Effect of Liuwei Dihuang Tang on ferroptosis pathway in retinal tissue of aging model rats / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To observe the effects of Liuwei Dihuang Tang on the expression levels of ferritin, recombinant solute carrier family 7 member 11(SLC7A11), glutathione(GSH), and glutathione peroxidase 4(GPX4)in retinal of aging model rats.METHODS: A total of 45 SD rats were randomly divided into a blank group, a model group, and a traditional Chinese medicine(TCM)group, with 15 rats in each group. The blank group was intraperitoneally injected with physiological saline, while the model group and TCM group were intraperitoneally injected with D-galactose [500 mg/(kg·d)]. At the same time, the TCM group was orally administered with Liuwei Dihuang Tang [6.75 g/(kg·d)], while the blank group and model group were orally administered with equal volume of physiological saline for 8 consecutive wk. The expression levels of ferritin, SLC7A11, GSH, and GPX4 in the retinal tissues of rats in each group were detected.RESULTS: The expression of ferritin in the retinal tissue of the model group was increased compared to the blank group(P<0.05), while the expression of GSH, SLC7A11, and GPX4 was reduced(P<0.05). The expression of GSH, SLC7A11, and GPX4 in the retina tissue of rats in the TCM group was higher than that in the model group(P<0.05).CONCLUSIONS: Liuwei Dihuang Tang may exert a delaying effect on retinal aging by regulating the ferroptosis pathway.

Effect of Liuwei Dihuang decoction on β-catenin and EMT in kidney tissue of UUO rats / 中国药理学通报

Aim To explore the effect of Liuwei Dihuang decoction ( LWDHD) on the expression of β-catenin, E-cadherin,α-SMA, the pathological changes of renal tissue, and the changes of an epithelial-mesen-chymal transformation ( EMT) in renal tissue of rats with unilateral ureteral obstruction ( UUO ) . Methods Forty-eight SPF grade SD rats were randomly divided into sham group ( Sham), model group ( UUO), Liuwei Dihuang decoction low, medium, and high groups ( LWDHD 3. 375, 6. 75, 13. 5 g · kg

Mechanism of Liuwei Dihuang Pills in treatment of mice with diminished ovarian reserve based on proteomics / 中国中药杂志

This study aims to investigate the efficacy and possible mechanism of Liuwei Dihuang Pills in the treatment of diminished ovarian reserve(DOR) by using proteomic techniques. Firstly, cyclophosphamide(60 mg·kg~(-1)) combined with busulfan(6 mg·kg~(-1)) was injected intraperitoneally to establish the mouse model of DOR. After drug injection, the mice were continuously observed and the success of modeling was evaluated by the disturbance of the estrous cycle. After successful modeling, the mice were administrated with the suspension of Liuwei Dihuang Pills by gavage for 28 days. At the end of the gavage, four female mice were selected and caged together with males at a ratio of 2∶1 for the determination of the pregnancy rate. Blood and ovary samples were collected from the remaining mice on the next day after the end of gavage. Hematoxylin-eosin(HE) staining and transmission electron microscopy(TEM) were then employed to observe the morphological and ultrastructural changes in the ovaries. The serum levels of hormones and oxidation indicators were measured by enzyme-linked immunosorbent assay. Quantitative proteomics techniques were used to compare the ovarian protein expression before and after modeling and before and after the intervention with Liuwei Dihuang Pills. The results showed that Liuwei Dihuang Pills regulated the estrous cycle of DOR mice, elevated the serum levels of hormones and anti-oxidation indicators, promoted follicle development, protected the mitochondrial morphology of ovarian granulosa cells, and increased the litter size and survival of DOR mice. Furthermore, Liuwei Dihuang Pills negatively regulated the expression of 12 differentially expressed proteins associated with DOR, which were mainly involved in lipid catabolism, inflammatory response, immune regulation, and coenzyme biosynthesis. These differentially expressed proteins were significantly enriched in sphingolipid metabolism, arachidonic acid metabolism, ribosomes, ferroptosis, and cGMP-PKG signaling pathway. In summary, the occurrence of DOR and the treatment of DOR with Liuwei Dihuang Pills are associated with multiple biological pathways, mainly including oxidative stress response, inflammatory response, and immune regulation. "Mitochondria-oxidative stress-apoptosis" is the key to the treatment of DOR by Liuwei Dihuang Pills. YY1 and CYP4F3 may be the key upstream targets that trigger mitochondrial dysfunction and ROS accumulation, and the metabolism of arachidonic acid is the main signaling pathway of drug action.

