Dryness-alleviating effect of processed Aurantii Fructus from Lingnan based on correlation of fecal metabolomics and dryness / 中国中药杂志

With the approach of untargeted metabolomics and correlation analysis, this study aimed to explore the mechanism of Aurantii Fructus from Lingnan region in alleviating dryness by analyzing the different effects of raw Aurantii Fructus(RAF) and processed Aurantii Fructus(PAF) on fecal endogenous metabolism in normal rats. Eighteen Sprague-Dawley(SD) rats were randomly divided into a control group(C), an RAF group(10 g·kg~(-1)), and a PAF group(10 g·kg~(-1)). After seven days of administration, the effects of RAF and PAF on dryness-related indexes were compared, including water intake, fecal water content, salivary secretion, the expression of AQP5, VIP, and 5-HT in the submandibular gland, as well as the expression of AQP3, VIP, and 5-HT in the colon. The fecal samples in each group were determined by LC-MS. Multivariate statistical analysis and Pearson correlation coefficient were used for screening the differential metabolites and metabolic pathways in alleviating dryness of RAF. The results indicated that both RAF and PAF showed certain dryness, and the dryness of RAF was more significant. Moreover, PAF could alleviate dryness of RAF to a certain extent by reducing the water intake, fecal water content, and the expression of AQP3, VIP, and 5-HT in the colon and increasing the salivary secretion and the levels of AQP5, VIP, and 5-HT in the submandibular gland. According to the analysis of fecal metabolomics, 99 and 58 metabolites related to dryness were found in RAF and PAF respectively, where 16 of them played an important role in alleviating dryness of RAF. Pathway analysis revealed that the mechanism of PAF in alleviating dryness of RAF was presumably related to the regulation of riboflavin metabolism, purine metabolism, arginine biosynthesis, pyrimidine metabolism, alanine metabolism, aspartate metabolism, glutamate metabolism, and retinol metabolism pathways. This study suggested that PAF might alleviate dryness of RAF by affecting the metabolic levels of the body, which provides a new basis for further clarifying the processing mechanism of PAF.

One case of bloodstream infection caused by Ureaplasma urealyticum / 中国热带医学

@#Abstract: Objective To analyze a case of bloodstream infection caused by Ureaplasma urealyticum after abortion in Anxi County Hospital, so as to provide basis for the clinical diagnosis and treatment. Methods　The diagnosis of Ureaplasma urealyticum in this patient with bloodstream infection was retrospectively analyzed. The basic clinical data and laboratory diagnosis data were collected, including the characteristics of blood culture curve, Wright staining of culture medium, drug sensitivity of Mycoplasma liquid identification, colony characteristics of solid medium, and the conclusion of targeted DNA sequencing. Through the comprehensive analysis of the above data, the rapid diagnosis of this case can be realized by optimizing the detection and diagnosis process. Results　The clinical manifestations of this patient were fever of 38.5 ℃, CRP:14.85 mg/L, WBC:14.33×109/L, NET: 85.40%, PCT: 0.12 ng/mL, IL-6: 665.6 pg/mL, positive after 3 days of blood culture, no bacteria were found in Gram stain, and sand-like purple bacteria were observed after adding Wright's stain. After inoculation in blood agar, Mycoplasma solid and liquid medium, no colonies were grown in blood agar, after 48 h and 5 d. On Mycoplasma A7 agar, the edge of brown fried egg colony was striature, and it could be identified as Ureaplasma urealyticum with the Mycoplasma ID & AST panel, which was resistant to quinolones and spectinomycin, but sensitive to macrolides, tetracyclines and lincomycin. Subsequent targeted DNA sequencing results were also confirmed for Ureaplasma urealyticum. Before receiving the report, clinical experience treatment with ceftriaxone metronidazole was used to fight infection with negative bacilli and anaerobic bacteria. Mycoplasma was not treated with targeted treatment. After 3 days, the patient's body temperature returned to normal, inflammation index decreased, and the patient asked to be discharged. Conclusions　At present, there are few reports of bloodstream infection caused by Ureaplasma urealyticum, and the lack of clinical understanding can easily lead to misdiagnosis and missed diagnosis. In order to improve the detection rate of Mycoplasma in blood culture, it is necessary to optimize the detection procedure of blood culture and provide accurate diagnosis and treatment basis for clinical practice. However, it is clear from this case that Mycoplasma bloodstream infection cases are self-limited infection and can recover by themselves without targeted treatment in patients with normal immunity. Therefore, it is very important to protect the immunity of patients.

