BACKGROUND: Arsenic trioxide (ATO) is indicated as a broad-spectrum medicine for a variety of diseases, including cancer and cardiac disease. While the role of ATO in hepatic ischemia/reperfusion injury (HIRI) has not been reported. Thus, the purpose of this study was to identify the effects of ATO on HIRI. METHODS: In the present study, we established a 70% hepatic warm I/R injury and partial hepatectomy (30% resection) animal models in vivo and hepatocytes anoxia/reoxygenation (A/R) models in vitro with ATO pretreatment and further assessed liver function by histopathologic changes, enzyme-linked immunosorbent assay, cell counting kit-8, and terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) assay. Small interfering RNA (siRNA) for extracellular signal-regulated kinase (ERK) 1/2 was transfected to evaluate the role of ERK1/2 pathway during HIRI, followed by ATO pretreatment. The dynamic process of autophagic flux and numbers of autophagosomes were detected by green fluorescent protein-monomeric red fluorescent protein-LC3 (GFP-mRFP-LC3) staining and transmission electron microscopy. RESULTS: A low dose of ATO (0.75 μmol/L in vitro and 1 mg/kg in vivo ) significantly reduced tissue necrosis, inflammatory infiltration, and hepatocyte apoptosis during the process of hepatic I/R. Meanwhile, ATO obviously promoted the ability of cell proliferation and liver regeneration. Mechanistically, in vitro  studies have shown that nontoxic concentrations of ATO can activate both ERK and phosphoinositide 3-kinase-serine/threonine kinase (PI3K-AKT) pathways and further induce autophagy. The hepatoprotective mechanism of ATO, at least in part, relies on the effects of ATO on the activation of autophagy, which is ERK-dependent. CONCLUSION Low, non-toxic doses of ATO can activate ERK/PI3K-AKT pathways and induce ERK-dependent autophagy in hepatocytes, protecting liver against I/R injury and accelerating hepatocyte regeneration after partial hepatectomy.
Animals ; Arsenic Trioxide ; Autophagy/physiology* ; Reperfusion Injury/prevention & control* ; Mice ; Male ; Proto-Oncogene Proteins c-akt/physiology* ; Arsenicals/therapeutic use* ; Oxides/therapeutic use* ; Liver/metabolism* ; Extracellular Signal-Regulated MAP Kinases/metabolism* ; Mice, Inbred C57BL

