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This paper compared the differences between two kinds of Bufonis Venenum produced by Bufo gargarizans gargarizans and B. gararizans andrewsi, and verified the rationality of the market value orientation of Bufonis Venenum based on the zebrafish mo-del. Twenty batches of Bufonis Venenum from Jiangsu province, Hebei province, Liaoning province, Jilin province, and Liangshan, Sichuan province, including B. gargarizans gargarizans and B. gararizans andrewsi, were collected. The UHPLC-LTQ-Orbitrap-MS combined with principal component analysis was used to compare the differences between two kinds of Bufonis Venenum. According to the limiting conditions of VIP>1, FC<0.5 or FC>2.0, and peak total area ratio>1%, 9 differential markers were determined, which were cinobufagin, cinobufotalin, arenobufagin, resibufogenin, scillaredin A, resibufagin, 3-(N-suberoylargininyl)-arenobufagin, 3-(N-suberoylargininyl)-marinobufagin, and 3-(N-suberoylargininyl)-resibufogenin. The content of 20 batches of Bufonis Venenum was determined according to the Chinese Pharmacopoeia(2020 edition) by high-performance liquid chromatography, and the 2 batches of Bufonis Venenum, CS7(8.99% of total content) and CS9(5.03% of total content), with the largest difference in the total content of the three quality control indexes of the Chinese Pharmacopoeia(bufalin, cinobufagin, and resibufogenin) were selected to evaluate their anti-liver tumor activity based on the zebrafish model. The tumor inhibition rates of the 2 batches were 38.06% and 45.29%, respectively, proving that only using the quality control indexes of the Chinese Pharmacopoeia as the value orientation of Bufonis Venenum market circulation was unreasonable. This research provides data support for the effective utilization of Bufonis Venenum resources and the establishment of a rational quality evaluation system of Bufonis Venenum.
Sujets)
Animaux , Danio zébré , Bufanolide/analyse , Bufonidae , Chromatographie en phase liquide à haute performance , Contrôle de qualité , Lignée cellulaire tumoraleRésumé
ObjectiveTo establish blood stasis models in zebrafish using three inducers and select the optimal model for evaluating the activity of Notoginseng Radix et Rhizoma in promoting blood circulation. MethodArachidonic acid (AA), ponatinib, and isoprenaline (ISO) were used to induce blood stasis models in zebrafish. A normal group, a model group, a positive drug group, and Notoginseng Radix et Rhizoma water extract freeze-dried powder groups at different concentrations were set up. The staining intensity of cardiac erythrocytes and the fluorescence intensity of cardiac apoptotic cells were calculated, the anti-thrombotic effect and anti-myocardial hypoxia activity of Notoginseng Radix et Rhizoma were evaluated. The activities of water extract and 70% methanol extract of Notoginseng Radix et Rhizoma were compared based on the preferred AA- and ISO-induced blood stasis models in zebrafish and the difference in the chemical composition was analyzed by UHPLC LTQ-Orbitrap MS/MS. ResultAfter induction by AA and ponatinib, the staining intensity of cardiac erythrocytes was reduced (P<0.01), and the fluorescence intensity of cardiac apoptotic cells increased after the induction by ISO (P<0.01). The freeze-dried powder of the water extract of Notoginseng Radix et Rhizoma could antagonize the thrombosis in the AA-induced model (P<0.01) and the myocardial apoptosis in the ISO-induced model (P<0.05), while no significant improvement in the thrombosis was observed in the ponatinib-induced model. The freeze-dried powder of 70% methanol extract of Notoginseng Radix et Rhizoma could inhibit myocardial apoptosis in the ISO-induced blood stasis model (P<0.01), and the effect was stronger than that of the freeze-dried powder of Notoginseng Radix et Rhizoma water extract. The difference in chemical composition lay in some saponins (such as ginsenoside Re), amino acids, and acetylenic alcohols. ConclusionAA, ponatinib, and ISO all can serve as inducers for the blood stasis model in zebrafish. AA- and ISO-induced models can be used to evaluate the activity of freeze-dried powder of Notoginseng Radix et Rhizoma water extract in promoting blood circulation. The chemical compositions of the freeze-dried powders of Notoginseng Radix et Rhizoma extracted with water and 70% methanol are quite different. For the ISO-induced blood stasis model, the freeze-dried powder of Notoginseng Radix et Rhizoma extracted with 70% methanol has a stronger ability against myocardial hypoxia. Saponins and acetylenic alcohols may be closely related to the effects of promoting blood circulation and resolving blood stasis.
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Objective To identify the potential prognostic biomarkers of the immune-related genes signature for patients with hepatocellular carcinoma (HCC). Methods Original HCC data were downloaded from TCGA, and the immune activity of each sample was calculated by ssGSEA. HCC samples were divided into high and low immune cell infiltration groups by "GSVA" package and "hclust" package. The ESTIMATE algorithm scored the tumor microenvironment in each HCC sample. The "limma" package and Venn diagram identified effective immune-related genes. Univariate Cox, Lasso regression and multivariate Cox regression analyses were used to explore key genes. The "rms" package was used to create nomograms and draw calibration curves. Results Compared with the high immune cell infiltration group, the tumor purity of the samples in the low immune cell infiltration group was higher, the immune score, ESTIMATE score and stromal score were lower. In the high immune cell infiltration group, the immune components were more abundant, and the expression levels of TIGIT, PD-L1, PD-1, LAG3, TIM-3, CTLA4 and HLA family were higher. Multivariate Cox regression analysis showed that four immune-related genes (S100A9, HMOX1, IL18RAP and FCER1G) were used to construct the prognosis model. Compared with other clinical features, the risk score of this prognostic model was recognized as an independent prognostic factor. Conclusion This study identified the immune-related core genes which may be used in targeted therapy and immunotherapy of HCC.
