Amygdalin Ameliorates Liver Fibrosis through Inhibiting Activation of TGF-β/Smad Signaling / 中国结合医学杂志

OBJECTIVE@#To observe the effect of amygdalin on liver fibrosis in a liver fibrosis mouse model, and the underlying mechanisms were partly dissected in vivo and in vitro.@*METHODS@#Thirty-two male mice were randomly divided into 4 groups, including control, model, low- and high-dose amygdalin-treated groups, 8 mice in each group. Except the control group, mice in the other groups were injected intraperitoneally with 10% carbon tetrachloride (CCl4)-olive oil solution 3 times a week for 6 weeks to induce liver fibrosis. At the first 3 weeks, amygdalin (1.35 and 2.7 mg/kg body weight) were administered by gavage once a day. Mice in the control group received equal quantities of subcutaneous olive oil and intragastric water from the fourth week. At the end of 6 weeks, liver tissue samples were harvested to detect the content of hydroxyproline (Hyp). Hematoxylin and eosin and Sirius red staining were used to observe the inflammation and fibrosis of liver tissue. The expressions of collagen I (Col-I), alpha-smooth muscle actin (α-SMA), CD31 and transforming growth factor β (TGF-β)/Smad signaling pathway were observed by immunohistochemistry, quantitative real-time polymerase chain reaction and Western blot, respectively. The activation models of hepatic stellate cells, JS-1 and LX-2 cells induced by TGF-β1 were used in vitro with or without different concentrations of amygdalin (0.1, 1, 10 µmol/L). LSECs. The effect of different concentrations of amygdalin on the expressions of liver sinusoidal endothelial cells (LSECs) dedifferentiation markers CD31 and CD44 were observed.@*RESULTS@#High-dose of amygdalin significantly reduced the Hyp content and percentage of collagen positive area, and decreased the mRNA and protein expressions of Col-I, α-SMA, CD31 and p-Smad2/3 in liver tissues of mice compared to the model group (P<0.01). Amygdalin down-regulated the expressions of Col-I and α-SMA in JS-1 and LX-2 cells, and TGFβ R1, TGFβ R2 and p-Smad2/3 in LX-2 cells compared to the model group (P<0.05 or P<0.01). Moreover, 1 and 10 µmol/L amygdalin inhibited the mRNA and protein expressions of CD31 in LSECs and increased CD44 expression compared to the model group (P<0.05 or P<0.01).@*CONCLUSIONS@#Amygdalin can dramatically alleviate liver fibrosis induced by CCl4 in mice and inhibit TGF-β/Smad signaling pathway, consequently suppressing HSCs activation and LSECs dedifferentiation to improve angiogenesis.

Study on the comparative analysis of the efficacy of transmesenteric vein extrahepatic portosystemic shunt and transjugular intrahepatic portosystemic shunt in the treatment of cavernous transformation of portal vein / 中华肝脏病杂志

Objective: To compare the safety and efficacy of transmesenteric vein extrahepatic portosystemic shunt (TEPS) and transjugular intrahepatic portosystemic shunt (TIPS) in the treatment of cavernous transformation of the portal vein (CTPV). Methods: The clinical data of CTPV patients with patency or partial patency of the superior mesenteric vein treated with TIPS or TEPS treatment in the Department of Vascular Surgery of Henan Provincial People's Hospital from January 2019 to December 2021 were selected. The differences in baseline data, surgical success rate, complication rate, incidence rate of hepatic encephalopathy, and other related indicators between TIPS and TEPS group were statistically analyzed by independent sample t-test, Mann-Whitney U test, and Chi-square test. Kaplan-Meier survival curve was used to calculate the cumulative patency rate of the shunt and the recurrence rate of postoperative portal hypertension symptoms in both groups. Results: The surgical success rate (100% vs. 65.52%), surgical complication rate (6.67% vs. 36.84%), cumulative shunt patency rate (100% vs. 70.70%), and cumulative symptom recurrence rate (0% vs. 25.71%) of the TEPS group and TIPS group were statistically significantly different (P < 0.05). The time of establishing the shunt [28 (2141) min vs. 82 (51206) min], the number of stents used [1 (12) vs. 2 (15)], and the length of the shunt [10 (912) cm vs. 16 (1220) cm] were statistically significant between the two groups (t = -3.764, -4.059, -1.765, P < 0.05). The incidence of postoperative hepatic encephalopathy in the TEPS group and TIPS group was 6.67% and 15.79% respectively, with no statistically significant difference (Fisher's exact probability method, P = 0.613). The pressure of superior mesenteric vein decreased from (29.33 ± 1.99) mmHg to (14.60 ± 2.80) mmHg in the TEPS group and from (29.68 ± 2.31) mmHg to (15.79 ± 3.01) mmHg in TIPS group after surgery, and the difference was statistically significant (t = 16.625, 15.959, P < 0.01). Conclusion: The best indication of TEPS is in CTPV patients with patency or partial patency of the superior mesenteric vein. TEPS improves the accuracy and success rate of surgery and reduces the incidence of complications.

