ObjectiveTraditional Chinese medicine， namely Dahuang Zhechongwan （DHZCW） was used to treat myocardial fibrosis in model rats， observe its effect on myocardial fibrosis in rats， and explore its action mechanism. MethodThirty-six SPF male Kunming rats were divided into blank group， model group， low-， medium-， high-dose groups of DHZCW （0.056， 0.084， 0.168 g·kg^-1）， captopril group （10 mg·kg^-1）， with six rats in each group. Except for the blank group， the other groups were intraperitoneally injected isoproterenol solution of 5 mg·kg^-1 for 15 consecutive days to replicate the myocardial fibrosis model. At the beginning of modeling， the rats in each group took drugs， and they were sacrificed 28 days after administration. Serum and heart tissue were collected for the corresponding detection. Hematoxylin-eosin （HE） staining and Masson staining were used to observe tissue inflammation， cellular degeneration， necrosis， and fibrosis. The contents of hydroxyproline （HYP）， tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， interleukin-6 （IL-6）， hyaluronic acid （HA）， laminin （LN）， type-Ⅲ procollagen （PC Ⅲ） in serum of rats and rats were determined by enzyme-related immunosorbent assay （ELISA）. The expression levels of key pathway proteins transforming growth factor-β₁ （TGF-β₁）， α-smooth muscle actin （α-SMA）， Smad2， Smad3， and Smad7 were detected by Western blot. The expression levels of key pathway genes TGF-β₁， α-SMA， Smad2， Smad3， Smad7， miR-29a-5p， miR-29b-2-5p， and miR-29c-5p were detected by Real-time quantitative polymerase chain reaction （Real-time PCR）. ResultCompared with the blank group， the pathological changes of fibrosis in the model group were obvious， the contents of serum HYP， TNF-α， IL-1β， IL-6， HA， LN， and PCⅢ were increased （P<0.01）， the protein expression levels of TGF-β₁， α-SMA， Smad2， and Smad3 were increased； the protein expression level of Smad7 was decreased （P<0.01）. The mRNA expression levels of TGF-β₁， α-SMA， Smad2， and Smad3 were increased （P<0.05， P<0.01）， while those of Smad7， miR-29a-5p， miR-29b-2-5p， and miR-29c-5p were decreased （P<0.01）. Compared with the model group， after 28 days of administration， serum HYP， TNF-α， IL-1β， IL-6， HA， LN， and PCⅢ in high-， medium-， and low-dose groups of DHZCW and captopril groups were decreased （P<0.01）. Except for the low-dose group， the protein contents of TGF-β₁， α-SMA， Smad2， and Smad3 were decreased， while the protein content of Smad7 was increased （P<0.01）. The mRNA expression levels of TGF-β₁， Smad2， α-SMA， and Smad3 in high-dose group of DHZCW were decreased （P<0.05，P<0.01）， while those of Smad7， miR-29a-5p， miR-29b-2-5p， and miR-29c-5p were increased （P<0.05）. The mRNA expressions of TGF-β₁， Smad2， and Smad3 in the medium-dose group of DHZCW were decreased （P<0.05， P<0.01）， while mRNA expression of Smad7 was increased （P<0.01）. The mRNA levels of TGF-β₁ and Smad2 in the low-dose group of DHZCW were decreased （P<0.01）. ConclusionDHZCW can improve myocardial fibrosis in rats， and its action mechanism may be related to the regulation of the TGF-β₁/Smads/miR-29 pathway. In addition， there is dose dependence in the range of 0.056-0.168 g·kg^-1， and the effect of the high-dose group is more stable.

