OBJECTIVE To study the protective effect of saikosaponin b2(SSb2)on corticosterone(CORT)induced PC12 cell injury and its mechanism.METHODS ① PC12 cells were divided into the cell control group(24 h of culture with RPMI-1640 medium),CORT group(24 h of culture with CORT 100-800 μmol·L-1)and SSb2 group(24 h of culture with SSb2 1.5625,3.125,6.25,12.5,25,50 and 100 μmol·L-1).MTT assay was used to detect the cell survival rate.②PC12 cells were divided into the cell control group(24 h of culture with RPMI 1640 medium),model group(24 h of culture with CORT 400 μmol·L-1),and model+SSb2 group(3 h pretreatment with SSb2 1.5625,3.125,6.25,12.5 and 25 μmol·L-1,removal of the supernatant before cells were co-incubated with CORT 400 μmol·L-1 and corresponding concentrations of SSb2 for 24 h).MTT assay was used to detect the cell survival rate while micro-plate assay was used to detect the lactate dehydrogenase(LDH)leakage rate of PC12 cells.③PC12 cells were divided into the cell control group,model group and model+SSb2 12.5 μmol·L-1 group.AnnexinV-FITC/PI flow cytometry assay was used to detect PC12 cell apoptosis,ultra-perfor-mance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS)cell metabonomics was used to detect metabolic profile changes and colorimetric assay was employed to detect the glutamic acid content and glutaminase activity in PC12 cells.RESULTS Compared with the cell control group,the cell viability decreased to(55±6)%(P<0.01)when the concentration of CORT was 400 μmol·L-1.When the concentration of SSb2 was higher than 50 μmol·L-1,there was significant toxicity to PC12 cells(P<0.01).②Compared with the cell control group,the cell survival rate was signif-icantly decreased(P<0.01),while the release rate of LDH was significantly increased(P<0.01)in the model group.Compared with the model group,the cell survival rate significantly increased(P<0.05,P<0.01),while the LDH release rate significantly decreased(P<0.01)in the model+SSb2 group.③ Com-pared with the cell control group,cell apoptosis was significantly increased in the model group(P<0.05).Compared with the model group,cell apoptosis was significantly decreased(P<0.05)in the model+ SSb2 group.Metabolomics results show that SSb2 significantly back-regulated nine differential metabo-lites of glutamate,creatine,N-acetylaspartate,L-tyrosine,citric acid,L-isoleucine,lactic acid,glutamine and choline.Further network analysis of the key metabolites regulated by SSb2 yielded five major metabolic pathways:D-glutamine and D-glutamate metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,alanine,aspartate and glutamate metabolism,tyrosine metabolism and arginine biosynthesis.Compared with the cell control group,the content of glutamate and activity of glutaminase were significantly decreased in the model group(P<0.01).Compared with the model group,the content of glutamate(P<0.01)and activity of glutaminase(P<0.05)were significantly increased in the model+SSb2 group.CONCLUSION SSb2 has a neuroprotective effect on CORT-injured PC12 cells,and the mechanism of which is related to inhibition of apoptosis and regulation of metabolic disorders.

Chaigui granules(CG)are a compound composed of six herbal medicines with significant antidepressant effects.However,the antidepressant mechanism of CG remains unclear.In the present study,we attempted to elucidate the antidepressant mechanism of CG by regulating purine metabolism and purinergic signaling.First,the regulatory effect of CG on purine metabolites in the prefrontal cortex(PFC)of chronic unpredictable mild stress(CUMS)rats was analyzed by ultra high-performance liquid chro-matography tandem mass spectrometry(UHPLC-MS/MS)targeted quantitative analysis.Meanwhile,purinergic receptors(P2X7 receptor(P2X7R),A1 receptor(A1R)and A2A receptor(A2AR))and signaling pathways(nod-like receptor protein 3(NLRP3)inflammasome pathway and cyclic adenosine mono-phosphate(cAMP)-protein kinase A(PKA)pathway)associated with purine metabolism were analyzed by western blotting and enzyme-linked immunosorbent assay(ELISA).Besides,antidepressant mecha-nism of CG by modulating purine metabolites to activate purinergic receptors and related signaling pathways was dissected by exogenous supplementation of purine metabolites and antagonism of puri-nergic receptors in vitro.An in vivo study showed that the decrease in xanthine and the increase in four purine nucleosides were closely related to the antidepressant effects of CG.Additionally,purinergic re-ceptors(P2X7R,A1R and A2AR)and related signaling pathways(NLRP3 inflammasome pathway and cAMP-PKA pathway)were also significantly regulated by CG.The results of exogenous supplementation of purine metabolites and antagonism of purinergic receptors showed that excessive accumulation of xanthine led to activation of the P2X7R-NLRP3 inflammasome pathway,and the reduction of adenosine and inosine inhibited the A1R-cAMP-PKA pathway,which was significantly ameliorated by CG.Overall,CG could promote neuroprotection and ultimately play an antidepressant role by inhibiting the xanthine-P2X7R-NLRP3 inflammasome pathway and activating the adenosine/inosine-A1R-cAMP-PKA pathway.