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OBJECTIVE Cerebral ischemia or ischemic stroke is due to insufficient blood supply to the brain, which causes hypoxia or ischemia in some areas. This work aimed to quantify the minerals and heavy metals in Qishiwei Zhenzhu pill in vivo and in vitro, analyze its effect on the types and abundance of intestinal flora, and study its mechanism on inflammation and apoptosis pathways as a treatment for cerebral ischemia. METHODS Microwave digestion and induc?tively coupled plasma mass spectrometry (ICP-MS) were used to determine the minerals and heavy metals in 10 batches of Qishiwei Zhenzhu pill in vitro. With the use of the middle cerebral artery obstruction (MCAO) model, ICP-MS was applied to determine the content of minerals and heavy metals in hepatic portal vein blood, abdominal aortic blood, brain, liver, kidney, hair, urine and feces at different time periods. On this model, the ileum, cecum, and colon tissues were tested for intestinal pathology, and 16S rRNA was used for sequencing. Species taxonomy, α diversity, and spe?cies microbial composition and structure analysis were also performed. Polymerase chain reaction and Western blotting were employed to determine the mRNA and protein expression of p38 MAPK, caspase-3, IL-1β and TNF-α in the isch?emic brain tissues of rats. RESULTS The average content of heavy metals in the 10 batches of Qishiwei Zhenzhu pill samples is in the descending order Hg>Cu>Pb. Significant differences in the metal elements are found among Qishiwei Zhenzhu pill from different manufacturers but not among the different batches of the same manufacturer. An extremely low content of heavy metals are absorbed into the blood or accumulated in the brain, liver, kidney, and other tissues. Stool is the main excretion route of minerals and heavy metals from Qishiwei Zhenzhu pill. This medicine helps repair the intestinal mucosa in MCAO rats. At the phylum level, it can regulate the abundance of Firmicutes and Proteobacteria in the intestinal flora of rats with cerebral ischemia. At the genus level, it can adjust the abundance of Escherichia Shigella. At the species level, it can adjust the abundance of Lactobacillus yoelii and Lactobacillus reuteri. Cluster classification results show that Qishiwei Zhenzhu pill can improve the intestinal flora of rats with cerebral ischemia, reduce the mRNA and protein expression of caspase-3 and IL-1βin rat brain tissues, and have a tendency to decrease the mRNA expres?sion of p38 MAPK and TNF-α. CONCLUSION Quantifying the minerals and heavy metals in Qishiwei Zhenzhu pill in vivo and in vitro will help improve their quality standards. Minerals and heavy metals are mainly excreted in feces, accumu?late in extremely low levels in various tissues, and do not damage the intestinal mucosa. The effective material basis of Qishiwei Zhenzhu pill in treating cerebral ischemia may be related to their Li, Cr, and Cd elements. These pills can improve the environment of intestinal flora, and their mechanism of treatment for cerebral ischemia may be related to the down-regulation of IL-1βinflammatory factor and inhibition of cell apoptosis.
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Objective:To investigate the application value of three-dimensional visualization technology in management of middle hepatic vein (MHV) processing in associating liver partition and portal vein ligation for staged hepatectomy(ALPPS).Methods:The retrospective and descriptive study was conducted. The clinical data of 40 patients with right massive liver cancer or multiple right liver lesions who underwent ALPPS in the First Affiliated Hospital of Guangxi Medical University from November 2017 to August 2019 were collected. There were 34 males and 6 females, aged (44±9)years, with a range from 26 to 64 years. All patients underwent multi-slice computed tomography (CT) plain and enhanced scan of superior abdominal region before operation, and the data were transmitted to the liver visualization analysis software IQQA system with 1.5 mm thin-layer images to complete the three-dimensional reconstruction of the liver and its blood vessels. Patients were performed ALPPS based on results of three-dimensional reconstruction and intraoperative findings. Observation indicators: (1) results of preoperative three-dimensional reconstruction; (2) surgical situations; (3) follow-up. Follow-up was conducted using outpatient examinations and telephone interview to detect postopeartive survival of patients up to March 2020. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were described as M (range). Count data were represented as absolute numbers. Results:(1) Results of preoperative three-dimensional reconstruction: 40 patients underwent three-dimensional reconstruction successfully, of which 37 clearly showed MHV, tumor location and relationship between them, 3 patients showed unclearly MHV and were classified based on two-dimensional images. Of the 40 patients, 12 had MHV classified as type A, 13 as type B, 9 as type C, and 6 as type D. Three-dimensional reconstruction of vessels showed 22 with umbilical veins and 9 with anterior veins. Of the 40 patients, 35 were predicted to preserve MHV, and 5 were predicted to resect MHV. Total estimated liver volume, tumor volume, and reserved liver volume were (1 012±119)cm 3, 600 cm 3(8-2 055 cm 3), (346±80)cm 3. The ratio of future liver remnant to standard liver volume was 34%±8%. (2) Surgical situations : 40 patients underwent the first-stage ALPPS, including 35 with preservation of MHV and 5 with resection of MHV, which was accorded with preoperative prediction. Thirty-four patients underwent the second-stage ALPPS, and 6 patients had failure to receive the second-stage ALPPS due to undificiency future liver remnant. The operation time and volume of intraoperative blood loss for 40 patients undergoing first-stage ALPPS were (350±79)minutes and 300 mL(range, 100-2 600 mL). Three patients received blood transfusion and no perioperative death occurred. There were 24 patients with grade A heptic insufficiency according to criteria of International StudyGroup of Liver Surgery (ISGLS) and 16 patients with grade B heptic insufficiency after the first-stage ALPPS. Twenty-eight patients had grade Ⅰ complications of Clavien-Dindo classification, including 17 with a small pleural effusion, 10 with a small pleural and abdominal effusion, 1 with hypoproteinemia; 8 patients had grade Ⅱ complications of Clavien-Dindo classification, including 5 with pneumonia, 1 with pneumonia combined with pleural and abdominal effusion, 1 with coagulation disorders, 1 with biliary fistula; 3 patients had grade Ⅲ complications of Clavien-Dindo classification, including 2 with pneumothorax and pneumonia, 1 with pneumothorax, pneumonia and coagulation disorders; 1 patient had grade Ⅳ complications of Clavien-Dindo classification as systemic inflammatory response syndrome. All patients with complications were improved after symptomatic treatment, anti infection, transfusion of fresh frozen plasma or drainage. For the 34 patients undergoing the second-stage ALPPS, the operation time and volume of intraoperative blood loss were (320±83)minutes and 500 mL(range, 200-6 000 mL). Twelve patients received blood transfusion. There were 12 patients with grade A heptic insufficiency according to criteria of ISGLS and 22 with grade B heptic insufficiency after the second-stage ALPPS. Eighteen patients had grade Ⅰ complications of Clavien-Dindo classification, including 11 with a small pleural effusion, 7 with a small pleural and abdominal effusion; 12 patients had grade Ⅱ complications of Clavien-Dindo classification, including 4 with pneumonia, 4 with coagulation disorders, 3 with massive abdominal effusion, 1 with biliary fistula; 3 patients had grade Ⅲ complications of Clavien-Dindo classification, including 1 with pneumothorax and pneumonia, 1 with massive pleural effusion, 1 with obstructive jaundice; 1 patient had grade Ⅳ complications of Clavien-Dindo classification as pneumonia and anemia. All patients with complications were improved after symptomatic treatment, anti infection, transfusion of fresh frozen plasma or drainage. (3) Follow-up: 40 patients were followed up for 2-35 months, with a median follow-up time of 17 months. The 6-month, 1-, and 2-year survival cases were 35, 26, 21 cases. Conclusion:Three-dimensional visualization technology can clearly show the MHV classification and its relationship with tumor location, which has an important guiding significance in the decision-making of MHV management in ALPPS.
