AIM:To investigate the effect of cis-dichlorodiamine platinun ( cDDP) resistance on proliferation , apoptosis, migration and angiogenesis of esophageal cancer cell line KYSE 150.METHODS:Using the method of increa-sing concentration of cDDP in culture for 10 months, the human esophageal carcinoma cDDP-resistant cell line named KYSE150/cDDP was established successfully .The drug sensitivity was measured by MTT assay .The changes of the biolog-ical behaviors between the parental cell line and resistant cell line were determined by morphological observation assay , MTT assay, colony formation assay , DAPI staining, wound healing assay and tube formation experiment .RESULTS: No significant morphological difference between KYSE 150 cells and KYSE150/cDDP cells was observed .Compared with KYSE150 cells, the drug resistance index of KYSE150/cDDP cells was 6.35, and the viability of KYSE150/cDDP cells was decreased.The colony formation rate of KYSE150/cDDP cells was (15.00 ±3.05)%, while the colony formation rate of KYSE150 cells was (86.70 ±6.57)%.The apoptotic rate of KYSE150/cDDP cells was (0.63 ±0.09)%, and that of KYSE150 cells was (8.46 ±1.33)%.Compared with KYSE150 cells, KYSE150/cDDP cells showed a stronger healing ability of scratch, and the migration rate was higher than that of KYSE 150 cells.The results of tube formation experiment showed that the vessel number in KYSE150/cDDP group was 76.20 ±3.18, while the vessel number in KYSE150 group was 50.60 ±1.33.The protein expression of MMP-2 and VEGFR2 in KYSE150/cDDP cells was higher than that in KYSE150 cells.CONCLUSION: KYSE150/cDDP cells present drug-resistant phenotype and show a slow growth rate . The ability of apoptosis is decreased , and the abilities of cell migration and angiogenesis are increased .This may be an im-portant reason for the failure of clinical chemotherapy for esophageal cancer .

OBJECTIVETo explore the role of P38 signaling pathway in neonatal rat astrocyte swelling and the expression of aquaporin-4 (AQP4) after oxygen-glucose deprivation (OGD) and recovery (OGD/R).

METHODSPrimarily cultured neonatal rat astrocytes were subject to OGD for 5 h followed by oxygen-glucose recovery in the presence or absence of the P38 inhibitor SB203580 (10 µmol/L). The astrocytes were investigated at 0.5, 2, 8 and 24 h after oxygen-glucose recovery for morphological changes and cell injuries using lactate dehydrogenase (LDH) assay. The expressions of P38, P-P38, and AQP4 mRNAs and proteins in the astrocytes were detected using RT-PCR and Western blotting.

RESULTSOGD/R caused significantly enhanced expression of P-P38 protein, and this effect was blocked by SB203580. AQP4 mRNA and protein expression declined transiently at 0.5 h after OGD and increased gradually to reach the peak level at 8 h (P<0.05). Application of the SB203580 significantly lowered OGD-induced AQP4 mRNA and protein up-regulation (P<0.05). Astrocyte swelling occurred after OGD/R but was obviously lessened by SB203580. LDH release increased markedly after OGD/R, and was attenuated by treatment with SB203580 (P<0.01).

CONCLUSIONP38 signaling pathway participates in astrocyte swelling after OGD/R, and blocking this pathway can attenuate AQP4 up-regulation and ameliorate the cell swelling.

Animals ; Animals, Newborn ; Aquaporin 4 ; metabolism ; Astrocytes ; metabolism ; pathology ; Brain Edema ; metabolism ; pathology ; Cell Hypoxia ; Glucose ; pharmacology ; MAP Kinase Signaling System ; physiology ; Oxygen ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism

ObjectiveTo investigate clinical application of intraoperative intraperitoneal hyperthermic chemotherapy using sustained-release fluorouracil in radical gastrectomy for advanced gastric cancer.MethodsThe clinical data of 280 advanced gastric cancer patients admitted from September,2002 to September,2010 were analyzed retrospectively.They were divided into three groups randomly and followed up.The postoperative morbidity,the mortality and the overall survival rates were evaluated.ResultsThere were no significant differences in these three groups with respect to postoperative morbidity ( P ＞ 0.05 ).The incidence of recurrence in intraperitoneal chemotherapy using sustained-release fluorouracil ( treatment group) was significantly lower than those of intraperitoneal chemotherapy and operative treatment( 16.18％,37.61％ and 41.28％,P ＜0.05).The 1,3- and 5-year overall survival rates of treatment group were 85.51％,61.28％ and 53.67％,respectively,and the 1-,3- and 5-year overall survival rates were 84.11％,39.98％ and 28.12％,and 81.28％,29.88％ and 25.21％ respectively in intrapeitoneal chemotherapy group and operative group.1-year overall survival rate had no significant differences among three groups with respect to ( P＞0.05).3-and 5-year overall survival rates in treatment group were higher signfficantly than those of intraperitoneal chemotherapy and operative treatment( P＜0.05).Conclusions Intraoperative intrapeitoneal hyperthermic chemotherapy using sustained-release fluorouracil is a kind of convenient,safe,and highly effective comprehensive treatment method,and it can kill isolated intraperitoneal cancer cells.It may reduce postoperative recurrence and improve survival rates.