Catalpol inhibits cell apoptosis through alleviating endoplasmic reticulum stress in nonalcoholic fatty liver disease / 药学学报

This study was designed to explore the protective effect and underlying mechanism of catalpol on hepatocyte apoptosis in nonalcoholic fatty liver disease (NAFLD). High fat diet (HFD) was used to establish NAFLD model in the in vivo experiment, and the procedures of the experiments and animal care protocol were approved by the Animal Care and Use Committee of Jianghan University. Human liver cancer cell line HepG2 was treated with palmitate (PA) to establish a lipid toxicity model in the in vitro experiments. The results showed that catalpol significantly decreased the contents of serum total glyceride (TG), total cholesterol (TC), alanine aminotransferase (ALT), and aspartate transaminase (AST) in HFD-fed mice. Results of TUNEL staining and flow cytometry analyses revealed that catalpol significantly inhibited hepatocytes apoptosis in HFD-fed mice and PA-treated HepG2 cells. Moreover, catalpol treatment significantly reduced the endoplasmic reticulum stress-related protein expression levels of binding immunoglobulin protein (BiP), phosphorylated PKR-like endoplasmic reticulum kinase (p-PERK), inositol-requiring kinase 1α (IRE1α), and transcriptional factor activating transcription factor 6 (ATF6), and apoptosis-related protein expression levels of C/EBP homology protein (CHOP), phosphorylated c-Jun N-terminal kinase (p-JNK), and cleaved cysteinyl aspartate specific proteinases (caspases)-12, -9, and -3 in HFD-fed mice and PA-treated HepG2 cells. Furthermore, endoplasmic reticulum stress agonist tunicamycin (TM) significantly reversed the inhibitory effect of catalpol on protein expression levels of BiP, p-PERK, IRE1α, and ATF6, subsequently the inhibitory effect of catalpol on expression levels of CHOP, p-JNK, Bcl-2, Bax, and cleaved caspases (-12, -9, and -3) was also attenuated in PA-treated HepG2 cells. Taken together, these findings demonstrated that catalpol could inhibit hepatocytes apoptosis and had a significant protective effect on liver injury, and its mechanism might be related to the relief of endoplasmic reticulum stress.

Advances in Immunotherapy for Drug Addiction / 中国医学科学院学报

Drug addiction is a major worldwide medical and social problem.Cocaine,nicotine,methamphetamine,heroin and other psychoactive substances,with small molecular weight,can easily cross the blood-brain barrier and eventually lead to addiction and other serious neuropsychological damage.There is no effective cure for addiction currently.The drug-antibody complex formed on the basis of active or passive immunotherapy could not cross the blood-brain barrier,which reduces the concentration of the free active drug and prevents its distribution in the brain,thereby weakening the drug addiction-related reward effects.It provides a promising way for the treatment of drug addiction.This article reviews the progress of immunotherapy against psychoactive substances such as cocaine,nicotine,methamphetamine and heroin in the past 50 years from the aspects of active immunity,passive immunity,drug metabolism-related enzymes,adjuvants and so on.The goal is to provide some ideas for the development of agents for the treatment of psychoactive substance addiction.

Roles of G Protein-gated Inward Rectifier Potassium Channels in Substance Addiction / 中国医学科学院学报

G protein-gated inward rectifier potassium(GIRK)channels are widely distributed in the central nervous system and play important roles in maintaining the resting membrane potential of neurons,adjusting neuronal excitability,and regulating the release of neurotransmitter.Studies have shown that addictive behavior is closely related to the expression and activity of the GIRK channels in the brain reward system and the GIRK channels may be a potential target for addiction treatment.This article summarizes the recent research advances in GIRK channels in terms of structure,intracranial tissue distribution,and especially substance addiction.

