AIM To establish the HPLC characteristic chromatograms for Yixin Fumai Granules,and to determine the contents of sodium danshensu,protocatechualdehyde,chlorogenic acid,calycosin-7-O-β-D-glucoside,ferulic acid,rosalinic acid,salvianolic acid A,salvianolic acid B,schisandrol A.METHODS The analysis was performed on a 35 ℃ thermostatic Acutfex PA-C18 column(4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％phosphoric acid flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelengths were set at 210,250,280,320 nm.Subsequently,cluster analysis and principal component analysis were performed.RESULTS There were 11 characteristic peaks in the characteristic chromatograms for 15 batches of samples with the similarities of more than 0.980.Nine constituents showed good linear relationships within their own ranges(r≥0.999 6),whose average recoveries were 97.60％-107.02％with the RSDs of 0.78％-1.87％.Various batches of samples were clustered into 4 categories,2 principal components demonstrated the accumulative variance contribution rate of 89.454％.CONCLUSION This sensitive and reproducible method can provide a reference for the quality evaluation and control of Yixin Fumai Granules.

AIM To establish a quantitative analysis of multi-components by single-marker(QAMS)method for the simultaneous content determination of baicalin,baicalin,hesperidin,naringin,neohesperidin,rosinic acid,glycyrrhizin,ammonium glycyrrhizate and prehumetin in Tongxuan Lifei Pills.METHODS The analysis was performed on a 30 ℃ thermostatic SVEA C18 column(4.6 mm × 250 mm,5 μm),with the mobile phase comprising of acetonitrile-water(containing 0.1％phosphoric acid)flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 250 nm.Baicalin was used as an internal standard to calculate the relative correction factors of the other eight constituents,after which the content determination was made.RESULTS Nine constituents showed good linear relationships within their own ranges(r ≥ 0.999 1),whose average recoveries were 96.15％-101.58％with the RSDs of 0.75％-1.74％.The result obtained by QAMS approximated those obtained by external standard method.CONCLUSION This accurate,stable and reproducible method can be used for the quality control of Tongxuan Lifei Pills.

AIM To evaluate the quality of Tibetan medicine"Sangdi"based on HPLC fingerprints combined with chemometrics.METHODS The analysis was performed on a 30 ℃ thermostatic Welch Ultimate AQ-C18 column(250 mm × 4.6 mm,5 μm),with the mobile phase comprising of acetonitrile-0.2％phosphoric acid flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 245 nm,after which cluster analysis,principal component analysis and orthogonal partial least squares discriminant analysis were performed,the contents of gentiopicroside,sweroside,mangiferin,isoorientin,8-hydroxy-1,3,5-trimethoxyxanthone(R2)and 1,8-dihydroxy-3,7-dimethoxyxanthone(R3)were determined.RESULTS There were 18 common peaks in the fingerprints for 15 batches of samples with the similarities of more than 0.90.Six constituents showed good linear relationships within their own ranges(R 2 ≥ 0.999 2),whose average recoveries were 96.93％-103.58％with the RSDs of 0.82％-2.9％.Various batches of samples were clustered into 2 categories,4 principal components demonstrated the accumulative variance contribution rate of 86.404％,mangiferin,gentiopicroside and isoorientin were taken as quality difference markers.CONCLUSION This stable,reliable and reproducibe method can provide a reference for the comprehensive quality evaluation of"Sangdi".

AIM To prepare anisamide-modified ursolic acid self-assembled nanoparticles,and to evaluate their anti-drug resistance effect of enzalutamide on prostate cancer.METHODS Nanoparticle precipitation method was adopted in the preparation of anisamide-modified and non-anisamide-modified self-assembled nanoparticles,respectively,after which the particle size,Zeta potential and encapsulation efficiency were determined,and the morphology was observed under transmission electron microscope.The intake of cancer-associated fibroblasts(CAFs)was investigated,after which the model for enzalutamide resistance in prostate cancer was established,CCK8 assay was applied to analyzing the sensitization effect of self-assembled nanoparticles on enzalutamide,and Western blot was used for the detection of NRG1,HER3,AKT expressions.RESULTS The anisamide-modified self-assembled nanoparticles demonstrated the average particle size,Zeta potential and encapsulation efficiency of(195.13±8.06)nm,(-29.07±0.55)mV and(94.58±0.84)％,respectively.CAFs displayed higher intake in the anisamide-modified self-assembled nanoparticles than that in the non-modified preparation and free Cy5(P＜0.05).Meanwhile,anisamide-modified self-assembled nanoparticles were able to inhibit enzalutamide resistance caused by CAFs,reduce NRG1 expression on CAFs,and anisamide-modified self-assembled nanoparticles-treated conditioned medium of CAFs could reduce HER3 and AKT expression on LNCaP cells(P＜0.05,P＜0.01).CONCLUSION Anisamide-modified ursolic acid self-assembled nanoparticles can enhance the targeting of CAFs,alleviate the drug resistance effect of enzalutamide on prostate cancer caused by CAFs,and reduce NRG1 expression in CAFs.

