Fufang E’jiao Jiang’s effect on immunity,hematopoiesis,and angiogenesis via a systematic“compound-effect-target”analysis

Fufang E’jiao Jiang(FEJ)as a healthy food consisting of medicine food homology materials approved by China’s Ministry of Health has been extensively applied to replenish qi and nourish blood,and it has a positive impact on women’s health.To find out the material basis and mechanism of FEJ,a systematic“compoundeffect-target”analysis including chemical composition resolution,zebrafish,network pharmacology,molecular docking,transcriptome,and bibliometric analysis was adopted.124 chemical components including ginsenosides,and phenylethanoid glycosides in FEJ were discovered,and effects of FEJ on promoting the generation of immune cells,erythropoiesis and angiogenesis in zebrafish were exhibited.Based on network pharmacology,molecular docking and in vivo activity assay,6 compounds including jionoside A1,isoacteoside,echinacoside,acteoside,lobetyolin,and rehmannioside D were identified as active components of FEJ.Transcriptome data showed that several pathways such as complement and coagulation cascades,ECM-receptor interaction,and PI3K-Akt signaling pathway were associated with proangiogenic effect of FEJ.19 common targets were obtained through combined analysis of network pharmacology and transcriptomics,and 5 targets of them were verified by PCR.The bibliometric analysis of these common targets revealed that FEJ was related to energy metabolism,pathway in cancer,etc.,which was consistent with the results of network pharmacology and transcriptome.The studies suggested that FEJ could replenish qi and nourish blood through multi-compound and multi-targets.

The components transitive regularity of three dosage forms of Liuwei Dihuang Fufang

Objective To explore the transitive regularity of holistic constituents from the crude slices of the medicinal raw materials(MCS)to the formula granules(FG),fufang decoction(FD),and finally,the concentrated pills(CP)of Liuwei Dihuang Fufang(六味地黄复方,LWDHF).Methods Samples for MCS,FG,FD,and CP of LWDHF were obtained,and a fingerprint data-base was established using high-performance liquid chromatography(HPLC),by separating the samples in an XB-C18 column and analyzing the transitive regularity of components us-ing the total quantum statistical moment(TQSM),including total quantum zero moment(AUCT),total quantum first moment(MRTT),total quantum second moment(VRTT),and its similarity approach.The AUCT,MRTT,and VRTT were calculated based on the representative HPLC chromatograms of FG,FD,and CP of LWDHF.Results AUCT of FG,FD,and CP of LWDHF was 71804,46553,and 144646μV·s,respectively;MRTT was 14.43,14.54,and 18.85 min,respectively;and VRTT was 106.98,112.84,and 269.12 min2,respectively.Comparing the similarity of FG/FD,FG/CP and FD/CP of LWDHF,the TQSM similarity values were 98.66%,76.62%,and 75.37%,respectively,whereas the tradi-tional similarity evaluation values were 98.68%,85.43%,and 85.60%,respectively.Conclusion The results perform little distinction in the total composition between FG and FD,whereas some distinction existed between FD and CP.Experimental evidence,therefore indicates that FG could be used as the alternative of MCS in clinical applications.

Effect of Fufang Danshen Pill on Bone Marrow Stem Mobilization when Myocardial Scathe

Objective：To investigate the effect of Fufang Danshen pill on bone marrow stem mobilization during myocardial scathe. Methods：Rat models with expansionary myocardial disease were established by Pituitrin and Furazolidone. Experimental rats were divided into the contrast group, the myocardial scathe group （MS group）, the myocardial scathe and Fufang Dansben pill group （ MS ＋ FD group） and the myocardial scathe and fluvastatin group （ MS ＋ FT group）. The ratio of CD34^＋ cells was examined at the 1^st, 3^nl and 6^th weekend. Index of heart structure and function including LVESD, LVEDD. LYEF, LVEDP and dp/dtmax were evaluated at the 6^th weekend. The HW/BW index was calculated. Results：In the MS group, the index of HW/BW, LVESD, LVEDD and LVEDP were obviously increased （P 〈 0.01 ） and index of dp/ dtmax and LVEF were obviously decreased （P 〈 0.05 ）. The ratio of CD34^＋ cells was significantly improved at the 1^at weekend and then reduced slowly with no difference from that of the contrast group at the 6th weekend. Compared the MS ＋ FD group and the MS ＋ FT group with the MS group, the index of HW/BW, LYESD, LYEDD and LYEDP of were signifi cantly decreased （ P 〈 0.05 ） and index of dp/dtmax and LVEF were increased （P 〈 0.01 ）. The ratio of CD34^＋ cells was significantly higher at the 1^st, 3^nl and 6^th weekend, but had no statistic meaning at 3^nl and 6^th weekend （P 〉 0.05 ）. Conclusion：Pituitrin and Furazolidone can be used to establish rat models with expansionary myocardial disease. There has bone marrow stem mobilization during the early period of myocardial scathe. Fufang Danshen pill has effect on improving bone marrow stem mobilization, lightening the expansionary degree of heart and protecting the heart function. The effect of Fufang Danshen pill is as same as that of fluvastatin.