Radix Bupleuri(RB)is commonly used to treat depression,but it can also lead to hepatotoxicity after long-term use.In many anti-depression prescriptions,RB is often used in combination with Radix Paeoniae Alba(RPA)as an herb pair.However,whether RPA can alleviate RB-induced hepatotoxicity remain unclear.In this work,the results confirmed that RB had a dose-dependent antidepressant effect,but the optimal antidepressant dose caused hepatotoxicity.Notably,RPA effectively reversed RB-induced hepatotoxicity.Afterward,the mechanism of RB-induced hepatotoxicity was confirmed.The results showed that saiko-saponin A and saikosaponin D could inhibit GSH synthase(GSS)activity in the liver,and further cause liver injury through oxidative stress and nuclear factor kappa B(NF-KB)/NOD-like receptor thermal protein domain associated protein 3(NLRP3)pathway.Furthermore,the mechanisms by which RPA attenuates RB-induced hepatotoxicity were investigated.The results demonstrated that RPA increased the abundance of intestinal bacteria with glycosidase activity,thereby promoting the conversion of saikosaponins to sai-kogenins in vivo.Different from saikosaponin A and saikosaponin D,which are directly combined with GSS as an inhibitor,their deglycosylation conversion products saikogenin F and saikogenin G exhibited no GSS binding activity.Based on this,RPA can alleviate the inhibitory effect of saikosaponins on GSS activity to reshape the liver redox balance and further reverse the RB-induced liver inflammatory response by the NF-κB/NLRP3 pathway.In conclusion,the present study suggests that promoting the conversion of saikosa-ponins by modulating gut microbiota to attenuate the inhibition of GSS is the potential mechanism by which RPA prevents RB-induced hepatotoxicity.

OBJECTIVE To investigate the dynamic changes in urine metabolic profiles in rats induced by D-galactose (D-Gal),and to study the correlations between the differential metabolites and behavior indicators using the proton nuclear magnetic resonance (1H-NMR)-based metabonomics.METHODS Subcutaneous injection of D-Gal 100 mg· kg-1 for 10 weeks was adopted in the model group.The sample of urine was collected at day 0 (dO),d14,d28,d42,d56 and d70.NMR metabonomics technique was used for acquisition of data,which was analyzed by multivariate statistical analysis.The ability of learning and memory were measured by Morris water maze test from d70.After the behavioral test,the rats were sacrificed and the hippocampus was observed by hematoxylin-eosin staining.RESULTS Principal component analysis (PCA) results revealed that there was considerable difference between the model group and the normal control group at d70.According to the varible importance plot (VIP) calculation and S-plot scores,a total of 12 metabolites were screened and identified as potential biomarkers at d70.The differences of metabolites and Morris water maze test were subjected to correlation analysis,and the results showed that the levels of choline,lactate and dimethylglycine in the model group were significantly increased and negatively correlated with the times of crossing the platform (r =-0.90,-0.50 and-0.52;n=10).Formate was significantly negatively correlated with the time spent in the target area (r =-0.51,n=10),but choline and formate were significantly positively correlated with the escape latency (r =0.72 and 0.53;n=10).However,the levels of creatine and taurine decreased in the model group,which was significantly positively correlated to the times of acrossing platforms (r =0.89 and 0.71;n=10),while alanine was significantly positively correlated to the time spent in the target area(r =0.74,n=10).Taurine,alanine and creatine were significantly negatively correlated with the escape latency (r =-0.66,-0.50 and-0.85;n=10).The correlations between the differential metabolites and the behavioral indicators were further proved.CONCLUSION The metabolic profile changes in urine from D-Gal induced aging model rats are significantly correlated with impairement of ability in learning and memory.1H-NMR metabonomics in urinary metabolic profile changes may be used as an evaluation index in the D-Gal induced aging rats model.

