significant difference in OS of both groups ( P = 0.094). The 1-, 3-, 5-year overall survival rates of patients with stage N 1 and stage N 2-N 3 were 63.5%, 34.7%, 17.3% and 52.2%, 11.9%, 0, respectively, and there was no statistically significant difference in OS of both groups ( P = 0.083). The 1-, 3-, 5-year overall survival rates were 64.6%, 30.5%, 18.3%, respectively in radiotherapy group and 38.2%, 0, 0, respectively in non-radiotherapy group, and there was a statistically significant difference in OS of both groups ( P = 0.008); the progression-free survival in radiotherapy group was better than that in non-radiotherapy group ( P = 0.028). The 1-, 3-, 5-year overall survival rates were 70.8%, 35.5%, 21.3% and 33.3%, 0, 0 and 35.4%, 0, 0, respectively in concurrent chemoradiotherapy group, radiotherapy group and chemotherapy group, and there was a statistically significant difference in overall survival among three groups ( P = 0.004). The results of univariate analysis showed that radiotherapy ( χ2 = 7.112, P = 0.008) and concurrent chemoradiotherapy ( χ2 = 10.940, P = 0.004) were the main factors affecting the prognosis. Conclusions:Lymph node and distant metastasis could occur in esophageal carcinoma patients with stage T 1 and T 2. Radiotherapy can prolong the progression-free survival time and concurrent chemoradiotherapy could benefit overall survival of these patients.

The urothelium is a transitional epithelium which lines on the renal pelvis, ureter, bladder and proximal urethra adjacent to the bladder. It’s a permeability barrier. In the process of urothelial development or injury repair, the mature urothelial cells expressing uroplakin appears after the hierarchical transcription of a series of transcription factors in basal stem cells. Understanding normal urothelial differentiation process of the urothelial cells could be beneficial to uncover the development of urothelial carcinoma so that to provide targeted therapy options and the study of the urinary tract repair process after injury. In addition, the study of stem cell differentiation microenvironment also provides important theoretical support for tissue engineering and tissue reconstruction. In this review, we discuss the latest findings on urothelial cell differentiation and its clinical significance.

Objective:To compare the prognosis of fluoroucil combined with cisplatin and paclitaxel combined with cisplatin regimens with concurrent radiotherapy in treatment of esophageal squamous cell carcinoma.Methods:A total of 120 patients with esophageal squamous cell carcinoma who were admitted to Anyang Tumor Hospital of Henan Province from December 2012 to November 2018 were randomly divided into group A and group B by using a random number generator. Group A was given cisplatin combined with 5-fluorouracil, and group B was given cisplatin combined with paclitaxel. Both groups had the same radiotherapy regimen, and both used intensity-modulated radiation therapy (IMRT). Completions of 50 Gy radiotherapy and at least one cycle of chemotherapy were considered to be in line with the plan. Survival data was analyzed in the term of intention-to-treat (ITT) and per-protocol (PP) set.Results:Of the 120 patients, 114 patients were treated and the adverse reactions could be evaluated, including 55 cases in group A and 59 cases in group B. The incidence of grade Ⅲ-Ⅳ leukopenia in group B was higher than that in group A [49.2% (29/59) vs. 25.5% (14/55)], and the difference was statistically significant ( χ2 = 6.805, P = 0.012), and there were no statistical differences in the other adverse reactions between the two groups (All P > 0.05). A total of 113 cases can be analyzed for survival. According to ITT analysis, the median progression-free survival (PFS) time in group A and group B was 28.0 months (95% CI 15.5-34.5 months) and 27.0 months (95% CI 17.0-41.0 months), the median overall survival (OS) time was 28.0 months (95% CI 15.8-34.2 months) and not reached, the differences were not statistically significant (both P > 0.05). According to PP analysis, the median PFS time in group A and group B was 28.0 months (95% CI 15.8-34.2 months) and 29.0 months (95% CI 14.9-45.1 months), the median OS time in group A and group B was 28.0 months (95% CI 3.7-52.3 months) and not reached, the differences were not statistically significant (both P > 0.05). Conclusions:The fluorouracil combined with cisplatin regimen and paclitaxel combined with cisplatin regimen with concurrent radiotherapy have similar PFS and OS time in treatment of esophageal squamous cell carcinoma, the adverse reactions are different, but they are all tolerable. In individualized clinical practice, the toxicities and costs of the two regimens can be comprehensively considered.

Objective To assess and compare the incidence,clinical characteristics,treatment,and prognosis of acute heart failure patients from different grades hospitals in Beijing.Methods In this prospective internet prognosis registered study (Beijing AHF Registry),a total of 3 335 consecutive patients admitted to 14 emergency departments in Beijing from January 1st 2011 to September 23rd 2012 were enrolled.According to hospital grade,these patients were divided into two groups,349 patients were from secondary hospitals,and 2 956 patients were from tertiary hospitals.Results Among the 3 335 patients,the medium age was 71 (58,79) years,and male accounted for 53.16％.The most common underlying disease were coronary disease (43.27％),hypertension (17.73％),cardiomyopathy (16.07％) etc.The average treatment time in Emergency Department was 66.82 h.The emergency department mortality rate was 3.81％ (127 cases).The 30-day and 1-year cumulative all-cause mortality were 15.3％ and 32.27％,respectively.The 30-day and 1-year cumulative all-cause readmission were 15.64％ and 46.89％,respectively.Compared with patients in tertiary hospitals,patients in secondary hospitals had more onset acute heart failure patients (63.64％ vs.49.93％),shorter emergency department treatment time (12 h vs.41 h),lower discharge rate (3.43％ vs.37.45％) and emergency department mortality(1.58％ vs.4.09％).Compared with those in tertiary hospitals,1-year cumulative all-cause mortality (25.6％ vs.33.2％),cardiovascular disease mortality (20.2％ vs.26.0％),aggravated heart failure mortality (22.4％ vs.28.8％) were lower in secondary hospitals.Following propensity score matching,compared to tertiary hospitals,patients in secondary hospitals showed lower utilization rate of beta-blockers and ACEFARB (4.51％ vs.28.17％,1.41％ vs.9.58％),except the pironolactone.Conclusion Acute heart failure in emergency department is associated with a high mortality rate and readmission rate.There is still a big gap between guidelines recommend medication current treatments for acute heart failure.

