Objective To observe the traditional Chinese medicine (TCM) and umbilical treatment on diarrhea-predominant irritable bowel syndrome(IBS-D)with mental disorders. Methods Thirty-seven cases with IBS-D were randomized into a treatment group(33 cases) and a control group(31 cases). The treatment group was given Shugan-Jianpi TCM decoction and pepper umbilical dressing. The control group was given Pinaverium Bromide Tablets and Flupentixol and Melitracen tablets. After having been treated for 4 weeks, Hamilton Anxiety Scale(HAMA), Hamilton Depression Scale(HAMD), IBS Symptom Severity Scale(IBS-SSS), IBS quality of life questionnaire (IBS-QOL) were determined. Results After the treatment, HAMD, HAMA, IBS-SSS in the treatment group and the control group all significantly decreased (the treatment group t=14.280, 13.530, 18.847,the control group t=11.514, 9.424, 13.829, P<0.01), and IBS-QOL increased markedly(t=-16.848, -11.712, P<0.01). Compared with HAMA, IBS-SSS and IBS-QOL in treatment group, the control group showed statistical difference(t=-3.183, -2.549, 3.904, P<0.01 or 0.05). The total effective rate of the treatment group was 94%higher than 75%of the control group(χ2=4.727, P<0.05). Conclusion Shugan-Jianpi TCM decoction and pepper umbilical dressing can improve the clinical symptoms and psychological symptoms and improve the quality of life of IBS-D patients.

[Objective] To investigate the expression of NF-κB and IL-lra in gastric cancer, and to explore their correlation with the clinicopathological parameters and prognosis of gastric cancer (GC). [Methods] Expression of NF-κB and IL-1ra was detected by immunohistochemistry in tissue microarry of 359 cases of GC. [Results] The expression rates of NF-κB in the patients with metastasis of lymph node, TNM Ⅲ-Ⅳ stage, and poorly differentiated of histology were 80.2%, 80.0%, and 79.2%, respectively. The expression rates of IL-1ra in the patients with tumor size ≤3 cm, early stage, non-metastasis of lymph node, and TNM Ⅰ-Ⅱ stage were 61.7%, 75.0%, 66.4%, and 61.9%, respectively. The 5-year survival rate of the cases with non-expressed NF-κB was statistically higher than that of the cases with expressed NF-κB(P=0.036). The 5-year survival rate of the patients with negative expression of NF-κB and positive expression of IL-1ra was statistically higher than the others (P=0.021). [Conclusions] NF-κB is the adverse predictors of prognosis of gastric cancer. IL-1ra maybe play a protective role in early gastric cancer stage, but it is necessary to study deeply and get more evidences.

Objective To establish a method for rapid preparation of antiserum against influenza virus (H7N9) hemagglutinin,and to study the possibility of using it in single radial immunodiffusion (SRID) assay for quantitative detection of antigen in H7N9 influenza vaccine.Methods Hemagglutinin proteins expressed in eukaryotic cells were used to immunize sheep.Serum samples were collected to detect antibody titers by ELISA and double immunodiffusion assay.Different concentrations of antiserum were used in SRID assay to get the optimized concentration.Results After 4 times of immunization,the antiserum titers achieved 1 ∶ 1 000 000 and 1 ∶ 32 as indicated by ELISA and double immunodiffusion assay,respectively.The antiserum could form a clear precipitation line in SRID assay.The detection of antigen in the range of 10 to 40 μg/ml showed good linearity in the standard curve.The antigen titers in six batches of H7N9 vaccine detected by this SRID assay were identical with those by SDS-PAGE assay.Conclusion The antiserum against H7N9 hemagglutinin for SRID assay was developed successfully,and could be used as a reagent for the quantitative detection of antigen in H7N9 influenza vaccine.

Objective To study the possibility of using heterogeneous antiserum in single radial immunodiffusion (SRID) for quantitative detection of HA in H7N9 influenza vaccine product when H7N9-specific antiserum is not available in order to establish a testing method for the detection of H 7N9 antigen in any urgent situation.Methods Antisera specific for H7N1, H7N2, H7N3 and H7N7 were obtained from NIBSC and used for SRID assay .Amino acid sequences of hemagglutinins were comparatively analyzed be-tween H7N9 virus and other viruses used to prepare heterogeneous antiserum .The titers of antisera against H7N9 and their homogenous antigens were detected by double immunodiffusion method .Based on the results of homology analysis and cross-reaction, a suitable antiserum was selected and its applicability was further validated by the SRID assay using H7N9 antigen.Results Influenza A virus subtype H7N3 that used for preparation of 07/278 antiserum showed the highest HA homology with H7N9 (97.14%).The titer of 07/278 antiserum against H7N9 antigen was 1 ∶8 as detected by double immunodiffusion assay .The H7N9 anti-gen and the 07/278 antiserum could form a clear precipitation line in SRID assay .The detection of H7N9 antigen in the range of 10 to 40μg/ml showed a good linearity in the standard curve .Conclusion The 07/278 antiserum from NIBSC can be used as an alternative reagent for the quantitative detection of hemaggluti -nin in H7N9 influenza virus vaccine .

