A novel TNKS/USP25 inhibitor blocks the Wnt pathway to overcome multi-drug resistance in TNKS-overexpressing colorectal cancer

Modulating Tankyrases (TNKS), interactions with USP25 to promote TNKS degradation, rather than inhibiting their enzymatic activities, is emerging as an alternative/specific approach to inhibit the Wnt/β-catenin pathway. Here, we identified UAT-B, a novel neoantimycin analog isolated from Streptomyces conglobatus, as a small-molecule inhibitor of TNKS-USP25 protein-protein interaction (PPI) to overcome multi-drug resistance in colorectal cancer (CRC). The disruption of TNKS-USP25 complex formation by UAT-B led to a significant decrease in TNKS levels, triggering cell apoptosis through modulation of the Wnt/β-catenin pathway. Importantly, UAT-B successfully inhibited the CRC cells growth that harbored high TNKS levels, as demonstrated in various in vitro and in vivo studies utilizing cell line-based and patient-derived xenografts, as well as APCmin/+ spontaneous CRC models. Collectively, these findings suggest that targeting the TNKS-USP25 PPI using a small-molecule inhibitor represents a compelling therapeutic strategy for CRC treatment, and UAT-B emerges as a promising candidate for further preclinical and clinical investigations.

The expression and function of PD-L1 in CD133(+) human liver cancer stem-like cells / 中华肿瘤杂志

Objective: To investigate the expression of programmed death protein-ligand 1 (PD-L1) in liver cancer stem-like cells (LCSLC) and its effect on the characteristics of tumor stem cells and tumor biological function, to explore the upstream signaling pathway regulating PD-L1 expression in LCSLC and the downstream molecular mechanism of PD-L1 regulating stem cell characteristics, also tumor biological functions. Methods: HepG2 was cultured by sphere-formating method to obtain LCSLC. The expressions of CD133 and other stemness markers were detected by flow cytometry, western blot and real-time quantitative polymerase chain reaction (RT-qPCR) were used to detect the expressions of stemness markers and PD-L1. The biological functions of the LCSLC were tested by cell function assays, to confirm that the LCSLC has the characteristics of tumor stem cells. LCSLC was treated with cell signaling pathway inhibitors to identify relevant upstream signaling pathways mediating PD-L1 expression changes. The expression of PD-L1 in LCSLC was down regulated by small interfering RNA (siRNA), the expression of stem cell markers, tumor biological functions of LCSLC, and the changes of cell signaling pathways were detected. Results: Compared with HepG2 cells, the expression rate of CD133 in LCSLC was upregulated [(92.78±6.91)% and (1.40±1.77)%, P<0.001], the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were also higher than those in HepG2 cells (P<0.05), the number of sphere-formating cells increased on day 7 [(395.30±54.05) and (124.70±19.30), P=0.001], cell migration rate increased [(35.41±6.78)% and (10.89±4.34)%, P=0.006], the number of transmembrane cells increased [(75.77±10.85) and (20.00±7.94), P=0.002], the number of cloned cells increased [(120.00±29.51) and (62.67±16.77), P=0.043]. Cell cycle experiments showed that LCSLC had significantly more cells in the G(0)/G(1) phase than those in HepG2 [(54.89±3.27) and (32.36±1.50), P<0.001]. The tumor formation experiment of mice showed that the weight of transplanted tumor in LCSLC group was (1.32±0.17)g, the volume is (1 779.0±200.2) mm(3), were higher than those of HepG2 cell [(0.31±0.06)g and (645.6±154.9)mm(3), P<0.001]. The expression level of PD-L1 protein in LCSLC was 1.88±0.52 and mRNA expression level was 2.53±0.62, both of which were higher than those of HepG2 cells (P<0.05). The expression levels of phosphorylation signal transduction and transcription activation factor 3 (p-STAT3) and p-Akt in LCSLC were higher than those in HepG2 cells (P<0.05). After the expression of p-STAT3 and p-Akt was down-regulated by inhibitor treatment, the expression of PD-L1 was also down-regulated (P<0.05). In contrast, the expression level of phosphorylated extracellular signal-regulated protein kinase 1/2 (p-ERK1/2) in LCSLC was lower than that in HepG2 cells (P<0.01), there was no significant change in PD-L1 expression after down-regulated by inhibitor treatment (P>0.05). After the expression of PD-L1 was knockdown by siRNA, the expressions of CD133, Nanog, Oct4A and Snail in LCSLC were decreased compared with those of siRNA-negative control (NC) group (P<0.05). The number of sphere-formating cells decreased [(45.33±12.01) and (282.00±29.21), P<0.001], the cell migration rate was lower than that in siRNA-NC group [(20.86±2.74)% and (46.73±15.43)%, P=0.046], the number of transmembrane cells decreased [(39.67±1.53) and (102.70±11.59), P=0.001], the number of cloned cells decreased [(57.67±14.57) and (120.70±15.04), P=0.007], the number of cells in G(0)/G(1) phase decreased [(37.68±2.51) and (57.27±0.92), P<0.001], the number of cells in S phase was more than that in siRNA-NC group [(30.78±0.52) and (15.52±0.83), P<0.001]. Tumor formation in mice showed that the tumor weight of shRNA-PD-L1 group was (0.47±0.12)g, the volume is (761.3±221.4)mm(3), were lower than those of shRNA-NC group [(1.57±0.45)g and (1 829.0±218.3)mm(3), P<0.001]. Meanwhile, the expression levels of p-STAT3 and p-Akt in siRNA-PD-L1 group were decreased (P<0.05), while the expression levels of p-ERK1/2 and β-catenin did not change significantly (P>0.05). Conclusion: Elevated PD-L1 expression in CD133(+) LCSLC is crucial to maintain stemness and promotes the tumor biological function of LCSLC.