Research Progress of Exosomes and Their Forensic Significance / 法医学杂志

Exosomes are membranous tiny vesicles secreted by cells, which are widely found in the extracellular matrix and various body fluids and carry a variety of biologically functional molecules such as proteins, lipids, messenger RNA (mRNA) and microRNA (miRNA). Exosomes not only play important biological roles in the field of immunology and oncology, but also have potential application value in the field of forensic medicine. This article reviews the discovery, production and degeneration mechanism, biological functions, isolation and identification methods of exosomes, summarizes the research on exosomes and their significance in the field of forensic science, and discusses their applications in body fluid identification, individual identification, postmortem interval estimation to provide ideas for the application of exosomes in forensic work.

Early risk factors for death in neonates with persistent pulmonary hypertension of the newborn treated with inhaled nitric oxide / 中国当代儿科杂志

OBJECTIVES@#To evaluate the early risk factors for death in neonates with persistent pulmonary hypertension of the newborn (PPHN) treated with inhaled nitric oxide (iNO).@*METHODS@#A retrospective analysis was performed on 105 infants with PPHN (gestational age ≥34 weeks and age <7 days on admission) who received iNO treatment in the Department of Neonatology, Children's Hospital of Nanjing Medical University, from July 2017 to March 2021. Related general information and clinical data were collected. According to the clinical outcome at discharge, the infants were divided into a survival group with 79 infants and a death group with 26 infants. Univariate and multivariate Cox regression analyses were used to evaluate the risk factors for death in infants with PPHN treated with iNO. The receiver operating characteristic (ROC) curve was used to calculate the cut-off values of the factors in predicting the death risk.@*RESULTS@#A total of 105 infants with PPHN treated with iNO were included, among whom 26 died (26/105, 24.8%). The multivariate Cox regression analysis showed that no early response to iNO (HR=8.500, 95%CI: 3.024-23.887, P<0.001), 1-minute Apgar score ≤3 points (HR=10.094, 95%CI: 2.577-39.534, P=0.001), a low value of minimum PaO2/FiO2 within 12 hours after admission (HR=0.067, 95%CI: 0.009-0.481, P=0.007), and a low value of minimum pH within 12 hours after admission (HR=0.049, 95%CI: 0.004-0.545, P=0.014) were independent risk factors for death. The ROC curve analysis showed that the lowest PaO2/FiO2 value within 12 hours after admission had an area under the ROC curve of 0.783 in predicting death risk, with a sensitivity of 84.6% and a specificity of 73.4% at the cut-off value of 50, and the lowest pH value within 12 hours after admission had an area under the ROC curve of 0.746, with a sensitivity of 76.9% and a specificity of 65.8% at the cut-off value of 7.2.@*CONCLUSIONS@#Infants with PPHN requiring iNO treatment tend to have a high mortality rate. No early response to iNO, 1-minute Apgar score ≤3 points, the lowest PaO2/FiO2 value <50 within 12 hours after admission, and the lowest pH value <7.2 within 12 hours after admission are the early risk factors for death in such infants. Monitoring and evaluation of the above indicators will help to identify high-risk infants in the early stage.

Research Progress on the Role of Mitochondrial DNA in the Pathogenesis of Chronic Liver Disease / 中华肝脏病杂志

Mitochondrial DNA is the mitochondria's own genetic material located within the mitochondrial matrix and is involved in cellular metabolism and energy supply. Mitochondrial DNA damage exacerbates oxidative stress by increasing the release of reactive oxygen species, while mitochondrial DNA release also triggers apoptosis and activates immune inflammatory responses through damage-related molecular patterns. Mitochondrial autophagy regulates mitochondrial DNA damage and release through a negative feedback mechanism to maintain intracellular homeostasis. Recent studies have shown that the occurrence and development of chronic liver disease are closely related to mitochondrial DNA-mediated immune inflammatory responses and oxidative stress.

Pathological Mechanism of HSOS in Rats Induced by Different Doses of Monocrotaline / 中国实验方剂学杂志