Objective:To investigate the incidence rate of upper digestive tract cancer in Cixian, China, a particular area with high inci-dence of esophageal cancer. Methods:Statistical analysis was performed on the 2003-2012 incidence data of upper digestive tract can-cer in Cixian. The annual incidence rate, Chinese population standardized incidence rate (the bid rate), and structure of world popula-tion standardized incidence rate (referred to as the world standard rate) were calculated. The incidence data were divided into two groups according to period (2003-2007 and 2008-2012), and different age groups were compared. Results:From 2003 to 2012, the in-cidence of upper gastrointestinal cancer was 165.36/10 million. The 2003-2007 crude incidence rate was 171.55/10 million), whereas 2008-2012 crude incidence rate was 151.41/10 million which has reduced over the last five years. Esophageal cancer incidence from 2003 to 2012 had a crude incidence rate of 108.05/10 million during the two periods (from 2003 to 2007, the incidence rate was 116.87/10 million;and from 2008 to 2012, 99.58/10 million), the crude incidence rate of the latter 5-year period has declined obvious-ly. From 2003 to 2012, the overall crude incidence rate of cardia cancer was 31.21/10 million, comparison of two peaiods (from 2003 to 2007 was 29.11/10 million, and 2008-2012, 33.23/10 million) indicated that the level of measurement of the latter period in-creased. At the same period, the overall incidence rate of gastric cancer was 26.10/10 million, comparison of the two periods (2003-2007 the crude incidence rate was 25.57/10 million, 2008-2012 was 26.60/10 million) indicated that the level of the parameter in the latter 5 years increased slightly. Conclusion:The incidence of esophageal cancer in Cixian decreased significantly, but the area remains to have the highest incidence rate of cardiac cancer morbidity. The incidence rate of distal gastric cancer increased significantly in males but decreased slightly in females, which suggests that early diagnosis and treatment of gastric cardia and distal stomach cancer is extremely important.

Objective:To explore the expressions of long non-coding RNA LINC00673 and ISG15 protein in pancreatic cancer and their clinical significances.Methods:The clinical data of 57 patients diagnosed as pancreatic ductal carcinoma (PDAC) at the Affiliated Cancer Hospital of Xiangya Medical College of Central South University from January 2014 to December 2018 were retrospectively analyzed. The relative expressions of LINC00673 in pancreatic cancer tissues and paracancerous normal tissues (within 3 cm from the edge of cancer tissues) were examined by using quantificational reverse transcription-polymerase chain reaction (qRT-PCR). The ISG15 protein expressions in pancreatic cancer tissues and paracancerous normal tissues were examined by using immunohistochemistry. The difference in LINC00673 expression between ISG15 protein positive and negative patients was compared. The correlation between LINC00673 and ISG15 protein expressions in pancreatic cancer was analyzed by Spearman rank correlation analysis. Moreover, the correlations of LINC00673 and ISG15 protein expressions with clinical stage and pathological classification of pancreatic cancer patients were analyzed.Results:The positive expression of ISG15 protein in pancreatic cancer tissues was 40.4% (23/57), which was higher than that in paracancerous normal tissues [15.8% (9/57)] ( χ2 = 7.90, P = 0.004), and the relative expression of LINC00673 in pancreatic cancer tissues was 0.99±0.36, which was lower than that in paracancerous normal tissues (1.26±0.41) ( t = 4.80, P < 0.001). For 23 (40.4%) ISG15-positive patients and 34 (59.7%) ISG15-negative patients, the relative expression of LINC00673 was 0.77±0.46 and 0.45±0.27 ( P < 0.001). Spearman analysis showed that there was a correlation between LINC00673 and ISG15 protein expressions ( ρ = -0.429, P = 0.001). The relative expression of LINC00673 decreased in patients with low differentiated or undifferentiated tumor, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between LINC00673 expression and patients' age, tumor site, preoperative CA199 level, and TNM stage (all P > 0.05); ISG15 protein expression increased in patients with low differentiated or undifferentiated tumor, TNM stage Ⅲ-Ⅳ, vascular invasion and lymph node metastasis (all P < 0.05), but there was no correlation between ISG15 protein expression and patients' gender, age, tumor site, and preoperative CA199 level (all P > 0.05). Conclusions:The expression of LINC00673 in pancreatic cancer is related to vascular invasion, tumor differentiation degree and lymph node metastasis, and the expression of ISG15 in pancreatic cancer is related to vascular invasion, tumor differentiation degree, lymph node metastasis and TNM stage. The combined detection of LINC00673 and ISG15 protein could be a valuable prognostic indicator for pancreatic cancer. The therapies targeting LINC00673 and ISG15 protein signaling pathways are expected to be a potential option for immunotherapy of pancreatic cancer.