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Objective:To study the clinical characteristics of patients with severe abdominal infection and the epidemiological characteristics of pathogenic bacteria in a hospital, to provide a basis for rational use of antibiotics and reduce the drug resistance rate of pathogens.Methods:A retrospective analysis was performed on 237 patients with abdominal disease as the primary disease admitted to the surgical intensive care unit (ICU) of the First Affiliated Hospital of Guangxi Medical University from January 1st, 2017 to December 31st, 2019. They were divided into two groups according to whether abdominal infection occurred or not. The clinical features of patients in both groups were analyzed, including gender, age, acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score, chronic underlying diseases, primary abdominal site, abdominal trauma or bleeding, multiple organ dysfunction syndrome (MODS) involving organs and surgical treatment. At the same time, the bacterial origin, bacterial distribution and antibiotics sensitivity test results of patients with abdominal infection were recorded.Results:Abdominal infection occurred in 141 of the 237 patients and did not occur in the remaining 96 patients. There were no statistically significant differences between the abdominal infection group and the non-abdominal infection group in terms of gender, age, chronic underlying diseases, etiology and trauma. The APACHE Ⅱ score in the abdominal infection group was obviously higher than that of the non-abdominal infection group (24.0±8.1 vs. 17.1±5.8, P < 0.01). Incidences of abdominal bleeding, MODS involving four or more organs, surgery and the times of surgery ≥ 3 in the abdominal infection group were significantly higher than those in the non-abdominal infection group (36.2% vs. 17.7%, 20.6% vs. 1.0%, 84.4% vs. 21.9%, 9.3% vs. 0%, all P < 0.05). Among the 141 patients with abdominal infection, 107 obtained positive microbial culture results, and a total of 133 pathogenic strains were detected, including 115 strains of bacteria (86.5%) and 18 strains of fungi (13.5%). The main source of bacteria was abdominal drainage (46.1% of non-bloody specimens and 13.9% of bloody specimens). Among the 115 bacteria, Gram-negative (G -) bacteria were the most common (72.2%) and Gram-positive (G +) bacteria accounted for 27.8%. Escherichia coli and Acinetobacter baumannii were the top two G - bacteria (40.9% and 13.9%, respectively), and enterococcus faecalis accounted for the largest proportion of G + bacteria (7.8%). The pathogenic bacteria of abdominal infection were sensitive to tigacycline. Conclusions:The patients with abdominal infection in our hospital had high APACHE Ⅱ score, more organs failure and were easily complicated with intraperitoneal hemorrhage and required surgical intervention and even repeated surgery. The pathogenic bacteria in patients with abdominal infection in ICU were mainly G - bacteria, and the rate of multi-drug resistance of Acinetobacter baumannii was high. Empirical anti-infective treatment should be started as soon as possible according to the microbial spectrum of the region until the pathogenic bacteria results are obtained. Broad-spectrum antimicrobial therapy and combined antimicrobial therapy are recommended for the healthcare acquired abdominal infection in hospital.
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Objective:To compare the effect and mechanisms of modified Erchentang and Xuefu Zhuyutang on high-fat diet-induced apolipoprotein-E knockout (ApoE-/-) mice nonalcoholic fatty liver disease (NAFLD). Method:Ten C57/BL6J mice were taken as normal control group and fed with normal feed. Totally 30 ApoE-/- mice were fed with high-fat diet to establish a disease model for 4 weeks. After 4 weeks, the 30 ApoE-/- mice were divided into model group, Xuefu Zhuyutang group (hereinafter referred to as Huoxue group) and modified Erchentang group (hereinafter referred to as Huatan group) by random number table method, with 10 in each group. The normal group and the model group were intragastrically administered with normal saline. The drug-administered group was intragastrically administered at a dosage that was ten times of the adult dose, once a day, for 8 weeks. Serum and liver were collected after the end of the 12-week experiment. The serum lipid and liver function levels of each group were measured, and the liver pathological morphology was observed. Protein and mRNA expressions of liver inflammatory mediators interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), matrix metalloproteinase-9 (MMP-9), monocyte chemotactic factor-1 (MCP-1) were detected by Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot. Result:The results of serum lipids and liver function showed that compared with the normal group, serum total cholesterol (TC), triglyceride (TG), low-density lipoprotein (LDL), aspartate aminotransferase (AST), and alanine aminotransferase (ALT) in the model group were significantly increased, while serum high-density lipoprotein (HDL) was significantly reduced (P<0.01). Compared with the model group, serum TG ,LDL and ALT were significantly decreased, HDL was significantly increased in the Huoxue group (P<0.05). The serum levels of TC, TG, LDL, AST and ALT in the Huatan group were significantly decreased,HDL was significantly increased (P<0.05,P<0.01), and TG was decreased. The mice serum HDL in the Huatan group was higher than that in the Huoxue group. The serum ALT in the Huoxue group was lower than that in the Huatan group. The pathological observation showed that compared with the normal group, hepatocytes in the model group had severe steatosis with many lipid droplet vacuoles, suggesting that the mouse NAFLD model was successful. Compared with the model group, each administration group alleviated hepatocyte steatosis, with no significant difference between the two administration groups. Western blot and Real-time PCR results showed that compared with the normal group, protein and mRNA expression levels of TNF-α, IL-1β, MCP-1, and MMP-9 in the model group were significantly increased (P<0.05,P<0.01). Compared with the model group, the Huoxue group significantly down-regulated the expressions of IL-1β, MCP-1 protein and MCP-1 mRNA(P<0.05,P<0.01). The Huatan group significantly reduced the expressions of IL-1β, TNF-α, MMP-9, MCP-1 protein, TNF-α and MMP-9 mRNA(P<0.05,P<0.01). Conclusion:Modified Erchentang and Xuefu Zhuyutang can alleviate the therapeutic effect of NAFLD mice to a certain extent, modified Erchentang has a better therapeutic effect.