Research progress on anti-tumor and tumor immunity of marine natural products / 药学学报

In recent years, the oceans have provided an important source of highly promising new anti-tumor drugs for innovation and screening, with approximately 56% of biologically active compounds being discovered to have anti-tumor effects each year. In this study, we classified and summarized the approved drugs of marine origin in terms of anti-tumor therapy, and firstly, we briefly overviewed the role of the immune system in cancer pathogenesis and discussed the current dilemma of cancer immunotherapy and highlighted the main anti-tumor targets of marine drugs. Further, with a focus on tumor immunity, we classified and outlined the history of currently approved marine original drugs by species origin, structural features, relevant pathways, and clinical application and therapy. Lastly, the limitations of current marine drug research were discussed, as well as prospects and trends in new drug development.

Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification / 法医学杂志

OBJECTIVES@#To establish a system for simultaneous detection of miR-888 and miR-891a by droplet digital PCR (ddPCR), and to evaluate its application value in semen identification.@*METHODS@#The hydrolysis probes with different fluorescence modified reporter groups were designed to realize the detection of miR-888 and miR-891a by duplex ddPCR. A total of 75 samples of 5 body fluids (including peripheral blood, menstrual blood, semen, saliva and vaginal secretion) were detected. The difference analysis was conducted by Mann-Whitney U test. The semen differentiation ability of miR-888 and miR-891a was evaluated by ROC curve analysis and the optimal cut-off value was obtained.@*RESULTS@#There was no significant difference between the dual-plex assay and the single assay in this system. The detection sensitivity was up to 0.1 ng total RNA, and the intra- and inter-batch coefficients of variation were less than 15%. The expression levels of miR-888 and miR-891a detected by duplex ddPCR in semen were both higher than those in other body fluids. ROC curve analysis showed that the AUC of miR-888 was 0.976, the optimal cut-off value was 2.250 copies/μL, and the discrimination accuracy was 97.33%; the AUC of miR-891a was 1.000, the optimal cut-off value was 1.100 copies/μL, and the discrimination accuracy was 100%.@*CONCLUSIONS@#In this study, a method for detection of miR-888 and miR-891a by duplex ddPCR was successfully established. The system has good stability and repeatability and can be used for semen identification. Both miR-888 and miR-891a have high ability to identify semen, and the discrimination accuracy of miR-891a is higher.

