Objective:To explore the effects of miR-21-5p exosomes derived from human umbilical cord mesenchymal stem cells on apoptosis of human granular cells.Methods:A granular cell apoptosis model was constructed by treating KGN cells with different concentrations (0 μmol/L, 30 μmol/L, 60 μmol/L, and 100 μmol/L) of phosphoramide nitrogen mustard for 48 h. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. The apoptosis rate was measured using flow cytometry to screen for the optimal concentration of phosphoramide nitrogen mustard for constructing an apoptosis model. Hsa-miR-21-5p overexpression plasmid was used for instantaneously transfecting human umbilical cord mesenchymal stem cells, and the expression level of hsa-miR-21-5p was detected by qPCR. The miR-21-5p exosomes were separated and identified by flow cytometry and electron microscope. Different concentrations (5 μg/mL, 10 μg/mL and 15 μg/mL) of miR-21 exosomes were added into successful KGN cell apoptosis model to overexpress miR-21. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. PKH-26 was used to trace the position of human granular cells. Results:The levels of bax mRNA and protein in KGN cells treated with 60 μmol/L phosphoramide nitrogen mustard were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group (all P<0.001), while the levels of bcl2 mRNA and protein were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group ( P=0.005, P<0.001). The apoptosis rate of KGN cells after 60 μmol/L phosphoramide nitrogen mustard intervention was (38.10±2.90)%, while the apoptosis rate of KGN cells after 30 μmol/L phosphoramide nitrogen mustard intervention was (16.75±2.55)%, they were all significantly higher than that of the 0 μmol/L phosphoramide nitrogen mustard intervention group ( P=0.020, P=0.006). Hsa-miR-21-5p was transiently transfected into human umbilical cord blood mesenchymal stem cells, and the expression of has-miR-21-5p was higher than that in control group detected by qPCR ( P<0.001). The positive rate of surface protein CD9, CD63 and CD81 was 14.9%, 16.4% and 31.4%. The exosome was observed as "tea tray" or "concave hemisphere" by electron microscope. The exosome labeled by PKH-26 entered the granular cells and exerted biological effects. There was no statistically significant difference in bax mRNA expression levels between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty plasmid exosomes groups and the 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05). However, the expression levels of bax mRNA in the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups were lower than those in the 60 μmol/L phosphoramide nitrogen mustard group ( P=0.008, P=0.003, P<0.001). However, there was no statistically significant difference in the expression of bcl2 mRNA among the groups (all P>0.05). From the perspective of protein levels, there was no statistically significant difference in BAX protein expression between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty exosomes groups and 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05), while the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups showed a decrease in BAX protein expression compared with the 60 μmol/L phosphoramide nitrogen mustard group, and the differences were statistically significant (all P<0.001). However, there was no statistically significant difference in the expression of BCL2 protein among the intervention groups (all P>0.05). Conclusion:Hsa-miR-21-5p exosomes derived from human umbilical cord blood mesenchymal stem cells can effectively exert the anti-apoptotic effect.

