Objective:To investigate the surgical efficacy and prognosis influencing factors of hilar cholangiocarcinoma based on multidisciplinary diagnosis and treatment.Methods:The retrospective cohort study was conducted. The clinicopathological data of 91 patients with hilar cholangiocarcinoma who underwent surgery in Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from April 2004 to April 2021 were collected. There were 59 males and 32 females, aged (61±10)years. Patients who were admitted from April 2004 to March 2014 underwent traditional surgical diagnosis and treatment, and patients who were admitted from April 2014 to April 2021 underwent multidisciplinary diagnosis and treatment. Observation indica-tors: (1) surgical situations; (2) postoperative situations; (3) postoperative pathological examina-tions; (4) postoperative prognosis analysis; (5) influencing factors of postoperative prognosis. Follow-up was conducted using telephone interview and outpatient examination. Patients were followed up once every 6 months after surgery to detect survival. The follow-up was up to April 2023. Measure-ment data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the independent sample t test. Measurement data with skewed distribution were represented as M(range), and comparison between groups was conducted using the Mann-Whitney U test. Comparison of ordinal data was conducted using the rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher exact probability. The Kaplan-Meier method was used to draw survival curve and calculate survival rate. The Log-Rank test was used for survival analysis. Univariate and multivariate analyses were conducted using the COX proportional hazard model. Results:(1) Surgical situations. Of the 91 patients, there were 65 cases receiving hemi- or expanded hemi-hepatectomy, 13 cases receiving tri-hepatectomy, 9 cases receiving partial hepatectomy, 4 cases receiving extrahepatic bile duct resection. There were 24 cases receiving combined vein resection and reconstruction, 8 cases receiving combined pancreaticoduodenectomy, 6 cases receiving com-bined hepatic artery resection and reconstruction, including 24 cases receiving extended radical surgery (tri-hepatectomy, hepatic artery resection and reconstruction, hepatopancreaticoduodenec-tomy). The operation time, volume of intraoperative blood loss and intraoperative blood transfusion rate of 91 patients was (590±124)minutes, 800(range, 500?1 200)mL and 75.8%(69/91), respectively. Of the 91 patients, cases receiving extended radical surgery, the volume of intraoperative blood loss were 4, 650(range, 300?1 000)mL in the 31 patients who were admitted from April 2004 to March 2014, versus 20, 875 (range, 500?1 375)mL in the 60 patients who were admitted from April 2014 to April 2021, showing significant differences between them ( χ2=4.39, Z=0.31, P<0.05). (2) Post-operative situations. The postoperative duration of hospital stay and cases with postoperative infectious complications were (27±17)days and 50 in the 91 patients. Cases with abdominal infection, cases with infection of incision, cases with bacteremia and cases with pulmonary infection were 43, 7, 5, 8 in the 91 patients. One patient might have multiple infectious complications. Cases with bile leakage, cases with delayed gastric emptying, cases with chylous leakage, cases with liver failure, cases with pancreatic fistula, cases with intraperitoneal hemorrhage, cases with reoperation, cases dead during the postoperative 90 days were 30, 9, 9, 6, 5, 3, 6, 3 in the 91 patients. Cases with abdominal infection was 10 in the 31 patients who were admitted from April 2004 to March 2014, versus 33 in the 60 patients who were admitted from April 2014 to April 2021, showing a significant difference between them ( χ2=4.24, P<0.05). Cases dead during the postoperative 90 days was 3 in the 31 patients who were admitted from April 2004 to March 2014, versus 0 in the 60 patients who were admitted from April 2014 to April 2021, showing a significant difference between them ( P<0.05). (3) Post-operative pathological examinations. Of the 91 patients, cases with Bismuth type as type Ⅰ?Ⅱ, type Ⅲ, type Ⅳ, cases with T staging as Tis stage, T1 stage, T2a?2b stage, T3 stage, T4 stage, cases with N staging as N0 stage, N1 stage, N2 stage, cases with M staging as M0 stage, M1 stage, cases with TNM staging as 0 stage, Ⅰ stage, Ⅱ stage, Ⅲ stage, ⅣA stage, ⅣB stage, cases with R 0 radical resection, cases with R 1 or R 2 resection were 15, 46, 30, 1, 9, 25, 30, 26, 49, 36, 6, 85, 6, 1, 7, 13, 58, 6, 6, 63, 28. Cases with R 0 radical resection, cases with R 1 or R 2 resection were 15, 16 in the 31 patients who were admitted from April 2004 to March 2014, versus 48, 12 in the 60 patients who were admitted from April 2014 to April 2021, showing a significant difference between them ( χ2=9.59, P<0.05). (4) Postoperative prognosis analysis. Of the 91 patients, 3 cases who died within 90 days after surgery were excluded, and the 5-year overall survival rate and median overall survival time of the rest of 88 cases were 44.7% and 55 months. The 5-year overall survival rate was 33.5% in the 28 patients who were admitted from April 2004 to March 2014, versus 50.4% in the 60 patients who were admitted from April 2014 to April 2021, showing a significant difference between them ( χ2=5.31, P<0.05). Results of further analysis showed that the corresponding 5-year overall survival rate of cases without lymph node metastasis was 43.8% in the 16 patients who were admitted from April 2004 to March 2014, versus 61.6% in the 31 patients who were admitted from April 2014 to April 2021. There was a significant difference in the 5-year overall survival rate between these patients without lymph node metastasis ( χ2=3.98, P<0.05). The corresponding 5-year overall survival rate of cases with lymph node metastasis was 18.5% in the 12 patients who were admitted from April 2004 to March 2014, versus 37.7% in the 29 patients who were admitted from April 2014 to April 2021. There was no significant difference in the 5-year overall survival rate between these patients with lymph node metastasis ( χ2=2.25, P>0.05). (5) Influencing factors of postoperative prognosis. Results of multivariate analysis showed that poorly differentiated tumor and R 1 or R 2 resection were inde-pendent risk factors influencing prognosis after surgical treatment of hilar cholangiocarcinoma ( hazard ratio=2.62, 2.71, 95% confidence interval as 1.30?5.29, 1.30?5.69, P<0.05). Conclusions:Compared with traditional surgical diagnosis and treatment, treatment of hilar cholangiocarcinoma based on multidisciplinary diagnosis and treatment can expand surgical indications, reduce proportion of dead patients within 90 days after surgery, improve proportation of radical resection and long-term survival rate. Poorly differentiated tumor and R 1 or R 2 resection are independent risk factors influencing prognosis after surgical treatment of hilar cholangiocarcinoma.