Intervention of Catalpol on High-fat Diet Induced Nonalcoholic Fatty Liver Disease in Mice / 中国医学科学院学报

To investigate the effect of catalpol on high-fat diet(HFD)-induced nonalcoholic fatty liver disease(NAFLD)and its underlying molecular mechanisms. Sixty C57BL/6J male mice were randomly divided into six groups:control group;HFD group;HFD+catalpol(100 mg/kg)group;HFD+catalpol(200 mg/kg)group;HFD+catalpol(400 mg/kg)group;and HFD+atorvastatin calcium(ATC)(30 mg/kg)group.The control group was fed a normal diet containing 4.4 kJ/g fat,whereas the other five groups were fed a high-fat diet containing 19.8 kJ/g fat.Mice in the catalpol or ATC treatment groups were administered by gavage for different doses of catalpol or ATC,whereas other mice were treated with saline.Body weight was measured once a week.Experiments were terminated after 18 weeks,and blood and liver samples were collected after an overnight fast(12 hours)for analysis.The body weight and liver weight were measured and the levels of serum total cholesterol(TC),triglyceride(TG),high-density lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),and aspartate transaminase(AST)as well as inflammatory factors tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,and IL-6 were determined by commercially available kits.Liver sections were stained with Oil Red O and HE to investigate the lipid accumulation and histopathological changes.The protein expressions of nuclear factor kappa-B(NF-κB)p65,inhibitor of nuclear factor kappa-B α(IκBα),B-cell lymphoma-2(Bcl-2),Bcl-2 associated x protein(Bax),and Caspase-3 were determined by Western blot. Compared to the model group,the body weight gains(all =0.001),liver index(=0.008,=0.001,=0.001),ALT(=0.004,=0.001,=0.001),and AST(=0.008,=0.001,=0.001)were significantly decreased in catalpol treatment groups,and the serum levels of TC(=0.005,=0.001),TG (all =0.001),and LDL-C(all =0.001)were also significantly decreased in middle and high dose groups,and the serum level of HDL-C was significantly increased in high group(=0.009).Moreover,compared to the model group,the degree of liver injury and lipid accumulation were obviously decreased in the catalpol treatment groups according to the pathology.Similarly,the release of inflammatory factors was significantly inhibited by the treatment with catalpol.The results of Western blot showed that the protein levels of NF-κB p65(=0.014,=0.001,=0.001)and Caspase-3(all =0.001)in the livers of HFD-fed mice were significantly reduced by catalpol treatment.In addition,the protein level of IκBα(=0.028,=0.001,=0.001)and the ratio of Bcl-2/Bax in high dose group(=0.003)was increased by treatment with catalpol. Catalpol can effectively improve the body weight gains,liver index,dyslipidemia,and lipid accumulation in HFD-fed mice and inhibit the release of inflammatory factors and hepatocyte apoptosis,thereby preventing the development of NAFLD induced by HFD.