Effect of Arsenic in Realgar on Rat Ability in Novel Objects Recognition Test / 中国实验方剂学杂志

Objective:To determine the contents of inorganic arsenic（iAs），monomethylarsonic acid（MMA） and dimethylarsinic acid（DMA） in brain tissues and blood by using hydride generation-cold trap-atomic absorptionspectrometry（HG-CT-AAS）， and to explore the toxic effects of Realgar on central nervous system of rats. Method:The 96 Wistar rats were randomly divided into 4 groups：normal control group，0.3，0.9 and 2.7 g·kg^-1 Realgar groups. They then received intragastric administration for 14，28 and 42 days respectively， so a total of 12 groups were formed， with 8 animals in each group. The normal group was given the same dose of sodium carboxymethyl cellulose （CMC-Na） by gavage. The contents of iAs，MMA and DMA in blood and brain tissues were determined by HG-CT-AAS. The novel object recognition test was conducted to observe the learning and memory ability of rats. The changes of hippocampal neuron ultrastructure were observed by transmission electron microscopy. Result:There was no difference in the growth，weight and hippocampal coefficient of the experimental animals. The method of HG-CT-AAS showed a good linearity，precision，accuracy and recovery in content determination of arsenic （at various forms） in rat brain and blood. MMA and DMA were detected in the brain of realgar groups at time-dose-effect relationship. iAs，MMA and DMA were detected in the blood of Realgar groups. The nuclear membrane， mitochondria and endoplasmic reticulum in hippocampus neurons of rats were gradually damaged with the increase of Rhubarb exposure dose and time. After 14 days of exposure to Realgar，compared with the normal control group，there was no significant difference in the novel object recognition index among Realgar groups. After 28 days of exposure，only 2.7 g·kg^-1 Realgar group showed statistically significant difference with the control group （<italic>P</italic><0.05）. After 42 days of exposure， the novel object recognition index of 0.9 and 2.7 g·kg^-1 Realgar groups was significantly lower than that in normal control group（<italic>P</italic><0.05）. Conclusion:The metabolites of Realgar in rats are iAs，MMA and DMA. MMA and DMA can be accumulated in the brain tissue through the blood-brain barrier，causing the decline of the ability of learning and memory and leading to damage of hippocampal neurons.

Cloning，Bioinformatics and Expression Analysis of UGDH1 Gene in Glycyrrhiza uralensis / 中国实验方剂学杂志

Objective:To clone uridine diphosphate （UDP）-glucose dehydrogenase （<italic>UGDH</italic>） gene of <italic>Glycyrrhiza uralensis</italic> and analyze its bioinformatics and expression. Method:Total RNA was extracted from roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， the complementary deoxyribonucleic acid （cDNA） sequence of <italic>GuUGDH</italic>1 gene （Gu was short for <italic>G. uralensis</italic>） was cloned by reverse transcription polymerase chain reaction （RT-PCR）， then sequencing and bioinformatic analysis were performed， and the specificity of the tissue was analyzed by real-time fluorescence quantitative PCR （Real-time PCR）. Result:The open reading frame（ORF）of <italic>GuUGDH</italic>1 gene was 1 443 bp in length and encoded 480 amino acid residues （GenBank accession number of MT968993）. Bioinformatics analysis showed that GuUGDH1 was a stable acidic hydrophilic protein with a relative molecular weight of 53.056 kDa， an isoelectric point of 5.89， no signal peptide and no transmembrane helix， and all of them were outside the membrane. There were three typical conserved domains， which belonged to the UDP-glucose/guanosine diphosphate （GDP）-mannose dehydrogenase family. Phylogenetic analysis showed that the <italic>GuUGDH</italic>1 gene was closely related to <italic>Glycine max</italic> and <italic>Spatholobus suberectus</italic>. The results of Real-time PCR showed that the expression of <italic>GuUGDH</italic>1 gene could be detected in the roots， stems， and leaves of 6-week-old seedlings of <italic>G. uralensis</italic>， and the expression level in the roots was significantly higher than that in the stems and leaves. Conclusion:In this study， the <italic>UGDH</italic>1 gene of <italic>G. uralensis</italic> was cloned and its protein sequence characteristics were systematically analyzed， which can provide theoretical basis for further research on the catalytic function of UGDH1 protein.