Comparison of the fingerprint and multi-component contents between Liuwei dihuang powder decoction pieces and traditional decoction pieces with different decocting time / 中国药房

OBJECTIVE To compare the change law of multi-components in the extraction process between Liuwei dihuang powder decoction pieces and traditional decoction pieces （hereinafter referred to as powder decoction pieces and traditional decoction pieces）， and to provide scientific basis for the modern technology research of Liuwei dihuang formula. METHODS Taking powder decoction pieces and traditional decoction pieces as samples， the samples were taken when soaking for 60 min， at 0， 5， 10， 15， 20， 25， 30， 40， 50， 60 min of the first decocting and at 5， 10， 20， 30， 40 min of the second decocting， respectively. HPLC method was used to establish the fingerprints of 2 kinds of decoction pieces with different decocting time. The similarity evaluation and peak identification were performed. The contents of 8 components including 5-hydroxyfurfural， catechin， monoglycoside， loganin， swertin glycoside， dihydroquercetin， paeonol and benzoyl paeoniflorin were all determined. RESULTS With different decocting time， the similarties between 2 kinds of decoction pieces and their respective control fingerprints R were all greater than 0.98. In the fingerprints of traditional decoction pieces， five chromatographic peaks were identified， namely， 5- hydroxyfurfural， monetin， swertiaoside， dihydroquercetin and paeonol； in the fingerprints of powder decoction pieces， six chromatographic peaks were identified， namely， 5-hydroxyfurfural， monoglycoside， swertiamarin， dihydroquercetin， paeonol and benzoyl paeoniflorin. The results of content determination showed that in the first 5 minutes of the first decocting， the decocting rate of almost all the ingredients in the powder decoction pieces was faster than that of the traditional decoction pieces； after 40 min， the contents of other active ingredients were lower than those of traditional decoction pieces except for 5-hydroxyfurfural and paeonol. In the process of second decocting， except for paeonol and loganin， the contents of other ingredients in powder decoction pieces were higher than that in traditional decoction pieces； catechin was completely decocted from the traditional decoction pieces in the first decocting， while it could still be detected in the powder decoction pieces in the second decocting. There was little difference in the total decocted amount of the 8 ingredients in the two decoction pieces. CONCLUSIONS The chemical composition of powder decoction pieces of Liuwei dihuang formula has no obvious advantages compared with traditional decoction pieces， and can not save the decocting time and the amount of medicinal materials.

Comparative proteomic profiling of hippocampi in mice treated with Jingui Shenqi Pills and Liuwei Dihuang Pills / 中国中药杂志

The effects of Jingui Shenqi Pills(Jingui) and Liuwei Dihuang Pills(Liuwei) which respectively tonify kidney Yang and kidney Yin on brain function have attracted great attention, while the differences of protein expression regulated by Jingui and Liuwei remain to be studied. This study explored the difference of protein expression profiles in the hippocampi of mice orally administrated with the two drugs for 7 days. The protein expression was quantified using LC-MS/MS. The results showed that among the 5 860 proteins tested, 151, 282 and 75 proteins responded to Jingui alone, Liuwei alone, and both drugs, respectively. The ratio of up-regulated proteins to down-regulated proteins was 1.627 in Jingui group while only 0.56 in Liuwei group. The proteins up-regulated by Jingui were mainly involved in membrane transport, synaptic vesicle cycle, serotonergic synapse, dopaminergic synapse and so on, suggesting that Jingui may play a role in promoting the transport of neurotransmitter in the nervous system. The proteins down-regulated by Liuwei were mainly involved in membrane transport, synapse, ion transport(potassium and sodium transport), neurotransmitter transport, innate and acquired immune responses, complement activation, inflammatory response, etc. In particular, Liuwei showed obvious down-regulation effect on the members of solute carrier(SLC) superfamily, which suggested that Liuwei had potential inhibitory effect on membrane excitation and transport. Finally, consistent results were obtained in the normal mouse and the mouse model with corticosterone-induced depressive-like behavior. This study provides an experimental basis for understanding the effect of Jingui and Liuwei on brain function from protein network.