Effect of Ginsenoside Rg2 and Its Stereoisomers on Oxygen-Glucose Deprivation and Reperfusion Induced Cortical Neuronal Injury Model / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the effect of natural type ginsenoside Rg2 (Rg2) and its stereoisomers [20 (R)-Rg2 and 20 (S)-Rg2] at different concentrations on oxygen-glucose deprivation/ reperfusion (OGD/R) induced cortical neuronal injury model in vitro, and to explore the mechanism, and compare their differences of action.</p><p><b>METHODS</b>Cortical neurons after 7-day culture were randomly divided into 5 groups, i.e., the control group, the model group, the Rg2 group, 20 (R) -Rg2 group, and 20 (S) - Rg2 group. Cortical neurons in the Rg2 group, 20 (R)-Rg2 group, and 20(S)-Rg2 group were pretreated with 20, 40, and 80 μmol/L Rg2, 20 (R) -Rg2, and 20 (S) -Rg2 for 24 h to prepare OGD/R model. The cell survival rate, the activity of Caspase-3, the intracellular Ca2+ concentration, contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected 24 h later.</p><p><b>RESULTS</b>Compared with the control group, cell survival rates and activities of SOD obviously decreased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly increased with statistical difference (P < 0.05). Compared with the model group, cell survival rates and activities of SOD obviously increased, the activity of Caspase-3, Ca2+ fluorescent optical gray value, and contents of MDA significantly decreased in 20 μmol/L Rg2 group, 40 μmol/L 20 (R) -Rg2 group, and 80 μmol/L 20 (S) -Rg2 group (P < 0.05). Compared with 20(S)-Rg2 group, cell survival rates increased and contents of MDA significantly decreased in 20, 40, and 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). The activity of Caspase-3 decreased and contents of SOD increased in 80 μmol/L 20 (R)-Rg2 group, and 40, 80 μmol/L Rg2 groups (P < 0.05). Ca2+ fluorescent optical gray value decreased in 40, 80 μmol/L Rg2 and 20 (R)-Rg2 groups (P < 0.05). Compared with 20 (R)-Rg2 group, Ca2+ fluorescent optical gray value decreased in 80 μmol/L Rg2 group (P < 0.05); contents of SOD increased in 40 and 80 μmol/L Rg2 groups (P < 0.05); contents of MDA decreased in 20, 40, and 80 μmol/L Rg2 groups (P < 0.05).</p><p><b>CONCLUSIONS</b>Rg2 and its stereoisomers could improve cell vitality of cortical neurons against OGD/R induced injury. This might be related to improving anti-apoptotic capacities and antioxidant abilities, and reducing Ca2+ inflow. Besides, the neuroprotective effect of 20 (R) -Rg2 was better than that of 20 (S) -Rg2, but inferior to that of Rg2.</p>

Effect of P2X7R agonist BzATP on cell growth and apoptosis in non-small cell lung cancer A549 cells / 国际肿瘤学杂志

Objective To investigate the expression of P2X7 receptor (P2X7R) and the effect of P2X7R agonist 2'-3'-O-(4-benzoyl-benzoyl) ethane adenosine triphosphate three amine salt (BzATP) on cell growth and apoptosis in non-small cell lung cancer A549 cells,and to explore the related mechanism.Methods The expression of P2X7R in A549 cells was detected by immunofluorescence.Cells were treated with different concentrations (150,300,600 μmol/L) of BzATP.Cells untreated with BzATP were used as control group.3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide (MTF) assay and Hoest33342 staining were respectively used to detect cell viability and apoptosis.Enzyme-linked immunosorbent assay (ELISA) was uesd to detect the concentration of tumor necrosis factor-α (TNF-α) of cell culture supernatants.The expressions of nuclear factor-κB (NF-κB) p65,inhibitor of α of NF-κB (IκBα) and the phosphorylation of inhibitor of α of NF-κB (phospho-IκBα) were detected by Western blotting.Results P2X7R was expressed on the cell membrane of A549 cells.Survival rate of A549 cell was significantly decreased with the concentrations of BzATP at 300 and 600 μmol/L [(67.87 ± 8.98) ％,(44.73 ± 6.92) ％],compared with the control group (98.60 ± 1.44) ％,the differences were statistically significant (t =4.481,P =0.027;t =3.920,P =0.038).BzATP promoted apoptosis,and increased the concentration of TNF-α of supernatant at 300 and 600 μmol/L [(57.35 ±6.41) pg/ml,(78.63 ± 11.33) pg/ml],compared with the control group (42.56 ±0.37) pg/ml,the differences were statistically significant (t =6.410,P =0.035;t =11.330,P =0.005).In addation,the expressions of NF-κB p65 and IκBα were respectively downregulated and upregulated by BzATP,while the expression of phospho-IκBα was not significantly altered.Conclusion P2X7R is expressed on A549 cell membrane.BzATP can inhibit cell proliferation and induce the apoptosis of A549 cells,and the mechanism of action may be related to promoting the release of TNF-α and inhibition of NF-κB pathway.