Objective: To investigate the incidence of postoperative pancreatic fistula (POPF) and its risk factors after radical gastrectomy. Methods: The prospective study was conducted. The clinicopathological data of 2 089 patients who underwent radical gastrectomy in 22 medical centers between December 2017 and November 2018 were collected, including 380 in the Zhongshan Hospital of Fudan University, 351 in the Renji Hospital of Shanghai Jiaotong University School of Medicine, 130 in the Ruijin Hospital of Shanghai Jiaotong University School of Medicine, 139 in the Peking University Cancer Hospital, 128 in the Fujian Provincial Cancer Hospital, 114 in the First Hospital Affiliated to Army Medical University, 104 in the First Affiliated Hospital of Nanchang University, 104 in the Affiliated Hospital of Qinghai University, 103 in the Weifang People′s Hospital, 102 in the Fujian Medical University Union Hospital, 99 in the First Affiliated Hospital of Air Force Medical University, 97 in the Sir Run Run Shaw Hospital Affiliated to Zhejiang University School of Medicine, 60 in the Hangzhou First People′s Hospital Affiliated to Zhejiang University School of Medicine, 48 in the Fudan University Shanghai Cancer Center, 29 in the First Affiliated Hospital of Xi′an Jiaotong University, 26 in the Lishui Municipal Central Hospital, 26 in the Guangdong Provincial People′s Hospital, 23 in the Jiangsu Province Hospital, 13 in the First Affiliated Hospital of Sun Yat-Sen University, 7 in the Second Hospital of Jilin University, 4 in the First Affiliated Hospital of Xinjiang Medical University, 2 in the Beijing Chao-Yang Hospital of Capital Medical University. Observation indicators: (1) the incidence of POPF after radical gastrectomy; (2) treatment of grade B POPF after radical gastrectomy; (3) analysis of clinicopathological data; (4) analysis of surgical data; (5) risk factors for grade B POPF after radical gastrectomy. Measurement data with normal distribution were represented as Mean±SD, and comparison between groups was analyzed using ANOVA. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Univariate analysis was conducted using the t test or chi-square test based on data excluding missing data of clinico-pathological and surgical data. Multivariate analysis was conducted using the Logistic regression model based on factors with P<0.20 in univariate analysis. Results: There were 2 089 patients screened for eligibility, including 1 512 males, 576 females and 1 without sex information, aged (62±11)years. The body mass index (BMI) was (23±3)kg/m². (1) The incidence of POPF after radical gastrectomy: the total incidence rate of POPF in the 2 089 patients was 20.728%(433/2 089). The incidence rates of biochemical fistula, grade B pancreatic fistula, and grade C pancreatic fistula were 19.627%(410/2 089), 1.101%(23/2 089), 0, respectively. (2) Treatment of grade B POPF after radical gastrectomy: 2 of 23 patients with grade B POPF after radical gastrectomy had drainage tube placed for more than 21 days and received anti-infective therapy. Four of 23 patients with grade B POPF after radical gastrectomy had ascites detected by imaging examination, of which 2 received peritoneal drainage guided by ultrasound, 1 received failed puncture drainage, 1 received no puncture drainage, and they were given anti-infective therapy. Eleven of 23 patients with grade B POPF after radical gastrectomy had no ascites detected by imaging examinations, and they were given anti-infective therapy and inhibitors of pancreas secretion for clinical manifestation as fever or elevated white blood cells. Six patients with no typical clinical manifestations were given somatostatin to inhibite pancreas secretion and prolonged duration of abdominal drainage tube placement (with a median time of 7 days). All the 23 patients recovered well after treatment, without reoperation. (3) Analysis of clinicopathological data: for the 2 089 patients, BMI, cases with or without neoadjuvant therapy were (23±3)kg/m^2, 1 487, 160 of patients without pancreatic fistula, (23±3)kg/m^2, 386, 22 of patients with biochemical fistula, and (24±3)kg/m^2, 22, 1 of patents with grade B pancreatic fistula, showing significant differences between the three groups (F=5.787, χ²=8.269, P<0.05). (4) Analysis of surgical data: for the 2 089 patients, cases with open surgery, laparoscopic assisted surgery, totally laparoscopic surgery (surgical method), cases with D₁ lymph lode dissection, D₂ lymph lode dissection, and other lymph lode dissection (range of lymph lode dissection), cases with no omentectomy, partial omentectomy, and total omentectomy (range of omentectomy), cases with no usage of energy facility, usage of CUSA, LigaSure, LigaSure+ CUSA as energy facility, cases with or without biological glue, the number of lymph node dissection were 737, 624, 292, 24, 1 580, 51, 418, 834, 381, 63, 1 530, 23, 16, 1 431, 201, 33±14 of patients without pancreatic fistula, 146, 189, 74, 11, 389, 9, 110, 171, 128, 35, 359, 6, 9, 378, 31, 31±14 of patients with biochemical fistula, and 14, 5, 4, 0, 20, 3, 6, 13, 4, 2, 18, 1, 2, 22, 1, 37±16 of patients with grade B pancreatic fistula, showing significant differences between the three groups (χ²=15.578, 9.397, 15.023, 28.245, 8.359, F=4.945, P<0.05). (5) Risk factors for grade B POPF after radical gastrectomy: results of univariate analysis showed that usage of energy facility was a related factor for grade B POPF after radical gastrectomy (χ²=9.914, P<0.05). Results of multivariate analysis showed that laparoscopic assisted surgery, combined evisceration, application of LigaSure + CUSA, the number of lymph lode dissection were independent factors for for grade B POPF after radical gastrectomy (odds ratio=0.168, 3.922, 9.250, 1.030, 95% confidence interval: 0.036-0.789, 1.031-14.919, 1.036-82.602, 1.001-1.059, P<0.05). Conclusions The incidence of grade B POPF after radical gastrectomy is relatively low. Laparoscopic assisted surgery, combined evisceration, application of LigaSure + CUSA, and the number of lymph lode dissection are independent risk factors for grade B POPF. Trial Registration: This study was registrated at ClinicalTrial.gov in United States with the registration number of NCT03391687.