Effects of Fufang Biejia Ruangan Pills on hepatic fibrosis in vivo and in vitro

AIM:To explore the protective effect and the relevant mechanisms of Fufang Biejia Ruangan Pills(FFBJRGP)on hepatic fibrosis in vivo and in vitro.METHODS:Hepatic fibrosis was induced by carbon tetrachloride composite factors.Adult Wistar rats were randomly divided into four groups:normal control group;hepatic fibrosis model group;FFBJRGP-treated group at a daily dose of 0.55 g/kg;and colchicinetreated group at a daily dose of 0.1 g/kg.The effects of FFBJRGP on liver function,serum levels of hyaluronic acid(HA),typeⅣcollagen(CⅣ),typeⅢprocollagen(PCⅢ),laminin(LN),histopathology,and expression of transforming growth factor(TGF-β1)and Smad3 in hepatic fibrosis were evaluated in vivo.The effects of FFBJRGP on survival rate,hydroxyproline content and cell cycle distribution were further detected in vitro.RESULTS:Compared with the hepatic fibrosis model group,rats treated with FFBJRGP showed a reduction in hepatic collagen deposition and improvement in hepatic lesions.Compared with those of the model group,the activities of alanine aminotransferase(62.0±23.7 U/L)and aspartate aminotransferase(98.8±40.0 U/L)in the FFBJRGP-treated group were decreased(50.02±3.7 U/L and 57.2±30.0 U/L,respectively,P<0.01).Compared with those in the model group,the levels of PCⅢ(35.73±17.90 g/mL),HA(563.82±335.54 ng/mL),LN(89.57±7.59 ng/mL)and CⅣ(29.20±6.17ng/mL)were decreased to 30.18±9.41,456.18±410.83,85.46±7.51 and 28.02±9.45 ng/mL,respectively.Reverse-transcriptase polymerase chain reaction and Western blotting also revealed that expression of TGF-β1 and Smad3 were down-regulated in vivo.Cell proliferation was inhibited,the level of hydroxyproline was decreased compared with the control group(P<0.01),and the cell cycle was redistributed when exposed to FFBJRGP in vitro.CONCLUSION:FFBJRGP inhibits hepatic fibrosis in vivo and in vitro,which is probably associated with downregulation of fibrogenic signal transduction of the TGF-β-Smad pathway.

Protective effects of Fufang Ejiao Jiang against aplastic anemia assessed by network pharmacology and metabolomics strategy

Objective To elucidate the mechanisms underlying the therapeutic effects of Fufang Ejiao Jiang(复方阿胶浆,FFEJJ)on aplastic anemia(AA)using integrated network pharmacology and serum metabolomics.Methods Traditional Chinese Medicine Systems Pharmacology(TCMSP),Pubmed,integrative pharmacology-based research platform of traditional Chinese medicine(TCMIP),and Bioinformatics Analysis Tool for Molecular mech ANism of Traditional Chinese Medicine(BATMAN-TCM)were used to identify the constituents and putative targets of FFEJJ.Gene Cards and DisGeNET databases were used to identify AA-associated targets.We constructed a herb-component-target network and analyzed the protein-protein interaction(PPI)network.Potential mechanisms were determined using Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analyses.In addition,an AA model was established using acetylphenylhydrazine(APH)and cetylphenylhydrazine(CTX).Ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-QTOF/MS)-based serum metabolomics was applied to screen potential metabolites and the related pathways associated with AA and the potential anti-anemic effects of FFEJJ.Results A total of 30 active components of FFEJJ and 24 targets were related to AA.PPI network analysis showed that VEGFA,AKT1,IL-6,CASP3,and ICAM1 were key nodes overlapping with proteins known to be related to AA.KEGG pathway enrichment analysis revealed that the presumed targets of FFEJJ were mainly associated with pathways linked to the promotion of hematopoiesis and improvement of the hematopoietic microenvironment.A total of 423 metabolite biomarkers were identified between the control and AA models,which are involved in the development of AA.In contrast,FFEJJ reversed the 79 differential metabolites altered by AA.Pathway analysis suggested that the synergistic effects of FFEJJ were mainly enriched in 24 metabolic pathways.Among them,sphingolipid metabolism,glycerophospholipid metabolism,and arachidonic acid metabolism were related to promoting hematopoiesis and improving the hematopoietic microenvironment,which partially conforms with network pharmacology.The interaction network formed by three key differential metabolites,including hydroxy-eicosatetraenoic acid(HETE),sphingosine 1-phosphate(S1 P),and lysophosphatidylcholine(lyso PC),and three predicted network targets(VEGFA,CASP3,and ICAM1)may be the potential mechanism underlying the anti-AA action of the multi-component of FFEJJ.Conclusion FFEJJ could be an alternative treatment option for AA.It acts by promoting hematopoiesis and improving the hematopoietic microenvironment.Network pharmacology-integrated metabolomics makes it possible to analyze TCMs from a systems perspective and at the molecular level.

