This study aims to evaluate the preventive effect of the exogenous protein chIFN-γ-chCD154 against Salmonella typhimurium(S.typhimurium)infection in White Leghorn chick-ens,and the potential mechanism.In this study,Escherichia coli was used to express the proteins chIFN-γ,chCD154 and chIFN-γ-chCD154.Before S.typhimurium infection,the White Leghorn chickens were pre-immunized via drinking water for three consecutive days,and infected with S.typhimurium by gavage.The results from Western blot,quantitative real-time PCR(qRT-PCR)analysis and histopathology analysis showed that compared to chIFN-γ and chCD154,chIFN-γ-chCD154 pre-treatment could synergistically increase the survival rate of infected chickens,reduce the bacterial load in the liver and cecum,and attenuate the pathological damage of liver and cecum.Moreover,chIFN-γ-chCD154 significantly decreased the mRNA expression of Toll-like receptor-4(TLR-4)and myeloid differentiation primary response gene 88(MyD88)in the cecum,and then inhibited the mRNA expression of NF-κB and pro-inflammatory cytokines(TNF-α and IL-6),and maintained the integrity of the intestinal tight junction proteins(zo-1,claudin-1,occlu-din).Compared with single protein pretreatment,chIFN-γ-chCD154 pretreatment significantly up-regulated the mRNA expression of the genes related to the vitamin D(VD)pathway(cyp27b1,VD receptor VDR,antimicrobial peptide AvBD7 and cathelicidin-b1)in S.typhimurium-infected peripheral blood mononuclear cells(PBMCs).The results of colony counting showed that the num-ber of S.typhimurium in the chIFN-γ-chCD154 group were the lowest.Also,chIFN-γ-chCD154 could up-regulate the relative abundance of beneficial bacteria such as Blautia,Ruminococcus,En-terococcus and Faecalibacterium,while down-regulate the relative abundance of harmful bacteria such as Staphylococcaceae in the cecum and improve the intestinal dysbiosis.In conclusion,chIFN-γ-chCD154 could activate the VD-antimicrobial peptide pathway and inhibit the TLR-4/MyD88/NF-κB inflammatory signaling pathway in S.typhimurium-infected chickens,which significantly improve the intestinal barrier function,reduce the inflammatory damage of liver and cecum,im-prove the structure of cecum microbial,promote the health of intestinal tract,and provide theoreti-cal basis for the development of chIFN-γ-chCD154 as a safe and effective alternative antibiotic.

This study aims to evaluate the preventive effect of the exogenous protein chIFN-γ-chCD154 against Salmonella typhimurium(S.typhimurium)infection in White Leghorn chick-ens,and the potential mechanism.In this study,Escherichia coli was used to express the proteins chIFN-γ,chCD154 and chIFN-γ-chCD154.Before S.typhimurium infection,the White Leghorn chickens were pre-immunized via drinking water for three consecutive days,and infected with S.typhimurium by gavage.The results from Western blot,quantitative real-time PCR(qRT-PCR)analysis and histopathology analysis showed that compared to chIFN-γ and chCD154,chIFN-γ-chCD154 pre-treatment could synergistically increase the survival rate of infected chickens,reduce the bacterial load in the liver and cecum,and attenuate the pathological damage of liver and cecum.Moreover,chIFN-γ-chCD154 significantly decreased the mRNA expression of Toll-like receptor-4(TLR-4)and myeloid differentiation primary response gene 88(MyD88)in the cecum,and then inhibited the mRNA expression of NF-κB and pro-inflammatory cytokines(TNF-α and IL-6),and maintained the integrity of the intestinal tight junction proteins(zo-1,claudin-1,occlu-din).Compared with single protein pretreatment,chIFN-γ-chCD154 pretreatment significantly up-regulated the mRNA expression of the genes related to the vitamin D(VD)pathway(cyp27b1,VD receptor VDR,antimicrobial peptide AvBD7 and cathelicidin-b1)in S.typhimurium-infected peripheral blood mononuclear cells(PBMCs).The results of colony counting showed that the num-ber of S.typhimurium in the chIFN-γ-chCD154 group were the lowest.Also,chIFN-γ-chCD154 could up-regulate the relative abundance of beneficial bacteria such as Blautia,Ruminococcus,En-terococcus and Faecalibacterium,while down-regulate the relative abundance of harmful bacteria such as Staphylococcaceae in the cecum and improve the intestinal dysbiosis.In conclusion,chIFN-γ-chCD154 could activate the VD-antimicrobial peptide pathway and inhibit the TLR-4/MyD88/NF-κB inflammatory signaling pathway in S.typhimurium-infected chickens,which significantly improve the intestinal barrier function,reduce the inflammatory damage of liver and cecum,im-prove the structure of cecum microbial,promote the health of intestinal tract,and provide theoreti-cal basis for the development of chIFN-γ-chCD154 as a safe and effective alternative antibiotic.

