Gastric cancer is the second most common malignancy in China.In recent years,with the development of laparoscopic technology and improved skills of gastrointestinal surgeons,total laparoscopic radical gastrectomy for gastric cancer has been developed rapidly.The digestive tract reconstruction is the key procedure and one of the difficulties of total laparoscopic radical gastrectomy.Each method of digestive tract reconstruction has its own characteristic,however,there has not yet reached a unified consensus until today.In this article,the advantages and disadvantages of these reconstruction methods and technical points were reviewed based on relative literatures and our application experiences.

Background and purpose: Peroxiredoxin Ⅱ (PrxⅡ) has the activity of peroxidase. The relevant studies found it played an important role in gastric cancer. This study aimed to investigate the expression of PrxⅡ in human gastric cancer tissues and cells, analyze its relationship with clinicopathological characteristics, and explore the relationship between PrxⅡ and the prognosis and the development of gastric cancer. Methods: The expression of PrxⅡmRNA and protein in gastric cancer tissues and the paired adjacent normal tissues from 45 patients was detected by real-time fluorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot. The same methods were used to detect the expression of PrxⅡ mRNA and protein in GES-1, MGC-803, MKN-45 and MKN-28. Tissue mi-croarray and immunohistochemistry were used to detect the expression of PrxⅡ protein in gastric cancer tissues and the paired adjacent normal tissues from 116 patients. The relationship between the results and clinicopathological char-acteristics was analyzed. The prognosis was analyzed. Results: According to results of RTFQ-PCR and Western blot, we found that PrxⅡ mRNA and protein in gastric cancer tissues were significantly higher than that in adjacent normal tissues (P<0.05). PrxⅡ mRNA and protein in gastric cancer cells were higher than that in normal gastric cells (P<0.01).Immunohistochemistry revealed that the expression of PrxⅡ protein in gastric cancer tissues (76.7％) was also significantly higher (P<0.01) than that in adjacent normal tissues (30.1％). The expression of PrxⅡ protein is significantly related to tumor size, histological differentiation, depth of invasion, TNM stage and lymph node metastasis (P<0.05), but had no significant relationship with the gender, age, tumor location and distant metastasis. Survival in patients with higher PrxⅡ expression significantly shorter than in those with lower expression (P<0.01). PrxⅡ is an independent prognostic factor of gastric cancer (P<0.05). Conclusion: PrxⅡ promotes the development of gastric cancer. It is one of the adverse prognostic factors of gastric cancer and may serve as a new therapeutic target for gastric cancer.

Objective To explore the expression of Ras-related protein 11(Rab11)in hypoxia, the effect of Rab11 on the invasion and migration of cervical cancer cell line SiHa and its possible mechanism. Methods SiHa cells were divided into 4 groups, the normoxic blank group (normal culture in normoxia), the hypoxic blank group (normal culture in hypoxia), the negative control group [transfection of negative control small interfering RNA(siRNA)in hypoxia], the Rab11-siRNA group (transfection of Rab11 siRNA in hypoxia). Western blot was used to examine the expression of Rab11, integrin α5, integrin β3, phosphorylated focal adhesion kinase(p-FAK), phosphorylated phosphatidylinositol 3 kinase(p-PI3K) protein, together with the expression of Ras correlative C3 creotoxin substrate 1(Rac1), which was critical in regulating cell invasion. The mRNA expression of Rab11 in the 4 groups was detected by realtime-qPCR. The cell invasion was detected by matrigel assay, while the cell migration was detected by transwell assay. Immunofluorescence was used to identify intracellular location of Rac1 in SiHa cell. Results (1) The expression of Rab11, intergrin α5, intergrin β3, p-FAK, p-PI3K and Rac1 in the normoxic blank group were 0.56±0.04, 0.33±0.03, 0.32±0.03, 0.36±0.03, 0.35±0.03 and 0.47±0.03, respectively. In the hypoxic blank group, they were 0.73±0.03, 0.74±0.03, 0.61±0.03, 0.62±0.03, 0.60±0.03 and 0.73±0.03, respectively. In the negative control group, their expressions were 0.72±0.03, 0.73±0.03, 0.59±0.03, 0.61±0.03, 0.59±0.03 and 0.72±0.03, respectively. While in the Rab11-siRNA group, they were 0.44±0.03, 0.30±0.03, 0.29±0.03, 0.30±0.03, 0.30±0.03 and 0.34±0.04, respectively. The expressions of Rab11, α5, β3, p-FAK, p-PI3K and Rac1 were significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and were significantly lower in the Rab11-siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (2) The expressions of Rab11-mRNA were 1.000±0.000, 1.454±0.114, 1.442±0.101, 0.570± 0.046 in the normoxic blank group, the hypoxic blank group, the negative control group and the Rab11- siRNA group, respectively. It was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11-siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (3) By Matrigel, the invasion cell number in the normoxic blank group, the hypoxic blank group,the negative control group and the Rab11-siRNA group were 65±12, 106±16, 104± 17 and 50±11, respectively. The invasion capacity was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11- siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (4) By transwell assay, the migration cells in the normoxic blank group, the hypoxic blank group, the negative control group and the Rab11-siRNA group were 127±12, 169±15, 161±13 and 77±13, respectively. The capacity of invasion was significantly higher in the hypoxic blank group than in the normoxic blank group(P<0.05), and was significantly lower in the Rab11- siRNA group than in the hypoxic blank group and the negative control group(P<0.05). (5) The immunofluorescence showed that the red fluorescence intensity around nucleus was significantly increased in the normoxic blank group, the hypoxic blank group and the negative control group than in the Rab11- siRNA group. Conclusions Hypoxia could promote the invasion and migration of SiHa cells. In hypoxia, the down regulation of Rab11 expression could inhibit the invasion and migration of SiHa cells. This might be due to the decreased expression of the intergrin α5, intergrin β3, p-FAK, p-PI3K and Rac1 protein.