Active fraction combination from Liuwei Dihuang decoction improves adult hippocampal neurogenesis and neurogenic microenvironmentin cranially irradiated mice / 中国药理学与毒理学杂志

OBJECTIVE Cranial radiotherapy is clinically used in the treatment of brain tumors;however, the conse?quent cognitive and emotional dysfunctions seriously impair the life quality of patients. LW-AFC, an active fraction combi?nation extracted from classical traditional Chinese medicine prescription Liuwei Dihuang decoction, can improve cogni?tive and emotional dysfunctions in many animal models;however, the protective effect of LW-AFC on cranial irradiation-induced cognitive and emotional dysfunctions has not been reported. Recent studies indicate that impairment of adult hippocampal neurogenesis (AHN) and alterations of the neurogenic microenvironment in the hippocampus constitute crit?ical factors in cognitive and emotional dysfunctions following cranial irradiation. Here, our research further investigated the potential protective effects and mechanisms of LW-AFC on cranial irradiation-induced cognitive and emotional dys?functions in mice. METHODS LW-AFC (1.6 g·kg-1) was intragastrically administered to mice for 14 d before cranial irra?diation (7 Gyγ-ray). AHN was examined by quantifying the number of proliferative neural stem cells and immature neu?rons in the dorsal and ventral hippocampus. The contextual fear conditioning test, open field test, and tail suspension test were used to assess cognitive and emotional functions in mice. To detect the change of the neurogenic microenvi?ronment, colorimetry and multiplex bead analysis were performed to measure the level of oxidative stress, neurotrophic and growth factors, and inflammation in the hippocampus. RESULTS LW-AFC exerted beneficial effects on the contex?tual fear memory, anxiety behavior, and depression behavior in irradiated mice. Moreover, LW-AFC increased the num?ber of proliferative neural stem cells and immature neurons in the dorsal hippocampus, displaying a regional specificity of neurogenic response. For the neurogenic microenvironment, LW-AFC significantly increased the contents of superox?ide dismutase, glutathione peroxidase, glutathione, and catalase and decreased the content of malondialdehyde in the hippocampus of irradiated mice, accompanied by the increase in brain-derived neurotrophic factor, insulin-like growth factor-1, and interleukin-4 content. Together, LW-AFC improved cognitive and emotional dysfunctions, promoted AHN preferentially in the dorsal hippocampus, and ameliorated disturbance in the neurogenic microenvironment in irradiated mice. CONCLUSION LW-AFC ameliorates cranial irradiation-induced cognitive and emotional dysfunctions, and the underlying mechanisms are mediated by promoting AHN in the dorsal hippocampus and improving the neurogenic micro?environment. LW-AFC might be a promising therapeutic agent to treat cognitive and emotional dysfunctions in patients receiving cranial radiotherapy.

Discussion for Quality Control of Chinese Patent Drugs from Quality Difference Among Different Dosage Forms of Liuwei Dihuang Preparations / 中国实验方剂学杂志

Objective:Through comprehensive evaluation and analysis of the quality of Liuwei Dihuang （LWDH） preparations from different manufacturers and combining factors such as production technology， the key factors in the quality control of LWDH preparations are explored to provide a reference for improving the quality control level of LWDH preparations. Method:Morroniside， loganin and paeonol as quality control markers of LWDH products were determined by high performance liquid chromatography （HPLC）， the mobile phase was acetonitrile （A） -0.3% phosphoric acid aqueous solution （B） for gradient elution （0-5 min， 5%-8%A； 5-20 min， 8%A； 20-35 min， 8%-20%A； 35-45 min， 20%-60%A； 45-55 min， 60%A）， the detection wavelength of paeonol was at 274 nm， and the detection wavelengths of morroniside and loganin were at 240 nm. The quality characteristics of LWDH preparations with different dosage forms （big candied pills， water-honeyed pills， concentrated pills， hard capsules and soft capsules） from different manufacturers were analyzed. Combined these results with their actual production processes， the key-points of quality control in the whole production process were discussed. Result:The contents of three index ingredients in 128 batches of LWDH preparations were all in conformity with the standards of the 2015 edition of <italic>Chinese Pharmacopoeia</italic>， however， the content limit of some dosage forms in the current standard was unreasonable. For example， although the daily dose of crude drugs for big candied pills were almost twice the dose of water- honeyed pills （15.00， 8.57 g， respectively）， they got exactly the same daily limits of the contents for both the quality markers. What′s more， these two formulations had the same process， so the differences between the process obviously could not be the reason of these differences. Conclusion:It is recommended that for the products with different dosage forms should have a similar content limits， if there are no obvious distinctions between their production processes. Which may benefit the quality control of the products with multi-dosage forms. The research on the quality standards of proprietary Chinese medicines should deeply study the existing characteristics of the quality standards， and fully respect the laws of the quality attributes of traditional Chinese medicines and the rules of the production process of Chinese patent medicines.