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Objective:To explore the effect of Ershiwuwei Zhenzhu tablets (EZT) on microvascular function. Method:The zebrafish models of thrombosis, microvascular defect and vascular endothelial injury were induced by using arachidonic acid, simvastatin and ponatinib respectively, and treated with EZT, astragaloside or asprin. To evaluate the protective effect of EZT on vascular endothelium and its effect on thrombus formation, zebrafish heart output and blood flow velocity were counted, and the vascular area of the zebrafish intestine and the intervascular diameter were calculated. The thrombus in the tail vein was observed under microscope. Result:Compared with model group,EZT improved the cardiac output (PP-1, and promoted angiogenesis in zebrafish at concentrations of 0.11, 0.33, 1 mg·L-1. Compared with the model group, the vascular diameter of the zebrafish internode was significantly increased at the concentrations of 33 mg·L-1 (P-1 (P-1(PConclusion:EZT could improve microvascular dysfunction, and its mechanism may be related to the reduction of vascular endothelial damage to promote its angiogenesis and the improvement of microvascular hemodynamics to reduce thrombus formation.
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Objective To investigate the clinical efficacy of hepatic artery ringed and restriction operation-associating liver partition and portal vein ligation for staged hepatectomy (HARO-ALPPS) in the treatment of giant hepatocellular carcinoma.Methods The retrospective and descriptive study was conducted.Clinical data of a 45-year-old male patient with giant hepatocellular carcinoma who was admitted to the First Affiliated Hospital of Guangxi Medical University in July 2018 were collected.In the first stage operation,right portal vein ligation+ in situ liver partition + right hepatic artery ringed operation was performed on the patient.In the second stage operation,right hemihepatectomy was performed on the patient.Observation indicators:(1) surgical and postoperative situations of the first stage operation;(2) surgical and postoperative situations of the second stage operation;(3) postoperative pathological examination;(4) changes in future liver remnant (FLR) and tumor volume;(5) perioperative hemodynamic changes of right hepatic artery,proper hepatic artery and left hepatic artery;(6) perioperative hemodynamic changes of left-portal vein and main portal vein;(7) follow-up.Follow-up using outpatient examination was performed to detect the prognosis of patients up to February 2019.Count data were described as absolute number.Results (1) Surgical and postoperative situations of the first stage operation:the patient underwent right portal vein ligation+ in situ liver partition+ right hepatic artery ringed operation successfully.The operation time and volume of intraoperative blood loss were 376 minutes and 400 mL.Inflammatory indicators including body temperature,white blood cells (WBC),C-reactive protein,procalcitonin,and liver function indices including total bilirubin (TBil),albumin (Alb),alanine aminotransferase (ALT),aspartate aminotransferase (AST),ascites,indocyanine green retention rate at15 min (ICG R15),score of model for end-stage liver disease (MELD) before the second stage operation were improved after symptomatic supportive treatment.Prothrombin time (PT) was in the normal range after the first stage operation.There was no complication of Clavien-Dindo classification Ⅱ or above.(2) Surgical and postoperative situations of the second stage operation:the patient underwent right hemihepatectomy successfully.The operation time and volume of intraoperative blood loss were 322 minutes and 900 mL.The patient received 300 mL of fresh frozen plasma infusion.Inflammatory indicators including body temperature,C-reactive protein,and liver function indices including Alb,ALT,AST,ascites,were recoved to normal level after symptomatic supportive treatment.WBC,procalcitonin,TBil,and PT were in the normal range.There was no complication of Clavien-Dindo classification Ⅱ or above.(3) Postoperative pathological examination:① Ⅱ stage hepatocellular carcinoma was confirmed,mass-like type,with tissue necrosis and microvascluar invasion.There was no distal metastasis and tumor did not invade liver capsule or surgical margin.Ishak score of surrounding tissues was 3 in the inflammation and 2 in the fibrosis.② Chronic inflammation was detected in the gallbladder mucosa.③ Reactive hyperplasia was found in the 2 lymph nodes of the group 8.④ One in the group 12 lymph nodes showed reactive hyperplasia.Immunohistochemistry showed positive Glypican-3,Hepatocyte,Arginase-1,NM23,weakly positive vascular endothelial growth factor,and negative Ki-67,vascular endothelial cell marker CD34,biliary epithelial marker CK19 and CK 7,tumor suppressor gene P21 and P23.(4) Changes in FLR and tumor volume:FLR was 565 mL and 580 mL at the 5th and 14th day after the first stage operation respectively,accounting for 54% and 57% of total liver volume.The FLR to body weight ratio was 0.96 and 0.98,and the growth rate of FLR was 194% and 202%.Tumor volume was 1 210 mL and 1 297 mL at the 5th and 14th day after the first stage operation respectively.Tumor necrosis volume was 635 mL and 500 mL at the 5th and 14th day after the first stage operation respectively.At the 5th and 14th day after the first stage operation,the liver CT examination showed that HARO was successfully underwent and the blood supply of remnant liver was good.Preoperative CT aothgraphy (CTA) examination showed that the right hepatic tumor had rich arterial blood supply.At the 5th day after the first stage operation,the CTA examination confirmed the ringed site of right hepatic artery,and the blood supply of the tumor gradually decreased.At the 14th day after the first stage operation,CTA examination showed significant decrease in the blood supply of liver tumors.Liver CT examination showed rich supply of the remnant liver and the liver volume of 829 mL at the 7th day after the second stage operation.(5) Perioperative hemodynamic changes of the right hepatic artery,proper hepatic artery and left hepatic artery.① Blood flow of right hepatic artery was 224.3,574.7,827.5,222.7,153.0,282.5,279.1,247.9 and 150.2 mL/min before the first stage operation,before right portal vein ligation,after right portal vein ligation and before right hepatic artery ringed and restriction,after right portal vein ligation and right hepatic artery ringed and restriction,at the 1st,3th,5th and 7th day after the first stage operation.Blood flow of right hepatic artery in the second stage operation was 505.0 mL/min.