Objective:To identify and screen the differential methylation genes in patients with cholangiocarcinoma and to predict the prognosis of patients with CCA.Methods:Cholangiocarcinoma tissues and paracancerous tissues of 8 patients with cholangiocarcinoma in Fujian Provincial Hospital from October 2019 to May 2020 were selected for 850K methylation sequencing analysis to obtain differentially methylated genes. The 2018 genome-wide methylation data and clinical information of 36 patients with cholangiocarcinoma were download from The Cancer Genome Atlas (TCGA) database, the 2012 cholangiocarcinoma methylation data (GSE32879) were download from the Gene Expression Omnibus (GEO) database, and the 2018 TCGA database differential survival genomic data of overall survival (OS) and disease-free survival (DFS) of cholangiocarcinoma were download from the GEPIA2 database. The differentially methylated positions (DMP) and differentially methylated regions (DMR) results of 850K methylation sequencing analysis of submitted samples, methylated genes in TCGA and GEO databases, and cholangiocarcinoma survival genes of samples were jointly submitted for testing, multi-data set analysis was performed by the Sangerbox VENN tool, and common differentially methylated genes were obtained by intersection screening. The minimum P value method was used to determine the cut-off value of gene expression in Sangerbox, and the patients were divided into high and low expression groups of differentially methylated genes. The OS, DFS, disease-specific survival (DSS), disease-free interval (DFI) and progression-free interval (PFI) of cholangiocarcinoma patients were compared between the two groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed. Results:A total of 121 954 DMP were identified by 850K methylation sequencing of cholangiocarcinoma tissues and paracancerous tissues of 8 patients; a total of 1 399 differentially methylated genes were identified in DMR, and the common prognosis related genes glucosaminyl (N-acetyl) transferase 1 (GCNT1) and neurotrophic receptor tyrosine kinase 3 (NTRK3) were identified by intersection identification. The expression of GCNT1 in the cholangiocarcinoma tissues was higher than that in the paracancerous tissues, and the difference was statistically significant ( P = 0.040). The expression of NTRK3 in cholangiocarcinoma tissues was higher than that in the paracancerous tissues, but the difference was not statistically significant ( P = 0.790). The minimum P value method was used to predict the prognosis of patients with cholangiocarcinoma based on the combined expression of GCNT1 and NTRK3, and the order was based on the sum of the expression levels of the two genes. When 30% of the ranking was taken as the cut-off value, the difference in DFS between the high expression group and the low expression group in cholangiocarcinoma was the most significant ( P < 0.001); there was no significant difference in OS between the two groups ( P = 0.065). The results of GO functional analysis showed that GCNT1 was involved in protein glycosylation, macromolecule glycosylation, glycosylation, glycoprotein biosynthetic process, glycoprotein metabolic process, transferase activity and transferring glycosyl groups, protein O-linked glycosylation, O-glycan processing, etc., and NTRK3 was involved in neurotrophin signaling pathway, Ras signaling pathway, EGFR tyrosine kinase inhibitor resistance, ErbB signaling pathway, phospholipase D signaling pathway, central carbon metabolism in cancer, natural killer cell mediated cytotoxicity, etc. The results of KEGG analysis showed that GCNT1 was mainly associated with system functions such as mucin-type O-glycan biosynthesis and metabolic pathways, and NTRK3 was mainly associated with cell surface receptor pathways, intracellular signal transduction, positive regulation of stimulatory responses, transmembrane receptor protein tyrosine kinase signaling pathway, enzyme-linked receptor protein signaling pathway, MAPK signaling pathway cascade and regulation, protein phosphorylation signal transduction and other system functions. Conclusions:The expressions of differentially methylated genes GCTNT1 and NTRK3 in cholangiocarcinoma have certain predictive effects on the prognosis of patients with cholangiocarcinoma.

