Characterization of the anti-fatigue effect of Astragali Radix from the Shanxi Hengshan area and its association with taurine metabolism based on 1H NMR metabolomic technology combined with network pharmacology / 药学学报

The objective of this work was to evaluate the anti-fatigue efficacy of Astragali Radix (AR) from the Shanxi Hengshan area and to reveal possible mechanisms by which it relieves fatigue. Efficacy differences between Guangling (GL) and Hunyuan (HY) AR preparations were compared and evaluated, and an 1H NMR metabolomic technique combined with statistical methods was used to identify the metabolites in different groups of mouse gastrocnemius muscle tissues. The differential metabolites after AR treatments were identified according to VIP and P values and the upstream targets were predicted with the help of Metscape. Cytoscape software was utilized to construct a network map of AR potential anti-fatigue targets. Key differential metabolites were identified based on shared targets and entered into the Metaboanalyst website for pathway enrichment analysis, which led to the preliminary elucidation of the molecular mechanisms. The results showed that intervention with AR can significantly improve the swimming-to-exhaustion time, increase liver glycogen, and reduce urea-nitrogen levels in mice. The difference between GL and HY ARs was relatively small, indicating that the quality of AR produced in the Hengshan area is consistent and stable. The metabolic fingerprints of mouse gastrocnemius muscle tissue extracts were composed of 34 metabolites, and the statistical results showed that 19 differential metabolites were significantly reversed after the Hengshan AR intervention. We found that the anti-fatigue effects of AR in the Shanxi Hengshan area were mainly associated with taurine and hypotaurine metabolism through regulation of GAD1, based on network pharmacological analysis. In conclusion, 1H NMR metabolomic techniques were combined with network pharmacology to compare and evaluate the quality of Hengshan ARs, and further associate the fatigue relieve with the regulation of taurine metabolism. This provides a theoretical basis for the resource utilization of Hengshan ARs and the development of anti-fatigue-related products. The animal experiments in this study followed the regulations of the Animal Ethics Committee of Shanxi University and passed the ethical review of animal experiments (Approval No. SXULL2021028).

The key role of gut-liver axis in pyrrolizidine alkaloid-induced hepatotoxicity and enterotoxicity

Pyrrolizidine alkaloids (PAs) are the most common phytotoxins with documented human hepatotoxicity. PAs require metabolic activation by cytochromes P450 to generate toxic intermediates which bind to proteins and form protein adducts, thereby causing cytotoxicity. This study investigated the role of the gut-liver axis in PA intoxication and the underlying mechanisms. We exposed mice to retrorsine (RTS), a representative PA, and for the first time found RTS-induced intestinal epithelium damage and disruption to intestinal barrier function. Using mice with tissue-selective ablation of P450 activity, we found that hepatic P450s, but not intestinal P450s, were essential for PA bioactivation. Besides, in RTS-exposed, bile duct-cannulated rats, we found the liver-derived reactive PA metabolites were transported by bile into the intestine to exert enterotoxicity. The impact of gut-derived pathogenic factors in RTS-induced hepatotoxicity was further studied in mice with dextran sulfate sodium (DSS)-induced chronic colitis. DSS treatment increased the hepatic endotoxin level and depleted hepatic reduced glutathione, thereby suppressing the PA detoxification pathway. Compared to RTS-exposed normal mice, the colitic mice displayed more severe RTS-induced hepatic vasculature damage, fibrosis, and steatosis. Overall, our findings provide the first mode-of-action evidence of PA-induced enterotoxicity and highlight the importance of gut barrier function in PA-induced liver injury.

Hepatic and intestinal biotransformation gene expression and drug disposition in a dextran sulfate sodium-induced colitis mouse model

We examined the impact of gut inflammation on the expression of cytochrome P450 (P450) and other biotransformation genes in male mice using a dextran sulfate sodium (DSS)-induced colitis model. Several P450 isoforms, including CYP1A, CYP2B, CYP2C, and CYP3A, were down-regulated, accompanied by decreases in microsomal metabolism of diclofenac and nifedipine, in the liver and small intestine. The impact of the colitis on clearance of oral drugs varied for four different drugs tested: a small decrease for nifedipine, a relatively large decrease for lovastatin, but no change for pravastatin, and a large decrease in the absorption of cyclosporine A. To further assess the scope of influence of gut inflammation on gene expression, we performed genome-wide expression analysis using RNA-seq, which showed down-regulation of many CYPs, non-CYP phase-I enzymes, phase-II enzymes and transporters, and up-regulation of many other members of these gene families, in both liver and intestine of adult C57BL/6 mice, by DSS-induced colitis. Overall, our results indicate that gut inflammation suppresses the expression of many P450s and other biotransformation genes in the intestine and liver, and alters the pharmacokinetics for some but not all drugs, potentially affecting therapeutic efficacy or causing adverse effects in a drug-specific fashion.