Simultaneous Determination of Four Amino Acids in Fufang Ejiao Buxue Granule by HPLC

[Objectives]To establish an High Performance Liquid Chromatography(HPLC)method for simultaneous determination of four amino acids in Fufang Ejiao Buxue Granule.[Methods]The quantitative analysis was carried out with a Waters Sunfire C 18 column(4.6 mm×250 mm,5μm).A binary mobile solvent was used:mobile solvent A was acetonitrile-0.1 mol/L sodium acetate solution(adjusting pH to 6.5 with 36%acetic acid)(7∶93)and mobile solvent B was acetonitrile-H 2O(4∶1).The mobile phase was delivered at a flow rate of 1.0 mL/min with a gradient elution profile(0-13 min,100%A→93%A;13-17.9 min,93%A→88%A;17.9-29.0 min,88%A→85%A;29-39 min,85%A→66%A;39-45 min,66%A→0%A).The column temperature was at 43℃.The detection wavelength was 254 nm.[Results]The injection volume of L-hydroxyproline,glycine,alanine,and L-proline showed a good linear relationship with the chromatographic peak area in the range of 0.012 to 0.117,0.022 to 0.218,0.010 to 0.097,0.016 to 0.160μg,separately.The average recovery rate(n=6)was 96.4%,97.3%,97.1%,and 99.4%,respectively;the relative standard deviations were 1.2%,1.9%,1.7%,and 0.9%,respectively.[Conclusions]This method is simple in operation and good in reproducibility,and provides a reliable method for controlling the quality of Fufang Ejiao Buxue Granules.

Protective Effect of Fufang Yatongding on Periodontitis Model Rats

[Objectives]The paper was to investigate the protective effect of Fufang Yatongding on experimental periodontitis in rats.[Methods]Experimental periodontitis rats were randomly divided into blank group(5 rats),model group,control group and experimental group,with 8 rats in each group.The rats in the blank group were fed with normal diet,and those in the model group,control group and experimental group were administered intragastrically with normal saline,minocycline hydrochloride solution and Fufang Yatongding solution,respectively.After 4 weeks,alveolar bone resorption was measured.Serum matrix metalloproteinases(MMPs)and inflammatory factors were detected by ELISA,and the changes in gingival tissue were observed by HE staining.[Results]Compared with the control group,the distance from enamel cementum to alveolar crest in the experimental group was decreased(P<0.05).Compared with the control group,the levels of serum MMPs and inflammatory factors in the experimental group were decreased(P<0.05).The results of HE staining showed that the cells in the gingival tissue of rats in the blank group were normal in morphology and intact in structure,and the cells in the gingival tissue of rats in the model group were damaged and out of order,while the cells in the control group were slightly intact and arranged orderly,and the pathological damage of rats in the experimental group was less than that in the control group.[Conclusions]Fufang Yatongding has protective effect on experimental periodontitis in rats by inhibiting the release of MMPs and inflammatory factors.