OBJECTIVE To explore the optimal process of extraction and purification of total phenols from deoiled Cinnamomum longepaniculatum leaves, and to evaluate the antioxidant activity in vitro before and after purification, so as to provide a theoretical basis for rational development and utilization of Cinnamomum longepaniculatum. METHODS The L18(37) orthogonal experiment was carried out on the basis of five-single factor and three-level experiments, such as concentration of ethanol(A), ratio of liquid-material(B), extraction temperature(C), extraction time(D), and extraction times(E). The extraction process of total phenols was optimized, by taking the amount of total phenols extraction as the process investigation index. Taking the adsorption and desorption performance under dynamic and static conditions as the investigation index of total phenols purification process, the purification process of total phenols was optimized by macroporous resin. With vitamin C(Vc) as the positive control, the antioxidant activity in vitro of total phenols before and after purification were evaluated by DPPH radical scavenging, ABTS+ radical scavenging and determination of total reducing capacity. RESULTS The best extraction technology conditions of total phenols were as follows: ratio of liquid-material 30 mL·g−1, concentration of ethanol 50%, extraction time 2 h, extraction temperature 90 ℃, extraction 2 times. Under such conditions , the extraction amount of total phenols from deoiled Cinnamomum longepaniculatum leaves was 15.63 mL·g−1. HPD-600 type macroporous resin showed the best purifying profile. The best purification technology conditions were as follows: the concentration of sample solution was 0.5 g·L−1, the sample solution pH value was 3, the sample volume was 2.5 BV, the sample flow rate 1 mL·min−1, the impurity was removed by 4.5 BV distilled water, the elution concentration of ethanol was 60%, the eluate pH value 7, the elution flow rate 1 mL·min−1, 4.5 BV eluent was collected and the total phenols assay was increased from 12.51% to 26.40%. The purified total phenols showed excellent antioxidant activity in vitro. The scavenging effect of total phenols after purification on DPPH and ABTS+ free radicals increased with the increase of total phenols concentration. The IC50 values of DPPH radical scavenging activity of total phenols before and after purification were (68.31±1.96)mg·L−1 and (38.07±0.66)mg·L−1, respectively. The IC50 values of ABTS+ radical scavenging activity of total phenols before and after purification were (23.88±1.66)mg·L−1and (14.28±0.19)mg·L−1, respectively. The absorbance values of the total reducing capacity of total phenols was 1.68 times higher than that before purification. The scavenging effect of total phenols before and after purification on DPPH, ABTS+ free radical scavenging and total reducing capacity was weaker than that of Vc. CONCLUSION The process is stable and feasible, suitable for industrial production, and has the advantages of simplicity, rapidity and accuracy, which lays a foundation for further research on Cinnamomum longepaniculatum and the development of natural antioxidants.