Identification of 24 Rana Species Including Rana dybowskii and Rana chensinensis Based on COⅠ Sequences / 中国实验方剂学杂志

Objective:To identify 24 <italic>Rana</italic> species such as <italic>Rana dybowskii</italic> by mitochondrial cytochrome C oxidase subunit I （<italic>CO</italic>Ⅰ） gene-based DNA barcoding and build the neighbour-joining （NJ） tree for hierarchical cluster analysis， so as to provide a basis for the identification and classification of <italic>Rana</italic> species as well as the discovery of new species. Method:<italic>R. dybowskii</italic>， <italic>R. chensinensis</italic>， <italic>R. amurensis</italic>， <italic>R. culaiensi</italic>s， and <italic>R. huanrenesis</italic>， ten for each species， were collected for DNA extraction and polymerase chain reaction （PCR） amplification<italic> </italic>and sequencing. A total of 50 <italic>CO</italic>Ⅰ gene sequences were obtained. Then 163 <italic>CO</italic>Ⅰ gene sequences for 24 species of <italic>Rana</italic> and one <italic>CO</italic>Ⅰ gene sequence for <italic>Pelophylax</italic>，<italic> Odorrana</italic>， <italic>Nidirana</italic>， <italic>Hylarana</italic>， and <italic>Amolops</italic> were harvested from GenBank. After sequence alignment by MEGA X， the parsimony-informative sites of <italic>CO</italic>Ⅰ gene sequences were analyzed and the intraspecific and interspecific genetic distances were calculated， followed by the built of NJ tree and hierarchical cluster analysis. Result:The <italic>CO</italic>Ⅰ gene sequences of 24<italic> Rana</italic> species including <italic>R. dybowskii</italic> were 554 bp in length and there were 210 parsimony-informative sites in total. The intraspecific genetic distance of each species was smaller than 2%. Except that the interspecific genetic distance between <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> was 0.004， the genetic distances between the other species ranged from 0.024 to 0.228. <italic>R. sangzhiensis</italic> and <italic>R. zhengi</italic> were clustered into one branch and some <italic>R. dybowskii</italic> and <italic>R. uenoi</italic> into one branch. There were two separate branches for <italic>R. chensinensis</italic> and the other species were all clustered independently. Conclusion:<italic>CO</italic>Ⅰ-based DNA barcoding enabled the identification of 24 species of <italic>Rana</italic> including <italic>R.dybowskii</italic>. The findings supported that <italic>R. sangzhiensis</italic>， <italic>R. zhengi</italic>， <italic>R. coreana</italic>， and <italic>R. kunyuensis</italic> were the same species. One branch of <italic>R. chensinensis </italic>might be one of the four undownloaded species in Ranidae or a new species. The results have demonstrated that <italic>CO</italic>Ⅰ-based DNA barcoding allows not only the identification of 24 species of Rana including <italic>R. dybowskii </italic>but also the classification of ranidae species and the discovery of new species or subspecies.

Fat distribution in Zhuang-Dong ethnic groups and its relationship with age in China / 解剖学报

Objective To reveal the relationship between lipid distribution and age in Zhuang-Dong ethnic group in China. Methods By bioelectrical impedance analysis method of Zhuang-Dong 13 ethnic body composition in China, u inspection method, for the inspection of body composition differences between the sexes, the indicators by adopting the method of correlation analysis on body composition and age related analysis, variance analysis method was used to explore three body composition differences between age groups. Results The body fat rate was not high and did not reach the obesity level. Half of the men and more than half of the women had fat rates in the standard range. The body fat rate of Dong nationality and Bouyei nationality was higher, but that of Kelao nationality and Kelao nationality was lower. With age, there was no significant change in the upper limb fat rate of males, while the visceral fat rate and trunk fat rate increased, and the lower limb fat rate decreased. There was little change in the total fat rate and the lower limb fat rate. With age, there was no significant change in body mass index (BMI), total fat percentage, trunk fat percentage, and limb fat percentage. Women had a significantly higher percentage of body fat than men. There was no significant correlation between left and right upper limb fat rate and age in males, body fat rate, visceral fat level and trunk fat rate were significantly positively correlated with age, and left and right lower limb fat rate and age were significantly negatively correlated. The left upper limb fat rate, left and right lower limb fat rate were negatively correlated with age, and the trunk fat rate was positively correlated with age. There was no significant correlation between age and total female lipid. Conclusion The body fat of Zhuang-Dong ethnic group in China is much thinner than that of north Asian ethnic group, and it has the characteristic of sebum development level of southern Chinese ethnic group.

Countermeasures and treatment for aortic acute syndrome with novel coronavirus pneumonia / 中华外科杂志

The novel coronavirus pneumonia (NCP) has cost a great loss to the health and economic property of Chines people. Under such a special circumstance, how to deal with such patients with acute aortic syndrome has become a serious challenge. Rapid diagnosis of concomitant NCP, safe and effective transportation, implementation of the interventional procedure, protection of vascular surgical team and postoperative management and follow-up of such patients have become urgent problems for us. Combined with the latest novel government documents, the literature and the experiences from Wuhan, we answered the above questions briefly and plainly. It also hopes to inspire the national vascular surgeons to manage critical emergencies in vascular surgery and even routine vascular diseases with NCP, as a final point to limit the severe epidemic situation, and minimize the damage of NCP.