Objective To observe the clinical efficacy and safety of Chushi Huowei Decoction in treating inflammation of gastric relict of spleen-stomach damp heat type. Methods Totally 62 patients with inflammation of gastric relict were divided into treatment group and control group with random number table, 31 patients in each group. The control group was given hydrotalcite and the treatment group was treated by adding of Chushi Huowei Decoction. After eight-week treatment, the clinical efficacy, gastric mucosal lesions efficacy, and gastric mucosal pathology efficacy of the two groups were compared. Results Eight-weeks later, traditional Chinese medicine symptom scores of both groups were obviously lower than those before treatment (P<0.01). And the decrease in the treatment group was more obvious than that in the control group (P<0.01). Curative effects of gastric mucosal lesions and pathologic condition in treatment group were better than those in the control group (P<0.05, P<0.01). Neither groups had any adverse reaction. Conclusion Chushi Huowei Decoction has reliable therapeutic effect and safety in treating inflammation of gastric relict of spleen-stomach damp heat type.

Objective To further understand the role of growth regulation of human ovarian cancer cells by transforming growth factor (TGF) β1.Methods The cell proliferation, cAMP synthesis, gene expression, and induction of programmed cell death (PCD) in human epithelial ovarian cancer cell line HO-8910 cells exposed to TGFβ1 in vitro were studied.Results TGFβ1 inhibited cell growth and DNA synthesis, and induced G0/G1 arrest in cell cycle. It could also trigger PCD in cells. This induction of PCD may occur within G0/G1 phase. Meanwhile, the assay also showed that TGFβ1 could inhibit the mRNA expression of c-myc, EGFR and TGFβ1 genes in cells.Conclusions TGFβ1 can not only act as an autocrine to inhibit cell proliferation, but also trigger PCD in HO-8910 cells. These functions may be fulfilled through some specific signal transduction pathways.

Objective To explore the Schistosoma japonicum cercariae collected method and the quantity of cercariae obtained from a great quantity artificial infected snails. Methods In laboratory condition, Oncomelania snails were infected with schistosome miracidia. Sixty days post-infection all snails were divided into 7 shares. Cercariae shed from 1 share snails every day and the number of all shedding days were 40. Cercariae shed from snails were collected with low-velocity centrifuge and the cercariae sediment were weighted. Results One thousand and nine hundred g snails bred for 120 days post-infection, the infection and survival rates were 36. 00% and 51. 58%. Cercariae col-lected were 10. 5 g from 40 days collection. Cercariae quantity of shedding from 1 000 positive snails per day was 0. 257 4 g.

Objective To investigate the value of mix recombinant antigen in schistosomiasis diagnosis. Methods The recombinant antigens of SjC23 (HD),SjC21.7 and SjCMP10 were expressed in vitro and purified by the affinity chromatography method. The efficacies of soluble egg antigen (SEA),single recombinant antigen and mix recombinant antigen for schistosomiasis diagnosis by Enzyme-linked Immunosorbent Assay were compared. Results The diagnostic efficacy was the same when the antibody IgG of the same group sera of schistosomiasis was detected by different quantities of 2.5 ?g/ml and 7.5 ?g/ml of SEA immobilized on microplate, and their absorbency A was the same, but there was a significant difference in the diagnostic efficacies between single recombinant antigen and mix recombinant antigen when the antibody IgG of the same group sera of schistosomiasis was detected by the same quantity of single recombinant antigen or mix recombinant antigen immobilized on microplate, the absorbency A of mix antigen reacted with the sera of schistosomiasis was significant higher than that of the single recombinant. The positive rates were very similar when 39 sera of acute schistosomiasis,80 sera of chronic schistosomiasis and 27 sera of advanced schistosomiasis were detected by SEA or mix recombinant antigen by ELISA in the same time. No cross-reaction presented when 20 clonorchiasis sera were detected by the mix recombinant antigen and no false positive presented when 40 of healthy sera were detected by the mix recombinant antigen. Conclusion The schistosomiasis diagnostic method by using the mix recombinant antigen has been established, which is helpful for improving the efficacy of schistosomiasis diagnosis.

This study reported the primary application of FA-ELISA from 107-121kDa fration antibody of Schistosoma Japonicum for detection of the short-term antibody in patients with schistosomiasis. The result showed that this method provided high sensitivity (with positive rate of 93. 33% with 30 cases of schistosomiasis) and good specificity (no false postive in 60 health objects). When one group of schistosome patients were examined with FA -ELISA and with the routine SEA-ELISA before chemotherapy and at 8 months,1 year and 1. 5 year after treatment,it showed good property for evaluation of chemotherapy efficacy with FA -ELISA,which was much better than the routine method.