ObjectiveTo observe the pathological changes of hepatic sinusoidal obstruction syndrome （HSOS） induced by different doses of monocrotaline （MCT） in rats， investigate the dose and duration of modeling， and elucidate the mechanism. MethodA total of 72 male SD rats were randomized into normal group （n=12）， and low-， medium-， and high-dose MCT groups （n=20 per group， 80，120，160 mg·kg-1， respecctively）. In the model groups， different doses of MCT were intragastrically administered to induce the HSOS in rats. After 48 h and 120 h separately， rats in each group were sacrificed and sampling was performed. The survival rate of rats in each group was calculated， and the body weight， liver weight， and and serum liver function indexes of the rats were examined. The histopathological changes of the liver were observed based on scanning electron microscopy， hematoxylin and eosin （HE） staining， and Sirius red （SR） staining. Glutathione S-transferase （GST） activity， total superoxide dismutase （T-SOD） activity， and malondialdehyde （MDA） content of liver tissue homogenate were measured with microplate method. The expression of liver tissue-related indexes was detected by real-time polymerase chain reaction （PCR）， Western blot， and immunohistochemistry. ResultThe activity of serum alanine aminotransferase （ALT） and aspartate aminotransferase （AST） in MCT groups rose with the increase in MCT dose （P<0.05， P<0.01） compared with that in the normal group. With the extension of modeling time， the activity of serum ALT and AST in the low-dose group decreased （P<0.01）， while the activity of them in the medium-dose and high-dose groups increased （P<0.01）. HE staining showed that hepatocyte necrosis， inflammatory cell infiltration， and erythrocyte accumulation in MCT groups. Electron microscopy demonstrated that fenestrae of liver sinusoidal endothelial cells widened and the sieve plates disappeared. Morever， the injury was worsened with the increase in MCT dose. In addition， the expression of CD44 in MCT groups was significantly reduced compared with that in the normal group （P<0.05， P<0.01）. SR staining showed that no positive staining was found in model groups after 48 h， while collagen deposition in portal areas and liver sinusoids could be seen in model groups after 120 h. MCT groups showed increase in MDA content and GST activity and decrease in T-SOD activity compared with the normal group， particularly the medium-dose and high-dose groups （P<0.01）， and the changes were dose-dependent after 120 h （P<0.01）. The protein expression of CD68 （pro-inflammatory macrophage marker） was raised with the increase in dosage， which was consistent with the results of immunohistochemistry （P<0.01）， while CD163 （anti-inflammatory macrophage marker） protein and mRNA expression was significantly decreased with the increase in dosage （P<0.01）. Western blot results showed that the expression of phosphorylated nuclear factor-κB/nuclear factor-κB （p-NF-κB/NF-κB） and phosphorylated protein kinase B/protein kinase B （p-Akt/t-Akt） was significantly increased in medium-dose and high-dose MCT groups （P<0.05，P<0.01）. The protein expression of α-smooth muscle actin （α-SMA） in liver tissues in MCT groups was significantly increased over time and with the increase in dose， and the mRNA expression of α-SMA， collagen type I α1 （Col1a1）， and collagen type Ⅳ α1 （Col4a1） showed the same trend （P<0.05， P<0.01）. The results of TUNEL staining showed that apoptotic cells were increased with the rise of MCT dose， while B-cell lymphoma-2（Bcl-2） /Bcl-2 associated X protein （Bax） was remarkably decreased （P<0.01）. ConclusionHSOS in rats induced by intragastric administration of different doses of MCT was aggravated with the increase of dosage. In the low-dose （80 mg·kg-1） MCT group， the liver healed spontaneously over time. However， liver damage caused by MCT of 120 mg·kg-1 and 160 mg·kg-1 aggravated over time， and even fibrosis and death occurred. The pathological mechanism of MCT-induced HSOS in rats may be that MCT triggered intense oxidative stress in liver tissue， thus activated pro-inflammatory macrophages to secrete large amounts of inflammatory factors， and further activated the NF-κB/Akt signalling pathway， leading to severe cell damage and death.

Molecular Mechanism Analysis of Jiangtang Xiaozhi Tablets in Treatment of NAFLD / 中国实验方剂学杂志

Objective:To explore the molecular mechanism of Jiangtang Xiaozhi tablets （JTXZT） in the treatment of non-alcoholic fatty liver disease （NAFLD） by means of network pharmacology and molecular docking. Method:With the help of traditional Chinese medicine （TCM） Systems Pharmacology Database and Analysis Platform （TCMSP）， TCMs Integrated Database （TCMID）， Encyclopedia of TCM （ETCM） and Bioinformatics Analysis Tool for Molecular Mechanism of TCM （BATMAN-TCM）， the chemical compositions of medicinal materials in JTXZT were obtained， the compound targets were predicted in SwissTargetPrediction database and STITCH database. The targets of NAFLD were searched by The Human Gene Database （GeneCards）， Online Mendelian Inheritance in Man （OMIM）， Therapeutic Target Database （TTD） and DisGeNET， and intersection analysis was performed with the targets of the active ingredients to obtain the targets of JTXZT for treatment of NAFLD. Based on STRING 11.0 database， the protein-protein interaction （PPI） network of therapeutic targets was constructed， and the enrichment analysis of therapeutic targets was carried out by DAVID 6.8. Finally， the interaction characteristics of key components and core therapeutic targets of JTXZT for treatment of NAFLD were verified based on molecular docking. Result:The key components of JTXZT for treatment of NAFLD were quercetin， luteolin， kaempferol， berberine， isorhamnetin， betulinic acid， oleanolic acid， ursolic acid. formononetin and hexitol， and the core targets of JTXZT for treatment of NAFLD were mitogen-activated protein kinase 1 （MAPK1）， Jun proto-oncogene， activator protein-1 （AP-1） transcription factor subunit （JUN）， MAPK3， protein kinase B1 （AKT1 or Akt1）， tumor protein p53 （TP53）， E1A binding protein p300 （EP300）， Fos proto-oncogene， AP-1 transcription factor subunit （FOS）， tumor necrosis factor （TNF），amyloid beta precursor protein （APP） and cytochrome P450 family 2 subfamily E member 1 （CYP2E1）. Biological function and pathway enrichment analysis showed that JTXZT mainly through xenobiotic metabolic process， oxidation-reduction process， cholesterol metabolic process and other biological processes， regulating phosphatidylinositol 3-kinase/protein kinase B （PI3K/Akt） signaling pathway， MAPK signaling pathway， NAFLD and insulin signaling pathway to play a role in the treatment of NAFLD. The results of molecular docking showed that the active components of JTXZT had a good affinity with the core targets of JTXZT for the treatment of NAFLD. Conclusion:JTXZT treats NAFLD through multiple active components， multiple key targets and multiple action pathways.