Objective To set up a computer-assisted polyp detection system under colonoscopy,and to preliminarily verify its effectiveness.Methods Based on Faster R-CNN algorithm and the open source implementation of the open source framework tensorflow and Faster R-CNN,a computer-assisted polyp detection system under colonoscopy was constructed.According to the size and difficulty of the training set,five test groups were set up:test group one,two,three and four contained 1 000,2 000,4 000 and 6 000 training samples,respectively.Test group five increased the probability of selecting the difficult samples based on 6 000 training samples.In different training sets,the sensitivity,specificity,other classification evaluation parameters,and the evaluation parameters of target detection such as recall and precision of this polyps detection system were calculated.Results Classification evaluation parameters showed that the sensitivities of test group one,two,three,four and five were 90.1％,93.3％,93.3％,93.3 ％ and 93.5 ％,respectively,and the difference was statistically significant (x2 =25.324,P＜0.01).The sensitivities of test group two,three,four and five were all higher than that of test group one,and the differences were statistically significant (x2 =13.964,13.508,13.508 and 13.386,all P＜ 0.006 25).There were no significant differences in specificity and positive predictive value among test groups (both P＞0.05).The negative predictive values of test group one,two,three,four and five were 90.4％,93.3％,93.3％,93.3％ and 93.5％,respectively,and the differences were statistically significant (x2 =21.862,P＜0.01).The negative predictive values of test group two,three,four and five were higher than that of test group one,and the differences were statistically significant (x2=11.447,11.564,11.755,13.760;all P＜0.006 25).As the training sample size increased from 1 000 to 2 000,the area under curve (AUC) increased by 2％,and further increased the sample size to 6 000,AUC increased by less than 1 ％.At this point maintaining the same sample size while increasing the proportion of difficult samples,AUC increased by 0.4％.The results of evaluation parameters of target detection showed that the recall rate of each test group was 73.6％,79.8％,79.5％,79.8％ and 83.3％,respectively,and the differences were statistically significant (x2 =71.936,P＜0.01).Among them,the recall rates of test group two,three and four were higher than that of test group one,and the differences were statistically significant (x2 =25.960,23.492 and 25.960,all P＜0.006 25),and the recall rate of test group five was higher than those of test group one,two,three and four,and the differences were statistically significant (x2=67.361,9.899,11.527 and 9.899;all P＜0.006 25).In addition,the precision rates of test group one,two,three,four and five were 87.9％,85.3％,90.2％,91.4％ and 89.2％,respectively,and the difference was statistically significant (x2=48.194,P＜0.01).The precision rates of test group three and five were higher than that of test group two,and the differences were statistically significant (x2 =24.508 and 15.223,both P＜0.006 25),and the precision rate of test group four was higher than those of test group one and two,and the differences were statistically significant (x2=13.524 and 39.120,both P＜0.006 25).As samples size and training difficulty increased,the values of F1-score and mean average precision increased steadily.Conclusions This study initially constructed a computer-assisted polyp detection system under colonoscopy.Currently the maximum sensitivity reached 93.5％,and the maximum recall rate reached 83.3％.Increasing the training set size may improve the polyp detection result to a certain degree,however it will reach a bottleneck.At this time,increasing the training difficulty can further improve the detection scores,especially the recall rate.