Objective:To explore the effects of miR-21-5p exosomes derived from human umbilical cord mesenchymal stem cells on apoptosis of human granular cells.Methods:A granular cell apoptosis model was constructed by treating KGN cells with different concentrations (0 μmol/L, 30 μmol/L, 60 μmol/L, and 100 μmol/L) of phosphoramide nitrogen mustard for 48 h. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. The apoptosis rate was measured using flow cytometry to screen for the optimal concentration of phosphoramide nitrogen mustard for constructing an apoptosis model. Hsa-miR-21-5p overexpression plasmid was used for instantaneously transfecting human umbilical cord mesenchymal stem cells, and the expression level of hsa-miR-21-5p was detected by qPCR. The miR-21-5p exosomes were separated and identified by flow cytometry and electron microscope. Different concentrations (5 μg/mL, 10 μg/mL and 15 μg/mL) of miR-21 exosomes were added into successful KGN cell apoptosis model to overexpress miR-21. The mRNA and protein levels of bax and bcl2 were detected using qPCR and Western blotting, respectively. PKH-26 was used to trace the position of human granular cells. Results:The levels of bax mRNA and protein in KGN cells treated with 60 μmol/L phosphoramide nitrogen mustard were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group (all P<0.001), while the levels of bcl2 mRNA and protein were significantly higher than those in the 0 μmol/L phosphoramide nitrogen mustard group ( P=0.005, P<0.001). The apoptosis rate of KGN cells after 60 μmol/L phosphoramide nitrogen mustard intervention was (38.10±2.90)%, while the apoptosis rate of KGN cells after 30 μmol/L phosphoramide nitrogen mustard intervention was (16.75±2.55)%, they were all significantly higher than that of the 0 μmol/L phosphoramide nitrogen mustard intervention group ( P=0.020, P=0.006). Hsa-miR-21-5p was transiently transfected into human umbilical cord blood mesenchymal stem cells, and the expression of has-miR-21-5p was higher than that in control group detected by qPCR ( P<0.001). The positive rate of surface protein CD9, CD63 and CD81 was 14.9%, 16.4% and 31.4%. The exosome was observed as "tea tray" or "concave hemisphere" by electron microscope. The exosome labeled by PKH-26 entered the granular cells and exerted biological effects. There was no statistically significant difference in bax mRNA expression levels between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty plasmid exosomes groups and the 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05). However, the expression levels of bax mRNA in the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups were lower than those in the 60 μmol/L phosphoramide nitrogen mustard group ( P=0.008, P=0.003, P<0.001). However, there was no statistically significant difference in the expression of bcl2 mRNA among the groups (all P>0.05). From the perspective of protein levels, there was no statistically significant difference in BAX protein expression between the 5 μg/mL, 10 μg/mL, and 15 μg/mL empty exosomes groups and 60 μmol/L phosphoramide nitrogen mustard group (all P>0.05), while the 5 μg/mL, 10 μg/mL, and 15 μg/mL miR-21-5-p exosomes groups showed a decrease in BAX protein expression compared with the 60 μmol/L phosphoramide nitrogen mustard group, and the differences were statistically significant (all P<0.001). However, there was no statistically significant difference in the expression of BCL2 protein among the intervention groups (all P>0.05). Conclusion:Hsa-miR-21-5p exosomes derived from human umbilical cord blood mesenchymal stem cells can effectively exert the anti-apoptotic effect.

Objective To investigate the safety and short-term efficacy of π-shaped anastomosis and circular anastomosis(reverse puncture device)in reconstruction of esophagojejunostomy after laparoscopic total gastrectomy.Methods A retrospective study was used to collect the clinical and pathological data of 75 cases of gastric cancer from January 2019 to March 2021.According to the different reconstruction methods of esophagojejunal anastomosis,the patients were divided into a linear cutting obturator group(π-shaped anastomosis group,n = 27)and a circular anastomat anastomosis group(reverse puncture device group,n = 48).The general information of the two groups,operation time,esophagojejunostomy time,intraoperative bleeding volume,number of intraoperative lymph node dissection,intraoperative complications,and postoperative complications were compared and analyzed.Results The operation time and esophagojejunostomy time in the π-shaped anastomosis group were(221.5±8.8)and(34.7±3.7)min,and the reverse puncture device group were and(246.9±5.6)and(47.2±4.6)min,respectively,the differences were statistically significant(t = 15.19,t = 11.81,P＜0.05).There were no statistical significance in the comparison of intraoperative bleeding volume and number of intraoperative lymph node dissection between the two groups(P＞0.05).In the reverse puncture device group,there were two intraoperative complications,including one case of esophageal jejunal anastomosis atresia and one case of anastomosis tear,postoperative complications occurred in 3 cases,postoperative anastomotic stenosis occurred in 2 case,and anastomotic bleeding occurred in 1 case.Conclusion Laparoscopic total esophagojejunostomy with π-shaped anastomosis and reverse puncture device are safe and feasible.In terms of esophagojejunostomy time,π-shaped anastomosis reconstruction time is shorter.When the small intestine diameter is relatively small and it is difficult to extend into the 25 mm stapler,the advantage of π-shaped anastomosis is more obvious.When the tumor is Siewert type Ⅰ and type Ⅱ adenocarcinoma of gastroesophageal junction,which infiltrates into above the dentate line,reverse puncture device method is recommended for reconstruction.