Expression of microRNA-495-5p in preterm infants with bronchopulmonary dysplasia: a bioinformatics analysis / 中国当代儿科杂志

OBJECTIVE@#To study the expression of microRNA-495-5p (miRNA-495-5p) in the serum of preterm infants with bronchopulmonary dysplasia (BPD) based on a bioinformatics analysis, and to provide a theoretical basis for further research on the association between miRNA-495-5p and BPD.@*METHODS@#A total of 40 preterm infants who were admitted to the neonatal intensive care unit from January 2015 to December 2016 were enrolled. Among these infants, 20 with early clinical manifestations of BPD were enrolled as the BPD group, and 20 without such manifestations were enrolled as the control group. Peripheral blood samples were collected. The miRNA microarray technique was used to screen out differentially expressed miRNAs in serum between the two groups. RT-PCR was used for validation of results. TargetScan, miRDB, and miRWalk databases were used to predict the target genes of miRNA-495-5p. The DAVID database was used to perform gene ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the target genes.@*RESULTS@#Compared with the control group, the BPD group had a significant increase in the expression of miRNA-495-5p in serum (P<0.05). A total of 117 target genes of miRNA-495-5p were predicted by the above three databases and they were involved in several molecular functions (including transcriptional regulatory activity, transcriptional activation activity, and transcription cofactor activity), biological processes (such as metabolic regulation, DNA-dependent transcriptional regulation, and vascular pattern), and cell components (including nucleoplasm, membrane components, and insoluble components) (P<0.05). As for signaling pathways, these genes were significantly enriched in the mTOR signaling pathway (P<0.05).@*CONCLUSIONS@#MiRNA-495-5p may be involved in the development and progression of BPD by regulating angiogenesis, stem cell differentiation, apoptosis, and autophagy, which provides clues for further research on the role and functional mechanism of miRNA-495-5p in BPD.

Effect of edaravone combined with early enteral nutrition on intestinal mucosal barrier function and inflammatory factors in patients with severe craniocerebral trauma / 中国基层医药

Objective:To investigate the effect of edaravone combined with early enteral nutrition on intestinal mucosal barrier function and inflammatory factors in patients with severe craniocerebral trauma.Methods:From January 2017 to December 2018, 80 patients with severe craniocerebral trauma admitted to Lanxi People's Hospital were divided into observation group(40 cases) and control group(40 cases) according to the random digital table method.The control group was given early enteral nutrition treatment, and the observation group was treated with edaravone on the basis of the control group.The course of treatment in both two groups was 4 weeks.Glasgow coma index(GCS), intracranial pressure, NIHSS, intestinal mucosal barrier function and inflammatory factors before and after treatment were compared between the two groups.Results:The GCS score of the observation group[(12.32±1.02)points] was higher than that of the control group[(9.87±1.45)points], while the intracranial pressure[(169.84±10.19)mmH 2O] and NIHSS score[(10.73±1.98)points]of the observation group were lower than those of the control group [(203.24±15.69)mmH 2O and (16.52±3.07)points], the differences were statistically significant( t=8.740, 11.291, 10.024, all P<0.05). The levels of DAO [(0.64±0.12)U/L], endotoxin [(2.54±0.48)U/mL] and D-lactate [(3.64±1.09)μg/L] in the observation group were lower than those in the control group [(1.32±0.30)U/L, (3.64±0.61)U/mL and (5.73±1.18)μg/L] ( t=13.310, 8.963, 8.229, all P<0.05). The levels of IL-6[(27.39±5.64)pg/L], hs-CRP[(10.38±3.24)mg/L] and TNF-α[(7.83±1.79)μg/L] in the observation group were lower than those in the control group [(39.98±9.97)pg/L, (15.64±3.19)mg/L and (13.24±3.21)μg/L] ( t=6.951, 7.317, 9.310, all P<0.05). The good prognosis rate of the observation group(80.00%) was higher than that of the control group(57.00%)(χ 2=4.713, P<0.05). Conclusion:Edaravone combined with early enteral nutrition has good effect on the patients with severe craniocerebral trauma.It can improve the intestinal mucosal barrier function, reduce the inflammatory response and improve the prognosis.