② Blood flow of proper hepatic artery was 399.7,793.5,830.5,1 075.4,784.7.5,821.2,722.8,467.4 and 555.4 mL/min before the first stage operation,before right portal vein ligation,after right portal vein ligation and before right hepatic artery ringed and restriction,after right portal vein ligation and right hepatic artery ringed and restriction,at the 1st,3th,5th and 7th day after the first stage operation.Blood flow of proper hepatic artery was 505.0,473.3,158.5,627.0,103.8 and 139.8 mL/min before right hepatectomy in the second stage operation,after right hepatectomy,at the 1st,3th and 5th day after the second operation,respectively.③ Blood flow of left hepatic artery was 147.5,13.8,19.4,16.2,62.1,93.9,67.1,30.8 and 106.1 mL/min before the first stage operation,before right portal vein ligation,after right portal vein ligation and before right hepatic artery ringed and restriction,after right portal vein ligation and right hepatic artery ringed and restriction,at the 1st,3th,5th,7th and 10th day after the first stage operation.Blood flow ot left hepatic artery was 52.0,43.2,112.4,103.6,80.7 and 56.1 mL/min before right hepatectomy in the second stage operation,after right hepatectomy,at the 1st,3th and 5th day after the second operation,respectively.(6) Perioperative hemodynamic changes of left-portal vein and main portal vein.① Blood flow of left portal vein was 552.6,181.2,412.2,320.0,1 777.7,1 284.7,749.5 and 484.2 mL/min before the first stage operation,before right portal vein ligation,after right portal vein ligation and right hepatic artery ringed and restriction,at the 1st,3th,5th,7th and 10th day after the first stage operation,respectively.Blood flow of left portal vein was 793.3,979.0,485.2,1 042.5,803.5 and 548.3 mL/min before right hepatectomy in the second stage operation,after right hepatectomy,at the 1st,3th,5th and 7th day after the second operation respectively.② Blood flow of main portal vein was 1 186.0,696.7,833.7,431.7,1 319.1,668.4,890.7,550.8 mL/min before the first stage operation,before right portal vein ligation,after right portal vein ligation and right hepatic artery ringed and restriction,at the 1st,3th,5th,7th and 10th day after the first stage operation,respectively.Blood flow of main portal vein was 846.4,937.4,891.2,1 671.0,2 697.8,and 1 230.0 mL/min before right hepatotectomy in the second stage operation,after right hepatectomy,at the 1st,3th,5th and 7th day after the second stage operation,respectively.(7) Follow up:the patient was followed up for 6 months and survived well,with Child A of liver function and normal alpha fetoprotein level.Liver contrast CT examination showed increase in the remnant liver,good blood supply,and no tumor recurrence.The FLR was 727 mL at the 2 months after operation.Conclusion For patients with giant hepatocellular carcinoma,HARO-ALPPS can be performed to decrease blood supply of tumor,increase tumor necrosis area,and reduce the incidence of intrahepatic arteriovenous fistula,which ensure blood supply of remnant liver hyperplasia.
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OBJECTIVE Based on the methods of microdialysis,HPLC-MS/MS and gene chip tech-nology,the mechanism of Baicalin and Geniposide(BC/GP)against excitatory amino acid toxicity in ce-rebral ischemia was studied. This will provide guidance for the clinical application of BC/GP and the study of excitatory amino acid toxicity in cerebral ischemia.METHODS (1)Microdialysis technique and HPLC-MS/MS was performed to study the pharmacodynamics of BC/GP against cerebral ischemia. ①18 SD rats with body weight of(280±20)g were randomly divided into control group,treatment groups with BC/CP at low dose,medium dose and high dose(equal to the dosage of crude drugs for 30 mg·kg-1, 45 mg·kg-1and 60 mg·kg-1respectively).Rats in each group were given intragastric administration for seven days to establish cerebral ischemia model. Then, microdialysis probe was applied to collect cerebrospinal fluid from hippocampus before and after cerebral ischemia. ② First, we established the HPLC-MS/MS method for measuring drugs and excitatory amino acids.Then we detected the microdi-alysis samples and observed their changes in animals.(2)The mechanism of BC/GP against excitatory toxicity of cerebral ischemia were observed at gene level by chip technique. ① 16 SD rats with body weight of 240±20 g were randomly divided into sham group, model group, treatment group of BC(60 mg·kg-1),treatment group of GP(60 mg·kg-1)and treatment group of BC/GP(7:3)(60 mg·kg-1).Rats in eachgroup were given intragastric administration for seven days to establish cerebral ischemia model. Then the rats were sacrificed,and the hippocampus were rapidly harvested and stored at-80℃for further detection. ②After the quality inspection of the hippocampal,the qualified samples were subjected to detect the levels of neurotransmitter receptor gene in the ischemic of rats by gene chip technology.Finally,the results were analyzed by the method of Δ ΔCt.RESULTS (1)Only three compounds includ-ed GP,glutamic acid and aspartic acid were detected in microdialysis samples by HPLC-MS/MS.The concentration of GP increased and lasted for 120 min with a significant dose-dependent after cerebral ischemia.Compared with low dose group,the AUC(0-t),MRT(0-∞),Cmaxand t1/2zin high-dose group showed significant difference(P<0.01).Compared with the model group,the levels of glutamic acid and aspartic acid in the treatment groups decreased significantly,especially in the middle and high dose groups.(2) 89 genes in the neurotransmitter receptor gene signaling pathway were detected by gene chip technol-ogy. There were 22 genes with |Fold Regulation|>1.5 in the model group, compared with the sham group.Five of the 22 genes showed statistically significant differences,including Grin2c(2.9026),Chrna7 (-1.5877), and Tacr2 (-1.7695). Htr3a (-1.8172) and Grm6 (-2.3527). There were 5 genes with |Fold Regulation|>1.5 in the BC group, compared with the model group, Two of them exhibited statistically significant differences,including Brs3(1.797)and Grin2c(-1.7979).There were 14 genes with|Fold Reg-ulation|>1.5 in the GP group, compared with the model group. Three of them displayed statistically significant differences,including Hcrtr2 (-1.6584), Sctr (-3.8524) and Grin2c (-4.8408). Compared with model group, the genes of |Fold Regulation|>1.5 in BC/GP (7:3) group are 5, and only one of them showed a significant differences. CONCLUSION (1)After administration of BC and GP,GP can cross the blood-brain barrier and reduce the release of excitatory amino acids in the hippocampus. (2) BC/GP can inhibit the interaction between excitatory amino acids and excitatory amino acid receptors and attenuate the toxicity of excitatory amino acids by down-regulating the expression of glutamic acid receptor Grin2c gene.(3)BC/GP may exert their brain protection effect by reducing the release of excit-atory amino acids and inhibiting the expression of excitatory amino acid receptors.