Species Identification of Sarcosaprophagous Flies Based on Vein Digital Image Analysis / 法医学杂志

Objective To identify species of common sarcosaprophagous flies based on digital image analysis of veins, in order to provide new idea for fast and accurate species identification of sarcosaprophagous flies in forensic entomology. Methods Random trapping of 226 male and female sarcosaprophagous flies that comprised of 7 common species, including Sarcophaga peregrina, Parasarcophaga ruficornis, Sarcophaga dux, Seniorwhitea reciproca, Bercaea cruentata, Aldrichina grahami, and Synthesiomysia nudiseta with carrion in the field was conducted. The 17 landmarks on the right wing of each fly were digitally processed and the images were analyzed. The effects of allometry were evaluated using a permutation test. Wing shape variations among 7 sarcosaprophagous fly species and female species was analyzed using canonical variate analysis （CVA）. Additionally, cross-validation test was used to evaluate the reliability of classification. Results Among 7 sarcosaprophagous fly species and female species, the effect of allometry had statistical significance （P<0.05）. The CVA results showed that among 7 sarcosaprophagous fly species and female species, differences in the wing shape were significant, and the first two canonical variates accounted for 82.9% and 84.1% of the total variation of vein shape. Vein digital image analysis can be used to separate the 7 common sarcosaprophagous flies, with an overall species identification accuracy of 81.2%-100.0%, and with a species identification accuracy of 75.0%-100.0% to distinguish the female flies of the 7 sarcosaprophagous flies species. Conclusion Vein digital image analysis is a relatively convenient and reliable method for identification of insect species, which can be used for species identification of common sarcosaprophagous flies.

Progress and Application of Entomological Evidence in Forensic Science / 法医学杂志

Entomological evidence provides entry points and clues for cases detection, in terms of estimation of the postmortem interval （PMI）, and place and cause of death. In recent years, the feasibility of entomological evidence in practice has been proved by theories and cases. It especially plays an important role in the investigation of cases with unnatural death, no monitoring, and highly corrupt cadaver. However, there are still some key issues to be further studied and standardized before the application of entomological evidence to forensic practice, to improve the effect of entomological evidence in forensic investigation and trial. This paper retrospectively reviews key studies of the application of entomological evidence in forensic science, mainly including discussion of forensic entomology inspection standard, identification studies of sarcosaprophagous insect species, collection of sarcosaprophagous insect growth and succession data under different environments and forensic entomotoxicology. With the rapid development of information technology and biotechnology, applying artificial intelligence and whole genome sequencing technology in forensic entomology has become a new research direction, which can improve the application value and range of entomological evidence in forensic science.

Main components in butyl alcohol extract of Baitouweng Decoction inhibited neutrophil chemotaxis / 中国中药杂志

The present study aims to investigate the effects of the main components(aesculin, berberine hydrochloride, and anemoside B4) in the butyl alcohol extract of Baitouweng Decoction(BAEB) on the chemotaxis of neutrophils induced by dimethyl sulfoxide(DMSO). HL60 cells were cultivated in RPMI-1640 complete medium, and transferred into a 6-well plate(2 × 10~5 per mL) with 4 mL in each well, followed by incubation with DMSO at 1.3% for five days. The morphologic changes of cells were observed under an inverted microscope. The CD11 b expression after DMSO induction was analyzed by flow cytometry. The effects of aesculin, berberine hydrochloride, and anemoside B4 on the cell proliferation and migration were detected by CCK8 assay and Transwell assay, respectively. The effects of the main components on the production and polarization of F-actin protein were also examined by flow cytometry and laser confocal microscopy. PI3 K/Akt signaling pathway was checked by Western blot. As revealed by the results, neutrophil-like HL60 cells were observed after DMSO induction. The CD11 b expression in these cells increased significantly as indicated by the flow cytometry. Additionally, 100 μg·mL~(-1) aesculin, 8 μg·mL~(-1) berberine hydrochloride, and 80 μg·mL~(-1) anemoside B4 were potent in inhibiting the migration of neutrophils and reducing F-actin expression. Berberine hydrochloride was verified to be capable of diminishing phosphorylated PI3 K/Akt protein expression. The findings indicate that aesculin, anemoside B4, and especially berberine hydrochloride in the BAEB can inhibit the chemotaxis of neutrophils, which is possibly achieved by the inhibition of F-actin and PI3 K/Akt signaling pathway.