Analysis of Pharmacodynamic Substance Basis of Fufang Changtai in Treating Colorectal Cancer by UPLC-Q-TOF-MS Combined with Network Pharmacology

[Objectives] To systematically study the main active components of Fufang Changtai(FFCT) in the treatment of colorectal cancer(CRC), and to explore its mechanism of action. [Methods] The main chemical components of FFCT were analyzed by ultra-high performance liquid chromatography with quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF-MS) combined with automatic analysis platform, and the main pharmacodynamic substances of FFCT were studied by network pharmacology method and its mechanism of action was explored. The binding degree between the active components and the core targets were verified by molecular docking technology. [Results] A total of 86 compounds were identified from FFCT, among which 26 compounds were Ginsenoside Rg3, Ginsenoside Rb1, Astragaloside III, etc. The key target pathway enrichment analysis showed that FFCT played its role in the treatment of CRC mainly through the PI3K-Akt signaling pathway and MAPK signaling pathway. [Conclusions] This study comprehensively identified the FFCT components. Supplemented by network pharmacology and molecular docking technology, it is expected to provide a scientific theoretical basis and an important reference for FFCT therapeutic components identification, key target verification and mechanism of action in the treatment of CRC.

Association rules analysis of Fufang Kushen injection in combination with traditional Chinese medicine or modern medications in treating Cervical cancer: real-world retrospective study

Objective: The present study aimed to analyze the association rules of Fufang Kushen injection in combination with other traditional Chinese medicine ( TCM) or modern medications in treating cervical cancer (CC) based on the electrical medical records extracted from real-world hospital information system. Methods: The clinicians’ prescriptions regarding to the combination of with TCM or modern medications were from hospital information system electronic medical data integration warehouse established by the Institute of Basic Medical Research of Chinese Medicine, China Academy of Chinese Medical Sciences, which integrated the hospital information system data of 22 hospitals. The association rules of the drug characteristics were analyzed through Apriori algorithm. Results: A total of 839 patients with CC were included. We found that is often combined with prescriptions which could clear heat, remove toxicity, supplement Qi. also combined with chemotherapeutic drugs, immunomodulatory drugs, 5-HT receptor blockers, and glucocorticoids. The combination presents a specific law. Conclusion: Fufang Kushen injection combined with hepatoprotective drugs, immunomodulators and glucocorticoids is often used to treat cervical cancer.

Simultaneous determination of 15 bioactive compounds in Fufang Zhenzhu Tiaozhi capsules by HPLC-DAD-ELSD

Fufang Zhenzhu Tiaozhi capsules （FTZc）, which is consisted of eight traditional Chinese herbal medicines and contains multiple bioactive ingredients, is a patented and clinically approved herbal formulation for the treatment of dyslipidemia. A feasible HPLC-DAD-ELSD method was developed to simultaneously determine 15 bioactive compounds （salidroside, specneuzhenide, magnoflorine, rosmarinic acid, salvianolic acid B, columbamine, jatrorrhizine, epiberberine, coptisine, palmatine, berberine, 5,7-dimethoxycoumarin, ginsenoside Rgl, ginsenoside Rbl and oleanic acid ） in FTZc for its quality control. The multiple wavelength detection mode of DAD was used. The chromatographic separation was performed on an Ultimate XB Cls column with gradient elution. The mobile phase A （acetonitrile） and B （0.25% glacial acetic acid and 0.13% triethylamine in water, v/v） were run at a flow rate of 0.8 mL/min. The developed method showed good precision and accuracy with overall intra- and inter-day variations of 0.7%-1.9% and 0.6%-3.0%, respectively. The recoveries measured at three concentration levels, varied from 95.5% to 103.8%. The validated method was successfully applied for the simultaneous determination of 15 bioactive compounds in three batches of FTZc. The results suggested that the developed method was convenient and reliable, particularly suitable for the routine quality control of FTZc.

Effects of Fufang Shenhua Tablet(复方肾华片) on the Expression of Toll-Like Receptors during Acute Kidney Injury Induced by Ischemia-Reperfusion in Rats