Objective To analyze the expression feature and function of microRNAs in exosomes secreted by leukemia cells (LCEX). Methods The mice leukemia cell line L1210 was taken as the example, and LCEXL1210 was obtained by isolating supernate of L1210 cells through density gradient centrifugation. MicroRNAs isolated from LCEXL1210 were analyzed by microarray analysis, compared with miRNA from L1210 cell line, and then some of miRNAs with different expression were verified by real-time PCR and were analyzed by Gene Ontology (GO) database. Results The number of miRNAs identified in LCEXL1210 was 1 044, and that in L1210 cell line was 872. The number of shared miRNAs between LCEXL1210 and L1210 cell line was 732, accounting for 70.1 % of LCEXL1210 and 83.9 % of L1210 cell line, respectively, which indicated that 70 % of LCEXL1210 was derived from the parental cells. Interestingly, 312 miRNAs in LCEXL1210 were found to be underrepresented in the parental cells, indicating their specificity in LCEXL1210. Some miRNAs were significantly highly expressed in LCEXL1210 compared with those in L1210 cell line, including miR-16-1, miR-210, miR-195 and so on, which showed that miRNAs isolated from LCEXL1210 were differentially expressed with those from the parental cells. Some differentially expressed miRNAs from LCEXL1210 were verified by real-time PCR, and then were analyzed by GO database, which demonstrated that these highly expressed miRNAs participated in the processes of various biological function and signal transduction. Conclusions MiRNAs isolated from LCEXL1210 show a high similarity to miRNAs isolated from L1210 cells, whereas of which one-third are specific. The highly expressed miRNAs participate in the processes of various biological function and signal transduction.