AIM To explore the mercury accumulation in KM mice after being given Zuotai at different doses and time.METHODS KM mice were randomly divided into blank group,Zuotai low-,middle-and high-dose (6.07,60.70 and 606.97 mg/kg,42 d;606.97 mg/kg,14 d) groups.The mercury contents in brain (olfactory bulb,cortex,hippocampus,hypothalamus,brain stem,cerebellum),heart,lung,kidney,liver,spleen,serum,muscle of mice were measured after administration.RESULTS Compared with the blank group,Zuotai at low-dose significantly increased the mercury contents in hippocampus,cerebellum,lung,kidney,liver and serum of mice after 42-day treatment;Zuotai at middle-dose markedly increased the mercury contents in olfactory bulb,cortex,hippocampus,brain stem,cerebellum,heart,lung,kidney,liver,spleen and serum of mice after 42-day treatment;the mice treated with high-dose of Zuotai for 42,14 days significantly increased the mercury contents in olfactory bulb,cortex,hippocampus,hypothalamus,brain stem,cerebellum,heart,lung,kidney,liver,spleen,muscle and serum.CONCLUSION Mercury can be accumulated in different tissues of mice after intragastric administration of Zuotai in a dose-and time-dependent manner,which suggests that Zuotai and its compound preparations should not be used in high-dose and long-term.

The purpose of this study is to evaluate the anti-aging effects and reveal the underlying mechanism of Scutellaria baicalensis Georgi ethanol extract (SBG) in D-galactose-induced rats. Fifty rats were randomly divided into five groups: vehicle control group, D-galactose group, and D-galactose combined with 50, 100, 200 mg x kg(-1) SBG. A rat aging model was induced by injecting subcutaneously D-galactose (100 mg x kg(-1)) for ten weeks. At the tenth week, the locomotor activity (in open-field test) and the learning and memory abilities (in Morris water maze test) were examined respectively. The urine was collected using metabolic cages and analyzed by high-resolution 1H NMR spectroscopy combined with multivariate statistical analyses. The SBG at doses of 50, 100 and 200 mg x kg(-1) treatments groups could significantly ameliorate aging process in rats' cognitive performance. The 50, 100, 200 mg x kg(-1) SBG regulated citrate, pyruvate, lactate, trimethylamine (TMA), pantothenate, β-hydroxybutyrate in urine favorably toward the control group. These biochemical changes are related to the disturbance in energy metabolism, glycometabolism and microbiome metabolism, which is helpful to further understanding the D-galactose induced aging rats and the therapeutic mechanism of SBG.

OBJECTIVE:To investigate the effects of atorvastatin with different doses on related indicators and major adverse cardiovascular events(MACE)in patients with acute myocardial infarction(AMI)after percutaneous coronary intervention(PCI). METHODS:120 patients with AMI after PCI were randomly divided into conventional dose group(60 cases)and high dose group (60 cases). All patients were immediately given low molecular weight heparin,Aspirin enteric-coated tablet,Clopidogrel bisulfate tablet,Angiotensin-converting enzyme inhibitor,β-blockers,nitrates and other conventional treatment;based on it,conventional dose group was given 20 mg Atorvastatin calcium tablet,once every day before bedtime;high dose group was given 40 mg Atorv-astatin calcium tablet,once every day before bedtime. The treatment course for both groups was 1 month. Total cholesterol(TC), triglyceride(TG),low density lipoprotein cholesterol(LDL-C),high density lipoprotein cholesterol(HDL-C),C-reactive protein (CRP),OX40L,interleukin (IL)-6 levels in 2 groups before and after treatment were observed,the correlation among OX40L with CRP and IL-6 were analyzed,and the incidence of MACE in 2 groups was recorded after 1 month follow-up. RESULTS:Be-fore treatment,there were no significant differences in the TG,TC,LDL-C,HDL-C,OX40L,CRP and IL-6 between 2 groups (P>0.05). After treatment, TG, TC, LDL-C, OX40L, CRP and IL-6 were aignificantly lower than before, TC, LDL-C, OX40L,CRP and IL-6 in high dose group were lower than conventional dose group,HDL-C was significantly higher than before, the differences were statistically significant(P<0.01 or P<0.05);but there was no significant difference in the HDL-C between 2 groups(P>0.05). Before treatment,OX40L showed positive correlation with CRP(r=0.746,P<0.01)and IL-6(r=0.763,P<0.01);after treatment,OX40L also showed positive correlation with CRP (r=0.755,P<0.01) and IL-6 (r=0.760,P<0.01). The incidence of MACE in high dose group were significantly lower than conventional dose group,the difference was statistically significant(P<0.05). CONCLUSIONS:Based on the conventional treatment,atorvastatin can significantly reduce lipid,inflamma-tion levels and incidence of MACE,but the effect is more significant in high dose.