Objectives:To evaluate the role of pancreas multidisciplinary team(MDT) clinic in the diagnosis of pancreatic diseases,patient compliance with MDT advice,and the impact of MDT on the postoperative survival of patients with pancreatic cancer.Methods:The study included 927 patients(554 males,373 females,aged (58.1±13.3)years (range: 15 to 89 years)) that had visited the pancreas MDT clinic of Zhongshan Hospital from May 2015 to December 2021,and 677 patients(396 males, 281 females, aged (63.6±8.9)years(range: 32 to 95 years)) who underwent radical surgery and with pathologically confirmed pancreatic adenocarcinoma from January 2012 to December 2020,of whom 79 patients had attended the pancreas MDT. The clinical and pathological data were collected and analyzed retrospectively. Diseases were classified in accordance with 2010 WHO classification of tumors of the digestive system and usual clinical practices. The Kaplan-Meier method was used for drawing the survival curve and calculating the survival rate. The univariate analysis was done by Log-rank test and the multivariate analysis was done by COX proportional hazards model. Survival rates were compared using χ 2 test. Results:Among the 927 patients that had visited the MDT clinic,233 patients(25.1%) were referred due to undetermined diagnosis. A direct diagnosis was made in 109 cases (46.8%,109/233) by the MDT clinic, of which 98 were consistent with the final diagnosis,resulting in an accuracy of 89.9%(98/109). The direct diagnosis rate in the recent years(36.6%(41/112),from June 2019 to December 2021) decreased compared to that in the previous years(56.2%(68/121),from May 2015 to May 2019),yet the accuracy in the recent years(90.2%,37/41) was basically the same as before (89.7%,61/68). The rate of compliance of the entire cohort was 71.5%(663/927), with the compliance rate in the recent two and a half years(81.4%,338/415) remarkably higher than that in the previous four years(63.4%,325/512). Patients with pancreatic cancer that attended the MDT exhibited a trend toward longer median postoperative survival than patients that did not attend the MDT,but the difference was not statistically significant(35.2 months vs.30.2 months, P>0.05). The 1-year and 3-year survival rates of patients that attended the MDT were significanly higher than patients that did not attend the MDT(88.6% vs. 78.4%, P<0.05;32.9% vs. 21.9%, P<0.05,respectively),but the 5-year survival rate was not statistically different(7.6% vs. 4.8%, P>0.05). Conclusions:The pancreas MDT clinic is an accurate and convenient way to diagnose intractable pancreatic diseases,and in the recent years the patients′ compliance rate with MDT advice has increased. Pancreatic cancer patients that have attended the MDT have higher 1-year and 3-year postoperative survival rates,but the long-term survival benefits of MDT still needs to be proved by clinical studies on a larger scale.

Objectives:To evaluate the role of pancreas multidisciplinary team(MDT) clinic in the diagnosis of pancreatic diseases,patient compliance with MDT advice,and the impact of MDT on the postoperative survival of patients with pancreatic cancer.Methods:The study included 927 patients(554 males,373 females,aged (58.1±13.3)years (range: 15 to 89 years)) that had visited the pancreas MDT clinic of Zhongshan Hospital from May 2015 to December 2021,and 677 patients(396 males, 281 females, aged (63.6±8.9)years(range: 32 to 95 years)) who underwent radical surgery and with pathologically confirmed pancreatic adenocarcinoma from January 2012 to December 2020,of whom 79 patients had attended the pancreas MDT. The clinical and pathological data were collected and analyzed retrospectively. Diseases were classified in accordance with 2010 WHO classification of tumors of the digestive system and usual clinical practices. The Kaplan-Meier method was used for drawing the survival curve and calculating the survival rate. The univariate analysis was done by Log-rank test and the multivariate analysis was done by COX proportional hazards model. Survival rates were compared using χ 2 test. Results:Among the 927 patients that had visited the MDT clinic,233 patients(25.1%) were referred due to undetermined diagnosis. A direct diagnosis was made in 109 cases (46.8%,109/233) by the MDT clinic, of which 98 were consistent with the final diagnosis,resulting in an accuracy of 89.9%(98/109). The direct diagnosis rate in the recent years(36.6%(41/112),from June 2019 to December 2021) decreased compared to that in the previous years(56.2%(68/121),from May 2015 to May 2019),yet the accuracy in the recent years(90.2%,37/41) was basically the same as before (89.7%,61/68). The rate of compliance of the entire cohort was 71.5%(663/927), with the compliance rate in the recent two and a half years(81.4%,338/415) remarkably higher than that in the previous four years(63.4%,325/512). Patients with pancreatic cancer that attended the MDT exhibited a trend toward longer median postoperative survival than patients that did not attend the MDT,but the difference was not statistically significant(35.2 months vs.30.2 months, P>0.05). The 1-year and 3-year survival rates of patients that attended the MDT were significanly higher than patients that did not attend the MDT(88.6% vs. 78.4%, P<0.05;32.9% vs. 21.9%, P<0.05,respectively),but the 5-year survival rate was not statistically different(7.6% vs. 4.8%, P>0.05). Conclusions:The pancreas MDT clinic is an accurate and convenient way to diagnose intractable pancreatic diseases,and in the recent years the patients′ compliance rate with MDT advice has increased. Pancreatic cancer patients that have attended the MDT have higher 1-year and 3-year postoperative survival rates,but the long-term survival benefits of MDT still needs to be proved by clinical studies on a larger scale.