Study on absorbed components of Aconitum kusnezoffii under Yunnan Baiyao compatibility in effect of activating blood circulation and removing blood stasis / 中国中药杂志

Rat model of blood stasis syndrome was prepared by subcutaneous injecting of epinephrine hydrochlorid,then the model rats were administrated by Yunnan Baiyao for 15 days. Blood rheology,coagulation function and histopathology were chosen as indicators to evaluate the successful replication of blood stasis syndrome model and the treatment effect of Yunnan Baiyao. UPLC-Q-TOF-MS was used to rapidly analyze the serum samples of blood stasis syndrome rat after 15 days Yunnan Baiyao treatment,Progenesis QI software was employed to identify the alkaloids components. The results showed that Yunnan Baiyao reduced the plasma viscosity and whole blood viscosity of rats with blood stasis syndrome,prolonged thrombin and prothrombin time,reduced fibrinogen content,and effectively improved pathological state such as inflammatory cell infiltration,blood stasis,congestion and edema of various organs in rats with blood stasis syndrome. Seven alkaloids components from Aconitum kusnezoffii,including karacolidine,senbusine B,isotalatizidine,karakoline,denudatine,talatisamine and chasmanine were found in the rat serum after Yunnan Baiyao treatment. Based on the effectiveness of Yunnan Baiyao in the treatment of blood stasis syndrome induced by epinephrine hydrochloride in rats,alkaloids components from the root of A. kusnezoffii absorbed into blood after Yunnan Baiyao treatment were clarified rapidly and accurately with the help of UPLC-Q-TOF-MS. Karacolidine,senbusine B,isotalatizidine,karakoline,denudatine,talatisamine and chasmanine are the pharmacodynamic material basis of the root of A. kusnezoffii for activating blood circulation and removing blood stasis.

Pericollapse Stage of Osteonecrosis of the Femoral Head: A Last Chance for Joint Preservation / 中华医学杂志(英文版)

<p><b>Objective</b>To propose a new definition of the pericollapse stage of osteonecrosis of the femoral head (ONFH) and review its significance in disease diagnosis and treatment selection.</p><p><b>Data Sources</b>A search for eligible studies was conducted in three electronic databases including PubMed, Cochrane Library, and Embase up to August 10, 2018, using the following keywords: "osteonecrosis", "prognosis", and "treatment".</p><p><b>Study Selection</b>Investigations appraising the clinical signs, symptoms, and imaging manifestations in different stages of ONFH were included. Articles evaluating the prognosis of various joint-preserving procedures were also reviewed.</p><p><b>Results</b>The pericollapse stage refers to a continuous period in the development of ONFH from the occurrence of subchondral fracture to early collapse (<2 mm), possessing specific imaging features that mainly consist of bone marrow edema and joint effusion on magnetic resonance imaging (MRI), crescent signs on X-ray films, and clinical manifestations such as the sudden worsening of hip pain. Accumulating evidence has indicated that these findings may be secondary to the changes after subchondral fractures. Of note, computed tomography provides more information for identifying possible subchondral fractures than does MRI and serves as the most sensitive tool for grading the pericollapse lesion stage. The pericollapse stage may indicate a high possibility of progressive disease but also demonstrates satisfactory long- and medium-term outcomes for joint-preserving techniques. In fact, if the articular surface subsides more than 2 mm, total hip arthroplasty is preferable.</p><p><b>Conclusions</b>The pericollapse stage with distinct clinical and imaging characteristics provides a last good opportunity for the use of joint-preserving techniques. It is necessary to separate the pericollapse stage as an independent state in evaluating the natural progression of ONFH and selecting an appropriate treatment regimen.</p>