Objective To evaluate the effect of nucleolar protein 14 (NOP14) on angiogenesis in melanoma.Methods Melanoma tissues were collected from 40 patients with pathologically diagnosed melanoma in Guangzhou First People's Hospital from January 2016 to December 2018,and immunohistochemical study was conducted to determine the expression of NOP14 and CD31 (expressed as microvessel density [MVD]).Melanoma cell lines A375 and SK-MEL-1 were both divided into 4 groups:empty vector group transfected with the empty vector,NOPI4 group transfected with a NOP14-overexpressing vector,siNOP14 group transfected with the siRNA targeting NOP14,and siNC group transfected with a negative control siRNA.Fluorescence-based quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of NOP14 respectively,and Western blot analysis and enzyme-linked immunosorbent assay (ELISA) to measure the expression of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) in cells and their culture media.Coculture models of human umbilical vein endothelial cells (HUVECs) and A375/SK-MEL-1 cells in the above groups were established in Transwell chambers,and cell counting kit-8 (CCK8) assay,Transwell migration and invasion assays and Matrigel-based vasculogenic mimicry assay were performed to evaluate the cellular proliferative,migratory,invasive activity and tube formation capacity respectively.A linear regression model was used to analyze the relationship between NOP14 expression and MVD in melanoma tissues,multi-way analysis of variance to analyze the difference in cellular proliferative activity,and independent-sample t test to compare other experimental indices between 2 groups.Results The expression of CD31 (MVD) was 44 ± 13 in the group with high NOP14 expression (n =20),58 ± 16 in that with moderate NOP14 expression (n =17),and 62 ± 11 in that with low NOP14 expression (n =3).The NOP14 expression was negatively correlated with MVD (r =-0.525,P =0.017).Compared with the empty vector group,the expression of VEGF and VEGFR in A375 and SK-MEL-1 cells and their culture media significantly decreased in the NOP14 group (all P ＜ 0.05).Compared with the siNC group,the expression of VEGF and VEGFR in the A375 and SK-MEL-1 cells and their culture media significantly increased in the siNOP14 group(all P ＜ 0.05).In the co-culture models of A375 cells and HUVECs,the NOP14 group showed significantly decreased proliferative activity of HUVECs (F =131.85,P ＜ 0.05),and numbers of migratory cells (22 ± 5 vs.63 ± 8,t =7.07,P =0.002),invasive cells (14 ± 5 vs.45 ± 10,t =4.94,P =0.008) and branch points (8 ± 2 vs.14 ± 3,t =5.06,P ＜ 0.001) compared with the empty vector group;compared with the siNC group,the siNOP14 group showed significantly increased proliferative activity of HUVECs (F =79.92,P ＜ 0.01),and numbers of migratory cells (152 ± 30 vs.59 ± 4,t =5.36,P =0.006),invasive cells (134 ± 21 vs.50 ± 8,t =6.40,P ＜ 0.001) and branch points (27 ± 3 vs.15 ± 4,t =6.10,P ＜ 0.001).In the co-culture models of SK-MEL-1 cells and HUVECs,the 4 groups showed the same trend of changes in the cellular proliferative,migratory,invasive activity and tube formation capacity of HUVECs as the above groups in the co-culture models of A375 cells and HUVECs.Conclusion The NOP14 expression is negatively correlated with MVD in melanoma tissues,and NOP14 can inhibit angiogenesis in melanoma.

Objective To explore the safety and efficacy of laparoscopic cholecystectomy in different time points after endoscopic retrograde cholangiopancreatography in elderly patients with cholecystolithiasis and choledocholithiasis. Methods 99 elderly patients （≥60 years old）with cholecystolithiasis and choledocholithiasis treated by sequential LC after ERCP in the Department of Hepatobiliary and Pancreatic surgery in our hospital from January 2015 to June 2020 were analyzed. Aocording to different time points after ERCP, LC was divided into experimental group (within 1 day after ERCP) and control group (within 2-7 days after ERCP). The complications were followed up by telephone within 3 months after operation. Results After analysis，the difference between the experimental group and the control group in the total length of hospitalization [(13.0±4.6)d vs ( 15.1±4.4)d] and total hospitalization expenses (38 172.5±12 448.9) vs ( 42 535.4±8 814.6)] was statistically significant (P<0.05). However, there was no significant difference in the conversion rate of LC to laparotomy, the total postoperative complications , the length of hospital stay after LC，the amount of blood loss during LC and the time of LC operation between the two groups(P>0.05). Conclusion For elderly patients with cholecystolithiasis and choledocholithiasis, LC within 1 day after ERCP does not increase the difficulty of operation and postoperative complications，but shortens the hospitalization time, and it is economical and safe at the same time.

