The haplotype-resolved genome assembly of autotetraploid rhubarb Rheum officinale provides insights into its genome evolution and massive accumulation of anthraquinones.

Rheum officinale, a member of the Polygonaceae family, is an important medicinal plant that is widely used in traditional Chinese medicine. Here, we report a 7.68-Gb chromosome-scale assembly of R. officinale with a contig N50 of 3.47 Mb, which was clustered into 44 chromosomes across four homologous groups. Comparative genomics analysis revealed that transposable elements have made a significant contribution to its genome evolution, gene copy number variation, and gene regulation and expression, particularly of genes involved in metabolite biosynthesis, stress resistance, and root development. We placed the recent autotetraploidization of R. officinale at ∼0.58 mya and analyzed the genomic features of its homologous chromosomes. Although no dominant monoploid genomes were observed at the overall expression level, numerous allele-differentially-expressed genes were identified, mainly with different transposable element insertions in their regulatory regions, suggesting that they functionally diverged after polyploidization. Combining genomics, transcriptomics, and metabolomics, we explored the contributions of gene family amplification and tetraploidization to the abundant anthraquinone production of R. officinale, as well as gene expression patterns and differences in anthraquinone content among tissues. Our report offers unprecedented genomic resources for fundamental research on the autopolyploid herb R. officinale and guidance for polyploid breeding of herbs.

High-quality Fagopyrum esculentum genome provides insights into the flavonoid accumulation among different tissues and self-incompatibility.

Common buckwheat (Fagopyrum esculentum) and Tartary buckwheat (Fagopyrum tataricum), the two most widely cultivated buckwheat species, differ greatly in flavonoid content and reproductive mode. Here, we report the first high-quality and chromosome-level genome assembly of common buckwheat with 1.2 Gb. Comparative genomic analysis revealed that common buckwheat underwent a burst of long terminal repeat retrotransposons insertion accompanied by numerous large chromosome rearrangements after divergence from Tartary buckwheat. Moreover, multiple gene families involved in stress tolerance and flavonoid biosynthesis such as multidrug and toxic compound extrusion (MATE) and chalcone synthase (CHS) underwent significant expansion in buckwheat, especially in common buckwheat. Integrated multi-omics analysis identified high expression of catechin biosynthesis-related genes in flower and seed in common buckwheat and high expression of rutin biosynthesis-related genes in seed in Tartary buckwheat as being important for the differences in flavonoid type and content between these buckwheat species. We also identified a candidate key rutin-degrading enzyme gene (Ft8.2377) that was highly expressed in Tartary buckwheat seed. In addition, we identified a haplotype-resolved candidate locus containing many genes reportedly associated with the development of flower and pollen, which was potentially related to self-incompatibility in common buckwheat. Our study provides important resources facilitating future functional genomics-related research of flavonoid biosynthesis and self-incompatibility in buckwheat.

Gegen Qinliantang and Its Modification in Treatment of Ulcerative Colitis： A Review / 中国实验方剂学杂志