Objective:To evaluate the risk factors on prognosis after resection of huge hepatocellular carcinoma.Methods:The clinical and followup data of 146 patients undergoing radical resection at Fujian Province Hospital from Jan 2012 to Dec 2017 was analyzed retrospectively.Results:Independent risk factors for tumor recurrence were neutrophil/lymphocyte ratio ≥2.49, serum alpha-fetoprotein ≥400 ng/ml, non-anatomical hepatectomy, ruptured huge hepatocellular carcinoma, multiple tumor and microvascular invasion and macrovascular invasion. These seven factors were used to develop a risk prediction model, in which 1-year recurrence-free rates in patients with low, middle, high risk group were 68.5%, 23.5%, and 0, respectively, and 3-year recurrence-free rates were 34.2%,15.3% and 0, respectively. Independent risk factors for tumor overall survival were neutrophil/lymphocyte ratio≥2.49, serum alpha-fetoprotein ≥400 ng/ml, HBV-DNA ≥2 000 IU/ml, multiple tumor, microvascular invasion, macrovascular invasion and hepatic capsule invasion. These seven factors were used to develop a risk prediction model, in which 1-year survival rates in patients with low, middle, high risk group were 94.7%, 74% and 40%, respectively, and 3-year survival rates were 68.4%,30.1%, and 5.7%, respectively.Conclusions:The recurrence rate of patients with huge hepatocellular carcinoma is high. Independent risk factors affecting prognosis were high neutrophil/lymphocyte ratio, high AFP level, high HBV-DNA, non-anatomical hepatectomy,ruptured,multiple tumor, microvascular and macrovascular invasion.

Chinese mitten crab (Eriocheir sinensis) is an important aquaculture species in Crustacea. Functional analysis, although essential, has been hindered due to the lack of sufficient genomic or transcriptomic resources. In this study, transcriptome sequencing was conducted on 59 samples rep-resenting diverse developmental stages (fertilized eggs, zoea, megalopa, three sub-stages of larvae, juvenile crabs, and adult crabs) and different tissues (eyestalk, hepatopancreas, and muscle from juvenile crabs, and eyestalk, hepatopancreas, muscle, heart, stomach, gill, thoracic ganglia, intes-tine, ovary, and testis from adult crabs) of E. sinensis. A comprehensive reference transcriptome was assembled, including 19,023 protein-coding genes. Hierarchical clustering based on 128 differ-entially expressed cuticle-related genes revealed two distinct expression patterns during the early lar-val developmental stages, demonstrating the distinct roles of these genes in 'crab-like"cuticle formation during metamorphosis and cuticle calcification after molting. Phylogenetic analysis of 1406 one-to-one orthologous gene families identified from seven arthropod species andCaenorhabditis elegans strongly supported the hypothesis that Malacostraca and Branchiopoda do not form a monophyletic group. Furthermore, Branchiopoda is more phylogenetically closely related to Hexapoda, and the clade of Hexapoda and Branchiopoda and the clade of Malacostraca belong to the Pancrustacea. This study offers a high-quality transcriptome resource for E. sinensis and demonstrates the evolutionary relationships of major arthropod groups. The differentially expressed genes identified in this study facilitate further investigation of the cuticle-related gene expression networks which are likely associated with'crab-like"cuticle formation during metamor-phosis and cuticle calcification after molting.

Objective To investigate the technical and therapeutic effects of laparoscopic surgery for adult patients with Dong Type C bile duct dilatation.Methods A retrospective cohort study approach was used.The clinical data of 47 patients with Dong Type C adult biliary dilatation who underwent surgery in Fujian Provincial Hospital from January 2014 to December 2017 were collected.There were 12 males and 35 females.The age ranged from 16 to 68 years,with a median age of 30 years.According to different surgical methods,the patients were divided into the laparoscopic group (treated with laparoscopic cyst excision and Roux-en-Y hepaticojejunostomy,n =21) and the open group (treated with traditional open operation,n =26).The intraoperative blood loss,complication rate,operation time,postoperative intestinal function recovery time and postoperative hospital stay were observed.Measurement data with normal distribution were expressed as mean ± standard deviation (Mean ±SD) and analyzed by t test.Comparison of count data was done by the chi-square test.Results There was no perioperative deaths in either group.There was no significant difference in intraoperative blood loss and complication rate between the two groups (P ＞ 0.05).The operation time of the laparoscopic group was significantly longer than that of the open group[(333.7 ±61.1) min vs (235.9 ±64.3) min],with statistically significant difference between graps (P =0.000).The recovery time for the intestinal function of the laparoscopic group was significantly shorter than that of the open group [(2.2 ± 0.5) d vs (2.9 ± 0.6) d],with statistically significant difference between groups (P =0.000).The postoperative hospital stay in the laparoscopic group was significantly shorter than the open group [(7.1 ± 1.8) d vs (12.0 ± 5.9) d],with statistically significant difference between groups (P =0.001).Conclusion For adult biliary dilatation patiens with Dong Type C,laparoscopic surgery is safe and feasible with the advantages of mini-invasive and quick recovery.