Effects of Modified Liuwei Dihuang Decoction on Kidney/Bone Injury of CKD-MBD Model Rats / 中国药房

OBJECTIVE：To study the effects of Modified liuwei dihuang decoction on kidney/bone injury of chronic kidney disease-mineral and bone disorder（CKD-MBD）model rats. METHODS ：The male SD rats were randomly divided into normal group（n＝10），high phosphorus group （n＝30），model group （n＝30），calcitriol group （positive control ，0.09 μg/kg，n＝30）， Modified liuwei dihuang decoction group （10 g/kg by crude drug ，n＝30）. CKD-MBD model was established by high phosphorus and adenine diet for 6 weeks. After modeling ，normal group and model group were given normal diet/high phosphorus diet and intragastric administration of water. Administration groups were fed with normal diet and given corresponding solution intragastrically（water as solvent ），0.1 mL/kg，once a day ，for consecutive 6 weeks. Blood sample of rats in the normal group were collected ，and they were sacrificed after the last administration. Blood sample of 10 rats in each other group were collected ， and they were sacrificed at 2，4 and 6 weeks after administration. The contents of blood urea nitrogen （BUN），serum creatinine （Scr），calcium，phosphorus，iPTH，FGF-23，RANKL and osteocalcin in serum were detected in each group. The bone mineral density（BMD）of femoral was measured ，the morphological changes of renal tissue and bone tissue were observed ，and the percentage of renal tubular injury and the score of renal interstitial fibrosis were calculated. RESULTS ：Compared with normal group，above indexes in high phosphorus group had no significant change at different time points （P＞0.05）. There was no abnormal change in renal/bone tissue. Compared with high phosphorus group at the same time point ，the contents of BUN ，Scr， phosphorus，iPTH，FGF-23，RANKL and osteocalcin in serum ，the percentage of renal tubular injury and the score of renal interstitial fibrosis in the model group were significantly increased ，while the contents of calcium in serum and the BMD of femoral were significantly decreased （P＜0.05 or P＜0.01）. The renal tissue showed diffuse fibrosis. The width of trabecular bone was increased and the number of osteoblasts was decreased. Compared with the model group at the same time point ，the contents of BUN（except for Modified liuwei dihuang decoction group after 2 weeks of administration ），Scr，serum phosphorus ，iPTH， FGF-23，RANKL and osteocalcin ，the percentage of renal tubular injury and the score of renal interstitial fibrosis in Modified liuwei dihuang decoction group and calcitriol group were decreased significantly at each time point ；serum calcium content and BMD（except for 2 weeks of administration ）were significantly increased （P＜0.05 or P＜0.01），and the pathological changes of renal/bone tissue were significantly improved ；there was no statistical significance in above indexes between Modified liuwei dihuang decoction group and calcitriol group （P＞0.05）. CONCLUSIONS ：Modified liuwei dihuang decoction can improve kidney/ bone injury of CKD-MBD model rats ，and improve BMD and regulate disorder of calcium and phosphorus metabolism.

Evaluation and analysis of high-quality products of Liuwei Dihuang Pills / 中国中药杂志