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Objective:To establish an HPLC method to determine the contents of ephedrine hydrochloride, (R, S)-goitrin, lae-trile,chlorogenic acid,licorice glycosides and glycyrrhizic acid in Xiao'er Kechuanling granule.Methods:The chromatography condi-tions were as follows:an Agilent Eclipse XDB-C18(250 mm×4.6 mm,5 μm) column was used,and acetonitrile-0.1% phosphoric acid was applied as the mobile phase. The flow rate was 1.0 ml·min-1with gradient elution. The wavelength was 207 nm for ephed-rine hydrochloride and laetrile,237 nm for glycyrrhizic acid,licorice glycosides and chlorogenic acid and 245 nm for (R,S)-goitrin. Results:The linear range of ephedrine hydrochloride was 2.423-96.920 μg·ml-1(r =0.999 2), and the average recovery was 100.1% (RSD=0.30%,n=6). The linear range of (R, S) -goitrin was 1.920-76.798 μg·ml-1(r=0.999 9), and the average recovery was 99.86% (RSD=1.14%,n=6). The linear range of chlorogenic acid was 2.396-92.891 μg·ml-1(r=0.999 9),and the average recovery was 98.57% (RSD =0. 75%,n =6). The linear range of amygdalin was 1.982-79.279 μg·ml-1(r =0.999 8),and the average recovery was 99.67% (RSD=0.59%,n=6). The linear range of glycyrrhizin was 2.136-85.440 μg· ml-1(r=0.999 9),and the average recovery was 98.57% (RSD=0.69%,n=6). The linear range of glycyrrhizic acid was 2.432-97.260μg·ml-1(r=0.999 9),and the average recovery was 98.57% (RSD=0.11%,n=6). Conclusion: The method is accu-rate and reliable,which can be used for the quality control of Xiao'er Kechuanling granule.
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Objective To investigate the correlation between interleukin-6 (IL-6) and future liver remnant (FLR) growth after associating liver partition and portal vein ligation for staged hepatectomy (ALPPS).Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 15 patients who underwent ALPPS at the First Affiliated Hospital of Guangxi Medical University between March 2017 and May 2018 were collected.Observation indicators:(1) intraoperative situations in the first staged ALPPS and the second staged ALPPS;(2) postoperative situations:① postoperative complications and duration of hospital stay,② results of pathological examination;(3) IL-6 concentration in the peripheral blood before and after operation;(4)follow-up situations.Follow-up using outpatient examination,telephone interview and internet was performed to detect life quality and survival of patients.Imaging examination was done to detect tumor recurrence and metastasis.Follow-up was done up to May 2018.Measurement data with normal distribution or similar normal distribution were represented as (x)±s.Measurement data with skewed distribution were described as M (range).Repeated measures data were analyzed by the repeated measures ANOVA.Correlation comparison was done using Pearson bivariate correlation test.Results (1) Intraoperative situations.① The first staged ALPPS:15 patients had liver parenchymal transection via anterior approach combined with selective hepatic vascular exclusion,without allogenic blood transfusion.The operation time,volume of intraoperative blood loss,FLR at postoperative 16 days,interval time to the second staged ALPPS,growth rate of liver volume,ratio of FLR and standard liver volume (SLV) were respectively 324 minutes (range,240-387 minutes),356 mL (range,200-600 mL),(582± 134) cm3,24 days (range,9-34 days),35%±20% and 53%±7%.② The second staged ALPPS:of 15 patients,13 underwent the second staged ALPPS successfully including 11 undergoing middle hepatic vein preserved right hepatectomy and 2 undergoing expanded right hemihepatectomy or right trisegmentectomy,1 underwent transcatheter arterial chemoembolization (TACE) due to FLR/SLV =31%,1 was detected yellow-white nodules at left lobe and confirmed as hepatocellular carcinoma by frozen section pathological examination,and then improved and discharged after 5-FU abdominal local chemotherapy combined with postoperative TACE.The operation time,volume of intraoperative blood loss of 13 patients undergoing the second staged ALPPS were 324 minutes (range,140-515 minutes) and 639 mL(range,100-1 400 mL).Two patients had blood transfusion including 1 with 800 mL of fresh frozen plasma and 4.0 U of red cells and 1 with 600 mL of plasma and 9.5 U of de-leucocytes and red cells.(2) Postoperative situations.① Postoperative complications and duration of hospital stay:15 patients had no perioperative death,9 and 6 were detected grade A and grade B liver failure respectively,15 had grade Ⅰ complications of Clavien-Dindo classification and no patient had grade Ⅱ and above complications,10 had pleural effusion including 1 with volume of effusion >500 mL.Of 13 patients undergoing the second staged ALPPS,4 and 9 were detected grade A and grade B liver failure respectively,8 and 5 had grade Ⅰ and Ⅱ complications of Clavien-Dindo classification and no patient had grade Ⅲ and above complications,11 had few pleural effusion with volume of effusion <500 mL.Patients with grade B liver failure and grade Ⅱ complications were recovered and discharged after treatments of liver protection,gastric protection,reinforced dressing change,continuous use of Alb,fresh frozen plasma transfusion.The patient with volume of pleural effusion > 500 mL was improved after closed thoracic drainage and other patients with pleural effusion were improved after symptomatic and supportive treatment.Duration of total hospital stay was 31 days (range,22-49 days) in 15 patients.② Results of pathological examination:13 patients undergoing complete ALPPS were diagnosed as hepaticocellular carcinoma with R0 resection and without cancer cells involving surgical margin,including 7 with grade Ⅱ portal vein tumor thrombus.Ishak score for postoperative pathological fibrosis and liver cirrhosis was 7.7±1.4 in 15 patients,including 1 case of 5,1 case of 6,2 case of 7,6 case of 8,5 case of 9.