Objective： To investigate the impact of a traditional Chinese medicinal compound known as Fufang Shenhua Tablet （复方肾华片, SHP） on the expression of Toll-like receptors （TLRs） during renal ischemia-reperfusion injury （IRI）-induced acute kidney injury （AKI） in rats. Methods： A total of 28 Wistar rats were randomly divided into five groups： （1） pseudo-operation control group, （2） ischemia-reperfusion model group, （3） Astragaloside group, （4） high-dose SHP group, and （5） low-dose SHP group. There were four rats in the pseudo-operation group and six rats in each of the other groups. The accepted ischemia-reperfusion model was established after a 7-day gavage intervention, and pathological changes and renal function were observed, using an enzyme-linked immunosorbent assay （ELISA） to detect interleukin 8 （IL-8） and interferon gamma （IFN-r） levels, as well as immunohistochemical staining to detect altered levels of TLR2 and TLR4 expression in renal tissue. Results： After 24 h, renal pathological damage and the expression levels of serum creatinine （Scr）, IL-8, IFN- r, TLR2, and TLR4 were significantly higher in the model group as compared with the pseudo-operation group （P〈0.05）. In addition, at 24 h the above indicators decreased significantly in the Astragaloside group, high- dose SHP group and low-dose SHP group as compared with the ischemia-reperfusion model group （P〈0.05）. TLR2 and TLR4 expression levels were significantly reduced in the SHP treatment and Astragaloside group as compared with the pseudo-operation group （P〈0.05）. Further, the high-dose SHP group showed significantly less renal damage score and decreased levels of TLR expression than those of low-dose SHP group and Astragaloside group （all P〈0.05）. Conclusion： SHP can alleviate the renal structural and functional damage caused by IRI-induced AKI in rats by reducing the damage of renal pathology, which may reduce inflammatory cytokine levels by downregulating the expression of TLRs in renal tissue in a dose-dependent manner.

Treatment of retinal vein occlusion in rabbits with traditional Chinese medicine Fufang XueShuan Tong

Background Retinal vein occlusion （RVO） is one of the most common causes of visual loss. Many approaches have been tried to treat central retinal vein occlusion （CRVO）, and branch retinal vein occlusion （BRVO） with various results. However, there is no defined protocol and limited evidence to support the interventions currently used. The aim of this study was to assess the efficacy of the traditional Chinese medicine Fufang XueShuan Tong （FXST） in treating experimentally created RVO. Methods RVO model was first induced in forty-four pigmented rabbits through photocoagulation following injection of rose Bengal. The rabbits were divided into four groups based on the dose of FXST administered （212 mg/kg, 424 mg/kg, 848 mg/kg and control group）. The rabbits were observed for four weeks after the procedure, using color fundus photography, fundus fluorescein angiography and electroretinogram examination. Vascular endothelial growth factor （VEGF）, interleukin-6 and nitric oxide （NO） levels in the vitreous and histopathologic evaluation were monitored. Results The obstructed vessels in the treatment groups reopened or anastomosed faster than those in the control group （P〈0.05）. The amplitude of maximum b wave and the oscillatory potential were significantly higher in the treatment groups than in the control group （P 〈0.01）. At both two weeks and four weeks, VEGF and IL-6 levels in the vitreous were significantly decreased in the treatment groups （P 〈0.01）, while NO levels were significantly elevated （P 〈0.01）. At the same time, histopathologic evaluation showed different retinal neuroepithelium structures in the different groups. Immunoreactivity of VEGF was greater in the control group than in the treatment groups. Conclusion FXST was helpful in reconstructing retinal vessels in the RVO model, protecting retinal structures and improvinq visual function, and could inhibit the neovascular factor.

Effect of compatible herbs on the pharmacokinetics of effective components of Panax notoginseng in Fufang Xueshuantong Capsule

Fufang Xueshuantong （FXT） is a well-known Chinese herbal formula which has been used to treat car- diovascular and ophthalmic diseases, especially diabetic retinopathy. Panax notoginseng （Burkill） F.H. Chen （PN） is the main herb of FXT, whose major bioactive constituents are ginsenosides. However, the scientific basis of the compatibility of FXT is still ambiguous. The present study investigated the scientific basis of the compatibility of FXT by comparing the pharmacokinetics of marker compounds after oral administrations of PN and FXT. A high performance liquid chromatography-electrospray ionization tandem mass spectrometry （HPLC-ESI-MS/MS） method was devel- oped for simultaneous detection of notoginsenoside R1 （NR1）, ginsenoside Rgl （GRgl）, and ginsenoside Rbl （GRbl） in rat plasma. The pharmacokinetic studies of FXT and PN were performed using the established method with the pharmacokinetic parameters being determined by non-compartmental analysis. The results showed that the phar- macokinetic parameters （maximum concentration, area under the curve （AUC0-t）, clearance, and mean residence time） of NR1, GRgl, and GRbl were significantly different after oral administration of FXT （P〈0.05） compared with PN. The AUO0-t values of GRgl and GRbl were 1.7- and 3.4-fold greater, respectively, in FXT than in PN. The compatible herbs of FXT could prolong the retention time and increase the systemic exposure of NR1, GRgl, and GRbl compared with PN in vivo, providing some scientific basis for the compatibility and clinical use of FXT.