ObjectiveTo investigate the risk factors for open pancreatic necrosectomy (OPN), an effective treatment method for severe acute pancreatitis (SAP) after the failure of percutaneous catheter drainage (PCD), in patients with SAP. MethodsA retrospective analysis was performed for 156 patients with SAP who underwent surgical intervention based on the step-up approach in The Affiliated Hospital of Zunyi Medical University from January 1, 2010 to June 30, 2018, and according to whether OPN was performed, the patients were divided into PCD group with 126 patients and PCD+OPN group with 30 patients. Related clinical data were collected, including age, sex, etiology, blood calcium on admission, white blood cell count on admission, whether CTSI score was ＞7, APACHE-Ⅱ score, Ranson score, presence or absence of peripancreatic fluid accumulation, presence or absence of infection, presence or absence of multiple organ failure (MOF), and whether PCD was performed at more than 1 week after admission. The t-test was used for comparison of continuous data between groups, and the chi-square test was used for comparison of categorical data between groups; a multivariate logistic regression analysis was used to determine the independent predictive factors for OPN. ResultsThe probability of OPN was 19.2% for SAP patients in the later stage. Compared with the PCD+OPN group, the PCD group had a significantly lower proportion of patients with MOF on admission ［27.0% (34/126) vs 70.0% (21/30), χ2=19.642, P＜0.01］ and a significantly higher proportion of patients undergoing PCD at less than 1 week after admission ［61.9% (78/126) vs 20.0% (6/30), χ2=17.121, P＜0.01］. MOF on admission (odds ratio ［OR］=5.343, 95% confidence interval ［CI］: 1.832-15.583, P＜0.05), initial PCD performed at more than 1 week after admission (OR= 5.518, 95% CI: 1.742-17.477, P＜0.05), and infection on admission (OR=5.016, 95% CI: 1.322-19.378, P＜0.05) were independent risk factors for subsequent OPN in SAP patients. ConclusionSAP with MOF on admission, initial PCD performed at more than 1 week after admission, and SAP with infection on admission are independent risk factors for subsequent OPN in SAP patients undergoing PCD in the early stage based on the step-up approach. Timely identification of related risk factors helps to grasp the timing of OPN in clinical practice and improve the clinical prognosis of SAP patients.

Background and purpose:Limbs soft tissue sarcoma(STS)is a common malignant tumor,and surgical resection is the main treatment method for it.The concept of barrier made us realize the blocking effect of natural barrier on STS,and we aimed to search for tissues that can act as barrier,and to perform complete resection of surgical margins around the tissue barrier.This study aimed to investigate the feasibility,safety and prognosis of barrier resection in the treatment of limbs STS.Methods:From December 2013 to September 2016,data of 72 patients who underwent barrier resection of STS of extremities in department of oncosurgery,Minhang Branch,Fudan University Shanghai Cancer Center were retrospectively analyzed,and the resection margin was sampled.All 72 patients underwent preoperative magnetic resonance imaging(MRI)or computed tomography(CT)design,and the physiological barrier or at least 3 cm distance was found outward from the anatomical location of the tumor.And en bloc excision was performed outside this barrier or at a distance of 3 cm.The influence of postoperative pathological margin,musculoskeletal tumor society(MSTS)score and postoperative complications on the patients were analyzed.The 1-and 3-year locoregional recurrence-free survival(LRFS)rates and sarcoma-specific survival(SSS)rates were evaluated,and the influencing factors were analyzed.This study was approved by the Ethics Committee of Fudan University Shanghai Cancer Center(number:1212117-12&1212117-12-1301).Results:All patients had negative margins.The 1-and 3-year LRFS rates were 98.2%and 93.3%,respectively.The 1-and 3-year SSS rates were 98.4%and 94.2%,respectively.The mean MSTS scores were 28.3 preoperatively and 25.5 postoperatively.Surgical complications were grade 1 to 2 in 20 cases and grade 3 in 1 case,and there were no grade 4 to 5 complications.Conclusion:Based on the combination of clinical,imaging and pathology data,barrier resection,including tumor resection and functional reconstruction,can be applied to the surgical treatment of STS,with good feasibility and safety,reliable margin and satisfactory local control.

Objective:To develop a new method based on neuraminidase (NA) activity for detec-ting virus titers of cell culture-based influenza vaccines and preliminarily analyze its application.Methods:Reaction conditions including the substrate concentration for enzymatic reaction, stop solution, the number of initially infected target cells and cell lysis buffer were optimized. The titers of cell culture-based influenza vaccine strains were detected by the established method and the results were compared with those by the traditional viral titration test.Results:The optimal substrate concentration for enzymatic reaction was 25 μmol/L, and the optimal stop solution was 0.2 mol/L Na 2CO 3. In the detection of NA activity in infected cells, the maximum relative fluorescence value was obtained by infecting 4×10 4 cells/well with influenza virus for 48 h and using 0.5% TritonX-100 for lysis. The developed method showed no significant differences with the traditional virus titration test in detecting the titers of four batches of influenza vaccine virus strains ( P>0.05), indicating that the two methods had a good consistency. Conclusions:This study established a new method based on NA activity to detect virus titers of cell culture-based influenza vaccines. The method could be used for the detection of virus strains used in the production of cell cultured-based influenza vaccines.