Liuwei Dihuang Pills is a typical traditional Chinese medicine with "the same product made by many manufacturers". The quality and price of products made in various factories was different obviously. In this study, the quality differences of Liuwei Dihuang Pills were evaluated over multi-dimensions and throughout the whole production cycle involving raw materials, production process, quality control and post-marketing re-studies based on the "Chinese patent medicine evaluation standard with quality at the core" established earlier by our research group. The research results showed that the product quality of various manufacturers was significantly different, and the product quality was positively correlated with the market shares of enterprises, indicating that enterprises with more market shares paid more attention to product quality; and the sales determined the concern degree of enterprises on products, which was in line with general cognition. During the single-item evaluation of Liuwei Dihuang Pills, the score of raw material selection was relatively low, and the enterprises paid less attention to the initial raw materials. The whole production process was better, and the national compulsory implementation of "Production Quality Management Standard"(GMP) had a positive effect in improving product quality. Quality control could basically guarantee product quality. Intelligent manufacturing promoted by the government was beneficial to ensure product uniformity. The score rate of "quality evaluation" item was basically qualified, which indicated that the quality control level of Liuwei Dihuang Pills was acceptable as a whole, but there was still room for improvement. "Re-evaluation and Brand Construction" had the lowest scores, reflecting that enterprises did not pay enough attention to in-depth study and re-evaluation of "the same product made by many manufacturers". The evaluation results were in line with expectations, provided a reference example for the evaluation of high-quality Chinese patent medicine, a basis for the high-quality and good price of Chinese patent medicine, scientific data for improving medical insurance bidding, and thus facilitated promoting the healthy development of the traditional Chinese medicine industry.

Effects of Liuwei Dihuang Decoction (Yukmijihwang-tang) on Physical Fatigue by Regulating Neurotransmitters in Brain / 中国结合医学杂志

OBJECTIVE@#To study the effect of Liuwei Dihuang Decoction () or Yukmijihwangtang (YJT) on endurance exercise by in vivo experiment.@*METHODS@#ICR mice were randomly divided into the control group (distilled water) and the YJT groups (1, 10, 100 mg/kg), 5 animals per group. YJT and distilled water were orally administered. The anti-fatigue effect of YJT was evaluated by open fifiled test (OFT), forced swimming test (FST), and tail suspension test (TST).@*RESULTS@#In the OFT, YJT signifificantly increased the total movement distance in a dose-dependent manner. Additionally, treatment with YJT signifificantly decreased immobility time in the FST and the TST. Various neurotransmitters such as norepinephrine (NE), serotonin (5-HT), dopamine (DA) levels were increased by FST. Administration of YJT down-regulated the expression levels of NE, 5-HT, 5-hydroxyindole-acetic acid (5-HIAA), and DA in the brain stem and hypothalamus of mice. Moreover, protein expression of HSP70 in mice liver and heart muscles was signifificantly increased in the YJT groups.@*CONCLUSIONS@#YJT could ameliorate fatigue and enhance exercise tolerance through suppressing of brain monoamines including NE, 5-HT, 5-HIAA, and DA in FST mice model.

Structure characteristics and quality control of Liuwei Dihuang Concentrated Pill from different manufacturers / 中国中药杂志

In this paper, we aim to control and evaluate the quality of Liuwei Dihuang Concentrated Pill by using the model of fingerprint technique and "component structure" theory. Agilent 5 TC-C_(18) column(4.6 mm×250 mm,5 μm) was used, with 0.1% formic acid solution-acetonitrile as mobile phase at a flow rate of 1.0 mL·min~(-1). The column temperature was 30 ℃, with detection wavelength of 242 nm and the sample volume of 10 μL. The characteristic fingerprint of Liuwei Dihuang Concentrated Pill was established by high performance liquid chromatography(HPLC) for its quality control. Seventeen common peaks were identified, and the similarity was 0.550-0.997 in 29 batches of samples, indicating that the quality difference among batches of Liuwei Dihuang Concentrated Pills was significant. The structural characteristics of the Moutan Cortex components in Liuwei Dihuang Concentrated Pills were characterized. On this basis, combined with the structural characteristics of genuine components of Moutan Cortex, the structural characteristics of components in Liuwei Dihuang Concentrated Pills were further analyzed. The results showed that there were significant differences in the contents and quantity ratios of 9 representative components(components) of Moutan Cortex in Liuwei Dihuang Concentrated Pills from different manufacturers, indicating internal quality differences among different batches of products. The fingerprint technique and the "component structure" theory established by the above research provide an analytical method and a research foundation for the quality evaluation of Liuwei Dihuang Concentrated Pills.