The effects of borneol on the pharmacokinetics and brain distribution of tanshinone IIA,salvianolic acid B and ginsenoside Rg1 in Fufang Danshen preparation in rats

Fufang Danshen preparation(FDP)is consisted of Salviae Miltiorrhizar Radix et Rhizoma(Danshen),Notoginseng Radix et Rhizoma(Sanqi)and Borneolum Syntheticum(borneol).FDP is usually used to treat myocardial ischemia hypoxia,cerebral ischemia and alzheimer’s disease,etc.In the treatment of cerebrovascular diseases,borneol is usually used to promote the absorption and distribution of the bioactive components to proper organs,especially to the brain.The purpose of this study is investigating the effects of borneol on the pharmacokinetics and brain distribution of tanshinone IIA(TS IIA),salvianolic acid B(SAB)and ginsenoside Rg1 in FDP.Male healthy Sprague-Dawley(SD)rats were given Danshen extracts,Sanqi extracts(Panax notoginseng saponins)or simultaneously administered Danshen extracts,Sanqi extracts and borneol.Plasma and brain samples were collected at different points in time.The concentration of TS IIA,SAB and Rg1 was determined by UPLC-MS/MS method.The main pharmacokinetics parameters of plasma and brain tissue were calculated by using Phoenix WinNolin 6.1 software.In comparison with Danshen and Sanqi alone,there were significant differences in pharmacokinetic parameters of TS IIA,SAB and Rg1,and the brain distribution of SAB and TS IIA when Danshen,Sanqi and borneol were administrated together.Borneol statistically significant shortened tmax of TS IIA,SAB and Rg1 in plasma and brain,increased the bioavaiability of Rg1,inhibited metabolism of Rg1 and enhanced the transport of TS IIA and SAB to brain.These results indicated that borneol could affect the multiple targets components and produce synergistic effects.Through accelerating the intestinal absorption and brain distribution,borneol caused the effective ingredients of Danshen and Sanqi to play a quicker therapeutic role and improved the therapeutic effect.

Effect of Fufang Xueshuantong Capsule(复方血栓通胶囊) on A Rat Model of Retinal Vein Occlusion

Objective：To establish a retinal vein occlusion（RVO） animal model and observe the therapeutic effect of a Chinese herbal composition（Fufang Xueshuantong Capsule,复方血栓通胶囊,FXC） in ischemic retinal disease.Methods：Fifteen adult male Sprague-Dawley rats underwent laser photothrombosis to induce RVO on their right eyes and were subsequently randomized to receive FXC（the intervention group,n=7） or placebo treatment（the control group,n=8）.Fundus fluorescein angiography was performed after 2,4 and 8 weeks of treatment.Real-time reverse transcription-PCR was used to quantify the mRNA expression of vascular endothelial growth factor（VEGF） and stromal cell-derived factor-1（SDF-1）.The main outcomes were the mRNA copies of VEGF and SDF-1 and the counts of RVO signs.Results：Laser photothrombosis procedure induced typical lesions of RVO,including hemorrhage,leakage,retinal detachment,capillary non-perfusion,filling defect of retinal vessels,and lateral circulation/dilation of small vessels.The retinal lesions were associated with an increased expression of VEGF（P0.05）.No significant change of SDF-1 expression was noticed.Compared with the control group,the intervention group had numerically fewer RVO lesions at week 2（1.71±0.76 vs.3.50±1.51,t=-2.82,P0.05）.The benefit of intervention remained at weeks 4 and 8.Conclusions：A rat model of laser photothrombosis-induced RVO was established and an increase in the VEGF expression was observed in the retinal lesion.The FXC had therapeutic benefit in improving retinal lesions in the rat model of RVO.