Objective To explore the impact of early use of renal replacement therapy on the prognosis of severe acute pancreati-tis patients .Methods 60 cases of patients with severe acute pancreatitis were chosen between September 2015 and June 2016 in De-partment of General Surgery in our hospital ,and were randomly divided into treatment group (n=30) and control group (n=30) with the method of number table .The patients in the control group were treated with conventional therapy ,treatment group was given the early use of renal replacement therapy (within 12 h after onset) .The scores of Acute Physiology and Chronic Health E-valuation Ⅱ (APACHE Ⅱ) ,partial pressure of oxygen (PaO2 ) ,oxygenation index (PaO2/FiO2 ) ,lactate (Lac) ,C-reactive protein (CRP) ,interleukin 1 (IL-1 ) ,and interleukin-6 (IL-6) levels before treatment ,12 h ,24 h ,48 h and 72 h after treatment were com-pared between two groups ,and the incidence of abdominal compartment syndrome (ACS) was compared as well .Results The difference of the indicators before treatment between two groups was not significant different (P>0 .05);the scores of APACHEⅡ ,PaO2 ,PaO2/FiO2 and Lac levels at 12 h ,24 h ,48 h and 72 h after treatment in treatment group were all lower than those in con-trol group ,and the difference was statistically significant (P<0 .05);CRP ,IL-1 and IL-6 levels at 24 h ,48 h and 72 h after treat-ment in treatment group were lower than those in control group ,and the difference was statistically significant (P<0 .05) .In the control group ,5 cases had ACS ,the incidence rate was 16 .7% ,and there was no case having ACS in treatment group and the differ-ence was statistically significant (P<0 .05) .Conclusion The early application of renal replacement therapy can effectively improve the physiological indexes of patients with severe acute pancreatitis ,inhibit the inflammatory response rapidly ,reduce damage to the abdominal organs ,and is worthy of promotion and application in clinical practice .

Objective To investigate the diagnosis and treatment of mucin-producing intrahepatic biliary tumor (MPIBT).Methods We retrospectively analyzed the clinical,radiologic,surgical and pathologic findings of 16 MPIBT cases from January 2004 to December 2011.Results There were six men and ten women,age ranged from 44 to 69 years (mean 60 years).Clinical presentation included jaundice with abdominal dull pain in 5 patients,acute cholangitis in 4 patients,painless jaundice in 2 patients,upper abdominal dull pain in 3 patients,no obvious symptoms in 1 patient,body weight loss more than 5 kg within 3 months in 5 patients.The most characteristic appearance of MPIBT on magnetic resonance cholangiopancreatography were asymmetry of intrahepatic bile duct dilatation and the dilatation in both extraand intrahepatic bile duct distal to the hepatic mass and not sudden interruption in extrahepatic bile duct.The primary tumor located in the left hepatic bile duct in 15 cases,in the right hepatic bile duct in one.13 MPIBT cases received hemihepatectomy and extrahepatic bile duct resection and Roux-en-Y anastomosis was done in 8 cases,3 received palliative biliary drainage.Pathologically 13 was papillary adenocarcinoma and 3 was papillary adenoma.The 1-,2-,3-year survival rates for the 16 MPIBT patients were 81％,66％,56％,respectively.Conclusions MPIBT had no specific clinical manifestations,MRCP might be an effective means for the diagnositic strategy and assessment of tumor extension before surgery,radical resection was the first choice of treatment,palliative biliary drainage could prolong the survival time.