Objective:To analyze the clinical characteristics of severe adenovirus pneumonia with bacterial infection in children, so as to provide clinical evidences for early diagnosis and reliable treatment.Methods:A total of 72 pediatric cases with severe adenovirus pneumonia admitted to Pediatric Intensive Care Unit at the First People′s Hospital of Chenzhou from January 2018 to August 2019 were included.The clinical features, laboratory and imaging data, efficacy of the treatments and prognosis were investigated retrospectively.Patients were divided into with bacterial infection group and without bacterial infection group.The clinical characteristics were analyzed.Results:Among the 72 children, there were 54 males and 18 females, aging from 3 months to 5 years, including 37cases with bacterial infection and 35 cases without bacterial infection.Compared with the group without bacterial infection, the group with bacterial infection had longer heat duration and hospital stay [12.0 (10.0, 18.5) days vs.10.0 (9.0, 12.0) days; 6.0(4.0, 7.0) days vs.11.0(6.5, 16.0) days, P<0.05], the incidences of diarrhea and hepatomegaly were higher[35.1% (13/37) vs.14.3%(5/35); 45.9%(17/37)vs.8.6%(3/35), P<0.05], the proportion of five lobes involved in lung imaging was higher [91.8% (34/37) vs. 57.1%(20/35), P<0.05]. The incidences of complications with respiratory failure, hemophagocytic lymphohistiocytosis, and bronchiolitis obliterans in the group with bacterial infection were significantly higher than those of without bacterial infection group ( P<0.05). Conclusion:For children under 2 years old age with severe adenovirus pneumonia, there are prolonged high fever and extensive pulmonary lesions.We should be highly alert to the combination of bacterial infection and timely anti-infection therapy.Children with severe adenovirus pneumonia with bacterial infection have severe clinical manifestations and many complications with respiratory failure, hemophagocytic lymphohistiocytosis, and bronchiolitis obliterans.

