[摘 要] 目的：探究小核核糖核蛋白多肽A（SNRPA）在肝细胞癌（HCC）组织和细胞中的表达及其调控HCC细胞HepG2和Hep3B恶性生物学行为的作用及其机制。方法: 数据库分析SNRPA在泛癌组织中的表达及其与病理分期、HCC患者预后的相关性。常规培养HepG2和Hep3B细胞，将si-NC，si-SNRPA#1、si-SNRPA#2转染HepG2和Hep3B细胞，实验分为si-NC组、si-SNRPA#1组和si-SNRPA#2组；将SNRPA-vector和SNRPA-oe载体转染LO2细胞，分为SNRPA-vector组和SNRPA-oe组。qPCR法检测正常肝细胞和肝癌细胞以及转染各组HepG2和Hep3B细胞中SNRPA mRNA的表达，MTT法、Transwell法和WB法分别检测转染后各组HepG2和Hep3B细胞的增殖、迁移和侵袭能力以及EMT相关蛋白表达的变化。结果: 数据库分析显示，SNRPA mRNA在多数肿瘤组织中均呈高表达（均P<0.001）且与病理分期有关联（P<0.05或P<0.01）。SNRPA在HCC组织和细胞中均呈高表达（P<0.05或P<0.01），且与HCC患者的预后有关联（P<0.01）。敲减SNRPA表达明显抑制HepG2和Hep3B细胞增殖（P<0.05或P<0.01）而过表达SNRPA则能促进LO2细胞增殖（P<0.01），敲减SNRPA表达明显抑制HepG2和Hep3B细胞的迁移和侵袭能力（均P<0.01），明显促进E-cadherin的表达上调（P<0.01），而抑制N-cadherin、vimentin的表达（P<0.01）。结论: SNRPA在HCC组织及细胞中呈明显高表达，其可能通过调控上皮间质转化（EMT）进程进而促进HepG2和Hep3B细胞的增殖、迁移和侵袭。

【Objective】 To compare the diagnostic value of endoscopic ultrasonography (EUS) with conventional imaging in pancreatic occupying lesions so as to provide guidance for the diagnosis of pancreatic occupying lesions. 【Methods】 We selected patients who underwent EUS in the Digestive Endoscopy Room of The First Affiliated Hospital of Xi’an Jiaotong University from September 2012 to December 2019 and were strictly screened by ultrasound endoscopists and diagnosed as pancreatic space-occupying lesions as the study subjects, and collected routine imaging examination results and related information and data of the patients (age, gender, tumor markers, blood amylase, urine amylase, and serum lipase) at the same time. We compared the sensitivity, specificity and accuracy of EUS and conventional imaging methods in the diagnosis of pancreatic space-occupying lesions, and evaluated their diagnostic value. 【Results】 A total of 188 cases were included, with more male ones than female ones. The age of patients in the tumor group was higher than that in the non-tumor group, and the levels of tumor markers serum CA-199 and CEA were significantly higher than those in the non-tumor group. The diagnostic sensitivity of pancreatic space-occupying lesions supported EUS (95.5%), which was significantly better than that of transabdominal ultrasound (54.4%), plain CT (57.1%), enhanced CT (75%), MRI (66.7%), PET-CT (72.7%) and other conventional imaging examination methods. The diagnostic accuracy of pancreatic space-occupying lesions supported EUS (91.5%), better than transabdominal ultrasound (58.3%), plain CT (57.1%), enhanced CT (74.6%), MRI (62.5%), and PET-CT (66.7%). The negative predictive value of the diagnosis of pancreatic space-occupying lesions supported EUS (76.7%), better than transabdominal ultrasound (27.9%), plain CT (22.9%), and enhanced CT (38.1%). 【Conclusion】 In the diagnosis of pancreatic space-occupying lesions, EUS is significantly better than conventional imaging methods such as transabdominal ultrasound, plain CT, enhanced CT, MRI, and PET-CT, and the negative predictive value of EUS is significantly better than that of transabdominal ultrasound, plain CT and enhanced CT.