Current situation and consideration of patent protection in classical representative famous prescriptions in China / 中国中药杂志

Classical Representative Famous Prescription is the valuable cultural heritage of Chinese medicine. In November 2018,the State Council issued the " Intensive Implementation of the National Intellectual Property Strategy in 2018 to Accelerate the Construction of IP Strong Country",explicitly proposing to strengthen the intellectual property protection of Classical Representative Famous Prescription.How about the current situation of intellectual property protection of lassical Representative Famous Prescription in China? We selected Liuwei Dihuang Pills,Shengmai Powder and Guizhi Fuling Pills( three representative drugs on market) from Chinese Pharmacopoeia2015 Volume I issued by Chinese Pharmacopoeia Commission to analyze their patent layout,reflecting its status quo of patent protection as follows: first,in recent years,the number of related patent applications for Classical Representative Famous Prescriptions has declined,which was positively correlated with the drug registration and approval policies in recent years,but the policy dividend has not been reflected in the patent application,which may be related to the long period of pharmaceutical R&D; secondly,the patent applicant in the field of Chinese medicine is mainly based on individuals,but the applicant of Classical Representative Famous Prescription is mainly of enterprises,and in addition,the company applicants have the highest authorization rate; thirdly,the main technologies are to improve preparation method and the dosage form in the research and development of Classical Representative Famous Prescription,but these two types of authorized patents have much difficulty in further application on the market. Therefore,the innovative entities shall look for a new breakthrough in secondary development and utilization of Classical Representative Famous Prescriptions.

Effects of Liuwei Dihuang active fraction combination on learning and memory impairment of neuroinflammatory model mice induced by lipopolysaccharide / 国际药学研究杂志

Objective: To study the effect and action mechanism of Liuwei Dihuang(LW)active fraction combi- nation(LW-AFC)on learning and memory impairment of neuroinflammatory model induced by lipopolysaccharide(LPS). Methods: The C57 mice were randomly divided into control group, model group, positive drug group and test groups ac- cording to their autonomic activity and body mass. The mice in the test groups were orally given LW[1.17 g/(kg•d)]and LW-AFC[1.6 g/(kg•d)]for two weeks in advance. The spatial learning and memory ability of the mice in each group were evaluated by Morris water maze test;immunofluorescence was used to observe the activation of microglia in the brain;Luminex 200 was used to detect the level of inflammatory cytokines in peripheral blood and hippocampus. Results: LW-AFC could improve the spatial learning and memory impairment induced by a single intraperitoneal injection (ip)of LPS(P<0.05); the activation of glial cells in the brain was inhibited(P<0.01);the levels of IL-6 and MCP-1 in the hippocampus were reduced(P<0.05). Conclusion: LW-AFC could reduce the neuroinflammation caused by the activation of microglia in brain, but could not effectively ameliorate the spatial learning and memory impairment caused by the activation of microglia in hippocampus.

Methodology Study on Rapid Quality Control of Liuwei Dihuang Capsule by AOTF-near Infrared Spectroscopy / 中国药房

OBJECTIVE： To establish the method for the rapidly non-destructive quality control of Liuwei dihuang capsule. METHODS： AOTF-NIR spectrometry was adopted. Taking 80 batches of Liuwei dihuang capsule produced by a manufacturer in recent three years as samples， HPLC chromatogram was adopted to determine the contents of loganin， morroniside， paeonol， paeoniflorin and ursolic acid； the content of water was determined according to general principles stated in 2015 edition of Chinese Pharmacopeia （part Ⅰ）. Taking 70 batches of samples as correction set， the partial least square method and the cross-validation algorithm were used to establish the NIR quantitative model of 6 indexes in Liuwei dihuang capsules with the Unscrambler quantitative analysis software. Taking residual 10 batches of samples as validation set， external validation was conducted for the model. RESULTS： The correlation coefficients （R2） of internal and external validation of loganin， morroniside， paeonol， paeoniflorin， the content of water quantitative model were all greater than 0.9； the correction of standand deviation （RMSEC） were 0.372 8， 0.025 4， 0.263 3， 0.288 5， 0.186 7 and 0.037 7； the prediction of standard deviation （RMSEP） were 0.462 2， 0.077 5， 0.472 1， 0.634 9， 0.293 4 and 0.206 9； the external verification showed that mean deviations of preclicted value to actual value were 6.04％， 6.05％， 5.87％， 6.97％， 5.62％ and 4.83％， with the mean deviation less than 10％.CONCLUSIONS：The established method can achieve rapidly non-destructive analysis Liuwei dihuang capsule.