Correlation analysis and breeding application of agronomic traits and quality indexes in Dendrobium nobile / 中国中药杂志

The present study aimed to explore the correlation between agronomic traits and quality indexes of Dendrobium nobile and its application value in agricultural breeding. The cultivated strains of D. nobile in Hejiang-Chishui producing areas were extensively collected,and the main agronomic traits and quality indexes were measured. The agronomic traits with significant correlation with quality indexes were screened out by the correlation analysis,and then the parental lines and self-bred F_1 generation plants were furtherverified. Among 96 lines of D. nobile,the content of soluble polysaccharides showed a significant negative correlation with dendrobine( P < 0. 01),and no significant correlation with agronomic traits in stems and leaves. The content of dendrobine exhibited a significant positive correlation with the stem width-thickness ratio( at the largest cross section; P < 0. 01),and no significant correlation with other agronomic traits. Regression analysis further verified the positive correlation between dendrobine content and stem width-thickness ratio( R2> 0. 9). Two lines,JC-10 and JC-35,with significant differences in stem width-thickness ratio were screened out( P <0. 05). The corresponding F1 generation plants by self-pollination both showed that the dendrobine content was higher with greater stem width-thickness ratio( P < 0. 01). The experimental results suggested that within a certain range,the dendrobine content was higher in D. nobile with flatter stem. Therefore,in the breeding of D. nobile,this specific trait could be used for screening plants with high content of quality indexes such as dendrobine.

Effect of berberine hydrochloride on cell wall integrity of Candida albicans hypha / 中国中药杂志

The aim of this paper was to investigate the effect of berberine hydrochloride on the cell wall integrity of Candida albicans hypha. The minimal inhibitory concentration(MIC) of berberine hydrochloride against clinical and standard C. albicans strains was detected by micro liquid-based dilution method; the effect of berberine hydrochloride on the colony formation of C. albicans SC5314 was investigated by spot assay; the effect of berberine hydrochloride on the metabolism of C. albicans SC5314 hypha was checked by XTT reduction assay, and the viability of C. albicans SC5314 hypha was tested by fluorescent staining assay. The effect of berberine hydrochloride on the morphology of C. albicans SC5314 hypha was examined by scanning electron microscope. The changes in the cell wall of C. albicans SC5314 hypha after berberine hydrochloride treatment were detected by transmission electron microscopy. The effect of berberine hydrochloride on β-glucan from C. albicans SC5314 was detected by flow cytometry. The effect of berberine hydrochloride on hypha-specific gene ECE1 and β-glucan synthase genes FKS1 and FKS2 in C. albicans was examined by qRT-PCR. The results showed that berberine hydrochloride showed a strong inhibitory effect on both clinical and standard strains of C. albicans, and the MIC was 64-128 μg·mL~(-1). Spot assay, XTT redunction assay and fluorescent staining assay showed that with the increase of berberine hydrochloride concentration, the viability of C. albicans SC5314 gradually decreased. The transmission electron microscopy scanning assay showed that this compound could cause cell wall damage of C. albicans. The flow cytometry analysis showed the exposure degree of C. albicans β-glucan. The qRT-PCR further showed that berberine hydrochloride could significantly down-regulate hypha-specific gene ECE1 and β-glucan synthase-related gene FKS1 and FKS2. In conclusion, this compound can down-regulate C. albicans and β-glucan synthase-related gene expressions, so as to destroy the cell wall structure of C. albicans, expose β-glucan and damage the integrity of the wall.

Cytotoxicity and underlying mechanism of evodiamine in HepG2 cells / 北京大学学报(医学版)

OBJECTIVE@#To investigate evodiamine (EVO)-induced hepatotoxicity and the underlying mechanism.@*METHODS@#HepG2 cells were treated with EVO (0.04-25 μmol/L) for different time intervals, and the cell survival rate was examined by cell counting kit-8 (CCK-8) method. After HepG2 cells were treated with EVO (0.2, 1 and 5 μmol/L) for 48 h, the alanine transaminase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), alkaline phosphatase (ALP) activities and total bilirubin (TBIL) content of supernatant were detected. A multifunctional microplate reader was used to detect the intracellular superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in HepG2 cells to evaluate the level of cell lipid peroxidation damage. The interactions between EVO and apoptosis, autophagy or ferroptosis-associated proteins were simulated by molecular docking. The HepG2 cells were stained by mitochondrial membrane potential (MMP) fluorescent probe (JC-10) and annexin V-fluorescein isothiocyanate/propidium iodide (Annexin V-FITC/PI), and MMP and apoptosis in HepG2 cells were detected by flow cytometry. The protein expression levels of caspase-9, caspase-3, bile salt export pump (BSEP) and multidrug resistance-associated protein 2 (MRP2) were detected by Western blot.@*RESULTS@#The cell survival rate was significantly reduced after the HepG2 cells were exposed to EVO (0.04-25 μmol/L) in a time- and dose-dependent manner. The half maximal inhibitory concentration (IC50) of the HepG2 cells treated with EVO for 24, 48 and 72 h were 85.3, 6.6 and 4.7 μmol/L, respectively. After exposure to EVO (0.2, 1 and 5 μmol/L) for 48 h, the ALT, AST, LDH, ALP activities and TBIL content in the HepG2 cell culture supernatant, and the MDA content in the cells were increased, and SOD enzyme activity was decreased. Molecular docking results showed that EVO interacted with apoptosis-associated proteins (caspase-9 and caspase-3) better. JC-10 and Annexin V-FITC/PI staining assays demonstrated that EVO could decrease MMP and promote apoptosis in the HepG2 cells. Western blot results indicated that the protein expressions of cleaved caspase-9 and cleaved caspase-3 were upregulated in the HepG2 cell treated with EVO for 48 h. In contrast, the protein expressions of pro-caspase-3, BSEP and MRP2 were downregulated.@*CONCLUSION@#These results suggested that 0.2, 1 and 5 μmol/L EVO had the potential hepatotoxicity, and the possible mechanism involved lipid peroxidation damage, cell apoptosis, and cholestasis.