AIM To optimize the extraction process for Jiangzhi Ligan Decoction,and to investigate its effects on improving lipid deposition in HepG2 cells.METHODS With extraction time,extraction frequency and solvent consumption as influencing factors,comprehensive score for tanshensu,lotine,catechin contents and extract yield as an evaluation index,the extraction process was optimized by Box-Behnken response surface method on the basis of single factor test.Palmitic acid-induced HepG2 cells were adopted in the establishment of non-alcoholic fatty liver disease model,after which the effect of drug-containing serum on TC and TG levels was investigated.RESULTS The optimal conditions were determined to be 90 min for extraction time,three times for extraction frequency,and 11 times for solvent consumption.Compared with the control group,the model group demonstrated increased TG,TC levels(P＜0.01);compared with the model group,the Jiangzhi Ligan Decoction high-dose group displayed decreased TG,TC levels(P＜0.05,P＜0.01),especially for those after optimization(P＜0.05,P＜0.01).CONCLUSION Jiangzhi Ligan Decoction can alleviate lipid deposition in HepG2 cells induced by palmitic acid,whose efficacy after extraction process optimization is better than that reported in literature.

AIM To investigate the effects of Heixiaoyao Powder on neuroinflammation in APP/PS1 transgenic mice.METHODS The 16-week-old male APP/PS1 transgenic mice were randomly divided into the model group,the BBG group(P2X7R specific antagonist,30 mg/kg)and high,medium and low dose Heixiaoyao Powder groups of(22.10,11.05,5.53 g/kg),in contrast to the male C57BL/6J mice of the same age and the same strain of the blank groups,with 12 mice in each group.When normal saline by gavage was dosed upon the blank group and the model group,and the other groups were treated with corresponding drug by gavage.After 90 days of administration,the mice had their learning and memory ability detected by Morris water maze test;their hippocampal pathological changes observed with HE staining;their MyD88 expression detected by immunofluorescence staining;their hippocampal levels of pro-inflammatory factors(TNF-α,IL-6),anti-inflammatory factors(IL-10),ATP and amyloid protein β(Aβ)detected by ELISA;their hippocampal mRNA expressions of P2X7R,TLR4,MyD88 and NF-κB-P65 detected by RT-qPCR method;and their hippocampal protein expressions of P2X7R,TLR4,MyD88,NF-κB-P65 and p-NF-κB-P65 detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated prolonged escape latency and reduced frequency in crossing platform(P＜0.01);decreased number of hippocampal neurons,deranged neurons,and darker cytoplasm staining;increased immunofluorescence expression of hippocampal MyD88(P＜0.01);decreased IL-10 level(P＜0.01);increased levels of TNF-α,IL-6,ATP and Aβ(P＜0.01);increased mRNA and protein expressions ofP2X7R,TLR4 and MyD88(P＜0.01),and increased protein expression of p-NF-κB-P65(P＜0.01).Compared with the model group,the groups intervened with Heixiaoyao Powder or BBG demonstrated shortened escape latency and increased frequency in crossing platform(P＜0.01);more number of neatly arranged hippocampal neurons;increased hippocampal IL-10 level(P＜0.01),decreased levels of TNF-α,IL-6,ATP and Aβ(P＜0.05,P＜0.01);decreased mRNA and protein expressions of P2X7R,TLR4 and MyD88(P＜0.05,P＜0.01);and decreased protein expression of p-NF-κB-P65(P＜0.05,P＜0.01).Except the low dose Heixiaoyao Powder group,the other treatment groups shared decreased immunofluorescence expression of MyD88(P＜0.05,P＜0.01).CONCLUSION Heixiaoyao Powder can significantly improve the learning and memory ability of APP/PS1 model mice,and its mechanism may lie in its function in alleviating cerebral neuroinflammation by reducing the abnormal Aβ aggregation via inhibiting the activation of ATP/P2X7R/NF-κB signaling pathway.