Effect of MPST in the treatment of hypertensive intracerebral hemorrhage and its influence on the expression of MMP-9, IL-8 and NSE / 中国基层医药

Objective:To investigate the effect of microinvasive craniopuncture scavenging technique(MPST) in the treatment of hypertensive intracerebral hemorrhage, and its influence on the expression of serum matrix metalloproteinase-9 (MMP-9), interleukin-8 (IL-8) and neuron specific enolase (NSE).Methods:From January 2017 to December 2018, 60 cases with hypertensive cerebral hemorrhage in Lanxi People's Hospital of Zhejiang province were divided into observation group (30 cases) and control group (30 cases) according to the digital table method.The control group was treated with conventional therapy, while the observation group was treated with MPST on the basis of the control group.The therapeutic effects of the two groups were compared, and the changes of Barthel index (BI) score, NIHSS score, perihematoma edema, MMP-9, IL-8 and NSE levels were compared.Results:The total effective rate in the observation group(93.33%) was higher than that in the control group (70.00%) (χ 2=5.455, P<0.05). The BI score[(60.19±5.87)points] of the observation group was higher than that of the control group[(49.83±4.56)points], while the NIHSS score[(7.93±1.42)points] was lower than that of the control group[(12.87±2.10)points]( t=7.634, 10.673, all P<0.05). The amount of edema around hematoma in the observation group [(6.20±1.27)mL] was lower than that in the control group [(9.83±1.76)mL] ( t=9.161, P<0.05). The levels of MMP-9 [(103.24±17.38)μg/L], IL-8 [(137.28±25.46)μg/L] and NSE [(8.98±2.16)μg/L] in the observation group were lower than those in the control group [(168.39±15.42)μg/L, (195.31±39.71)μg/L and (13.13±2.63)μg/L] ( t=15.358, 6.738 and 6.679, all P<0.05). Conclusion:MPST is effective in the treatment of hypertensive intracerebral hemorrhage, and it can reduce the serum levels of MMP-9, IL-8 and NSE.

Four new polyhydroxylated steroids from the South Sea sponge Plakortis sp / 中国天然药物

Four new polyhydroxylated steroids plaksterols A-D (1-4), together with two known related steroids ergost-7,9(11),22-trien-3β,5α,6α-triol (5) and ergosta-6β-methoxy-7,22-diene-3β,5α-diol (6), were isolated from methanol extract of the South China Sea marine sponge Plakortis sp. Their structures were identified by spectroscopic analysis, including NMR, MS, and IR. The cytotoxicity of the polyhydroxylated steroids were evaluated, and compound 6 showed moderate inhibitory activities against K562, HL-60 and BEL-7402 cells.

Identification of Peripheral Blood and Menstrual Blood Based on the Expression Level of MicroRNAs and Discriminant Analysis / 法医学杂志

Objective To construct a discriminant analysis model based on the differential expression of multiple microRNAs （miRNAs） in two kinds of blood samples （peripheral blood and menstrual blood） and three non-blood samples （saliva, semen and vaginal secretion）, to form an identification solution for peripheral blood and menstrual blood. Methods Six kinds of miRNA （miR-451a, miR-144-3p, miR-144-5p, miR-214-3p, miR-203-3p and miR-205-5p） were selected from literature, the samples of five kinds of body fluids commonly seen in forensic practice （peripheral blood, menstrual blood, saliva, semen, vaginal secretion） were collected, then the samples were divided into training set and testing set and detected by SYBR Green real-time qPCR. A discriminant analysis model was set up based on the expression data of training set and the expression data of testing set was used to examine the accuracy of the model. Results A discriminant analysis statistical model that could distinguish blood samples from non-blood samples and distinguish peripheral blood samples from menstrual blood samples at the same time was successfully constructed. The identification accuracy of the model was over 99%. Conclusion This study provides a scientific and accurate identification strategy for forensic fluid identification of peripheral blood and menstrual blood samples and could be used in forensic practice.