Development of programmed death receptor-1 and programmed death receptor-1 ligand in oral squamous cell carcinoma / 华西口腔医学杂志

Oral squamous cell carcinoma (OSCC) is a common malignant tumor in the oral and maxillofacial region. At present, the treatment of OSCC is mainly based on surgical oriented comprehensive sequence therapy, especially the triple therapy of surgery, radiotherapy, and chemotherapy. However, the overall five-year survival rate is relatively low. Therefore, researching the pathogenesis and treatment methods of OSCC is important. The immune checkpoint of programmed death receptor-1 (PD-1) and programmed death receptor-1 ligand (PD-L1) have been the focus of research in recent years. Several studies have shown that the high expression of PD-1/PD-L1 in most OSCC microenvironments may contribute to the immune escape of tumors. In this study, the research status of immune checkpoint of PD-1/PD-L1 and its relevant inhibitors in OSCC were reviewed.

Clinical evaluation on three-dimensional laparoscopic splenectomy for massive splenomegaly secondary to liver cirrhosis / 中华普通外科杂志

Objective To investigate the clinical effect of three-dimensional laparoscopic splenectomy for massive splenomegaly secondary to liver cirrhosis.Methods The clinical data of 67 patients who underwent laparoscopic splenectomy for massive splenomegaly at Henan Province People's Hospital from Jan 2013 to Dec 2018 were collected.Results Operation time,volume of intraoperative blood loss and blood transfusion,number of patients with intraoperative blood transfusion,and conversion to laparotomy were in favor of 3D group,with statistically differences (t =12.900,18.255,19.711,x2 =10.747,0.685,P ＜ 0.05).Postoperative intraabdominal bleeding,pancreatic fistula,and postoperative hospital stay in 2D group were more than those in 3D group,with statistically differences (x2 =3.511,4.527,t =12.969,P ＜ 0.05).All patients were followed up for 5 to 60 months.Portal thrombosis occurred in 6 patients vs 5 patients,respectively (x2 =0.028,P ＞ 0.05) and resolved with oral coumarin.Conclusions Three-dimensional laparoscopic splenectomy can provide more realistic visual effects of surgical procedures and has an obvious advantage in laparoscopic splenectomy for massive splenomegaly secondary to liver cirrhosis.

Efficacy of intrahepatic cholangiocarcinoma treated with chemotherapy through portal vein pump after radical surgery / 中华肝胆外科杂志

Objective To evaluate the clinical application value of portal vein implantation pump for chemotherapy in patients with intrahepatic cholangiocarcinoma (ICC) after radical surgery.Methods The clinical data of 97 patients with ICC who underwent radical surgery in Henan People's Hospital from June 2012 to June 2016 were retrospectively analyzed.Results Among the 97 patients,14 patients received portal venous pump chemotherapy (portal group),33 patients received peripheral venous chemotherapy (peripheral group),and 50 patients did not receive postoperative chemotherapy (control group).There were no statistically significant differences in gender and age between the three groups.The results of survival analysis indicated that the disease-free survival (DFS) period and overall survival (OS) time in the portal group and the peripheral group were significantly better than that in control group (both P ＜ 0.05).In addition,despite the lack of statistical significance (P ＞ 0.05),for the control of intrahepatic metastasis,portal vein pump chemotherapy was better than that of systemic chemotherapy via peripheral vein,and almost all side effects of chemotherapy in the portal group were lower than those in the peripheral group.Conclusion Portal vein pump chemotherapy can improve the prognosis of intrahepatic bile duct patients,especially for the control of intrahepatic metastasis,and can reduce systemic side effects of chemotherapy.