Objective:To uncover the effect of circ0025847 on the proliferation of colorectal cancer cells and its molecular mechanisms.Methods:qRT-PCR was utilized to analyze the expression of circ0025847 and QK1 in human colorectal cancer cells (HCT116, SW480) and normal mucosa cells (NCM460) .CCK-8 was used to analyze the effect of circ0025847 and QK1 on proliferation in colorectal cancer cells. Bioinformatics method was used to screen RBP which could bind to circ0025847. RNA pulldown and RIP was utilized to confirm whether QK1 binds to circ0025847.Effects of circ0025847 over-expression on QK1 expression was analyzed by Western blot.NC group, circ0025847 overexpression group and circ0025847 overexpression+ QK1 inhibitor group were established and the proliferation effect was determined by CCK8.Results:circ0025847 (the expression levels in NCM460, HCT116 and SW480 cells were 1.01±0.05, 0.49±0.08, 0.45±0.10) and QK1 (the expression levels in NCM460, HCT116 and SW480 cells were 0.98±0.07, 0.50±0.07, 0.47±0.09) expression was significantly downregulated in colorectal cancer cells. Overpression of circ0025847 and QK1 suppressed colorectal cancer cells growth.RNA pull-down and RIP clarified that circ0025847 bind to QK1 and circ0025847 positively regulate QK1 expression (7 199.20±12.44 VS 3 889.80±11.03) . circ0025847 inhibiting the proliferation of colorectal cancer cells by promoting the expression of QK1 was confirmed by rescue experiment.Conclusion:circ0025847 inhibits colorectal cancer cells proliferation via positively regulating QK1 expression, indicating that circ0025847 may be potential therapeutic target of colorectal cancer.

Objective:To analyze of the main influecing factors of mosic embryos during the preimplantation genetic test for chromosomal structural rearrangement (PGT-SR) to avoid the increase of risk of abortion and genetic abnormalities and to improve the diagnostic rate of mosic embryos.Methods:We used a retrospective cohort study to analyze 94 cycles of infertile patients undergoing PGT-SR and 551 cycles of intracytoplasmic sperm injection (ICSI) from January 2018 to December 2019 in the Reproductive Medical Center of the Maternal and Child Health Hospital. The relationship of mosic embryos was analyzed between the age, the number of oocytes, gonadotropin (Gn)/oocyte, the grade of blastocysts and chromosome carrier of different genders by the SPSS21.0 software.Results:In the PGT-SR cycle, single factor analysis found that mosaic embryos were related to age and sperm concentration ( P=0.02, P=0.04), but multivariate logistic regression analysis showed that age ( OR=3.41, 95% CI=1.34-8.66, P=0.01), sperm concentration ( OR=0.41, 95% CI=0.17-0.96, P=0.04) and chromosome carrier of different genders ( OR=2.21, 95% CI=1.04-4.70, P=0.04) were the main factors of embryo mosaicism. Conclusion:Female age, sperm concentration and chromosome carrier of different genders maybe affect the formation of mosaic embryos, providing theoretical basis for selective transfer of mosaic embryos.

Objective:To analyze of the main influecing factors of mosic embryos during the preimplantation genetic test for chromosomal structural rearrangement (PGT-SR) to avoid the increase of risk of abortion and genetic abnormalities and to improve the diagnostic rate of mosic embryos.Methods:We used a retrospective cohort study to analyze 94 cycles of infertile patients undergoing PGT-SR and 551 cycles of intracytoplasmic sperm injection (ICSI) from January 2018 to December 2019 in the Reproductive Medical Center of the Maternal and Child Health Hospital. The relationship of mosic embryos was analyzed between the age, the number of oocytes, gonadotropin (Gn)/oocyte, the grade of blastocysts and chromosome carrier of different genders by the SPSS21.0 software.Results:In the PGT-SR cycle, single factor analysis found that mosaic embryos were related to age and sperm concentration ( P=0.02, P=0.04), but multivariate logistic regression analysis showed that age ( OR=3.41, 95% CI=1.34-8.66, P=0.01), sperm concentration ( OR=0.41, 95% CI=0.17-0.96, P=0.04) and chromosome carrier of different genders ( OR=2.21, 95% CI=1.04-4.70, P=0.04) were the main factors of embryo mosaicism. Conclusion:Female age, sperm concentration and chromosome carrier of different genders maybe affect the formation of mosaic embryos, providing theoretical basis for selective transfer of mosaic embryos.