Research Progress on MicroRNA in Forensic Medicine as Molecular Markers / 法医学杂志

MicroRNA （miRNA） belongs to a class of endogenous non-coding small RNA molecules with a length of 18-24 nucleotides. The expression of miRNA is highly conservative, has time sequence and is highly tissue-specific. MiRNA could not be easily degraded by ribonuclease, and is resistant to changes in environmental factors such as temperature and pH value. Moreover, miRNA can even be detected in corrupt tissue. As a result, miRNA has broad application prospects in many fields of forensic medicine such as source identification of body fluid and estimation of cause of death. This article briefly summarizes the application of miRNA in forensic practice, such as body fluid identification, determination of postmortem interval and cause of death analysis.

Triterpenoids from Galbanumofuygur medicine andtheir anticholinesterase activities / 药学学报

Eight triterpenes were isolated from the methanol extract of Galbanum by various chromatographic methods including silica gel, ODS opening column, recrystallization and semi-preparative HPLC. Their structures were determined by spectroscopic methods and physicochemical properties as 3β,19α,21α-trihydroxyl-12-en-28-oic acid (1), sumaresinolic acid (2), 3β,19α-dihydroxyl-12-en-28-oic acid (3), oleanolic acid (4), 3β,6β,19α-trihydroxyl-12-en-28-oic acid (5), 19α-hydroxy oleanonic acid (6), 6α-hydroxy oleanonic acid (7), and (11R,12R)-3α,6α-dihydroxy-epoxyolean-28α,13α-olide (8). Among them, compound 1 is a new compound, while compounds 2-8 were newly isolated from the Apiaceae family. The ability of compounds 1-8 to inhibit cholinesterase was determined with an improved Ellman method. Compound 1 showed strong inhibitory activity against butyrylcholinesterase. The molecular docking results indicated that Trp82, His438, Phe329 and Ala328 played an important role in the binding of compound 1 to butyrylcholinesterase.

Effect of long-term intake of ginseng extracts on gut microbiota in rats / 中国中药杂志

As a traditional Chinese medicine in China, ginseng has a wide range of medicinal and health value. At present, the nutritional value of ginseng as a medicinal food has been a hotspot in studies. Intestinal flora plays an important role in the organism, which has been confirmed by many researchers. In order to find out the effect of long-term intake of ginseng extracts on the gut microbiota structure of rats, MiSeq sequencing platform was applied in macro gene sequencing of cecal contents in the long-term use of ginseng extracts modelin rats. According to the findings, after long-term administration with ginseng extracts, probiotics such as Bifidobacterium, Lactobacillus, Allobaculum and Clostridium, in the intestinal flora of rats were significantly increased, suggesting that long-term intake of ginseng extracts could facilitate the growth of probiotics. Meanwhile, some pathogenic bacteria, such as Butyricimonas, Parabacteroides, Alistipes, Helicobacter, were significantly down-regulated, indicating that long-term intake of ginseng extracts may have a positive effect in inhibiting the colonization of pathogenic bacteria. In conclusion, this study provided an important basis for the research on the effect of long-term use of ginseng extracts on the intestinal flora of rats.

Increased Endogenous Sulfur Dioxide Involved in the Pathogenesis of Postural Tachycardia Syndrome in Children: A Case-Control Study / 中华医学杂志(英文版)

<p><b>Background</b>The pathogenesis of postural tachycardia syndrome (POTS) remains unclear. This study aimed to explore the changes and significance of sulfur dioxide (SO) in patients with POTS.</p><p><b>Methods</b>The study included 31 children with POTS and 27 healthy children from Peking University First Hospital between December 2013 and October 2015. A detailed medical history, physical examination results, and demographic characteristics were collected. Hemodynamics was recorded and the plasma SOwas determined.</p><p><b>Results</b>The plasma SOwas significantly higher in POTS children compared to healthy children (64.0 ± 20.8 μmol/L vs. 27.2 ± 9.6 μmol/L, respectively, P < 0.05). The symptom scores in POTS were positively correlated with plasma SOlevels (r = 0.398, P < 0.05). In all the study participants, the maximum heart rate (HR) was positively correlated with plasma levels of SO(r = 0.679, P < 0.01). The change in systolic blood pressure from the supine to upright (ΔSBP) in POTS group was smaller than that in the control group (P < 0.05). The ΔSBP was negatively correlated with baseline plasma SOlevels in all participants (r = -0.28, P < 0.05). In the control group, ΔSBP was positively correlated with the plasma levels of SO(r = 0.487, P < 0.01). The change in HR from the supine to upright in POTS was obvious compared to that of the control group. The area under curve was 0.967 (95% confidence interval: 0.928-1.000), and the cutoff value of plasma SOlevel >38.17 μmol/L yielded a sensitivity of 90.3% and a specificity of 92.6% for predicting the diagnosis of POTS.</p><p><b>Conclusions</b>Increased endogenous SOlevels might be involved in the pathogenesis of POTS.</p>