Clinical efficacy of laparoscopic radical resection of hilar cholangiocarcinoma / 中华消化外科杂志

Objective:To investigate the clinical efficacy of laparoscopic radical resection of hilar cholangiocarcinoma.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 25 patients with hilar cholangiocarcinoma who were admitted to Henan Provincial People′s Hospital from January 2017 to July 2019 were collected. There were 16 males and 9 females, aged from 51 to 75 years, with a median age of 64 years. All the 25 patients underwent laparoscopic radical resection of hilar cholangiocarcinoma. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up using out-patient examination and telephone interview was performed to detect local recurrence and distant metastasis of patients up to December 2019. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were described as M (range). Count data were described as absolute numbers. Results:(1) Surgical situations: of the 25 patients, 15 patients in Bismuth typeⅠunderwent laparoscopic radical resection of hilar cholangiocarcinoma+ regional lymph node dissection+ Roux-en-Y choledocho-jejunostomy, 2 patients in Bismuth type Ⅱ underwent laparoscopic radical resection of hilar cholangiocarcinoma+ perihilar resection+ regional lymph node dissection+ Roux-en-Y choledochojejunostomy, 2 patients in Bismuth type Ⅲa underwent laparoscopic radical resection of hilar cholangiocarcinoma+ regional lymph node dissection+ right hemihepatectomy+ hepatic caudate labectomy+ Roux-en-Y choledochojejunostomy, 3 patients in Bismuth type Ⅲb underwent laparoscopic radical resection of hilar cholangiocarcinoma+ regional lymph node dissection+ left hemihepatectomy+ hepatic caudate labectomy+ Roux-en-Y choledochojejunostomy, 3 patients in Bismuth type Ⅳ underwent laparoscopic radical resection of hilar cholangiocarcinoma+ regional lymph node dissection+ hepatic caudate labectomy+ Roux-en-Y choledochojejunostomy. The operation time was (388±118)minutes, and volume of intraoperative blood loss was 200 mL(range, 50-2 000 mL). Six patients were treated with blood transfusion intraoperatively. The operation time of 2 patients in Bismuth type Ⅲa was 375 minutes and 465 minutes, and the volume of intraoperative blood loss was 200 mL and 1 000 mL, respectively; 1 case received blood transfusion. The operation time of 3 patients in Bismuth type Ⅲb was 410 minutes, 465 minutes, 501 minutes, and the volume of intraoperative blood loss was 300 mL, 400 mL, 450 mL, respectively; neither had intraoperative blood transfusion. The operation time of 3 patients in Bismuth type Ⅳ was 415 minutes, 560 minutes, 600 minutes, and the volume of intraoperative blood loss was 300 mL, 600 mL, 800 mL, respectively; 1 case had intraoperative blood transfusion. (2) Postoperative situations: of the 25 patients, 4 patients had grade Ⅰ complications, including 2 cases of biliary fistula (1 case in Bismuth type Ⅰ and 1 case in Bismuth type Ⅲa), 1 case of pulmonary infection (Bismuth type Ⅳ), and 1 case of postoperative liver insufficiency (Bismuth type Ⅲa), all of them were improved after conservative treatment. Results of postoperative pathological examination: bile duct adenocarcinoma and high-grade intraepithelial neoplasia were detected in 23 and 2 patients; there were 8 cases with nerve invasion, 3 cases with lymph node metastasis and no vascular thrombus. The duration of hospital stay and hospitalization expenses were 24 days (range, 10-45 days) and 9.4×10 4 yuan [range, (5.3-18.7)×10 4 yuan] for all the 25 patients; the above indicators were 36 days, 45 days, 15.1×10 4 yuan, 18.7×10 4 yuan for the 2 patients in Bismuth type Ⅲa, 15 days, 26 days, 33 days, 7.3×10 4 yuan, 11.5×10 4 yuan, 15.9×10 4 yuan for 3 patients in Bismuth type Ⅲb, 24 days, 39 days, 41 days, 12.1×10 4 yuan, 15.2×10 4 yuan, 16.7×10 4 yuan for the 3 patients in Bismuth type Ⅳ, respectively. (3) Follow-up: 25 patients were followed up for 2-36 months, with a median follow-up of 16 months. Of the 25 patients, 18 had no recurrence or metastasis, 2 patients in Bismuth type Ⅳ had extensive intraperitoneal metastasis, 1 patients in Bismuth type Ⅲa had trocar hole metastasis, and 4 patients died. Conclusions:Laparoscopic radical resection of hilar cholangiocarcinoma is safe and feasible. Surgeries should be selected strictly based on surgical indications and the Bismuth type.

Exploring Research on Processing Method of Polygoni Multiflori Radix Praeparata / 中国实验方剂学杂志

Objective::To investigate the effects of black bean juice with different stewing times on the appearance character and the content changes of effective components of Polygoni Multiflori Radix Praeparata. Method::HPLC was employed with Agilent ZORBAX Extend-C18 column (4.6 mm×250 mm, 5 μm), a gradient mobile phase of methanol (A)-water (B) was eluted (0-30 min, 5%-100%A; 30-40 min, 100%A), the flow rate was 1.0 mL·min-1, the injection volume was 10 μL and the column temperature was 35 ℃, detection wavelength was set at 280 nm. The contents of stilbene glycoside, emodin, emodin methyl ether, emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside in samples prepared at different processing times were simultaneously determined by HPLC. Result::The content of stilbene glycoside decreased gradually with the increase of stewing time, compared with 8 h, its content decreased by 76% at 64 h. The contents of emodin-8-O-β-D-glucoside and emodin methyl ether-8-O-glucoside increased first, and then decreased, reaching the highest value at 24 h, and then decreased to the level similar to the content of 8 h after 40 h, and then fluctuated slightly. The contents of emodin and emodin methyl ether increased first, and then decreased, reached the maximum when stewed for 32 h, then decreased slowly and tended to be stable. Conclusion::The stewing time has significant influence on the content of various components in Polygoni Multiflori Radix Praeparata, and the changing trend is different, the processing time of Polygoni Multiflori Radix Praeparata shall be standardized. At the same time, it is not sufficient to take stilbene glycoside and anthraquinones as the indicator ingredients for this decoction pieces, the quality control indicators such as polysaccharides shall be considered.