Effects of novel Fufang Biejia Ruangan Tablets with sheep placenta as substitute for Hominis Placenta on CCl4-induced liver fibrosis

Objective:Fufang Biejia Ruangan Tablet(FBRT) is widely used for the treatment of liver fibrosis.However,Hominis Placenta(HP),as an important adjuvant of FBRT,has been restricted for medicinal using due to the limited availability,ethical controversy and safety issues.The present study aimed to investigate the therapeutic effects of novel FBRT(N-FBRT) with sheep placenta(SP) as substitute for HP on liver fibrosis and explore its possible mechanisms.Different dosages of SP in N-FBRT were also evaluated.Methods:Rats were subcutaneously injected with CCl_(4)to induce liver fibrosis and then treated with NFBRT and FBRT.The anti-hepatic fibrosis effect was determined based on biomarkers analysis of liver function and hepatic fibrosis,and the liver pathology was visualized by H&E staining and Masson staining.The oxidative stress and inflammatory cytokines were also detected.Immunohistochemical staining of a-SMA,real time PCR and Western blotting were performed to evaluate hepatic stellate cells(HSCs)activation and TGF-β1/Smad signaling pathway.Results:N-FBRT and FBRT could ameliorate CCl_(4)-induced liver fibrosis and improve liver function,as evidenced by lowering serum biomarkers levels of liver function and hepatic fibrosis,and decreasing hepatic Hyp content and collagen deposition,and improving the hepatic morphology and architecture changes.Moreover,the anti-liver fibrosis effect was better when the dosage of SP used in N-FBRT was 1/2 of HP in FBRT.Administration of N-FBRT markedly alleviated oxidative stress and inflammatory cytokines,and inhibited a-SMA expression.Furthermore,the mRNA expression of Col Ⅰ,Col Ⅲ,a-SMA and TGF-β1,and proteins expression of a-SMA,TGF-β1,Smad2/3 and p-Smad2/3 were significantly down-regulated by N-FBRT treatment.Conclusion:SP can be used as substitute for HP to prepare N-FBRT for the treatment of liver fibrosis and the anti-liver fibrosis effect of N-FBRT is achieved by eliminating oxidative stress and inflammation,and inhibiting HSCs activation and ECM production by blocking TGF-β1/Smad signaling pathway.

Anti-hypertensive and endothelia protective effects of Fufang Qima capsule(复方芪麻胶囊) on primary hypertension via adiponectin/adenosine monophosphate activated protein kinase pathway

OBJECTIVE:To investigate the potential mechanism of the vascular remodeling effect and provide additional information about anti-hypertension activity of Fufang Qima capsule(复方芪麻胶囊,QM).METHODS:Spontaneous hypertensive rats(SHRs)were used to study the underlying mechanism of the anti-hypertension activity of QM.In this study,SHRs were randomly divided into 5 groups:model group,Telmisartan group(7.2 mg/kg,p.o.),and three QM groups(0.9298,1.8596,and 3.7192 g/kg,p.o.).Wistar Kyoto rats(WKY)were used as normal control group.Blood pressure(BP),aorta,perivascular adipose tissue(PVAT)histology were investigated to evaluate the effect of QM.Nitric oxide(NO)and endothelial nitric oxide synthase(eNOS)phosphorylation were measured.Adiponectin(APN)secretion,as well as APN signal pathway proteins including APN,adiponectin receptors(R1 and R2)and adenosine 5’-monophosphate-activated protein kinase(AMPK)were all analyzed.RESULTS:QM significantly reduced BP and ameliorated the vascular pathological change,i.e.intima media thicken and collagen fiber hyperplasia.Meanwhile,QM increased concentration of NO and the phosphorylation of eNOS in the aorta.The anti-hypertensive and endothelia-protective effect of QM could be attributed to activating APN/AMPK pathway by up-regulating the expression of APN in PVAT and APN Receptor 2,AMPKαand phosphorylated AMPKαin the aorta.CONCLUSION:The QM alleviation effect mechanism for primary hypertension was via modulating the APN/AMPK signal pathway.

Effect of treatment with Fufang Huangqi decoction(复方黄杞汤剂) on dose reductions and discontinuation of pyridostigmine bromide tablets, prednisone, and tacrolimus in patients with type Ⅰ or Ⅱ myasthenia gravis