Objective To compare vein valve function following pharmacomechanical thrombolysis (PMT) with simple catheter-directed thrombolysis (CDT) for deep vein thrombosis. Methods We retrospectively analyzed the clinical data of sixty patients who suffered acute lower extremity deep vein thrombsis in our hospital between October 2016 and March 2017. All patients underwent contralateral preprocedural duplex and bilateral postprocedure duplex to access patency and valve function. The patients were divided into three groups including a group A with catheter-directed thrombolysis (CDT) alone (36 patients with 20 males and 16 females at average age of 56 years), a group B with PMT alone (15 patients with 8 males and 7 females at average age of 55 years), and a group C with PMT combined CDT (9 patients with 4 males and 5 females at average age of 56 years). The valve function was compared among the Group A, Group B and Group C. Results There were 40.0% (24/60) patients with bilateral femoral vein valve reflux, 40.0% (24/60) patients with unilateral femoral vein valve reflux (all in the treated limbs), 20% (12/60) patients had no reflux in both limbs. Of the limbs treated with CDT alone, PMT alone and PMT combined CDT, the rate of valve reflux was 38.9% (14/36), 33.3% (5/15), and 55.6% (5/9) respectively (P=0.077). Conclusion In the patients suffering acute DVT, PMT or PMT combined CDT does not hamper valve function compared with CDT alone.

Objective To assess the safety and efficacy of endoscopic retrograde cholangiopancreatography (ERCP) for children with pancreaticobiliary diseases. Methods Data of children under 14 years old who have underwent ERCP in Nanjing Drum Tower Hospital between September 2007 and August 2016 were reviewed for completion, complications and therapeutic methods. Results A total of 41 children underwent 68 ERCP, including 6(8. 8%) diagnostic and 62(91. 2%) therapeutic procedures. All procedures were performed under deep sedation. Cannulation failed in only 1 child with anomalous junction of pancreaticobiliary duct. The procedure success rate was 98. 5%( 67/68 ) . There were 8 adverse events, including 7 mild post?ERCP pancreatitis and 1 fever. Incidence of adverse event was 11. 8%( 8/68) . There was no such severe adverse event as bleeding, perforation, death, or other anesthesia related adverse event. Thirty?two children ( 78. 0%) had follow?up, ranging from 2 month to 6 years. Children followed lived well with no long?term adverse event. Conclusion ERCP is an effective and safe procedure for the diagnosis and treatment of pancreaticobiliary diseases in children.

The paper analyzes different expressions of indirectly associated strong-weak connection in the process of biomedical literature knowledge discovery,defines the effect and function of strong-weak connection on literature knowledge discovery,and indicates that the accuracy rate of the discovery of potential connection through strong connection is obviously higher than the accuracy rate obtained through weak connection,and strong connection can serve as an important factor for improving the application effect of literature knowledge discovery.

Objective To investigate the diagnostic value of intraductal ultrasonography ( IDUS ) with or without brush cytology on distinguishing malignant from benign biliary stricture. Methods The data of patients with biliary stricture, who underwent endoscopic retrograde cholangiography( ERC) and IDUS in Drum Tower Hospital from September 2007 to October 2015 were retrospectively analyzed. Features of IDUS images and results of brush cytology were compared with postoperative histological results to evaluate the diagnostic yield of IDUS and brush cytology on differential diagnosis of malignant biliary stricture. Results A total of 230 patients with biliary stricture underwent ERC and IDUS, and 206 patients meanwhile received brush cytology, including 84 cases of malignant biliary stricture. Of the 230 patients, 87 cases underwent surgery, and 75 revealed malignant results, including 65 cases of cholangiocarcinoma, 8 cases of pancreatic adenocarcinoma, and 2 cases of periampullary adenocarcinoma. Using postoperative pathologic results as the gold standard, 72 malignant and 9 benign biliary strictures were correctly diagnosed by IDUS. The sensitivity, specificity, positive predictive value, and negative predictive value of IDUS for diagnosis of malignant biliary stricture was 96. 0％, 75. 0％, 96. 0％, and 75. 0％, respectively. Sixty-five patients underwent IDUS with brush cytology during ERC before surgery, and 56 out of 65 showed malignant results. Thirty-one malignant and 9 benign biliary stricture were correctly diagnosed by brush cytology. The sensitivity, specificity, positive predict value, and negative predict value of brush cytology for diagnosis of malignant biliary stricture was 55. 4％, 100％, 100％, and 26. 5％, respectively. If brush cytology combined with IDUS to diagnose malignant biliary stricture, the sensitivity rose up to 94. 6％, while the specificity went down to 77. 8％, and the positive and negative predictive value was 96. 4％ and 70. 0％, respectively. Conclusion IDUS exhibited excellent sensitivity and specificity, while brush cytology revealed perfect specificity and positive predictive value on diagnosis of malignant biliary stricture. If IDUS combined with brush cytology, the sensitivity and negative predictive value could rise significantly, while the specificity and positive predictive value mildly went down, which suggested that IDUS remained a certain degree of over judgment and false positive rate.