Gegen Qinliantang composed of Puerariae Lobatae Radix， Scutellariae Radix， Coptidis Rhizoma， and Glycyrrhizae Radix et Rhizoma exhibits exterior-releasing and interior-clearing effects and can treat diarrhea caused by fever invasion. Therefore， it is highly praised by physicians for treating damp-heat dysentery. This article systematically reviews the clinical applications and basic research advances of Gegen Qinliantang in treating ulcerative colitis. Modern clinical research has demonstrated that modified Gegen Qinliantang has also been used to treat ulcerative colitis， and it is often used in combination with Western drugs （mesalazine， olsalazine， sulfasalazine， etc.）， other Chinese medicine prescriptions （Tongxie Yaofang， Baitouweng Tang， Shaoyaotang， Xianglianwan， etc.）， and acupuncture. Such therapties can significantly alleviate symptoms， such as abdominal pain， diarrhea， and mucopurulent bloody stools， lower traditional Chinese medicine symptom scores and Baron score， improve the quality of life and sleep， reduce adverse reactions， and decrease the relapse rate within one year. Pharmacological studies have demonstrated that Gegen Qinliantang can inhibit the secretion of pro-inflammatory cytokines， such as tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6）， and enhance the release of the anti-inflammatory cytokine interleukin-10 （IL-10） to alleviate intestinal inflammation. Additionally， it can modulate cellular signaling pathways and interfere with upstream and downstream connections in the pathways to inhibit oxidative stress response and prevent cell apoptosis. By regulating the balance of T-helper 17 （Th17） and regulatory T （Treg） cells and controlling the normal expression of immunoglobulins， Gegen Qinliantang can restore immune function. Moreover， this prescription can enrich the beneficial microorganisms in the intestine and stimulate intestinal epithelial cell regeneration to protect and repair the intestinal mucosal barrier. The active components in Gegen Qinliantang， such as puerarin， berberine， baicalin， and glycyrrhizin， can exert antibacterial， anti-inflammatory， and metabolic regulatory effects. This review provides a basis for treating ulcerative colitis with Gegen Qinliantang.

Identification and expression analysis of flavonoid O-methyltransferases gene family in Artemisia argyi / 药学学报

italic>Artemisia argyi (A. argyi) is a Chinese herbal medicine in China. The main active components are volatile oils, flavonoids, and other compounds, which have various pharmacological activities. Methoxylated flavonoids are the main active ingredients in A. argyi. Flavonoid O-methyltransferase (FOMT) is a key enzyme in the O-methylation of flavonoids. In order to further understand the function and characteristics of FOMT proteins, this paper carried out the whole genome mining and identification of FOMT genes in A. argyi and performed phylogenetic, chromosomal localization, gene sequence characterization, subcellular localization prediction, protein structure, gene structure analysis, and expression pattern analysis. The results showed that a total of 83 FOMT genes were identified in the genome of A. argyi. The phylogenetic tree shows that FOMT genes are divided into two subgroups, CCoAOMT (caffeoyl CoA O-methyltransferase) subfamily (32 genes) and COMT (caffeic acid O-methyltransferase) subfamily (51 genes). Gene sequence analysis showed that the number of amino acids encoded by FOMT was 70-734 aa, the molecular weight was 25 296.55-34 241.3 Da, and the isoelectric point was 4.51-9.99. Compared with 32 members of the CCoAOMT subfamily, nearly 1/3 of the 51 members of the COMT subfamily were hydrophobic proteins and 2/3 were hydrophilic proteins. Subcellular localization prediction showed that more than 80% of CCoAOMT subfamily members were located in the cytoplasm, and 96% of COMT subfamily members were located in the chloroplast. COMT subfamily members have more motifs than CCoAOMT subfamily members. The N-terminal motifs of COMT subfamily proteins are relatively variable, while the C-terminal motifs are relatively conserved. Expression pattern analysis showed that CCoAOMT subfamily members were mainly expressed in roots, while COMT members were mainly expressed in leaves. Some FOMTs showed the tissue expression specificity by real-time quantitative PCR analysis, especially in leaves. In this study, we identified and analyzed the FOMT gene family in A. argyi, and provided a theoretical basis for further research on the function of FOMTs and the biosynthesis of methylated flavonoids in A. argyi.

Exploring new catechin derivatives as SARS-CoV-2 Mpro inhibitors from tea by molecular networking, surface plasma resonance, enzyme inhibition, induced fit docking, and metadynamics simulations.