Objective To explore the relations between pulse pressure and LDL-C for elderly patients with hyperten-sion. Methods A total of 451 elderly patients with hypertension were selected and divided into four groups. Group A was 65 to 69 years of age, group B was 70 to74 years of age, group C was 75 to 79 years of age, and group D was ≥80 years of age. They were also divided into four groups according to PP levels: PP1 group ≤40 mm Hg, PP2 group 41 to 60 mm Hg, PP3 group 61 to 80 mm Hg, and PP4 group>80 mm Hg. They were divided into LDL-C group 1 (≥2.6 mmol/L) and LDL-C group 2 (<2.6 mmol/L) according to their LDL-C levels. Distribution of different age groups in four PP groups and the effects of different levels of LDL-C on pulse pressure were analyzed. Results (1) The propor-tions of PP2 group among the four age groups were 59.41%, 48.84%, 55.45% and 51.67%. The proportions of PP4 a-mong the four age groups were 4.95%, 3.88%, 3.96% and 7.50%. (2) Compared with PP1 group, the proportion of LDL-C ≥2.6 mmol/L in PP2 group, PP3 group and PP4 group was significantly increased. Conclusion (1) PP levels in different ages groups are concentrated on 41-60 mmHg. Among elderly patients with hypertension>80 mmHg, the pro-portion of patients with advanced age (≥years of age) is relatively higher. (2) LDL-C is the risk factor of increased pulse pressure for elderly patients with hypertension.

OBJECTIVETo explore the distribution of serum antibodies against human papillomavirus (HPV) 16/18 among women at high-risk for cervical cancer.

METHODSAll women when tested positive for anyone of the cervical cancer screening programs, from Xinmi county of Henan province in 2011, were recruited as the subjects of this study. Cervical exfoliated cells were collected, using cervical brush for HPV DNA testing, and 10 ml venous blood was drawn for HPV-16, 18 serum antibodies testing, by enzyme-linked immunosorbent assay.

RESULTSAmong the 952 women under study, 230 cases (24.2%)showed HPV DNA positive, with positivity rates of HPV16 and 18 L1 virus-like particle (VLP)antibodies as 23.2% and 6.5%, respectively. The overall positivity rate of any type of HPV16, 18 VLP antibodies was 26.8% . Geometric means of HPV16, 18 VLP antibody titers were 79.1 (Yangshengtang Unit,YU/ml) and 125.0(YU/ml). Positivity rate of HPV16 antibody was significantly associated with age, viral load of HPV DNA, and cervical lesion severity (P < 0.05). Seropositivity of HPV18 was also increasing with the increase of viral load (P < 0.01) with different cervical lesion significantly showing different titer of HPV18 antibody (P < 0.01). Based on the results of HPV DNA detection among the two years of study, women with HPV persistent infection showed significant higher positive rate of HPV16/18 antibodies than women who did not have HPV infection or emerging infection (P < 0.001). When comparing to those women without HPV infection, the ones with transient infection showed higher seropositivity rates on both HPV16 antibodies and titer of HPV16 antibody (P < 0.001).

CONCLUSIONSeroprevalence rates on HPV16 and 18 among the unvaccinated high-risk women in Henan were high. Prevalence of both HPV16 and 18 antibodies were correlated with age, viral load, cervical lesion and history of infection.Women with high viral load, high grade cervical lesion or history of infection would more likely to be seropositive.

Adult ; Aged ; Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Human papillomavirus 16 ; immunology ; Human papillomavirus 18 ; immunology ; Humans ; Middle Aged ; Papillomavirus Infections ; epidemiology ; Seroepidemiologic Studies ; Uterine Cervical Neoplasms ; virology

OBJECTIVETo determine the distribution of anti-hepatitis E virus (HEV) IgG antibody and anti-human papilloma virus (HPV) IgG antibody among female residents of Xinmi and investigate the risk factors of HEV infection.

METHODSA questionnaire was used to collect data on the demographic characteristics and suspected risk factors of HEV infection, including behavioral habits. All questionnaire responders also provided peripheral blood samples for investigation by enzyme-linked immunosorbent assay to detect HEV-IgG. Demographic data were statistically evaluated by t-test and univariate analysis, and HEV infection risk factors were statistically evaluated by a binary logistic regression model.

RESULTSThe average age of the 952 questionnaire responders was 47.16 + 8.09 years. The demographic parameters of education level, income, experience of stillbirth, and age were associated with HEV-IgG positivity (all P less than 0.05). Age, occupation, and income were identified as independent risk factors for HEV-IgG positivity (all P less than 0.05). No statically association was found between sexual behavior and anti-HEV or anti-HPV levels, or HEV infection.

CONCLUSIONThe female population surveyed in Xinmi, Henan Province showed a higher HEV-IgG positive rate than generally reported in the literature, and this rate shows an increasing trend with age, Risk factors for HEV infection among this group are age, income and occupation.

Adult ; Aged ; Antibodies, Viral ; blood ; China ; epidemiology ; Female ; Hepatitis E ; blood ; epidemiology ; immunology ; Hepatitis E virus ; Humans ; Immunoglobulin G ; blood ; Middle Aged ; Risk Factors