Objective:To investigate the short-term efficacy of totally laparoscopic distal gastrectomy (TLDG) after endoscopic submucosal dissection (ESD) versus direct TLDG for early gastric cancer.Methods:The propensity score matching and retrospective cohort study was conducted. The clinicopathological data of 623 patients with early gastric cancer who were admitted to the First Affiliated Hospital of Nanjing Medical University from March 2014 to December 2019 were collected. There were 405 males and 218 females, aged from 26 to 86 years, with a median age of 62 years. Of 623 patients, 25 cases undergoing TLDG after ESD were divided into ESD+TLDG group and 598 cases undergoing TLDG directly were divided into TLDG group. Observation indicators: (1) the propensity score matching conditions and comparison of general data between the two groups after propensity score matching; (2) intraoperative and postoperative situations of TLDG; (3) stratification analysis of the ESD+TLDG group. The propensity score matching was conducted by 1∶2 matching using the nearest neighbor method. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was done using the t test. Measurement data with skewed distribution were represented as M (range) and comparison between groups was done using the Mann-Whitney U test. Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test or Fisher exact probability. Comparison of ordinal data between groups was analyzed using the Mann-Whitney U test. Results:(1) The propensity score matching conditions and comparison of general data between the two groups after propensity score matching: 75 of 623 patients had successful matching, including 25 in the ESD+TLDG group and 50 in the TLDG group. Before propensity score matching, the body mass index (BMI), cases with tumor diameter ≤20 mm, 21 to 30 mm or>30 mm, cases with tumor classified as stage Ⅰ, stage Ⅱ or stage Ⅲ of clinical staging were (22.3±3.6)kg/m 2, 16, 6, 3, 24, 1, 0 of the ESD+TLDG group, respectively, versus (24.3±2.7)kg/m 2, 238, 125, 235, 312, 126, 160 of the TLDG group, showing significant differences in the above indicators between the two groups ( t=2.744, Z=?2.834, ?4.209, P<0.05). After propensity score matching, the BMI, cases with tumor diameter ≤20 mm, 21 to 30 mm or >30 mm, cases with tumor classified as stage Ⅰ or stage Ⅱ of clinical staging were (22.3±3.6)kg/m 2, 16, 6, 3, 24, 1 of the ESD+TLDG group, versus (23.6±2.9)kg/m 2, 29, 12, 9, 48, 2 of the TLDG group, showing no significant difference between the two groups ( t=1.542, Z=?0.597, 0.000, P>0.05). (2) Intraoperative and postoperative situations of TLDG: after propensity score matching, the operation time and time to postoperative drainage tube removal were 180 minutes(range, 124 to 289 minutes) and 6 days(range, 4 to 13 days) of the ESD+TLDG group,respectively,versus 170 minutes(range, 106 to 250 minutes) and 6 days (range, 4 to 9 days) of the TLDG group, showing significant differences between the two groups ( Z=-2.396, -3.039, P<0.05). Cases with the volume of intraoperative blood loss <50 mL, 50 to 100 mL or >100 mL, the number of lymph node dissected, duration of postoperative hospital stay, cases with perioperative complications as incision fat liquefaction, delayed gastric emptying, anastomotic bleeding or pulmonary infection were 7, 9, 9,34(range, 16 to 58), 8 days(range, 6 to 31 days), 1, 1, 0, 0 of the ESD+TLDG group,respectively,versus 18, 26, 6, 39 (range, 22 to 68), 8 days (range, 6 to 29 days), 0, 0, 1, 1 of the TLDG group, showing no significant difference between the two groups ( Z=-1.703, -1.958, -1.139, χ2=0.033, P>0.05). Cases with anastomotic bleeding were recovered after hemostasis under endoscopy and cases with other perioperative complications were recovered after conservative treatment. (3) Stratification analysis of the ESD+TLDG group. ① For 5 cases undergoing TLDG ≤14 days after ESD and 20 cases undergoing TLDG >14 days after ESD, the operation time of TLDG, cases with the volume of intraoperative blood loss <50 mL, 50 to 100 mL or >100 mL during TLDG, the number of lymph node dissected, time to postoperative drainage tube removal, duration of postoperative hospital stay, cases with perioperative complications were 200 minutes(range, 170 to 289 minutes), 0, 3, 2, 36(range, 9 to 57), 7 days(range, 5 to 9 days), 8 days(range, 7 to 9 days), 1 and 180 minutes (range, 124 to 253 minutes), 8, 6, 6, 34(range, 8 to 78), 6 days(range, 4 to 13 days), 8 days(range, 6 to 31 days), 1, respectively, showing no significant difference in the operation time of TLDG, volume of intraoperative blood loss during TLDG, the number of lymph node dissected, time to postoperative tube removal and duration of postoperative hospital stay between the two groups ( Z=?1.536, ?1.993, ?0.238, ?0.932, ?0.589, P>0.05), and no significant difference in cases with perioperative complications between the two groups ( P>0.05). ② For 13 cases undergoing TLDG ≤21 days after ESD and cases undergoing TLDG >21 days after ESD, the operation time of TLDG, cases with the volume of intraoperative blood loss as <50 mL, 50 to 100 mL or >100 mL during TLDG, the number of lymph node dissected, time to postoperative drainage tube removal, duration of postoperative hospital stay, cases with perioperative complications were 200 minutes(range, 145 to 289 minutes), 2, 6, 5, 34(range, 8 to 57), 6 days(range, 4 to 11 days), 8 days(range, 6 to 11 days), 1 and 179 minutes(range, 124 to 240 minutes), 6, 3, 3, 34(range, 16 to 78), 6 days(range, 5 to 13 days), 8 days(range, 6 to 31 days), 1, respectively, showing a significant difference in the operation time of TLDG between the two groups ( Z=?2.241, P<0.05), while showing no significant difference in the volume of intraoperative blood loss during TLDG, the number of lymph node dissected, time to postoperative drainage tube removal, duration of postoperative hospital stay between the two groups ( Z=?1.471, ?0.163, ?0.084, ?0.194, P>0.05) and no significant difference in cases with perioperative complications between the two groups ( P>0.05). ③ For 15 cases undergoing TLDG ≤28 days after ESD and 10 cases undergoing TLDG >28 days after ESD, the operation time of TLDG, cases with the volume of intraoperative blood loss <50 mL, 50 to 100 mL or >100 mL during TLDG, the number of lymph node dissected, time to postoperative drainage tube removal, duration of postoperative hospital stay, cases with perioperative complications were 190 minutes (range, 145 to 289 minutes), 2, 7, 6, 33(range, 8 to 57), 6 days(range, 4 to 11 days), 8 days(range, 6 to 31 days), 1 and 179 minutes(range, 124 to 240 minutes), 6, 2, 2, 37(range, 16 to 78), 6 days (range, 5 to 13 days), 8 days(range, 6 to 14 days), 1, respectively, showing no significant difference in the operation time of TLDG, volume of intraoperative blood loss during TLDG, the number of lymph node dissected, time to postoperative tube removal and duration of postoperative hospital stay between the two groups ( Z=?1.619, ?2.000, ?0.667, ?0.370, ?0.057, P>0.05), and no significant difference in cases with perioperative complications between the two groups ( P>0.05). Conclusions:Compared with cases undergoing TLDG directly, the operation time to TLDG and time to drainage tube removal after TLDG for cases undergoing ESD+TLDG are prolonged, but there is no difference in the short-term efficacy. For cases undergoing TLDG ≤21 days after ESD and cases undergoing TLDG >21 days after ESD, there is a significant difference in the operation time of TLDG.