AIM To investigate the improvement effects of Poria cocos polysaccharides(PCPs)on mouse nonalcoholic fatty liver disease(NAFLD).METHODS Forty-eight C57BL/6 mice were randomly divided into the blank group,the model group,the simvastatin group(4 mg/kg)and the high,medium and low dose PCPs groups(200,100 and 50 mg/kg),with 8 mice in each group.The NAFLD model was reproduced by 16 weeks feeding of high-fat and high-cholesterol diet,followed by 8 weeks administration of corresponding drug by gavage.The mice had their body mass and liver coefficient assessed;their levels of hepatic free fatty acid(FFA),and serum total cholesterol(TC),triglyceride(TG),high density lipoprotein cholesterol(HDL-C),low density lipoprotein cholesterol(LDL-C),aspartate aminotransferase(AST),alanine aminotransferase(ALT),γ-glutamyltransferase(γ-GT)and malondialdehyde(MDA)detected;their hepatic pathological changes and lipid deposition observed using HE staining,NAFLD activity score(NAS)and oil red O staining;and their hepatic protein expressions of Akt,mTOR,p-Akt,p-mTOR and SREBP-1c detected by Western blot.RESULTS Compared with the blank group,the model group demonstrated all increased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α.levels(P＜0.05,P＜0.01);decreased HDL-C level and activities of SOD and GSH-Px(P＜0.05,P＜0.01);more obvious hepatic pathological damage as revealed by increased NAS score(P＜0.01)and increased lipid deposition area(P＜0.01).Compared with the model group,the groups intervened with high or medium dose PCPs,or simvastatin displayed decreased body weight,liver coefficient,hepatic FFA level,and serum TC,TG,LDL-C,AST,ALT,γ-GT,MDA,IL-1β and TNF-α levels(P＜0.05,P＜0.01);increased HDL-C level and SOD,GSH-Px activities(P＜0.05,P＜0.01);decreased hepatic pathological damage as revealed by the decreased NAS score and lipid deposition area(P＜0.05,P＜0.01);and decreased hepatic protein expressions of p-Akt,p-mTOR and SREBP-1c protein(P＜0.05)as well.CONCLUSION PCPs can improve mouse NAFLD,and its mechanism may lie in their function in reversing abnormal lipid metabolism via Akt/mTOR/SREBP-1c signaling pathway.

AIM To investigate the repair effects of tauroursodeoxycholic acid(TUDCA)combined with bone marrow mesenchymal stem cells(BMSCs)transplantation on spinal cord injury(SCI)in rats.METHODS The rats were randomly divided into the sham operation group,the model group,the TUDCA group,the BMSCs transplantation group and the combination therapy of TUDCA and BMSCs transplantation group,with the SCI rat model established by Allen's method.The next day after modeling,the rats of TUDCA and combination therapy groups were given 200 mg/kg TUDCA by gavage.On the 3rd day after modeling,rats in BMSCs transplantation group and combination therapy group were injected with 1 mL tuned bone marrow BMSCs(the 3rd generation,1× 106/mL)via tail vein.Rats in the sham operation group and the model group were given gastric perfusion of normal saline and injection of 1 mL PBS through tail vein.On the 3rd,7th and 14th day after modeling,the rats had their motor function of hind limbs observed and BBB score determined.After the corresponding drug administration,the rats had their movement track of hind limbs recorded by footprint experiment;their the protein expressions of IL-6,IL-10,Arg-1,PI3K and Akt in spinal cord tissue detected by Western blot;their pathological changes of spinal cord tissue observed by HE staining and Nissl staining;and their expressions of MAP2,GAP43 and GFAP detected by immunofluorescence staining.RESULTS Compared with the model group,the groups intervened with TUDCA,or BMSCs transplantation,or combination therapy shared improved hind limb function and spinal cord histomorphology(P＜0.05);increased fluorescence intensity of MAP2 and GAP43,and protein expressions of IL-10,Arg-1,p-PI3K and p-Akt(P＜0.05);decreased fluorescence intensity of GFAP and IL-6 protein expressions(P＜0.05);among which the combination therapy group took the lead(P＜0.05).CONCLUSION The combination therapy of TUDCA and BMSCs transplantation may restore the function of the rat model of SCI by reducing inflammatory reaction,alleviating secondary injury,and promoting axon and myelin regeneration via PI3K/Akt signaling pathway.