Research progress on ion channels in human glioma / 肿瘤研究与临床

Glioma is a serious threat to human health.Increasing evidences indicate that abnormal expression and activities of a number of ion channels,e.g.voltage-gated K+,Ca2+,C1-channels,are involved in the growth,proliferation,migration and invasion of glioma cells.Ion channels may affect the occurrence and development of glioma cells through regulating the membrane potential,adjusting the cytoplasmic volume,ion concentration,and pH value.In-depth understanding the role of ion channels in formation,development and transfer process of glioma may provide new targets for glioma prevention and treatment.In this paper,the advanced studies of ion channels in glioma were briefly reviewed.

Establishment and stability of a hyperglycemic mouse model induced by alloxan / 中国比较医学杂志

Objective To explore the influence of drug dosage, solvent and other main influencing factors on the successful establishment of alloxan-induced hyperglycemia mouse model and the effect on the stability of this model. Methods 160 6-8-week-old Kunming mice ofSPF grade, (male:female=1:1) were used in this study.The influences of different dosages of alloxan and solvent combinations on the successful establishment rate of the model, survival rate, body weight, fasting blood glucose, blood glucose area under curve, serum insulin level and their stabilities were dynamically observed for six weeks.Results By single intraperitoneal injection of 160 mg/kg bw alloxan ( pH 4.5 citrate sodium as solvent) , we were able to obtain a stable experimental hyperglycemic mouse model with higher levels of successful establishment rate (70%), survival rate (75%), fasting blood glucose (15-20 mmol/L), glucose area under the curve (55-65 mmol/L) and a lower but not loss of serum insulin levels (21 mIU/L).Conclusions In the present study we have carefully considered the influence of main factors such as drug dosages, solvent, etc., on the alloxan-induced experimental hyperglycemic mouse model, and successfully established this model after 6-week period observation of its stability.This model may provide a useful tool in the research of experimental diabetes and hypoglycemic functional studies.

Effect of Tangzhiping granule on blood lipids and free fatty acids in rats with insulin resistant diabetes / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effects of Tangzhiping Granule (TZPG) on blood lipids and free fatty acids (FFA) in rats with insulin resistant diabetes (IRD).</p><p><b>METHODS</b>A blank control group consisted of randomly selected normal rats was set up. The remaining rats were established to IRD model by high-fat high-sugar diet feeding and streptozotocin injection. Then the 32 successfully modeled rats were randomized into the model group (treated by saline), the Tangmaikang group (treated with Tangmaikang Granule 1.35 g/kg), and the two TZPG groups treated with high dose (2.70 g/kg) and low dose TZPG (1.35 g/kg) respectively through intragastric infusion for 4 weeks. The body weight (BW), fasting blood glucose (FBG), insulin (INS), blood lipids including triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), and FFA were detected, and the insulin sensitivity index (ISI) calculated.</p><p><b>RESULTS</b>Compared with the blank control group, BW, FBG and INS increased while ISI decreased in the model group (P < 0.05 or P < 0.01). All the above-mentioned abnormal indices were improved in the three treated groups (Tangmaikang, high and low dose TZPG group), but the improvements in the high dose TZPG group were more significant than those in the other two groups (P < 0.05 or P < 0.01). Similar outcomes were also seen in blood lipids detection, in which TG, TC, LDL-C and FFA were higher and HDL-C were lower in model rats than those in blank controls, they were improved in the three treated groups (P < 0.05), and the best improvements were seen in the high dose TZPG group (P < 0.05).</p><p><b>CONCLUSION</b>TZPG could reduce levels of BW, FBG, INS, TC, TG, LDL-C and FFA, and increase levels of ISI and HDL-C in rat model of insulin resistant type 2 diabetes, so as to improve the insulin resistance in them.</p>