(3) IL-6 concentration in the peripheral blood before and after operation:IL-6 concentration in the peripheral blood before surgery was (8±3)ng/L in 15 patients,and (207±150)ng/L,(104±65)ng/L,(45±38)ng/L,(26±9)ng/L,(18±10)ng/L at 1,3,5,7,10 days after the first staged ALPPS,showing a statistically significant difference in changing trend before and after surgery (F=25.877,P<0.05) and statistically significant differences in paired comparison between 1,3,5,7,10 days after the first staged ALPPS and before surgery respectively (P<0.05).There was correlation between IL-6 concentration in the peripheral blood at 1,3 days after the first staged ALPPS and growth of FLR (r=0.766,0.881,P<0.05),and also between IL-6 concentration in the peripheral blood at 1,3 days after the first staged ALPPS and growth rate of FLR (r =0.810,0.879,P< 0.05).(4) Follow-up:15 patients were followed up for 1-14 months with a median time of 7 months.Of the 15 patients,1 without the second staged ALPPS died of multiple organ dysfunction syndrome at 7 months after the first staged ALPPS,14 survived and took care of theirselves in daily life during follow-up with improved life quality,including 1 detected multiple lung metastases at 12 months after complete ALPPS with mild increased AFP and 13 undetected new lesions in the remnant liver on contrast-enhanced CT and liver contrast-enhanced ultrasonography with normal AFP.Conclusion The peak of IL-6 concentration in peripheral blood at 1,3 days after the first staged ALPPS is significantly correlated with the hyperplasia of FLR,which may be used to predict the hyperplasia of FLR.
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This study aimed to assess the relationship of OAS2 rs739901 5'-flanking C/A polymorphisms with the susceptibility to Enterovirus-71 (EV71) infection.We investigated 294 hand-foot-mouth disease (HFMD) Chinese children with EV71 infection (165 mild cases and 129 encephalitis cases).The improved multiplex ligation detection reaction (iMLDR) technique was used to test the genotypes.In EV71-infected patients,the CA genotype distribution (P=0.007),A allele frequency (OR 1.32,95% CI 1.0-1.7,P=0.034)and CA+AA carriage frequency (P=0.003) of OAS2 rs739901 5'-flanking were obviously elevated as compared with controls,but there were no statistically significant differences between mild cases and encephalitis cases.In EV71-infected patients,the counts of white blood cells (P=0.034) and blood glucose concentrations (P=0.042) were raised in A carriers (CA+AA).Among different genotypes of encephalitis cases,the contents of cerebrospinal fluid (CSF) showed no significant differences.IFN-γ levels in EV71-infected patients were higher than those in controls (mild group vs.control group,P<0.01;encephalitis group vs.control group,P<0.001).In encephalitis cases,IFN-γ levels were reduced (P<0.05) in A carriers compared to CC genotype,however,there were no significant differences between genotypes CA and AA (P=0.226).These findings suggest that OAS2 rs739901 5'-flanking C/A genetic polymorphisms involve the susceptibility to EV71 infection,and A allele might be a risk factor of the susceptibility to EV-71 infection.
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This study aimed to assess the relationship of OAS2 rs739901 5'-flanking C/A polymorphisms with the susceptibility to Enterovirus-71 (EV71) infection.We investigated 294 hand-foot-mouth disease (HFMD) Chinese children with EV71 infection (165 mild cases and 129 encephalitis cases).The improved multiplex ligation detection reaction (iMLDR) technique was used to test the genotypes.In EV71-infected patients,the CA genotype distribution (P=0.007),A allele frequency (OR 1.32,95% CI 1.0-1.7,P=0.034)and CA+AA carriage frequency (P=0.003) of OAS2 rs739901 5'-flanking were obviously elevated as compared with controls,but there were no statistically significant differences between mild cases and encephalitis cases.In EV71-infected patients,the counts of white blood cells (P=0.034) and blood glucose concentrations (P=0.042) were raised in A carriers (CA+AA).Among different genotypes of encephalitis cases,the contents of cerebrospinal fluid (CSF) showed no significant differences.IFN-γ levels in EV71-infected patients were higher than those in controls (mild group vs.control group,P<0.01;encephalitis group vs.control group,P<0.001).In encephalitis cases,IFN-γ levels were reduced (P<0.05) in A carriers compared to CC genotype,however,there were no significant differences between genotypes CA and AA (P=0.226).These findings suggest that OAS2 rs739901 5'-flanking C/A genetic polymorphisms involve the susceptibility to EV71 infection,and A allele might be a risk factor of the susceptibility to EV-71 infection.
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Objective To evaluate the use of intercostal trocars (ICS) and transthoracic trocars in laparoscopic resection of liver segments 7 and 8.Method From November 2015 to June 2017,20 patients who underwent laparoscopic S7 or 8 segmentectomy for liver tumors in the Department of Hepatobiliary Surgery,the First Affiliated Hospital,Guangxi Medical University were analyzed retrospectively.Results Ports were inserted at the 8th or 9th ICS,respectively,in addition to the conventional abdominal ports.The mean operation time was 225.0 min (110.0 ~ 486.0 min).Anatomical resection was completed in 1 patient,and non-anatomical resection in 19 patients.The conversion rate was 0%.Pringle's maneuver was used in 9 patients.The mean blood loss was 85.0 (25.0 ~410.0) ml,and the mean length of hospital stay was 7.0 (5.0 ~ 12.0) days.The complication rate was 10.0%.Pathologic findings revealed that 17,2,1 patient(s) had HCC,hemangioma,and inflammatory nodule,respectively.The mean tumor size and tumor free margin were 33.8 (15.0 ~ 74.0) mm;and 15.0 (1.0 ~ 30.0) mm,respectively.There was no HCC recurrence on follow-up,expect for one patient who developed tumor recurrence at 20 months after laparoscopic liver resection.Conclusions In selected patients,laparoscopic liver resection using intercostal trocars was useful and safe for tumors located in liver segments 7 and 8.The long-term oncologic outcomes need to be further evaluated.