Objective:To study LIM kinase 1 (LIMK1) expressional condition, and its regulatory effects on the proliferation and metastasis of hepatocellular carcinoma cells and tissues.Methods:The online database starBase v3.0 and GEPIA were used to analyze the LIMK1 expression in hepatocellular carcinoma cells and normal liver tissues, and then the relevant survival analysis was performed. LIMK1 expression in hepatocellular carcinoma cell line was analyzed by Western blot. Hep3B and Huh7 cells were transiently transfected after LIMK1 protein expression was down-regulated by small interfering RNA (siRNA). LIMK1 effects on the proliferation of Hep3B and Huh7 cells were observed by MTT assay and colony formation assay. Transwell assay was used to detect the change in metastatic ability of hepatocellular carcinoma cell after the down-regulation of LIMK1 expression. Western blot was used to detect the changes of related indexes in the process of epithelial mesenchymal transition after the down-regulation of LIMK1 expression. Data were analyzed by one-way ANOVA.Results:The expression level of LIMK1 in liver cancer tissues was significantly higher than that of normal liver tissues, and was related with prognosis ( P ?< 0.01). Furthermore, LIMK1 expression in HCC cell lines was significantly higher than that of immortalized liver L02 cells ( P < 0.05). Functional correlated experiment showed that the proliferation and metastatic ability of liver cancer cells were significantly inhibited after LIMK1 expression down-regulation ( P < 0.05). Simultaneously, LIMK1 was also involved in the process of epithelial-mesenchymal transition. Conclusion:LIMK1 was overexpressed in HCC tissues and cells, and may regulate the proliferation and metastasis of HCC cells and participate in epithelial-mesenchymal transition process.

【Objective】 To explore the application value of preoperative gastroscopic carbon nanoparticles labeling in patients undergoing laparoscopic radical gastric cancer surgery. 【Methods】 We included cases of laparoscopic radical gastric cancer surgery at The First Affiliated Hospital of Xi’an Jiaotong University from January 2017 to December 2019. Some cases received submucosal injection of carbon nanoparticles under the gastroscope before surgery. The effects of carbon nanoparticles labeling on the number of lymph nodes detected， operation duration and surgical complications were compared and analyzed. 【Results】 A total of 397 patients undergoing laparoscopic radical gastric cancer surgery were enrolled. Among them， 78 cases underwent gastroscopic carbon nanoparticles tracer labeling before operation. No complications were observed. The total number of lymph nodes detected by pathology after surgery in the carbon nanoparticles group significantly increased ［（22.0（4.0） vs. 22.0（3.0）， P=0.033）］ while the operation time significantly reduced ［（185.0±37.48）min vs. （213.4±23.66）min， P<0.001］ compared with those in the control group. New gastric cancer lesions were revealed by gastroscopy in three cases （3.8%） of carbon nanoparticles labeling， and the original planned operation method was changed in two cases （2.6%）. 【Conclusion】 Preoperative endoscopic carbon nanoparticles tracer labeling can not only help shorten the time of laparoscopic radical surgery for gastric cancer and increase the number of total lymph nodes detected for more accurate TNM staging， but also provide an opportunity for the discovery of synchronous multiple gastric cancer.

Objective:To investigate the esophageal microecology in patients with esophageal carcinoma (EC), and to compare the difference in esophageal flora between patients with esophageal cancer and healthy people.Methods:From July 2018 to July 2019, at Taihe Hospital, 82 EC patients and 20 age-and gender-matched healthy controls during the same period were selected. The pathology of EC were divided into poorly differentiated (8 cases), moderately differentiated (9 cases) and well differentiated cancers (13 cases) according to the degree of differentiation. The esophageal tissue samples of EC patients and healthy individuals were collected. Sample DNA was extracted and the V4 region of bacterial 16S rRNA was amplified by polymerase chain reaction (PCR). Sequencing was performed by lllumina HiSeq 4000 sequencing platform. Alpha-diversity analysis and principal co-ordinates analysis (PCoA) were performed, and linear discriminant analysis (LDA) of linear discriminant analysis effect size (LEfSe) was used to screen different species. The random forest model was verified by receiver operating characteristic (ROC) curve and the esophageal bacterial phenotype was predicted by BugBase database. Non-parametric Kruskal-Wallis H test and Wilcoxon rank sum test were used for statistical analysis. Results:The Chao1 index of the EC patients was higher than that of healthy controls (362.51(284.29, 646.13) vs. 284.83(244.31, 344.74)), and the difference was statistically significant ( Z=-2.857, P=0.004). The results of PCoA showed that the distance between samples of EC patients and healthy control samples was relatively close, and there was no significant difference in the composition of microecology between the two groups ( P>0.05). The abundance of esophageal Cyanobacteria and Verrucomicrobia of EC patients were both higher than those of healthy controls (0.2% vs. 0.1%, 0.4% vs. 0), while the abundances of esophageal Proteobacteria, SR1 and TM7 phylum of EC patients were lower than those of healthy controls (21.9% vs. 34.2%, 0.1% vs. 0.2%, 0.2% vs. 0.5%), and the differences were statistically significant ( Q=0.090, 0.077, 0.010, 0.026 and 0.001, all P<0.05). The abundances of Clostridia, Elostridiales, Pasteurella, Pasteurellaceae, Eikenella, Actinobacillus and Haemophilus in poorly differentiated patients, moderately differentiated and higher differentiated patients were 28.3%, 24.2% and 17.0%, 28.3%, 24.2% and 17.0%, 3.2%, 0.3% and 5.0%, 3.2%, 0.3% and 5.0%, 0, 1.5% and 0.1%, 0.5%, 0 and 0.7%, 1.3%, 0.2% and 3.9%, respectively, and the differences were statistically significant ( Q=0.579, 0.557, 0.390, 0.711, 0.768, 0.768 and 0.768, all P<0.05). LEfSe analysis showed that the abundances of Fusobacterium, Ruminococcus, Odorbacterium and S24_7 of EC patients were higher than those of healthy controls (21.5% vs. 11.7%, 0.5% vs. 0.1%, 0.1% vs. 0 and 0 vs. 0), and the differences were statistically significant (LDA=2.591, 2.379, 2.790 and 2.927, all P<0.05). The ROC curve confirmed that the random forest model was reliable and the AUC value was 0.92. BugBase database phenotypic prediction showed that the phenotype of esophageal bacteria related to biofilm formation, pathogenic potential, mobile elements, oxygen demand (aerobic, anaerobic and facultative bacteria), and oxidative stress tolerance of EC patients were more abundant than those of healthy controls (all P<0.05). Conclusions:The esophageal flora of patients with esophageal cancer has changed. Fusobacterium, Ruminococcus, Odoribacterium and S24_7 may be potential biomarkers of esophageal flora.