OBJECTIVE: To investigate the clinical efficacy of Fufang Huangqi decoction(复方黄杞汤剂) in combination with pyridostigmine bromide tablets, prednisone, and tacrolimus in the treatment of type Ⅰ and Ⅱ myasthenia gravis(MG) through changes in the clinical symptom scores of 100 patients with type Ⅰ and Ⅱ MG. This study also aimed to examine dose reductions and discontinuation of these 3 Western medicines after administration of Fufang Huangqi decoction. METHODS: The clinical data on 100 patients with type I or II MG who were treated in the outpatient department of the Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, China, between June 2017 and June 2020 were collected. The patients were divided into 4 groups based on whether they had taken pyridostigmine bromide tablets, prednisone, and/or tacrolimus at the time of their hospital visit: the Fufang Huangqi decoction group(group A), the pyridostigmine bromide tablets + Fufang Huangqi decoction group(group B), the pyridostigmine bromide tablets + prednisone + Fufang Huangqi decoction group(group C), and the pyridostigmine bromide tablets + tacrolimus + Fufang Huangqi decoction group(group D). The average treatment time was(15.6 ± 11.5) months(range: 0.5-55 months). Changes in the clinical symptom scores of the 4 groups of patients after medication administration and dose reductions and discontinuation of the 3 Western medicines were analyzed. RESULTS: An overall effectiveness rate of 86.00% was achieved in the 100 patients after treatment for(15.6 ± 11.5) months(range 0.5-55 months). The effectiveness rates were 85.71% in group A, 88.24% in group B, 76.92% in group C, and 80.00% in group D. The dosage of pyridostigmine bromide was reduced for 69.12% of the patients in group B for the first time after(4.2 ± 4.1) months, and 45.59% of the patients in group B discontinued pyridostigmine bromide after(8.8 ± 6.1) months. The dosage of pyridostigmine bromide was reduced for 46.15% of the patients in group C for the first time after(5.3 ± 3.4) months, and 23.08% of the patients in group C discontinued pyridostigmine bromide after(19.8 ± 11.0) months;76.92% reduced hormone dosage after(2.8 ± 1.9) months, and 23.08% discontinued hormone treatment after(6.7 ± 2.9) months. The dosage of pyridostigmine bromide was reduced for 1 patient in group D after 1 month;this patient discontinued pyridostigmine bromide after 3 months and reduced tacrolimus dosage after 5 months. One patient in group D discontinued pyridostigmine bromide and tacrolimus on his own initiative at 0.5 months and took Fufang Huangqi decoction for 2 months without discontinuing Western medicine. CONCLUSION: Fufang Huangqi decoction is effective for the treatment of type Ⅰ and Ⅱ MG and improves the associated clinical symptoms. Moreover, this agent is conducive to dose reductions and discontinuation of basic Western medicines, thereby reducing the side effects experienced by patients.

Simultaneous Determination of Seven Alkaloids in Fufang Zhenzhu Tiaozhi Capsule by HPLC Coupled with DAD

Objective To establish a reverse-phase liquid chromatography method for the determination of seven alkaloids(magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine)in Fufang Zhenzhu Tiaozhi Capsule.Methods Chromatography was performed on a Dionex Acclaim C_(18)column(250 mm×4.6 mm,5.0μm)at 30℃.The mobile phase was composed of acetonitrile-potassium dihydrogen phosphate solution(0.015 mol/L,40:60,including 1.7 g/L sodium dodecyl sulfate and phosphoric acid used to regulate pH value to 3.0),with a flow rate of 1.0 mL/min.The detection wavelength was 270 nm.Results The calibration curves of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were linear in the range of 1.07-10.65,0.78-7.55,0.75-7.50,1.60-15.95,2.69-26.85,2.31-23.10,and 6.04-60.40 mg/mL.The average recoveries of magnoflorine,columbamine,jatrorrhizine,epiberberine,coptisine,palmatine,and berberine were 101.0%,101.2%,100.1%,100.0%,100.1%,101.1%,and 99.7%,respectively.Conclusion The method could be used for the quantitative determination of the preparation.

Simultaneous Determination of Four Substances in Plasma and Dermal Microdialysates of Guinea Pig after Different Acupoints Administration of Fufang Baijiezi Gel

Objective To study the pharmacokinetic properties of Fufang Baijiezi Gel（FBG） after different acupoints administration. Methods Sinapine thiocyanate, tetrahydropalmatine, 6-gingerol, and asarinin, which were four substances of FBG, were determined by a sensitive liquid chromatography tandem mass spectrometry method（LC-MS） both in plasma and dermal microdialysates of guinea pig simultaneously. Microdialysates were separated on an Ultimate？ XB-Phenyl analytical column（150 mm ×2.1 mm, 5μm） and detected by electrospray ionization（ESI） in selected ion monitoring（SIM） mode. The method was validated in terms of selectivity, linearity, sensitivity, and recovery. Result A significant difference was observed in main pharmacokinetic parameters of C max, t max, and AUC between acupoints administration and nonacupoints administration. Conclusion Acupoints administration resulted in a more obvious increase in bioavailability of sinapine thiocyanate, tetrahydropalmatine, 6-gingerol, and asarinin than nonacupoints administration.