Objective To explore the expression of miRNA-181a (miR-181a) in patients with multiple myeloma (MM) and its effect on biological features of MM cells. Methods CD138+cells of bone marrow from 25 MM patients and 10 patients with hematological non-malignancies were purified by using immunomagnetic separation, and the expression of miR-181a in CD138+cells and MM cell lines including RPMI 8226, H929 and U266 were detected by real-time quantitative PCR. The effects of down-regulation and up-regulation of miR-181a expression on the biological characteristics of MM cells were studied with miR-181a antagomir and agomir. Results Compared with patients with hematological non-malignant diseases, the expression of miR-181a in CD138+ cells was upregulated in MM patients. Compared with CD138+ cells in hematological non-malignancies, high expressions of miR-181a were observed in RPMI 8226 and U266 myeloma cell line, while low expressions of miR-181a were observed in H929 cells. Down-regulation of miR-181a with 100 nmol/L miR-181a antagomir could inhibit the proliferation of U266 cells at 24,48 and 72 h [(67.1 ± 3.3) %vs. (50.5 ± 4.1) %, (71.5 ± 3.6) % vs. (52.3 ± 2.2) %, (78.1 ± 5.4) % vs. (69.5 ± 4.3) %, P < 0.05 respectively], whereas up-regulation of miR-181a with 100 nmol/L miR-181a agomir could significantly promote the proliferation of H929 cells at 24 h and 48 h [(21.2 ± 2.4) %vs. (38.5 ± 3.6) %, ( 61.3 ± 5.4) %vs. (82.2 ±6.9)%, P<0.01 respectively]. Cell cycle analysis showed that miR-181a antagomir made U266 cell cycle arrest in the G0/G1 phase. Meanwhile, susceptibility test results indicated that the apoptosis of U266 cells induced by doxorubicin, paclitaxel and 5-fluorouracil was increased when the proliferation of miR-181a expression was down-regulated with miR-181a antagomir. In migration assay, the data showed that down-regulation of miR-181a with miR-181a antagomir could inhibit the migration of U266 cells, and the proportion of migrated cells in the experimental group (62 ± 10) %was lower than that in the control group (89 ± 12) %(P< 0.05), whereas up-regulation of miR-181a with miR-181a agomir could improve the migration of H929 cells, and the proportion of migrated cells in the experimental group (242 ± 9) % was higher than that in the control group (98 ± 8)%(P<0.01). Conclusions The high expression of miR-181a expressed highly by MM cells may promote the proliferation, migration and drug resistance of myeloma cells, indicating that miR-181a could be an important prognostic biomarker candidate, and the application of gene silencing may improve the prognosis of MM.