Objective To find out the valuable early diagnostic antigen of Schistosoma japonicum. Methods The sera of rabbits were collected at different time after the rabbits were infected with cercariae of Schitosoma japonicum. The fractions of the soluble cercaria antigen (SCA), soluble adult worm antigen (AWA) and soluble egg antigen (SEA) were separated by SDS-PAGE and recognized by Western blotting with rabbits' sera of different time of post-infection. Results In Western blotting, the bands of 94, 48, 41, 40 kDa and 38 kDa of SCA appeared the earliest and were recognized by the rabbits sera of 2-week post-infection, the bands of 71 kDa and 23 kDa of SCA reacted with the rabbits sera of 3-week post-infection strongly. The bands of 71 kDa and 58 kDa of AWA appeared the earliest and were recognized by rabbits sera of 3-week post-infection. The bands of SEA reacted earliestly to the rabbits sera of 4-week post-infection were 270, 151, 73, 69, 50 kDa and 24 kDa. Conclusion The fraction antigens of 94, 71, 48, 41, 40, 38 kDa and 23 kDa of SCA, the fraction antigens of 71 kDa and 58 kDa of AWA and the fraction antigens of 270, 151, 73, 69, 50 kDa and 24 kDa of SEA could be recognized by sera of acute infected rabbits and might have potential early immuno-diagnosis value for schistosomiasis.

Objective To develop a fast, simple immunodiagnosis assay for early diagnosis of schistosomiasis. Methods The soluble cercariae antigen(SCA) labeled colloidal dye was used as the detecting antigen for schistosomiasis. A dipstick dye immunoassay(SCA-DDIA) for early diagnosis of schistosomiasis was established. The antibodies in sera of infected rabbits in early stage of infection by SCA-DDIA were detected and compared with SEA-DDIA. The sera from people with acute and chronic schistosomiasis and other parasitic diseases and from healthy people were tested by SCA-DDIA and SEA-DDIA. Results In infected rabbits during early stage of infection, the average time of antibody detected by SCA-DDIA was 22 d , at day 30 post-infection all experimental rabbits were positive with SCA-DDIA, the detected time was earlier than that with SEA-DDIA. The sensitivity of SCA-DDIA for acute, chronic schistosomiasis japonica were 100.0% and 93.3% respectively. The specificity for healthy persons was 99.0%. The cross reaction rates with paragonimiasis westermani, clonorchiasis sinensis and fasciolopsiasis buski were 26.3%, 0 and 0 respectively. The results were similar to that by SEA-DDIA. Conclusion The SCA-DDIA is more useful for early diagnosis of schistosomiasis.

Objective To synthesize and fusion express the predicted T-cell epitopes of Schistosoma japonicum, and analyze their immunogenicities. Methods The plus and minus oligo-nucleic acid strands of epitopes P7, P17, P18 were synthesized following their DNA sequence, respectively. The Nco I restriction enzyme sites were added to the 5′ end of epitope gene and the Xho I restriction enzyme sites were added to the 3′ end of epitope gene. The plus and minus strand of each epitope gene was annealed to form double strand DNA fragments. Then the double strand DNA fragments encoding epitope peptide were cloned into the site between Nco I and Xho I of plasmid pET32c(+) to construct recombinant plasmid which was transformed into E.coli DH5?. The recombinant plasmid containing P7, P17, P18 genes respectively was identified by PCR, restriction digestion and DNA sequencing, and then transformed into E.coli BL21 (DE3) for expressing the fusion protein. The fusion protein of peptide-thioredoxin(Trx) was expressed by inducing with IPTG and analyzed with SDS-PAGE. The fusion proteins were purified with Ni2+ column affinity chromatography. Meanwhile, the peptides P7, P17, P18 were synthesized artificially following their amino acid se-quence. By using the purified epitope peptide fusion proteins and synthesized epitope peptides, the splenic cells of C57BL/6J mice immunized with ultraviolet-attenuated cercaria of Schistosoma japonicum were stimulated respectively. The stimulation activity of fusion proteins and synthesized peptides were assayed by detecting the incorporation rate of 3 H-thymidine. Results The double strand DNA fragments of epitopes P7, P17, P18 were successfully cloned to form recombinant plasmids, all of which could express a 20 kDa fusion protein. Both the fusion protein and synthesized epitope peptides of P7 and P17 were able to stimulate the lymphocyte cells to proliferation effectively. Conclusion The peptide P7 and peptide P17 are testified as T-cell epitopes of Schistosoma japonicum.