Objective To investigate kupffer cells (KCs) expressing indoleamine 2,3-dioxygenase(IDO)in the inhibition of allogeneic T-cell proliferation in vitro. Methods Real-time PCR was used to investigate the expression of IDO mRNA and FasL mRNA in KCs pretreated with or without IFNγ. High performance liquid chromatography was used to analyze the catabolism of tryptophan by IDO from KCs. Allogeneic T-cell response was used to confirm the inhibition of KCs in vitro. The proliferation of lymphocytes was detected using [3 H] thymidine incorporation. Cell cycle and lymphocyte apoptosis were evaluated by flow cytometric assay. Results Real-time PCR revealed IDO mRNA and FasL mRNA expression in KCs pretreated with IFN-γ. IDO catabolic effect was confirmed by a decrease in tryptophan and increase in kynurenine concentration. KCs expressing IDO and FasL from BABL/c mice acquire the ability to suppress the proliferation of T-cells from C57BL/6, which could be blocked by the addition of 1-methyl-tryptophan and anti-FasL antibody. The co-cultured T-cells with KCs expressing IDO and FasL could induce allogeneic T-cell apoptosis and exhibited cell-cycle arrest in G1. Conclusion In addition to the Fas/FasL pathway, IDO may also play an important role in KCs to inhibit allogeneic T-cell proliferation in vitro.

Objective To investigate the effect of postoperative anti-viral therapy using adefovir dipivoxil in recurrence of hepatocellular carcinoma HCC with HBV infection. Methods Sixty HCC patients with HBV infection were randomized into two groups:Group A (n=23) received hepatectomy only, and Group B (n=37) received hepatectomy and postoperative therapy using adefovir dipivoxil. The changes in liver func-tion, the suppression of HBV-DNA, HBeAg seroconversion rate, tumor recurrence rate and median survival in the two groups were observed and compared. Results In Group B, serum albumin, total bilirubin, AST and ALT were significantly improved compared with Group A (P < 0.05). Furthermore, the rate of 6-month and 1-year HBV-DNA suppression, the rate of 6-month and 1-year HBeAg seroconversion were significantly improved compared with Group A (P < 0.05). In Group A and Group B the tumor recurrence rate was 82. 6% (19/23) vs 78.4% (29/37) (P > 0.05), the recurrence time was 7.3 vs9.6 months (P > 0.05) and the median survival time was 17.4 vs 24.9 months (P < 0.01). Conclusion The results suggest that an-ti-viral therapy using adefovir dipivoxil postoperatively may improve the remnant liver function, suppress the HBV reaction, prolong the survival for patients with HCC with HBV infection.

OBJECTIVETo investigate the chemical constituents of Hypericum petiolulatum.

METHODThe chemical constituents were isolated and purified by column chromatography on silica gel, macroporous adsorbent resin, Sephadex LH-20, and preparative HPLC. Structures of the compounds were identified by NMR and MS spectroscopic methods.

RESULTNine lignans were obtained and their structures were elucidated as (-)-(2R, 3R)-1-O-feruloyl-8,8'-bisdihydrosiringenin (1), (-) -secoisolariciresinol 4-O-beta-D-glucopyrano-side (2), isolariciresinol-beta-4'-O-beta-D-glucopyranoside (3), 5-methoxy-9beta-xylopyra-nosyl-(+) -isolariciresinol (4), (+) -lyoniresinol 2alpha-O-beta-D-xylopyranoside (5), 5-methoxy-9-beta-xylopyranosyl-(-) -isolariciresinol (6), isolariciresinol 6a-O-beta-D-gluco-side (7), (+)-lyoniresinol 3alpha-O-beta-D-xylopyranoside (8) and 7-methoxy-5-benzofuranpropanol 4'-O-beta-D-glucopyranoside (9).

CONCLUSIONCoupound 1 was new and compounds 2-9 were obtained from the genus Hypericum for the first time.