[Abstract] Objective: To explore the regulatory effect of long non-coding RNA (lncRNA) SNHG5 on invasion and migration of hypoxia-induced hepatocellular carcinoma (HCC) cells. Methods: A total of 20 pairs of cancer and para-cancerous tissue specimens resected from HCC patients in the First Affiliated Hospital of Xi'an Jiaotong University from January 2017 to June 2018, and human HCC cell lines (HepG2, MHCC-97L, MHCC-97H , Huh7) as well as immortalized human liver LO2 cells were collected for this study. Bioinformatics methods were used to analyze the binding sites between hypoxia-inducible factor 1α (HIF-1α) and SNHG5. pCMVHIF-1α and shRNA-SNHG5 (sh-SNHG5) plasmids were transfected into HCC cells, respectively. qPCR was used to detect the expres‐ sion level of SNHG5 in HCC tissues and hypoxia-induced HCC cells. Western botting was used to detect the expression level of HIF-1α protein in HCC cells, and Transwell chamber method was used to detect the migration and invasion ability of HCC cells after SNHG5 si‐ lence under normoxia and hypoxia condition. Results: Compared with para-cancerous tissues and immortalized human liver LO2 cells, the expression of SNHG5 was significantly up-regulated in HCC tissues and cell lines (all P<0.01). Hypoxia promoted the expression level of SNHG5 in HCC cells, and its mechanism might be related to the combination of hypoxia-activated HIF-1α and SNHG5 promoter to promote its transcription. Hypoxia promoted the invasion and migration ability of HepG2 and MHCC-97L cells (all P< 0.01), but knockdown of SNHG5 significantly inhibited the invasion and migration ability of HepG2 and MHCC-97L cells under hy‐ poxic conditions (all P<0.01). Conclusion: SNHG5 is highly expressed in HCC tissues and cell lines and plays an important role in the invasion and migration of HCC cells induced by hypoxia.

Objective To investigate the expression of DNA methyltransferase 3b (DNMT3B) in hepatocellular carcinoma (HCC) and its effect and mechanism on the proliferation,invasion and migration of HCC cells.Methods The expression of DNMT3B gene was detected by qRT-PCR in 46 cases of HCC tissues and corresponding adjacent tissues;the results and clinical pathological parameters were analyzed.SiRNA targeting DNMT3B was transfected into MHCC97-H cells by RNA interference (RNAi) technique.The mRNA and protein expression levels of related genes were detected by qRT-PCR and Western blot.The cell proliferation was measured by MTT assay,and the invasion and migration abilities were measured by Transwell assay.Results In 46 HCC patients,the expression of DNMT3B (73.91％) was significantly higher in HCC than in adjacent normal tissue.The high expression of DNMT3B gene was associated with histological type and tumor size of HCC (all P＜0.05).Inhibition of DNMT3B gene expression decreased proliferation,invasion and migration of MHCC97-H cells.Interference with DNMT3B gene increased the expressions of tumor suppressor genes RASSFA1,APC and MTSS1 at mRNA and protein levels.Conclusion DNMT3B is associated with the progression of HCC.It may inhibit the proliferation,invasion and migration of HCC cells by regulating the methylation of downstream tumor suppressor gene.