SARS-CoV-2 Mpro (Mpro) is the critical cysteine protease in coronavirus viral replication. Tea polyphenols are effective Mpro inhibitors. Therefore, we aim to isolate and synthesize more novel tea polyphenols from Zhenghedabai (ZHDB) white tea methanol-water (MW) extracts that might inhibit COVID-19. Through molecular networking, 33 compounds were identified and divided into 5 clusters. Further, natural products molecular network (MN) analysis showed that MN1 has new phenylpropanoid-substituted ester-catechin (PSEC), and MN5 has the important basic compound type hydroxycinnamoylcatechins (HCCs). Thus, a new PSEC (1, PSEC636) was isolated, which can be further detected in 14 green tea samples. A series of HCCs were synthesized (2-6), including three new acetylated HCCs (3-5). Then we used surface plasmon resonance (SPR) to analyze the equilibrium dissociation constants (KD) for the interaction of 12 catechins and Mpro. The KD values of PSEC636 (1), EGC-C (2), and EC-CDA (3) were 2.25, 2.81, and 2.44 µM, respectively. Moreover, compounds 1, 2, and 3 showed the potential Mpro inhibition with IC50 5.95 ± 0.17, 9.09 ± 0.22, and 23.10 ± 0.69 µM, respectively. Further, we used induced fit docking (IFD), binding pose metadynamics (BPMD), and molecular dynamics (MD) to explore the stable binding pose of Mpro-1, showing that 1 could tightly bond with the amino acid residues THR26, HIS41, CYS44, TYR54, GLU166, and ASP187. The computer modeling studies reveal that the ester, acetyl, and pyrogallol groups could improve inhibitory activity. Our research suggests that these catechins are effective Mpro inhibitors, and might be developed as therapeutics against COVID-19.

Discovery of honokiol thioethers containing 1,3,4-oxadiazole moieties as potential α-glucosidase and SARS-CoV-2 entry inhibitors.

Honokiol, isolated from a traditional Chinese medicine (TCM) Magnolia officinalis, is a biphenolic compound with several biological activities. To improve and broaden its biological activity, herein, two series of honokiol thioethers bearing 1,3,4-oxadiazole moieties were prepared and assessed for their α-glucosidase and SARS-CoV-2 entry inhibitory activities. Among all the honokiol thioethers, compound 7l exhibited the strongest α-glucosidase inhibitory effect with an IC50 value of 18.9 ± 2.3 µM, which was superior to the reference drug acarbose (IC50 = 24.4 ± 0.3 µM). Some interesting results of structure-activity relationships (SARs) have also been discussed. Enzyme kinetic study demonstrated that 7l was a noncompetitive α-glucosidase inhibitor, which was further supported by the results of molecular docking. Moreover, honokiol thioethers 7e, 9a, 9e, and 9r exhibited potent antiviral activity against SARS-CoV-2 pseudovirus entering into HEK-293 T-ACE2h. Especially 9a displayed the strongest inhibitory activity against SARS-CoV-2 pseudovirus entry with an IC50 value of 16.96 ± 2.45 µM, which was lower than the positive control Evans blue (21.98 ± 1.98 µM). Biolayer interferometry (BLI) binding and docking studies suggested that 9a and 9r may effectively block the binding of SARS-CoV-2 to the host ACE2 receptor through dual recognition of SARS-CoV-2 spike RBD and human ACE2. Additionally, the potent honokiol thioethers 7l, 9a, and 9r displayed relatively no cytotoxicity to normal cells (LO2). These findings will provide a theoretical basis for the discovery of honokiol derivatives as potential both α-glucosidase and SARS-CoV-2 entry inhibitors.

The n-butanol fraction of the Xiao-Chai-Hu decoction alleviates the endocrine disturbance in the liver of mice exposed to lead.