Analysis on Medication Rules of Traditional Chinese Medicine for Treatment of Diabetic Peripheral Neuropathy Based on Clinical Literature / 中国实验方剂学杂志

Objective:To investigate the medication rules of traditional Chinese medicine (TCM) for the treatment of diabetic peripheral neuropathy (DPN).Method:The literature published in China Knowledge Resource Integrated Database (CNKI), Wanfang Database, China Biomedical Literature Service System (SinoMed), VIP Database and PubMeb from 2008 to 2019 were retrieved by setting the topics of diabetic peripheral neuropathy and TCM. After screening, a database was established to analyze the medication rules (efficacy, frequency, flavor and meridian tropism, common couplet medicinals and core medicines) of TCM by frequency statistics, association rules and data statistical methods of constructing complex networks.Result:A total of 461 papers for treatment of DPN were included in this study, including 275 kinds of TCM and a total frequency of 6 361 times. Astragali Radix had the highest frequency. Among all kinds of medicinal materials, activating blood circulation and removing stasis was the most commonly used medicine, followed by Qi-invigorating medicine. Flavor of medicines was mainly sugariness and warm, and most of their meridian tropism was liver meridian. After the analysis by association rules, the couplet medicinals with the highest support was Astragali Radix-Angelicae Sinensis Radix. The core medicines obtained by complex network analysis were Astragali Radix, Angelicae Sinensis Radix, Chuanxiong Rhizoma, Spatholobi Caulis, Cinnamomi Ramulus, Carthami Flos, Pheretima, Paeoniae Radix Rubra, Salviae Miltiorrhizae Radix et Rhizoma and Persicae Semen.Conclusion:This study comprehensively analyzes the medication rules of TCM clinical treatment of DPN. The main treatment methods of TCM for DPN are invigorating Qi and blood, activating blood circulation and removing stasis, activating meridians to stop pain, which can provide guidance for the TCM clinical use and new Chinese medicines research and development of DPN.

Effect of Huangqi Guizhi Wuwutang on AGEs/RAGE/NF-κB Signaling Pathway in Diabetic Peripheral Neuropathy Rats / 中国实验方剂学杂志

Objective:To study the effect of Huangqi Guizhi Wuwutang on receptor of advanced glycation end products(AGEs)/advanced glycation endproducts (RAGE)/nuclear transcription factor-kappa B p65 (NF-κB p65) signaling pathway in the diabetic peripheral neuropathy rats through an animal modeling experiment, and discuss the mechanism of Huangqi Guizhi Wuwutang in alleviating diabetic peripheral neuropathy. Method:Rat model of diabetic peripheral neuropathy was established by high-fat diet and intraperitoneal injection with streptozotocin (STZ). After successful modeling, Huangqi Guizhi Wuwutang intervention began in the fifth week. The patients in high-dose group (19.40 g∙kg-1∙d-1), middle-dose group (4.85 g∙kg-1∙d-1) and low-dose group (2.43 g∙kg-1∙d-1) were given by gavage continuously for 12 weeks. The western medicine control group was given 25 mg∙kg-1∙d-1 by gavage. After the experiment, serum interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected by enzyme-linked immunosorbent assay (ELISA). Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect RAGE and NF-κB p65 mRNA expressions in sciatic nerve tissue. The expressions of RAGE, NF-κB and phosphorylation(p)-NF-κB p65 proteins in sciatic nerve tissues were detected by Western blot (WB). Result:Compared with the normal group, serum IL-1β and TNF-α protein levels, RAGE mRNA and NF-κB p65 mRNA levels, RAGE protein, NF-κB p65 protein and p-NF-κB p65 protein levels were significantly increased in the model group (P<0.01), the ratio of p-NF-κB p65 to NF-κB p65 was increased, and the phosphorylation of NF-κB p65 was enhanced (P<0.01). After the intervention of Huangqi Guizhi Wuwutang, compared with the model group, serum IL-1β and TNF-α protein levels, RAGE and NF-κB p65 mRNA levels, RAGE protein, NF-κB p65 protein and p-NF-κB p65 protein levels were all decreased (P<0.01), the ratio of p-NF-κB p65 to NF-κB p65 was decreased in high-dose group (P<0.01). The effect was obvious with the increase of dose of astragalus cassia twig. Conclusion:Huangqi Guizhi Wuwutang can alleviate diabetic peripheral neuropathy, and its mechanism may be related to blocking the expression of RAGE on tissue cell surface in AGEs/RAGE/NF-κB signaling pathway, inhibiting the activation of NF-κB and inducing TNF-α triggered oxidative stress and excessive inflammatory response, so as to avoid cell damage and dysfunction.