Objective:To observe the efficacy of self-help cognitive behavioral therapy for insomnia (CBTI), trazodone hydrochloride and their combination in the treatment of depression and insomnia comorbidity in the elderly.Methods:90 elderly patients with insomnia and depression admitted to the 901th Hospital of the Joint Logistic Support Force from October 2019 to October 2021 were selected as the study subjects. They were divided into trazodone group, CBTI group and trazodone combined with CBTI group(combination group), with 30 cases in each group. Trazodone group was treated with oral trazodone hydrochloride, CBTI group was treated with self-help CBTI, and the combination group was treated with oral trazodone hydrochloride combined with self-help CBTI. All three groups were treated for 4 weeks. The sleep latency, total sleep time and sleep efficiency of each group were compared at the time of admission and after 4 weeks of treatment. Pittsburgh Sleep Quality Index (PSQI) and Epworth Sleepiness Scale (ESS) were used for sleep assessment before and after treatment, and Self-Rating Depression Scale (SDS) was used for depression assessment.Results:Before treatment, there was no significant difference among the three groups in terms of sleep latency, total sleep time, sleep efficiency, PSQI, ESS and SDS (all P>0.05). After treatment, the sleep latency of the three groups was shorter than that before treatment, and the total sleep time was longer than that before treatment (all P<0.05). The sleep efficiency of the trazodone group and the combination group was higher than that before treatment, with statistically significant difference (both P<0.05). The indexes of the combined group were better than those of the trazodone group and the CBTI group (all P<0.05). The sleep latency of the trazodone group was shorter than that of the CBTI group, and the total sleep time was longer than that of the CBTI group (all P<0.05), with statistically significant difference (all P<0.05). After treatment, the PSQI, except for the SDS of CBTI group, the ESS and SDS of the three groups were lower than those before treatment (all P<0.05). The PSQI, ESS and SDS of the combined group were lower than those of the trazodone group and the CBTI group, and the ESS and SDS of the trazodone group were lower than those of the CBTI group, with statistically significant difference (all P<0.05). Conclusions:For the elderly patients with depression and insomnia, the combination of self-help CBTI and trazodone can not only improve insomnia but also relieve depression symptoms, and the effect is better than that of trazodone and self -help CBTI alone.