Asymptomatic bacteriuria before arthroplasty / 中国组织工程研究

BACKGROUND: Hematogenous spread of urinary tract infection is one of the causes of infection after arthroplasty. The significance of a special portion of urinary tract infection, namely asymptomatic bacteriuria in arthroplasty is little reported. OBJECTIVE: To explore the correlation of preoperative asymptomatic bacteriuria and infection after arthroplasty and evaluate the benefits of screening and treating asymptomatic bacteriuria. METHODS: The researchers searched PubMed, Embase, the Cochrane Library and CNKI databases to collect the clinical studies on asymptomatic bacteriuria before arthroplasty. Then, data extraction, quality assessment of the literature, and result analysis were conducted. RESULTS AND CONCLUSION: Finally, seven eligible articles were included. (1) Preoperative asymptomatic bacteriuria is significantly associated with the occurrence of postoperative prosthetic infection, but it is not a causal relationship. (2) Patients with asymptomatic bacteriuria have a high rate of superficial wound infection but no evidence shows that urinary infection is the direct source of contamination. (3) Asymptomatic bacteriuria is an indicator of increased susceptibility to infection after arthroplasty. (4) Treating asymptomatic bacteriuria before arthroplasty cannot decrease the incidence of various postoperative infectious complications. (5) Current clinical evidence does not support the routine screening and treatment of asymptomatic bacteriuria before arthroplasty.

Development of Multiplex Immunoassay Based on Inductively Coupled Plasma Mass Spectrometry / 分析化学

Immunoassay is of great importance in the field of clinical detection. Our group proposes the inductively coupled plasma mass spectrometry ( ICP-MS) based immunoassay. Multiplex immunoassay can be realized by stable isotope tagging strategy combined with ICP-MS detection. Since then, large amount of researches in this field have been made at home and abroad, and the method has been proved applicable to the analysis of biological small molecules, proteins, nucleic acids and cells. In this article, the history and development of immunoassay, the characters of ICP-MS based immunoassay, single component and multiplex immunoassay, and the magnetic immunoassay that is recently fast developed are reviewed. The prospect of the developing tendency is also anticipated, hoping to provide references for the research in this field.

Effects of simvastatin on expression of Bcl-2 and Bax in myocardium of rats with renal ischemia-reperfusion injury / 中国病理生理杂志

AIM:To observe the effect of simvastatin on myocardial tissue after renal ischemia-reperfusion in-jury and its mechanism .METHODS:A rat model of renal ischemia-reperfusion injury was prepared by clamping the bilat-eral renal arteries for 45 min.The rats (n=36) were randomly divided into sham operation group , renal ischemia-reperfu-sion (I/R) group and simvastatin group with 12 rats in each group.The content of serum creatinine (SCr), blood urea ni-trogen ( BUN) and myocardial tissue malondialdehyde ( MDA) , the myocardial activity of lactate dehydrogenase ( LDH) , creatine kinase (CK) and superoxide dismutase (SOD), and the myocardial protein expression of Bcl-2 and Bax were de-tected.RESULTS:Compared with sham operation group , the content of SCr , BUN and myocardial MDA , and the myo-cardial activity of LDH and CK in I/R group were significantly increased (P<0.05), and the activity of SOD was signifi-cantly decreased (P<0.05).Compared with I/R group, the content of SCr, BUN and myocardial MDA, and the myocar-dial activity of LDH and CK in simvastatin group were significantly decreased ( P<0.05 ) , while SOD activity was en-hanced (P<0.05).The protein expression of Bcl-2 and Bax in sham operation group was less than that in I/R group (P<0.05), and the protein level of Bax in simvastatin group was significantly lower than that in I /R group (P<0.05), while the protein level of Bcl-2 was increased (P<0.05).CONCLUSION:Simvastatin has a protective effect on the my-ocardium of the rats with renal ischemia-reperfusion injury , and the protective mechanism may be related to the elimination of free radicals by simvastatin , increase in the protein expression of Bcl-2 and decrease in the protein expression of Bax .