Expression and role of programmed cell death ligand-1 in sorafenib-resistant hepatocellular carcinoma cells / 第二军医大学学报

Objective To explore the expression and role of programmed cell death ligand-1 (PD-L1) in sorafenib-resistant human hepatocellular carcinoma cells. Methods Sorafenib-resistant human hepatocellular carcinoma cell lines (Hep3B-SR, HepG2-SR) were established by the procedure of stepwise increase in sorafenib concentrations. CCK-8 assay was used to detect half inhibition concentration (IC50). The expressions of P-glycoprotein (P-gp), multidrug resistance-associated protein 1 (MRP1) and PD-L1 were examined by Western blotting and qPCR. The expression of PD-L1 was silenced by transfecting siRNA into the drug-resistant cells, and the silencing efficiency was tested. After silencing the expression of PD-L1 in the drug-resistant cells, CCK-8 assay was used to detect IC50, the drug-resistance index was calculated, and the expressions of P-gp and MRP1 were examined. Then cell proliferation assay, wound healing assay, colony formation assay and flow cytometry were applied to examine cell proliferation, migration, clone formation and apoptosis, respectively. Results The drug-resistance indexes of Hep3B-SR and HepG2-SR were 5.4 and 5.2, respectively. The expressions of P-gp, MRP1 and PD-L1 in the drug-resistant cells were significantly up-regulated in comparison with the parental cells (all P0.01). After inhibiting the expression of PD-L1, the drug-resistance indexes of Hep3B-SR and HepG2-SR decreased to 1.8 and 1.5, respectively, and the expressions of P-gp and MRP1 were down-regulated (all P0.01). Silencing the PD-L1 expression could significantly inhibit the proliferation, migration and colony formation of drug-resistant cells, and could significantly promote the apoptosis (all P0.01); and these effects were strengthened by combining sorafenib. Conclusion PD-L1 is highly expressed in sorafenib-resistant hepatocellular carcinoma cells. Inhibition of PD-L1 expression can partially reverse the drug-resistance of the cells and significantly enhance the anticancer effect of sorafenib. Inhibiting the expression of PD-L1 can effectively repress the growth of sorafenib-resistant hepatocellular carcinoma cells, which is more if combining with sorafenib.

Effects of acetylated HMGB1 in rats with sepsis associated encephalopathy / 临床麻醉学杂志

Objective To investigate the effects of acetylated HMGB1 on cognitive function in rats with sepsis associated encephalopathy (SAE) and the effect of HMGB1 inhibitor.Methods Forty-eight males mice were randomly assigned to three groups (n=16): sham group (group S),cecal ligation puncture group (group C),cecal ligation puncture+sodium butyrate group (group B).Cecal ligation puncture was applied to establish the SAE model,and group S received sham operation.Rats in groups S and C were injected with normal saline 5 ml/kg 30 min and 4 h after CLP,respectively.The rats in group B were intraperitoneally injected with sodium butyrate 500 mg/kg 0.5 h and 4 h after CLP,respectively.All animals were performed Morris water maze test on 4th day after operation,and the exploring time of space exploration experiments were assessed on 7th day after CLP surgery.IL-6,BDNF,HMGB1 and acetylated HMGB1 expression in hippocampus of all rats were determined by Western Blot.Results Compared with group S,the latency of rats in group C was longer and the exploring time was shorter (P<0.05).Compared with group C,the latency of rats in group B was shorter and the exploring time was longer (P<0.05).Compared with group S,the expression of IL-6,HMGB1 and acetylated HMGB1 in group C increased (P<0.05) and the level of BDNF decreased (P<0.05).Compared with group C,the expression of IL-6,HMGB1 and acetylated HMGB1 in group B decreased (P<0.05) and the level of BDNF increased (P<0.05).Conclusion HMGB1 inhibitor sodium butyrate can inhibit the expression of acetylated HMGB1 in the hippocampus of SAE rats,and reduce the cognitive impairment induced by sepsis.