Objective To predict and verify the upstream regulatory microRNA (miRNA)of protein kinase D1 (PKD1),and to investigate its role in cerulein induced acute pancreatitis (AP)in rats. Methods Potential up-stream regulatory miRNA of PKD1 was predicted by using bioinformatics software. Dual luciferase reporter gene system and Western blot were applied to verify the regulation of PKD1 by the selected miRNA. Experimental AP was induced by 6 intraperitoneal injection of cerulein (20 μg/ kg)at hourly intervals after administration of the CY5 - labeled notar-get control (AP group,n = 20)or selected miRNA (treatment group,n = 20),respectively by intraperitoneal injection into rats. Other rats were divided randomly into a normal control group (n = 10)without any treatment. Besides 10 rats in either AP or treatment group were sacrificed 6 hours after the first injection of cerulein,and the rats were all sacri-ficed 24 hours after the first injection. The blood samples and pancreatic tissues of each rat were collected to test serum amylase and lipase activities,or to make hematoxylin - eosin stain for AP pathological scores as well as PKD1 immuno-histochemical staining,respectively. Results TargetScan 7. 1 software analysis showed that miR - 128 - 3p was the po-tential upstream regulatory miRNA of PKD1,which was verified by dual luciferase reporter gene system and Western blot detection. Compared to the normal control group,serum amylase and lipase activities after 6 h exposure to cerulein increased in both AP group and the treatment group[13313. 00(9424. 00 - 15995. 00)U/ L,13552. 00(10399. 50 -18408. 25)U/ L vs. 1430. 50(1214. 25 - 1543. 25)U/ L;547. 00 (515. 00 - 627. 00)U/ L,857. 50(522. 00 -1222. 25)U/ L vs. 34. 00(32. 50 - 34. 75)U/ L],and the differences were significant(χ2 = 8. 715,P < 0. 05;χ2 =9. 115,P < 0. 05),which indicated that the rat models of AP were successfully established. The immunohistochemical scores of PKD1 after 24 h exposure to cerulein decreased in the treatment group[0. 50(0 - 2. 75)scores],compared with the normal control group [4. 00(4. 00 - 8. 00)scores]and the AP group [4. 00(3. 75 - 8. 00)scores],and difference was significant(χ2 = 18. 302,P < 0. 05). Accordingly,the total pathological scores of HE staining decreased significantly in the treatment group,as compared to the AP group (3. 80 ± 0. 85 vs. 6. 90 ± 1. 15,t = 4. 481,P < 0. 01). The results showed that the inflammatory cell infiltration and tissue necrosis were significantly improved after miR -128 - 3p treatment. Conclusions miR - 128 - 3p is the upstream regulatory microRNA of PKD1 which protects pan-creata from necrotic injury and inflammatory cell infiltration in PKD1 - mediated acute pancreatitis.

Objective:To explore the diagnosis, surgical methods and therapeutic effect of primary duodenal malignant tumor.Methods:The clinical data of 116 patients with primary duodenal malignant tumor from January 2010 to December 2018 were retrospectively analyzed.Results:Among 116 patients, adenocarcinoma was in 74 cases, interstitial tumor was in 25 cases, carcinoid was in 9 cases, the others was in 8 cases. Before operation, duodenoscopy was performed in 107 cases, and CT examination was performed in 76 cases. There were 57 cases of pancreaticoduodenectomy, 15 cases of duodenal segmental resection, 13 cases of subtotal gastrectomy and duodenal bulbar resection, 13 cases of duodenal partial resection, and 18 cases of palliative short circuit operation. The total incidence of postoperative complication was 31.9% (37/116), including pancreatic fistula in 8 cases (grade B 5 cases, grade C 3 cases), biliary fistula in 6 cases, abdominal infection in 5 cases, pulmonary infection in 4 cases, intestinal fistula in 3 cases, delayed gastric emptying in 3 cases, and hemorrhage in 8 cases. Four cases (3.4%) died during the perioperative period. Single factor Cox regression analysis result showed that the postoperative survival time was related to the tumor differentiation degree, operation method, tumor infiltration degree and lymphatic metastasis ( P<0.05 or <0.01); multi-factor Cox regression analysis results showed that the operation method, tumor infiltration degree and lymphatic metastasis were the independent risk factors for the postoperative survival time of patients with primary duodenal malignant tumor ( P<0.05). The patients were followed up until June 2021, and 9 cases were lost to follow-up. Kaplan-Meier survival curve analysis result showed that the postoperative overall 1-,3- and 5-year survival rates were 82.11%, 57.56% and 33.11%, respectively. Conclusions:Adenocarcinoma is the main primary malignant tumor of duodenum. Duodenoscopy and CT are the main examination methods. Radical resection is the most effective treatment for primary duodenal malignant tumor, and pancreaticoduodenectomy is the first choice. Surgical method, tumor infiltration degree and lymphatic metastasis are the independent risk factors affecting the prognosis of patients.