Corticosteroid-induced Osteonecrosis of the Femoral Head: Detection, Diagnosis, and Treatment in Earlier Stages / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>This review aimed to provide a current recommendation to multidisciplinary physicians for early detection, diagnosis, and treatment of corticosteroid-induced osteonecrosis of the femoral head (ONFH) based on a comprehensive analysis of the clinical literature.</p><p><b>DATA SOURCES</b>For the purpose of collecting potentially eligible articles, we searched for articles in the PubMed, Cochrane Library, Embase, and CNKI databases up to February 2017, using the following key words: "corticosteroid", "osteonecrosis of the femoral head", "risk factors", "diagnosis", "prognosis", and "treatment".</p><p><b>STUDY SELECTION</b>Articles on relationships between corticosteroid and ONFH were selected for this review. Articles on the diagnosis, prognosis, and intervention of earlier-stage ONFH were also reviewed.</p><p><b>RESULTS</b>The incidence of corticosteroid-induced ONFH was associated with high doses of corticosteroids, and underlying diseases in certain predisposed individuals mainly occurred in the first 3 months of corticosteroid prescription. The enhanced awareness and minimized exposure to the established risk factors and earlier definitive diagnosis are essential for the success of joint preservation. When following up patients with ONFH, treatment should be started if necessary. Surgical treatment yielded better results than conservative therapy in earlier-stage ONFH. The ideal purpose of earlier intervention and treatment is permanent preservation of the femoral head without physical restrictions in daily living.</p><p><b>CONCLUSIONS</b>Clinicians should enhance their precaution awareness of corticosteroid-induced ONFH. For high-risk patients, regular follow-up is very important in the 1st year after high-dose prescription of corticosteroids. Patients with suspected ONFH should be referred to orthopedists for diagnosis and treatment in its earlier stage to preserve the joint.</p>

An update on the role of intestinal cytochrome P450 enzymes in drug disposition

Oral administration is the most commonly used route for drug treatment. Intestinal cytochrome P450 (CYP)-mediated metabolism can eliminate a large proportion of some orally administered drugs before they reach systemic circulation, while leaving the passage of other drugs unimpeded. A better understanding of the ability of intestinal P450 enzymes to metabolize various clinical drugs in both humans and preclinical animal species, including the identification of the CYP enzymes expressed, their regulation, and the relative importance of intestinal metabolism compared to hepatic metabolism, is important for improving bioavailability of current drugs and new drugs in development. Here, we briefly review the expression of drug-metabolizing P450 enzymes in the small intestine of humans and several preclinical animal species, and provide an update of the various factors or events that regulate intestinal P450 expression, including a cross talk between the liver and the intestine. We further compare various clinical and preclinical approaches for assessing the impact of intestinal drug metabolism on bioavailability, and discuss the utility of the intestinal epithelium-specific NADPH-cytochrome P450 reductase-null (IECN) mouse as a useful model for studyingroles of intestinal P450 in the disposition of orally administered drugs.

DZNep raises miR-200c expression to delay the invasion and migration of MGC-803 gastric carcinoma cells / 生理学报

The aim of the present study was to investigate the regulatory effects of histone methylation modifications on the expression of miR-200c, as well as invasion and migration of gastric carcinoma cells. Gastric carcinoma cell line, MGC-803, were treated by 2.5 μmol/L histone methyltransferase inhibitor, DZNep. The expression of miR-200c was detected by real-time quantitative PCR (qRT-PCR). The epithelial-mesenchymal transition (EMT) indicators (ZEB1/2 and E/N-cadherin), EZH2, EED, SUZ12 and H3K27me3 expressions were detected by Western blot. Cell migration and invasion abilities were detected by Transwell and scratch tests. The result showed that, compared with DMSO (control) group, DZNep significantly increased the expression of miR-200c to about 2.1 times, inhibited ZEB1, ZEB2, and N-cadherin expressions, and activated E-cadherin expression; Also, DZNep decreased the protein expressions of EZH2, EED, SUZ12 and H3K27me3; Moreover, DZNep could inhibit MGC-803 cell invasive and migrative abilities, as well as MMP9 expression. These results suggest DZNep raises miR-200c expression to delay the invasion and migration of gastric carcinoma cells, and the underlying mechanisms involve the regulations of EMT-related proteins and polycomb repressive complex 2.