According to the training requirement of the biological-psycho-social medical model to the clinical medical students,in order to promote the comprehensive improvement of medical students' professional knowledge,hands-on ability and human qualities,we design the course of introduction to clinical medicine.Through six major functional modules such as basic professional quality,clinical diagnosis basic technology,basic skills related to the operation,the new progress in clinical medicine and technology,clinical nursing and medical relationship,and medical information management,we build up the core content of integrated course of introduction to clinical medical.In this way,the clinical and related basic knowledge and skills are integrated,the clinical course is closely connected with the basic curriculum,the medical and the humanities exchanges.Through this design,the foundation is laid out for the collaborative efforts of the organ-system of integrated curriculum reform.

Objective To detect the expression profile of transcription factor C/EBPβ in human immortalized normal hepatic cell lines and hepatocellular carcinoma cell lines so as to determine the correlation between C/EBP3 with cell death mediated by endoplasmic reticulum stress in hepatocellular cells.Methods We cultured the human immortalized normal hepatic cells lines HHL5 and HL7702 and hepatocellular carcinoma cell lines SMMC7721;Bel7402,HepG2 and Hep3B.Hep3B cells were used as the cell model in tunicamycin-induced endoplasmic reticulum stress.Cellular morphology was observed under an inverted optical microscope.MTT assay was used to assess the inhibition of cell growth.To detect cell apoptosis,the cells were dyed with Hoechst 33258 and observed using a fluorescence microscope.RToPCR and Western blotting were used to detect the expression of at mRNA and protein levels,respectively.Results We found that normally the mRNA and protein isoform of C/EBPβ,C/EBPβ-1,were both expressed in all of the four hepatocellular cell lines and the two immortalized normal hepatic cell lines,while C/EBPβ protein isoform C/EBPβ-3 was only expressed in the two immortalized normal hepatic cell lines.Tunicamycin increased the expressions of both mRNA and protein of C/EBPβ in Hep3B cells and the increase of protein isoform C/EBPβ-3 was the most remarkable.In Hep3B cells,cell death was induced by tunicamycin through endoplasmic reticulum stress activity.Apoptosis as well as paraptosis was observed in tunicamycin-induced cell death.Conclusion C/EBPβ-3,one of the protein isoforms of C/EBPβ,is only expressed in normal hepatic cell lines,but not in hepatocellular cell lines.C/EBPβ is involved in cell death mediated by endoplasmic reticulum stress activity in hepatocellular carcinoma cells.

OBJECTIVE: To compare the efficacy and safety of combined radiofrequency ablation (RFA) and transcatheter arterial chemoembolization (TACE) with RFA alone for hepatocellular carcinomas (HCC). MATERIALS AND METHODS: Randomized controlled trial (RCT) studies that compared the clinical or oncologic outcomes of combination therapy of TACE and RFA versus RFA for the treatment of HCC were identified through literature searches of electronic databases (Pubmed, Embase, Cochrane Library, China Biology Medicine disc, China National Knowledge Infrastructure, and Google Scholar). Hazard ratios (HRs) or odds ratios (ORs) with their corresponding 95% confidence interval (CI) were combined as the effective value to assess the summary effects. The strength of evidence was rated by the Grading of Recommendations Assessment, Development, and Evaluation system. RESULTS: Six RCTs with 534 patients were eligible for inclusion in this meta-analysis. The meta-analysis showed that the combination of TACE and RFA is associated with a significantly longer overall survival (HR = 0.62, 95% CI: 0.49-0.78, p < 0.001) and recurrence-free survival (HR = 0.55, 95% CI: 0.40-0.76, p < 0.001) in contrast with RFA monotherapy. The seemingly higher incidence of major complications in the combination group compared with RFA group did not reach statistical significance (OR = 1.17, 95% CI: 0.39-3.55, p = 0.78). CONCLUSION: In patients with HCC, the combination of TACE and RFA is associated with significantly higher overall survival and recurrence-free survival, as compared with RFA monotherapy, without significant difference in major complications.
Carcinoma, Hepatocellular/*surgery ; Catheter Ablation/adverse effects/*methods ; Chemoembolization, Therapeutic/adverse effects/*methods ; China ; Combined Modality Therapy ; Disease-Free Survival ; Humans ; Liver Neoplasms/*surgery ; Odds Ratio ; Proportional Hazards Models ; Treatment Outcome