Comparative Study on Glycosides Content in Liuwei Dihuang Decoction and Cornus Officinalis - Cortex Moutan Couple / 中国中医药信息杂志

Objective To establish an HPLC method for the content determination of morroniside, sweroside, paeoniflorin and loganin of Liuwei Dihuang Decoction and its Cornus Officinalis-Cortex Moutan couple; To discuss the relationship between the whole prescription and the couple of main pharmacodynamic components. Methods The HPLC method was used at Hypersile C18 column (4.6 mm × 250 mm, 5 μm); the mobile phase consisted of methanol-water (24:76); the detective wavelength was 236 nm; the flow rate was 1.0 mL/min; the column temperature was 30 ℃. Results The linear ranges of morroniside, sweroside, paeoniflorin and loganin were among 0.480–7.680 μg (r=0.999 3), 0.103–1.650 μg (r=0.999 5), 0.120–1.920 μg (r=0.999 1) and 0.227–3.630 μg (r=0.999 7), respectively. The average recovery rates and RSD were 102.79%, 102.29%, 100.99%, 102.48%, and 1.73%, 1.48%, 1.32%, 0.75%, respectively. The contents of morroniside, sweroside and paeoniflorin in Liuwei Dihuang Decoction were slightly higher than that in Cornus Officinalis - Cortex Moutan couple, and the contents of loganin were almost the same. Conclusion The method is simple, stable, accurate and reproducible. It can be used for content determinate of glycosides in Liuwei Dihuang Decoction and Cornus Officinalis-Cortex Moutan couple. Cornus Officinalis-Cortex Moutan couple has the glycosides with tonifying kidney effect of Liuwei Dihuang Decoction.

Construction of NIM sub-network regulated by Liuwei Dihuang decoction and its characteristics / 中国药理学与毒理学杂志

OBJECTIVE To construct the neuroendocrine immunomodulation (NIM) sub-network regulated by Liuwei Dihuang decoction (LW) and analyze its characteristics. METHODS We took the GSE57273 in GEO database and screened the differentially expressed genes (DEGs)(P<0.01) by the GEO2R tool as gene expression signature of LW. The global PPI network was constructed in the context of whole PPI network through direct interaction algorithm and forest algorithm respectively.Then the enrichment and the topological characteristics of NIM signaling molecules were evaluated by permutation test. Finally, we abstracted the NIM sub-network by NIMNT, which combined the NIM molecular network and forest algorithm, and analyzed the topological characteristics of it by the Network Analyzer(release 2.7)plugin in Cytoscape v3.5.1.RESULTS We got 2468 DEGs in the gene expression signature of LW.After analyzing the global PPI network of LW got by two kinds of algorithms,we found that the NIM signaling molecules significantly enriched and located in important positions in the global PPI network. The NIM sub-network regulated by LW contained 1099 nodes and 1056 edges. We screened out 22 hub nodes (Degree>10). Among them, there were 19 NIM signaling molecules in which only ESR1 changed significantly and 3 non-DEGs(NFATC2,RARA,TP53).However,the down-stream of the hub nodes were significantly changes. CONCLUSION The results suggested that LW might mainly regulate the non-hub nodes to recovery of the network imbalance of the body in the state of disease.

Liuwei Dihuang Decoction improves cognitive impairments via regulating immune system in APP/PS1 transgenic mouse,a mouse model of Alzheimer disease / 中国药理学与毒理学杂志

OBJECTIVE To investigate the effect and mechanisms of Liuwei Dihuang Decoction (LW)on cognition in PrP-hAβPPswe/PS1ΔE9(APP/PS1)transgenic mice.METHODS LW was adminis-trated with oral for 3 months.The locomotor activity test was performed to investigate the spontaneous motor activity of mice. The Morris water maze test and shuttle box test were performed to investigate the spatial learning and memory and active avoidance response respectively.The Αβ deposits and neuron loss in the hippocampus was detected by immunofluorescence staining and nissl staining respectively. The flow cytometry was employed to investigate the lymphocyte subsets of the mice.The 3H-thymidine incorporation was performed to investigate the splenocytes proliferation. RESULTS The treatment of LW ameliorated the impairments of spatial learning and memory and active and passive avoidance in APP/PS1 mice. The administration of LW alleviated neuron loss in the brain, suppressed amyloid-β (Αβ) deposits in the hippocampus of APP/PS1 mice. The treatment of LW significantly increased ConA-and LPS-induced proliferation of splenocytes,increased CD3+T cells and CD19+B cells in the spleen lymphocytes and reduced Gr1+cells in APP/PS1 mice.CONCLUSION This data indicated the adminis-tration of LW ameliorated behavioral and pathological deterioration via regulating immune function.