The incidence of adenocarcinoma of the esophagogastric junction (AEG) has been raised in recently years.Comprehensive treatment based on surgical treatment is currently a general strategy for the treatment of AEG.As a minimally invasive treatment,laparoscopic surgery has been gradually applied to the treatment of AEG.Because of the particularity of the anatomy and pathology of AEG,laparoscopic radical resection still has many difficulties and controversies.Up to now,there are a few high-level evidences for the range of lymph node dissection and gastrectomy and the selection of digestive tract reconstruction,and the treatment strategy of total laparoscopic surgery for AEG has not reached a consensus.Therefore,laparoscopic surgery for AEG has gradually become a hot topic in clinical research.Here,combined with the experience of laparoscopic surgery for AEG and the latest guidelines and literatures,authors presented the general strategies for the laparoscopic treatment of AEG in our center.

The incidence of the esophagogastric junction (EGJ) cancer tends to increasing in recent years.Comprehensive treatment based on surgical treatment is currently a general strategy for the treatment of EGJ cancer.Because of the particularity of the anatomy and pathology of EGJ cancer,there were difficulties and controversies existing in the surgical treatment of EGJ cancer.Medical researchers have attached great importance to the treatment of EGJ cancer and made remarkable progress in it.Therefore,the authors summarize the progress of surgical treatment of EGJ cancer,and present it in four aspects of surgical approach lymph,node dissection,esophagogastric resection and digestive tract reconstruction.

In recent years, with the development of laparoscopic technology, more and more totally laparoscopic gastrectomy has been applied. The reconstruction of digestive tract is the key procedure of laparoscopic gastrectomy and is associated with the postoperative quality of life. Each method of digestive tract reconstruction has its own characteristic, however, unified consensus on how to choose the optimal method for digestive tract reconstruction has not yet been reached till today. In this article, we will discuss and evaluate the advantages, disadvantages and indications of these reconstruction methods after totally laparoscopic distal gastrectomy, total gastrectomy and proximal gastrectomy combined with relative literatures and our practical experience in order to provide the reference to choose the reasonable reconstruction method at the premise of radical resection, which may decrease the morbidity of postoperative complication, increase the quality of life, and bring benefits to patients definitely.

Objective To evaluate the clinical improvements after autologous bone marrow mononuclear cells (BMMNCs) percutaneously injected into coronary artery in patients with heart failure due to non-ischemic cardiomyopathy using PET myocardial peffusion/metabolic imaging.Methods From February 2011 to October 2012,40 patients with heart failure due to non-ischemic cardiomyopathy were selected.The test group including 15 patients (13 males,2 females,average age (57.5±14.5) years) received the autologous BMMNCs intracoronary injection on the basis of drug treatment.The other 25 cases (21 males,4 females,average age (58.0±12.0) years) were taken as the control group and only received the drug treatment.All patients were followed up for 24 months,and the myocardial perfusion/metabolism imaging,echocardiography,brain natriuretic peptide (BNP) test,6-minute walking experiment were performed.The data were analyzed by two-sample t test.Results During the follow-up period,the test group had no ventricular arrhythmia and other serious complications,and the patients' symptoms had been improved.There was no change in myocardial perfusion after treatment of autologous BMMNCs,but the myocardial metabolic defect by volume reduced from (43.79± 17.99) cm3 to (28.19±9.27) cm3 (t =3.33,P＜0.01) 24 months after the treatment.The myocardial metabolic defect by volume at the baseline and after 24 months in the control group was (43.30±15.70) cm3,(48.51±15.77) cm3 respectively (t=1.01,P＞0.05).In the test group,the left ventricular end-diastolic diameter decreased from (64.0±8.0) mm to (59.0±7.0) mm 24 months after the treatment (t=2.04,P＜0.05),and the left ventricular ejection fraction was significantly higher than that before treatment:(45.0±4.0) ％ vs (27.0±6.0) ％ (t =10.81,P＜0.01).Conclusion PET myocardial perfusion/metabolic imaging can be used as tools in evaluating the therapeutic effect of autologous BMMNCs in patients with heart failure due to non-ischemic cardiomyopathy.