The beneficial effects of hypoxic preconditioning are abolished in the diabetes. The present study was designed to investigate the protective effects and mechanisms of repeated episodes of whole body hypoxic preconditioning (WBHP) in db/db mice. The protective effects of preconditioning were explored on diabetesinduced vascular dysfunction, cognitive impairment and ischemia-reperfusion (IR)-induced increase in myocardial injury. Sixteen-week old db/db (diabetic) and C57BL/6 (non-diabetic) mice were employed. There was a significant impairment in cognitive function (Morris Water Maze test), endothelial function (acetylcholineinduced relaxation in aortic rings) and a significant increase in IR-induced heart injury (Langendorff apparatus) in db/db mice. WBHP stimulus was given by exposing mice to four alternate cycles of low (8%) and normal air O2 for 10 min each. A single episode of WBHP failed to produce protection; however, two and three episodes of WBHP significantly produced beneficial effects on the heart, brain and blood vessels. There was a significant increase in the levels of brain-derived neurotrophic factor (BDNF) and nitric oxide (NO) in response to 3 episodes of WBHP. Moreover, pretreatment with the BDNF receptor, TrkB antagonist (ANA-12) and NO synthase inhibitor (LNAME) attenuated the protective effects imparted by three episodes of WBHP. These pharmacological agents abolished WBHP-induced restoration of p-GSK-3β/GSK-3β ratio and Nrf2 levels in IR-subjected hearts. It is concluded that repeated episodes of WHBP attenuate cognitive impairment, vascular dysfunction and enhancement in IRinduced myocardial injury in diabetic mice be due to increase in NO and BDNF levels that may eventually activate GSK-3β and Nrf2 signaling pathway to confer protection.

The beneficial effects of hypoxic preconditioning are abolished in the diabetes. The present study was designed to investigate the protective effects and mechanisms of repeated episodes of whole body hypoxic preconditioning (WBHP) in db/db mice. The protective effects of preconditioning were explored on diabetesinduced vascular dysfunction, cognitive impairment and ischemia-reperfusion (IR)-induced increase in myocardial injury. Sixteen-week old db/db (diabetic) and C57BL/6 (non-diabetic) mice were employed. There was a significant impairment in cognitive function (Morris Water Maze test), endothelial function (acetylcholineinduced relaxation in aortic rings) and a significant increase in IR-induced heart injury (Langendorff apparatus) in db/db mice. WBHP stimulus was given by exposing mice to four alternate cycles of low (8%) and normal air O2 for 10 min each. A single episode of WBHP failed to produce protection; however, two and three episodes of WBHP significantly produced beneficial effects on the heart, brain and blood vessels. There was a significant increase in the levels of brain-derived neurotrophic factor (BDNF) and nitric oxide (NO) in response to 3 episodes of WBHP. Moreover, pretreatment with the BDNF receptor, TrkB antagonist (ANA-12) and NO synthase inhibitor (LNAME) attenuated the protective effects imparted by three episodes of WBHP. These pharmacological agents abolished WBHP-induced restoration of p-GSK-3β/GSK-3β ratio and Nrf2 levels in IR-subjected hearts. It is concluded that repeated episodes of WHBP attenuate cognitive impairment, vascular dysfunction and enhancement in IRinduced myocardial injury in diabetic mice be due to increase in NO and BDNF levels that may eventually activate GSK-3β and Nrf2 signaling pathway to confer protection.

Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells (SGC-7901). Methods Different concentrations of coix seed oil (2, 4, 8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation, and flow cytometry for drug-induced cell apoptosis, scratch test for cell migration inhibited by coix seed oil, Transwell chamber for drug-inhibited cell invasion, and Western blot for expression of related proteins PRMT5, PI3 K and AKT. Results The results of MTT showed that 2, 4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells, and the inhibition rate of 2 mg/mL was (30. 02 ± 1. 56) ％, which showed significant difference compared to the control group (P ＜ 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were (16. 25 ± 2. 54) ％, (12. 60 ± 1. 12) ％, respectively, and which showed a significant difference compared with (2. 0 ± 1. 22) ％ in the control group (P ＜ 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2, 4 mg/mL of coix seed oil had significant difference when compared to control group (P ＜0. 01). The results of invasion experiments showed that 2, 4 mg/mL of coix seed oil could significantly inhibit cell invasion, and the number of cell invasion was (134. 00 ± 2. 86), (167. 00 ±0. 99), respectively, which showed significant difference compared to (268. 00 ± 2. 05) in the control group (P ＜ 0. 01). The migration number in 8 mg/mL coix seed oil group was (167 ± 0. 99), a significant difference was observed when compared to the control group (P ＜ 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5, PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation, migration and invasion of gastric cancer SGC-7901 cells, its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules, induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.

Objective To investigate the possible mechanism of coix seed in inducing apoptosis of gastric cancer cells (SGC-7901). Methods Different concentrations of coix seed oil (2, 4, 8 mg/mL) were applied to SGC-7901 cells. MTT assay was used to detect the effect of drugs on cell proliferation, and flow cytometry for drug-induced cell apoptosis, scratch test for cell migration inhibited by coix seed oil, Transwell chamber for drug-inhibited cell invasion, and Western blot for expression of related proteins PRMT5, PI3 K and AKT. Results The results of MTT showed that 2, 4 mg/mL of coix seed oil could significantly inhibit the proliferation of SGC-7901 cells, and the inhibition rate of 2 mg/mL was (30. 02 ± 1. 56) ％, which showed significant difference compared to the control group (P ＜ 0. 01). The results of flow cytometry showed that the apoptosis rates of coix seed oil cells at concentrations of 2 and 4 mg/mL were (16. 25 ± 2. 54) ％, (12. 60 ± 1. 12) ％, respectively, and which showed a significant difference compared with (2. 0 ± 1. 22) ％ in the control group (P ＜ 0. 01). The results of cell scratch test showed that the migration of SGC-7901 cells treated with 2, 4 mg/mL of coix seed oil had significant difference when compared to control group (P ＜0. 01). The results of invasion experiments showed that 2, 4 mg/mL of coix seed oil could significantly inhibit cell invasion, and the number of cell invasion was (134. 00 ± 2. 86), (167. 00 ±0. 99), respectively, which showed significant difference compared to (268. 00 ± 2. 05) in the control group (P ＜ 0. 01). The migration number in 8 mg/mL coix seed oil group was (167 ± 0. 99), a significant difference was observed when compared to the control group (P ＜ 0. 05). Western blot analysis showed that different concentrations of coix seed oil could significantly down-regulate the expression of PRMT5, PI3 K and AKT in SGC-7901 cells. Conclusion Coix seed oil can significantly inhibit the proliferation, migration and invasion of gastric cancer SGC-7901 cells, its possible mechanism is to down-regulate the signal pathway of PRMT5-PI3 K/AKT to inhibit the activation of various anti-apoptotic molecules, induce apoptosis and suppress invasion and metastasis of gastric cancer SGC-7901 cells by down-regulating the PRMT5-PI3 K/AKT signaling pathway.