Background and purpose:The previous research has found that the prostate stromal cells derived from different prostate zones have distinct effect on prostate epithelial cells. We also revealed that LMO2 protein was highly expressed in PZ stromal cells (PZSCs) and prostate cancer associated fibroblasts (CAFs) compared with TZ stromal cells. This study investigated the effect of LMO2 protein in prostate stromal cells on proliferation and invasion of prostate cancer PC-3 cells and its mechanisms. Methods:Lentivirus overexpression vectors were used to establish LMO2-overexpressed prostate WPMY-1 stromal cell line. shRNA plasmids were used to suppress LMO2 in CAFs. LMO2 mRNA and protein level of both WPMY-1 and CAFs were evaluated by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. Then, PC-3 cells were co-cultured with different prostate stromal cells and the in vitro proliferation and invasion of PC-3 were measured by CCK-8 and matrigel invasion assays respectively. Results:When co-cultured with LMO2-overexpressed prostate stromal cells, both proliferation and in-vasion of PC-3 were improved. However, when co-cultured with CAFs which have inhibited expression of LMO2, the proliferation and invasion of PC-3 were reduced. The protein array proifling found that both interleukin-11 (IL-11) and ifbroblast growth factor-9 (FGF-9) were enhanced extensively in the supernatant collected from LMO2-overexpressed WPMY-1 cells. Conclusion:The expression of LMO2 in prostate stromal cells could be responsible for development of prostate cancer. Paracrine of cytokines, such as IL-11 and FGF-9, from LMO2-overexpressed stromal cells had effects on the proliferation and invasion of prostate cancer cells.

Objective Gastrointestinal stromal tumor( GIST) is a rare mesenchymal tumor from gastroin-testinal tract,and surgery remains the only curative treatment.In order to improve the outcome of surgical treat-ment,reduce the risk of surgery,and increase the quality of life,preoperative imatinib( IM) treatment for GIST is investigated.Methods We retrospectively studied the multidisciplinary team model treatments for 8 GIST cases receiving preoperative IM treatment.The cases were prescribed IM of 400 mg daily for 12 to 40 weeks and exten-sively followed until surgery was considered feasibleg.The clinical significance and safety profile were analyzed. Results Partial responsive rate was 62.5% in this study.There was no intolerable sever adverse effects by IM preoperative treatment.All the cases received R0 dissection,with no intraoperative tumor rupture.The postopera-tive recovery was satisfied.Conclusion IM preoperative treatment brings significant clinical benefit to large stomach GISTs and cardiac region GISTs.The preoperative treatment should be monitored carefully under a multi-disciplinary team.Preoperative IM treatment is an evocative treatment strategy for high risk GIST.

Objective To construct four types of glucagon-like peptide-1 (GLP-1) and human serum albumin (HSA) fusion proteins that can be realeased at different rate in vivo by introducing protease cleavage sites between these two moieties.The therapeutic effect and release rate are studied to achieve balanced pharmacokinetics ( PK) and pharmacody-namics ( PD) of GLP-1 and HSA fusion proteins.Methods The gene with different polypeptide joint of GLP-1 and HSA fusion proteins were synthesized by overlap extension PCR amplification, cloned into expression vector pPIC9 and transformed into Pichia pastoris GS115.Then, fusion proteins were obtained by protein purification after being induced by methanol.The preliminary PK and PD of the fusion proteins were studied after purification.Results The fusion protein Gly2-GLP-1-GGGGG-HSA showed no release while Gly2-GLP-1-VTR-HSA, Gly2-GLP-1-SARSVRA-HSA, and Gly2-GLP-1-GRSRVTRSV-HSA showed a slow, medium and fast release rate, respectively, after incubation with furin.In vitro biological activity test results dispalyed that each type of fusion protein promoted insulin secretion of MIN6 cells.In vivo PK test indicated the half-life size of fusion proteins was the largest in Gly2-GLP-1-GGGGG-HSA, followed by Gly2-GLP-1-VTR-HSA, Gly2-GLP-1-SARSVRA-HSA, and Gly2-GLP-1-GRSRVTRSV-HSA.In vivo PD test exhibited hypoglycemic activity that was the highest in Gly2-GLP-1-VTR-HSA, followed by Gly2-GLP-1-SARSVRA-HSA, Gly2-GLP-1-GRSRVTRSV-HSA, and Gly2-GLP-1-GGGGG-HSA.Conclusion GLP-1 can be released from fusion proteins with full activity after the introduction of protease cleavage sites.Releasable fusion proteins at an appropriate release rate have the most balanced PK and PD.