ETHNOPHARMACOLOGICAL RELEVANCE: Lead is a toxic heavy metal that causes health risks globally. However, the mechanism of endocrine poisoning and detoxification of lead poisoning, especially in the liver, still needs to be studied. Xiao-Chai-Hu decoction (XCHD) is regarded as an antidote and an anti-hepatotoxic traditional prescription that has been recorded in the pharmacopeia of the People's Republic of China. AIM OF THE STUDY: The study aimed to probe the hepatoprotective activity of XCHD in the regulation of endocrine dysfunction in the liver and its molecular mechanism. MATERIALS AND METHODS: The mice from the Institute of Cancer Research (ICR) were exposed to different concentrations of XCHD and lead. Then, serum biochemical indices and liver pathology were analyzed. The key differential genes were detected by qRT-PCR and Western blot. RESULTS: According to the biochemical and histopathological analysis, XCHD-NBA was the most effective in attenuating lead-induced hepatotoxicity. From the transcriptome, we analyzed the key genes of XCHD-NBA in the regulation of lead toxicity, including Tubb2a, Stip1, Cyp4a12a, Cyp2c50, Ugt1a1, Cyp3a11, Cyp4a12b, Ahsa1, Cyp2c54, Tubb4b, Esr1, Hsp90aa1, Tuba1a, Tuba1c, and Hsph1. We also analyzed the main components of XCHD-NBA by LC-MS. Because of their extensive role in regulating the endocrine function, baicalin and glycyrrhizin were identified as the main active components of XCHD in regulating endocrine disorders caused by lead. CONCLUSIONS: Lead can disturb the endocrine regulatory process of the liver, while XCHD-NBA alleviates lead-induced liver injury by regulating the endocrine regulatory process.

Application of enzymatic probe sonication for selenium speciation in animal feeds.

Enzymatic probe sonication (EPS) was investigated as a novel and alternative technology for the extraction of five Se species SeCys2, MeSeCys, Se (IV), SeMet and Se (VI) from animal feeds. The critical parameters of EPS such as enzyme types, ultrasonic power, sample/enzyme ratio, extraction time and temperature were carefully optimized. Under current conditions, one sample can be extracted in 60s, in contrast to several hours required by the conventional enzymatic methods Moreover, the extraction is performed in pure water, no further temperature control or pH adjustment is needed, and therefore the risk of species interconvertion is drastically reduced or inhibited. The Se species were separated and quantitatively determined by high-performance liquid chromatography-hydride generation atomic fluorescence spectrometry (HPLC-HG-AFS), which allows the separation of organic and inorganic Se species in a single chromatographic run. The newly developed method was successfully applied to the determination of Se species in 18 feed samples collected from markets and local farms. Concerning the feed samples studied, the results of this work suggest that stricter measures should be taken to control the Se-enriched feed supplements in terms of Se species.

Different acupuncture and moxibustion methods at Heding (EX-LE 2) for knee osteoarthritis with-deficiency and cold-stagnation syndrome / 中国针灸

<p><b>OBJECTIVE</b>To observe the differences for knee osteoarthritis (KOA) with-deficiency and cold-stagnation syndrome among acupuncture plus ginger moxibustion at Heding (EX-LE 2) on the affected side, simple acupuncture and simple ginger moxibustion at the same acupoint.</p><p><b>METHODS</b>Ninety patients were randomly assigned into an acupuncture group, a ginger moxibustion group and a combination group, 30 cases in each one. Acupuncture was used in the three groups at bilateral Liangqiu (ST 34), Xuehai (SP 10), and Dubi (ST 35), Neixiyan (EX-LE 4), Yanglingquan (GB 34), Zusanli (ST 36), Yinlingquan (SP 9), Sanyinjiao (SP 6), Xuanzhong (GB 39), Taichong (LR 3) on the affected side. Acupuncture at the affected Heding (EX-LE 2) was applied in the acupuncture group; ginger moxibustion at the affected Heding (EX-LE 2) in the ginger moxibustion group; ginger moxibustion after acupuncture at Heding (EX-LE 2) in the combination group. The treatment was given once a day, and 10 treatments made one session. There were 2 days at the interval between 2 sessions. Before and after (22/30), which was significantly higher than 46.7% (14/30) of the acupuncture group and 20.0% (6/30) of the ginger moxibustion group (both<0.05), and the rate of the acupuncture group was higher than that of the ginger moxibustion group (<0.05).Based on acupuncture, ginger moxibustion at the affected Heding (EX-LE 2) after acupuncture at the same acupoint are more effective than simple acupuncture at the affected Heding (EX-LE 2) and simple ginger moxibustion at the acupoint for KOA with-deficiency and cold-stagnation syndrome. two-session treatment, knee function scores, visual analogue scale (VAS) score, and present pain intensity (PPI) score were compared in the three groups. The clinic effects were evaluated too.</p><p><b>RESULTS</b>After treatment, the knee function scores of the three groups were all higher than those before treatment (all<0.05), and the VAS score and PPI score were lower (all<0.05). The differences before and after treatment for knee function, VAS and PPI score of the combination group were higher than those of the other two groups (all<0.05), and the differences of the acupuncture group were higher than those of the ginger moxibustion group (all<0.05). The cured and remarkable effective rate of the combination group was 73.3% (22/30), which was significantly higher than 46.7% (14/30) of the acupuncture group and 20.0% (6/30) of the ginger moxibustion group (both<0.05), and the rate of the acupuncture group was higher than that of the ginger moxibustion group (<0.05).</p><p><b>CONCLUSIONS</b>Based on acupuncture, ginger moxibustion at the affected Heding (EX-LE 2) after acupuncture at the same acupoint are more effective than simple acupuncture at the affected Heding (EX-LE 2) and simple ginger moxibustion at the acupoint for KOA with-deficiency and cold-stagnation syndrome.</p>