Based on urine metabolomics to study effect of Liuwei Dihuang Wan on growth and development of rats / 药学学报

On base of traditional physiological studies, building a comprehensive evaluation system for the efficacy of Liuwei Dihuang Wan (LW) for regulating the growth and development of rats by indicators that the overall metabolic profile and biomarkers of urinary metabonomics. Sixty-seven day-old Sprague-Dawley rats were randomly divided into four groups that are the control group and low-dose, medium-dose, and high-dose groups of LW. At the age of 28 days, different doses of LW extracts were continuously administered for 16 days. On the basis of behavior, body weight, organ index and femur length, a UPLC-HDMS technique was used to establish urine metabolomics method to characterize the profile of urine metabolism before and after intervention of LW, analysis and identificate related to drug effects of differential metabolism markers, and study the effect of LW on the growth and development of rats. LW could improve the overall behavioral ability of rats, promote weight gain and the growth of organs and bones, and 36 differential metabolites and 13 target metabolic pathways of urine under the intervention of LW were identified, and 8 metabolic markers were regarded as a key marker that LW effect the growth and development of rats. LW has a significant regulatory effect on the urine metabolic network during the growth and development of rats. This study interpreted the scientific implications of the effect of "Nourishing Kidney Yin" on the growth and development of rats.

Investigation of herb-drug interaction between Liuwei Dihuang Pills and nifedipine based on rat liver microsomes and HPLC fingerprints / 中国药科大学学报

@#A new method based on rat liver microsomes and chromatographic fingerprint comparison was established to investigate the possible herb-drug interactions between Liuwei Dihuang Pills(LDP)and nifedipine(NIF). LDP and NIF were incubated simultaneously with rat liver microsomes at 37 °C for 60 min in a shaking water bath. The separation was achieved on a C18 column using 0. 1% formic acid solution and acetonitrile as mobile phase with a liner gradient program. The flow rate was set at 1. 0 mL/min. Detection was achieved by UV light at 240 nm. To evaluate the interactions between LDP and NIF, the similarity of the fingerprinting chromatograms before and after co-incubation was calculated by similarity evaluation system for chromatographic fingerprint of TCM. Furthermore, pharmacokinetic experiments in rat suggested that there was no significant difference in the pharmacokinetic parameters of the typical components in LDP.

The Clinical Experience of Professor HE Ruoping in the Treatment of Perimenopausal Syndrome / 浙江中医药大学学报

[Objective] To summarize the academic views and clinical experience of Professor HE Ruoping in the treatment of peri menopausal syndrome by using traditional Chinese medicine. [Method] From the clinical treatment, collecting the clinical information of the teacher, research and discuss the traditional Chinese medicine academic ideas and clinical experience of teacher HE in the treatment of perimenopausal syndrome, to summarize the traditional Chinese medicine of perimenopausal syndrome in dialectical characteristic.And further explain the characteristics of the treatment of teacher HE through case analysis. [Result] Teacher HE thinks that the main pathogenesis of perimenopausal syndrome is the deficiency of kidney Yin, and divides the patients into stagnation of liver-qi type, Qi stagnation and blood stasis, disharmony between heart and kidney, heart and spleen deficiency type, yin-yang deficiency type, a total of five kinds of syndrome types, according to their clinical manifestation.When confronted with such patients, teacher HE often chooses Liuwei Dihuang Pills to tonify kidney-yin and relieve liver for smooth Qi, promoting qi to activate blood, restoring normal coordination between heart and kidney, invigorating heart and spleen, tonify both Yin and Yang, gives patients more grooming solution at the same time to adjust their physical and mental state as a whole, so the patients could receive remarkable effect and avoid the potential risks of hormone replacement therapy.The case in this article has obtained good treatment results. [Conclusion] Teacher HE treats perimenopause syndrome by tonifying kidney-yin and taking accompanied symptoms and signs into account meanwhile, which has significant clinical effect and the value of popularization and application.