AIM To investigate the renal protective effects of Qizhi Tongluo Formula on a mouse model of diabetic nephropathy.METHODS The male db/db mice were randomly divided into the model group,the dapagliflozin group(0.76 mg/kg)and the low,medium and high dose Qizhi Tongluo Formula groups(7.83,15.65 and 31.3 g/kg),with 6 mice in each group,in contrast to the 6 db/m mice of the control group.When the mice of the control group and the model group were given distilled water by gavage,those of the other administration groups were dosed with the corresponding drug by gavage once daily for 8 weeks.After the drug administration,the mice had their levels of FBG,BUN,Scr and 24 h-UTP detected;their renal pathological changes observed by transmission electron microscopy(TEM)and HE staining;their levels of serum Nrf2,HO-1,Keap1 and renal oxidative stress assessed by ELISA;their renal Nrf2 protein expression observed by immunofluorescence(IF);their renal protein expressions of Nrf2,HO-1 and Keap1 detected by Western blot;and their renal Nrf2,HO-1,and Keap1 mRNA expressions detected by RT-qPCR.RESULTS Compared with the control group,the model group displayed increased levels of 24 h-UTP,Scr,FBG and renal MDA(P＜0.01);decreased renal activities of SOD,CAT and GSH-Px(P＜0.01);mild glomerular mesangial hyperplasia,vacuolated renal tubular epithelial cells,widely fused podocyte foot processes,disappearance of tear film,decreased secretion levels of serum Nrf2 and HO-1 and renal protein and mRNA expressions of Nrf2 and HO-1(P＜0.05,P＜0.01);and decreased secretion levels of serum Keap1 and renal Keap1 protein and mRNA expressions(P＜0.01).Compared with the model group,the high-dose Qizhi Tongluo Formula group demonstrated decreased levels of 24 h-UTP,Scr,FBG and renal MDA(P＜0.01);increased renal activities of SOD,CAT and GSH-Px(P＜0.01);alleviated renal pathological damage,increased secretion levels of serum Nrf2 and HO-1 and renal protein and mRNA expressions of Nrf2 and HO-1(P＜0.01);and increased level of serum Keap1 secretion and renal Keap1 protein and mRNA expressions(P＜0.01).CONCLUSION Qizhi Tongluo Formula can inhibit oxidative stress and alleviate kidney damage in db/db mice by activating Nrf2/Keap1/ARE signaling pathway.

AIM To explore the clinical effects of Jiawei Yanghe Decoction combined with Budesonide and Formoterol Fumarate Powder for Inhalation on patients with mild to moderate bronchial asthma in chronic and persistent period.METHODS One hundred and eighteen patients were randomly assigned into control group(59 cases)for 4-week administration of Budesonide and Formoterol Fumarate Powder for Inhalation,and observation group(59 cases)for 4-week administration of both Jiawei Yanghe Decoction and Budesonide and Formoterol Fumarate Powder for Inhalation.The changes in clinical effects,ACT score,bronchial asthma control rate,pulmonary function indices(FEV1,PEF,FEV1％,PEF％),inflammatory indices(EOS,EOS％,FeNO),TCM syndrome score and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed increased bronchial asthma control rate,ACT score,PEF(P＜0.05),and decreased TCM syndrome score(P＜0.05),especially for the observation group(P＜0.05);the observation group exhibited increased FEV1,FEV1％,PEF％(P＜0.05),among which FEV1,PEF％were higher than those in the control group(P＜0.05);the observation group showed decreased inflammatory indices(P＜0.05),among which FeNO was lower than that in the control group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with mild to moderate bronchial asthma in chronic and persistent period,Jiawei Yanghe Decoction combined with Budesonide and Formoterol Fumarate Powder for Inhalation can safely and effectively alleviate clinical symptoms,improve pulmonary functions,airway inflammatory reactions,and enhance bronchial asthma control rate.