A new anti-fibrinolytic hemostatic compound 8-O-acetyl shanzhiside methylester extracted from Lamiophlomis rotata.

BACKGROUND: Fibrinolysis prevents blood clots from growing and becoming problematic. Antifibrinolytics are used as inhibitors of fibrinolysis. Aprotinin was doubted after identification of major side effects, especially on kidney. Lysine analogues has their own defects and whether they are adequate substitutes for aprotinin is still under doubt. Lamiophlomis rotata (Benth.) Kudo. was previous found to have hemostatic activity. But the active compound in L. rotata and its hemostatic mechanism were unknown. OBJECTIVES: To find the major hemostatic compound in L. rotata and identify its haemostasis mechanism. METHODS: Traumatic hemorrhage model and coagulant activity assays were monitored in mice and platelets in drug treatment group and control group. Hyperfibrinolysis model was established by intravenous administration of urokinase in mice. Capillary blood clotting time (CBCT), activated partial thromboplastin time (APTT), prothrombin time (PT), thrombin time (TT), fibrinogen and euglobulin clot lysis time (ECLT) were measured. RESULTS: The anti-fibrinolytic activity come from 8-O-Acetyl shanzhiside methylester (ASM) one of the highest iridoid glycosides contents in TIG extracted from L. rotata. ASM significantly (P<0.05) shorten CBCT and reduced blood loss volume in vivo, but did not influence mice APTT, PT or TT. In particular, it significantly prolonged ECLT in hyperfibrinolysis mice. It indicated that ASM could inhibit fibrinolysis. ASM was also effective in CBCT, traumatic bleeding volume and ECLT in hyperfibrinolysis mice model. CONCLUSIONS: ASM was the major hemostatic compound in L. rotata. The haemostasis mechanism of ASM was achieved by anti-fibrinolytic activity. ASM was a new fibrinolysis inhibitor as iridoid glycoside compound.

[Protective Effect of Saussurea involucrata Alcohol Extract on Liver Mitochondria in Mice Under Hypoxia Condition].

OBJECTIVE: To study the protective effect of Saussurea involucrata alcohol extract on liver mitochondria in mice under hypoxia condition. METHODS: The hypoxia mice model was established, the BALB/c mice were randomly divided into four groups:normal group ,hypoxia model group, positive control group and Saussurea involucrata alcohol extract group. Mice were put into low pressure oxygen chamber and decompressed, adapted to hypobaric hypoxia environment of simulated altitude of 8,000 m for 12 h, and then recovered to normal altitude. The mice were sacrificed and the liver mitochondria was isolated, the mitochondrial membrane potential and the activity of malate dehydrogenase, aconitase, α-ketoglutarate dehydrogenase, pyruvate dehydrogenase and mitochondrial complex I, II and V were measured. RESULTS: Compared with hypoxia model group, Saussurea involucrata alcohol extract protected mitochondrial membrane potential, sustained the activities of aconitase, α-ketoglutarate dehydrogenase, pyruvate dehydrogenase, and mitochondrial complex I, II and V under hypoxia condition. CONCLUSION: Saussurea involucrata alcohol extract can protect the liver mitochondrial function in mice under hypoxia condtion.