Objective To identify the influence of chemotherapy-induced serum alkaline phosphatase (ALP) elevation on the tumor-free survival (TFS) in patients of gastric carcinoma after radical gastrectomy.Methods The clinical data of 189 gastric carcinoma patients receiving radical surgery and postoperative adjuvant chemotherapy between Jan,2010 and Dec,2015 were reviewed and statistically analyzed.Results The TFS of patients with serum ALP elevation was obviously inferior than those without ALP elevation (x2 =5.717,P =0.017),serum ALP elevation was an independent risk factor influencing patients' TFS (HR =2.178,P =0.032),the degree of serum ALP elevation was associated with patients' TFS (x2 =4.627,P =0.031).Conclusion Serum ALP elevation during postoperative chemotherapy indicates the increases of recurrence or metastasis rate of gastric cancer patients after radical gastrectomy.

Eight mouse hybridoma cell lines which stably secreted monoclonal antibodies ( McAbs ) against human prostate-specific antigen-α1-antichymotrypsin complex ( PSA-ACT ) were obtained through hybridoma technique. After purification, the immunological characters of 8 McAbs were identified and classified by epitopes analysis through indirect enzyme-linked immunosorbent assay ( ELISA) . A pair of McAbs was chosen from above 8 McAbs, based on which a highly sensitive, simple and rapid chemiluminescence enzyme immunoassay ( CLEIA) was developed for determination of PSA-ACT in human serums using the lumino-H2 O2 reaction catalyzed by horseradish peroxidase ( HRP) as the chemiluminescence system. Several experiment factors such as coating buffer, coating concentration, dilution ratio of PSA-ACT-HRP complex, incubation time, immunoreaction protocol and chemiluminescence reaction time were optimized. The results showed that the linear range of the proposed method for PSA-ACT determination was 0-40 ng/mL (R2=0. 9943), with the detection limit of 0. 53 ng/mL. The inter-assay relative standard deviations (RSDs) were 4. 6%-6. 6%, and intra-assay RSDs were 5 . 7%-8 . 0%. The recoveries of PSA-ACT at three spiked levels in serum samples were 95. 4%-104. 2%. The proposed method exhibited a cross-reactivity of 0. 6% with free-PSA. The proposed method is stable, sensitive, rapid and simple, and provides a foundation for the development of PSA-ACT CLEIA kit and shows great value in clinical auxiliary diagnosis of prostate cancer.

Interactions between FMDV and cardiac cells are multifaceted and complex ,these interactions leads to pro-teins alterations in cardiac cells inevitably .To understand the pathogenesis of myocarditis after FMDV infection in mice ,the suckling mouse model for myocarditis induced by foot-and-mouth disease virus (FMDV) was established in this study .Suckling mice within 3 days old was selected to infect by FMDV .Myocarditis caused by FMDV in suckling mice was confirmed with clinical symptom monitor .The observation of Hematoxylin and eosin stain (H&E stain) and transmission electron microscopy (TEM) were performed after samples processing .According to conventional polymerase chain reaction (PCR) methods ,prim-ers of VP1 gene was designed ,synthesised and specific FMDV VP1 gene was amplified from the heart muscle of suckling mice . The results indicated that suckling mice appeared low spirit condition ,dyspnea ,and dull reaction within 36 hours after chal-lenge with FMDV .Infiltration of inflammatory cells and dissolution of myocardial fibers were observed with H&E stain and TEM .Special target gene of FMDV was amplified from the heart of infected group .Obvious inflammation in the heart of suck-ling mice caused by FMDV was observed .It's suggested that suckling mouse model for myocarditis induced by FMDV was es-tablished successfully ,which would lay the foundation for researches of myocarditis mechanism in young cloven-hoofed ani-mals .

Porcine reproductive and respiratory syndrome is caused by the PRRS virus(PRRSV), which has six structural proteins(GP2, GP3, GP4, GP5, M and N). GP5 and N protein are important targets for serological detection by enzyme-linked immunosorbent assay(ELISA)and other methods. Toward this goal, we developed an indirect ELISA with recombinant GP5 antigens and this method was validated by comparison to the LSI PRRSV-Ab ELISA kit. The results indicated that the optimal concentration of coated recombinant antigen was 0.2 μg/well for a serum dilution of 1:40. The rate of agreement with the LSI PRRSV-Ab kit was 88.7%(266/300). These results support the potential use of recombinant GP5 as an antigen for indirect ELISA to detect PRRSV antibodies in pigs.