Objective:To investigate the efficacy and safety of plasma exchange(PE) in the treatment of autoimmune hemolytic anemia in children.Methods:The data from 8 hospitals in China during November 2014 to April 2017 were collected, and the clinical characteristics of PE in children with AHA were analyzed retrospectively.Results:A total of 21 children with AHA were included in the study, including 17 cases from PICU and 4 cases from pediatric kidney ward, with 11 boys and 10 girls, and the median age was 3.64(0.25, 11.10)years old, and median hospital stay was 12(4, 45)days.There were 15 cases(71.4%) with infection, 2 cases(9.5%)with autoimmune diseases, 4 cases(19.0%) with unknown.Consciousness disturbance occurred in 4 patients before replacement and recovered to normal after PE.The volume of blood decreased in two cases(9.5%) and completely relieved.There were 20 cases of anemia (95.2%), 15 cases were normal after PE, and 5 cases were improved.Jaundice occurred in 18 cases (85.7%), 12 cases were normal after PE, 6 cases were improved.Hepatosplenomegaly was found in 11 cases, 10 cases were normal after PE, 1 case was improved.After PE, the hemoglobin and red blood cell count increased, while the total bilirubin, indirect bilirubin, urea nitrogen and lactate dehydrogenase decreased, there were significant differences between pre-and post-replacement ( P<0.05). Only 1 case had allergic reaction, which was improved after symptomatic treatment, and PE was continued.After PE, 2 cases (9.5%) had complete remission, 16 cases (76.2%) had partial remission and 3 cases (14.3%) had been discharged. Conclusion:PE therapy can obviously improve the clinical symptoms and laboratory indexes of children with AHA who have failed to respond to conservative treatment.It can be used as a treatment measure for children with severe AHA and has a good safety.

Hyperuricemia/gout is a common metabolic disease in China, which is a serious threat to people′s health. In clinical practice, the standardization of prevention and diagnosis and the rate of treat-to-target need to be improved. There is still a lack of education for the patients about the understanding of clinical guidelines, the disease knowledge and the importance of cooperating with doctors to carry out diagnosis and treatment. From the most concerned issues of the patients, we established the hyperuricemia/gout patient practice guideline working group with multidisciplinary physicians and patients. Seventeen opinions, as the hyperuricemia/gout patient practice guidelines, are proposed in accordance with the relevant principles of the "WHO guidelines development manual" , and with the international normative process, aiming to improve the patients compliance, improve the level of health management of the disease.

Objective:To detect 11 kinds of illegally added chemicals in anti-fatigue health foods including sildenafil ,tadalafil, vardenafil , acetildenafil , homosildenafil , hydorxyhomosildenafil , norneosildenafil , aminotadalafil , pseudovardenafil , thioaildenafil and noracetildenafil.Methods:An HPLC-MS/MS method was used with LUNA C18(250 mm ×4.6 mm, 5μm)as the analysis column and triethylamine phosphate , acetonitrile and methanol as the mobile phase with gradient elution at the flow rate of 1ml.min-1 .An ESI+scan and an MS/MS mode were applied .The qualitative and quantitative analysis were carried out by HPLC-MS/MS.Results:Through the detection of 100 batches of samples , 85 batches were only found illegally added chemical substance sildenafil mainly at the concentration of 5-15 mg per granule .Conclusion:The method of HPLC-MS/MS can be used as a powerful screening tool for the rapid drug screening in basic level drug control laboratories to reduce chemical substances illegally added in anti -fatigue health foods .

Objective To investigate the risk factors of delayed gastric emptying after pancreaticoduodenectomy (PD) for periampullary carcinoma.Methods The clinical data of 96 periampullary carcinoma patients who received PD with or without pylorus preservation at Zhongshan Hospital Fudan University from October 2010 to September 2013 were retrospectively analyzed.The risk factors associated with delayed gastric emptying were analyzed.The univariate and multivariate analysis were done using the chi-square test and Logistic regression model respectively.Results A total of 17 patients had postoperative delayed gastric emptying, with the incidence of 18％.The results of univariate showed that female sex, pancreatogastrostomy, increased intraoperative blood loss and transfusions and postoperative abdominal infection were risk factors of delayed gastric emptying (x2 =4.512, 6.412, 6.245, 6.695, 6.816, all P ＜ 0.05).The results of multivariate analysis revealed that only pancreatogastrostomy and postoperative abdominal infection were independent risk factors of delayed gastric emptying (OR =4.031 and 5.281).Conclusions Pancreatogastrostomy and postoperative abdominal infection indicate a higher incidence of delayed gastric emptying after pancreaticodudenectomy for periampullary carcinoma.