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AIM To study the effects of baicalin and gardenoside pairing on 5-lox/CysLTs/CysLT pathway of rats with cerebral ischemia injury.METHODS SD rats randomly assigned into control group,model group,baicalin and gardenoside pairing groups (7 ∶ 3,dosed at 30,45,60 mg/kg,respectively) in their cerebral ischemia recovery period were simulated with permanent middle cerebral artery occlusion (pMCAO) if necessary.After a week of natural recovery,the drugs' pharmacodynamic effects were evaluated by the neurofunctional scoring and HE staining,their impact on the content of CysLTs was determined by ELISA,and the influence on the expression of 5-lox,CysLT1 and CysLT2 was detected by western blot.RESULTS One-week consecutive administration of baicalin and gardenoside pairing contributed to a reduction in infiltration and tissue edema of ischemic cells.If compared with pMCAO group,the baicalin and gardenoside pairing groups were observed with significantly lowered scores of neurologic functioning,inhibited microglia activation,decreased content of CysLTs and down-regulated expression of 5-LOX,CysLT1 and CysLT2 (P < 0.01).CONCLUSION Baicalin and gardenia pairing's alleviating effect to the damage of cerebral ischemia may be associated with its inhibition on microglia activation and 5-lox/CysLTs/CysLT signaling pathway,and thus an inflammatory impairment reduction.
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Objectives: To investigate the genetic mutation in a Chinese family with Pulmonary Arterial Hypertension (PAH). Methods: Whole exome sequencing was performed in two patients and two healthy family members in the PAH pedigree. Patient-specific variations were screened by bioinformatics methods and compared between groups. To further identify the association between these variations and PAH, Sanger sequencing was used to analyze the genotype of PAH patients and 100 healthy controls. Results: Two affected persons were found among the eight family members. The patients was presented as dyspnea after exercise, and right-heart catheterization was performed to measure the mean pulmonary arterial pressure (mPAP, 77 mmHg), cardiac output (CO, 4.92 L/min), and pulmonary vascular resistance (PVR, 13.4 Wood units). The hereditary characteristic in this family presented in mother and child, suggesting an autosomal dominant patter. Exome sequencing, mutation detection and sanger variants validation revealed a novel heterozygous frameshift mutation (c.747_748 insCCTTTGATGGAACATGA:p.V250fs) in the BMPR2 gene. Meanwhile, this heterozygous insertion mutation was absent in 100 ethnically matched control samples screened by direct sanger sequencing. Conclusions: Our study revealed a novel heterozygous frameshift mutation in the BMPR2 gene, expanding the BMPR2 mutation spectrums.
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Objective To determine the susceptibility genes and resistance genes in HLA-DRB1 alleles in Tibetan patients with cystic and alveolar hydatid diseases,so as to provide the references for the research of the genetic characteristics and infec-tion mechanism of Tibetan hydatid diseases. Methods The case control method was applied. The Tibetan patients with cystic and alveolar hydatid diseases(63 and 73 cases respectively)in Yushu and Guoluo Tibetan Autonomous Prefecture,and unrelat-ed healthy people(60 cases)in this area were selected as the study subjects. The polymerase chain reaction-sequence based typ-ing(PCR-SBT)technique was applied for genotyping of HLA-DRB1,and the comparison of the gene frequency. Results The frequency of HLA-DRB1*04 in the alveolar/cystic echinococcosis group was lower than that in the control group(χ2 =4.71, 4.31,both P<0.05). Conclusion HLA-DRB1*04 genotypes may be associated with the resistance of cystic and alveolar echi-nococcosis and its resistance genes.
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Objective To explore key gene and pathway of radioresistance in esophageal carcinoma and reveal its molecular mechanism of radioresistance. Methods The gene expression profiles of GSE61772 and GSE61620 were downloaded from the Gene Expression Omnibus (GEO). Differentially expressed genes ( DEGs ) were screened by GEO2R. Gene Ontology ( GO ) enrichment, Kyoto Encyclopedia of Genes and Genomes ( Kegg ) enrichment and protein-protein interaction ( PPI ) network construction were performed by DAVID and String softwares. RT-PCR was used to detect the differences in the expression of different genes in different radiosensitivity cells. Results A total of 49 differentially expression genes were screened. These genes were mainly involved in the regulation of multicellular biosynthesis, ion transport, DNA synthesis, metabolism, cell proliferation and so on. The major biological pathways included a Wnt signal pathway. 12 DEGs interacted with each other, and CHN2 may be a key node. The expression of CHN2 gene had no obvious difference between TE13R and TE13. Conclusions 49 differentially expressed genes, including CHN2, may be involved in radioresistance of esophageal carcinoma, and the Wnt signaling pathway may be an important pathway in this radioresistance.
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Objective To explore key gene and pathway of radioresistance in esophageal carcinoma and reveal its molecular mechanism of radioresistance. Methods The gene expression profiles of GSE61772 and GSE61620 were downloaded from the Gene Expression Omnibus (GEO). Differentially expressed genes ( DEGs ) were screened by GEO2R. Gene Ontology ( GO ) enrichment, Kyoto Encyclopedia of Genes and Genomes ( Kegg ) enrichment and protein-protein interaction ( PPI ) network construction were performed by DAVID and String softwares. RT-PCR was used to detect the differences in the expression of different genes in different radiosensitivity cells. Results A total of 49 differentially expression genes were screened. These genes were mainly involved in the regulation of multicellular biosynthesis, ion transport, DNA synthesis, metabolism, cell proliferation and so on. The major biological pathways included a Wnt signal pathway. 12 DEGs interacted with each other, and CHN2 may be a key node. The expression of CHN2 gene had no obvious difference between TE13R and TE13. Conclusions 49 differentially expressed genes, including CHN2, may be involved in radioresistance of esophageal carcinoma, and the Wnt signaling pathway may be an important pathway in this radioresistance.