Chronic toxicity of a novel recombinant human granulocyte colony-stimulating factor in rats / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To assess the severity and reversibility of the chronic toxicity of a novel recombinant human granulocyte colony-stimulating factor (rhG-CSFa) in rats and the dose-effect relationship.</p><p><b>METHODS</b>A total of 100 Sprague-Dawley rats (equal numbers of male and female) were randomly divided into five groups (20 rats in each group): four groups were treated with rhG-CSFa at 500, 100, 10, 1 µg/kg, respectively, and one group was treated with vehicle only to serve as the control. The rats were received subcutaneous injections of rhG-CSFa or vehicle daily for 13 weeks. During the course of the chronic toxicity study, the physical status, body weight, and food consumption were monitored. Half of the rats in each group (n = 10) were sacrificed after the last rhG-CSFa administration, and the other half were sacrificed at five weeks after the last rhG-CSFa administration. Urinalyses, blood biochemistry, hematological analysis, histopathological examination, and immunological tests were performed for each of the rats.</p><p><b>RESULTS</b>The hematological analyses revealed that the mean white blood cells count, neutrophils count, and neutrophils percentage were increased in male rats at the dose of 10 µg/kg or higher, and these were related with the biological activity of rhG-CSFa. Some small abnormalities were observed in the spleen of a few rats when used highest dose (500 µg/kg, a dosage of 200 folds higher than the normal clinical dosage), but these abnormalities were recovered within 5-week recovery period. No other rhG-CSFa-related abnormalities were observed in this chronic toxicity study.</p><p><b>CONCLUSION</b>No significant toxicity and immunogenicity are observed with rhG-CSFa administration to rats in the chronic toxicity studies.</p>

Construction of recombinant adenovirus vector expressing shRNAs targeting COX-2, AKT1 and PIK3R1 gene and its inhibition effect on proliferation of human gastric adenocarcinoma / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To construct a recombinant adenovirus vector that expresses small hairpin RNAs (shRNA) against COX-2, AKT1 and PIK3R1 gene and to evaluate its potential for suppressing the cell proliferation of human gastric adenocarcinoma SGC701 cell in vitro and in vivo, which will enable the development of a gene therapy protocol for the treatment of human gastric adenocarcinoma.</p><p><b>METHODS</b>Three strips of shRNA targeting AKT1, COX-2 and PIK3R1, was subcloned into adenovirus expression vector. After verification, it was amplified and titered. The recombinant adenovirus expression vector was infected into human gastric adenocarcinoma SGC7901 cells in vitro and the infected cells were injected in nude mice. The mRNA and protein expression levels of AKT1, COX-2 and PIK3R1 were determined by real-time PCR and Western blot respectively. Cell proliferation in vitro was determined by methyl thiazolyltetrazolium (MTT) assay and flow cytometry, tumor growth in vivo was measured by volume of tumor in nude mice.</p><p><b>RESULTS</b>Restriction digestion and sequencing analysis showed that the rAd5-C-A-P adenovirus expression vector was constructed successfully. It significantly inhibited the expression of AKT1, COX-2 and PIK3R1, and cell growth was inhibited over 70% as indicated by MTT assay and accompanied with G0/G1 phase arrest. Cell growth on matrigel matrix showed that the rAd5-C-A-P transfected cells were detached from the matrix or grew in a scattered clustering pattern, indicating poor cell growth activities in 2-D matrigel. Tumor growth in nude mice in the C + A + P group was inhibited (P<0.01).</p><p><b>CONCLUSION</b>shRNA targeting COX-2, AKT1 and PIK3R1 down regulated significantly the expression of the three genes in a sequence-specific manner, exerted proliferation inhibition effect on SGC7901 cells in vitro and in vivo.</p>

Characterization of on-road CO, HC and NO emissions for petrol vehicle fleet in China city / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Vehicle emissions are a major source of air pollution in urban areas. The impact on urban air quality could be reduced if the trends of vehicle emissions are well understood. In the present study, the real-world emissions of vehicles were measured using a remote sensing system at five sites in Hangzhou, China from February 2004 to August 2005. More than 48000 valid gasoline powered vehicle emissions of carbon monoxide (CO), hydrocarbons (HC) and nitrogen oxide (NO) were measured. The results show that petrol vehicle fleet in Hangzhou has considerably high CO emissions, with the average emission concentration of 2.71%+/-0.02%, while HC and NO emissions are relatively lower, with the average emission concentration of (153.72+/-1.16)x10(-6) and (233.53+/-1.80)x10(-6), respectively. Quintile analysis of both average emission concentration and total amount emissions by model year suggests that in-use emission differences between well maintained and badly maintained vehicles are larger than the age-dependent deterioration of emissions. In addition, relatively new high polluting vehicles are the greatest contributors to fleet emissions with, for example, 46.55% of carbon monoxide fleet emissions being produced by the top quintile high emitting vehicles from model years 2000-2004. Therefore, fleet emissions could be significantly reduced if new highly polluting vehicles were subject to effective emissions testing followed by appropriate remedial action.