[Chemical Constituents with Anti-hypoxia Activity from Saussurea involucrata].

OBJECTIVE: To investigate the chemical constituents with anti-hypoxia activity from Saussurea involucrata. METHODS: The chemical constituents, isolated and purified by column chromatography from Saussurea involucrata, were identified by several spectroscopic methods. The anti-hypoxic activities of these compounds were examined using the normobaric hypoxic model of mice. RESULTS: Twelve compounds were isolated from petroleum ether extract of Saussurea involucrata and identified as n-octacosane (1), 1-undecanol (2), heptadecan-l-ol(3), heptacosan-1-ol(4), myristicin (5), apiol(6), ß-sitosterol(7), lupeol(8), moslosooflavone (9), mosloflavone (10), negletein(11), and 5, 6-dihydroxy-7, 8-dimethoxyflavone(12). CONCLUSION: All compounds except 7 and 8 are isolated from this plant for the first time. Compound 1, 5 and 8 - 12 can significantly prolong the survival time of hypoxic mice.

Convergent synthesis of moslosooflavone, isowogonin and norwogonin from chrysin.

A convergent synthesis route of moslooflavone, isowogonin and norwogoninis reported,starting from chrysin, an easily available flavone, by methylation, bromination, methoxylation and demethylation procedures. This synthetic route is convenient and can give the three rare flavones in good yield.

[Effects and HPA Axis Related Mechanism of Kaixin-San and Danggui-Shaoyao-San on Glucose and Lipid Metabolism in Chronic Stress Rats with High-fat Diet].

OBJECTIVE: To study the effects and potential mechanism of Kaixin-San and Danggui-Shaoyao-San on glucose and lipid metabolism in chronic stress rats fed with high-fat diet. METHODS: 50 male Wistar rats were randomly divided into normal control group (distilled water), high-fat diet with chronic stress group (distilled water), melatonin group(20 mg/kg), Kaixin-San group (445 mg/kg) and Danggui-Shaoyao-San group (3360 mg/kg). All drugs were orally administered. In addition to the normal control group, each group of rats were fed with high-fat, diet. Simultaneously, stress were carried out after drugs administration 1 h daily. The duration was lasted for six weeks. The rat body weight daily was recorded, and the 24 h period urine was collected to detect the level of urine corticosterone (CORT) after three weeks. The level of plasma intraperitoneal glucose tolerance (IVGTT) was detected after six weeks. Finally, rats were executed, and serum fasting blood glucose (FBG), triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), insulin (INS), adrenocorticotropic hormone releasing hormone (CRH), adrenocorticotropic hormone (ACTH), CORT and melatonin ( MLT) were determined. The weight of adrenal gland, liver glycogen and muscle glycogen levels were detected. The adrenal gland index, Homeostasis Model Assessment of Insulin Resistance (HOMA-IR) and insulin sensitivity index( ISI) were calculated. RESULTS: Compared with normal group, model rats body weight, IVGTT (120 min), plasm CORT were decreased significantly. Serum TG, TC, LDL-C and urine CORT after three weeks were increased significantly. Kaixin-San and Danggui- Shaoyao-San could regulate the above indexes. CONCLUSION: Kaixin-San and Danggui-Shaoyao-San may regulate the activity of HPA axis, and improve glucose and lipid metabolism disorder in model rats by increasing melatonin secretion.

Protective effect of apigenin on ischemia/reperfusion injury of the isolated rat heart.

Apigenin (Api), a mainly bioactive component of Apium graveolens L. var. dulce DC. (a traditional Chinese medicinal herb), possesses a wide range of biological activities, including antioxidant effects. It also has been shown to associate with lower prevalence of cardiovascular diseases, but its mechanisms of action remain unclear. The aim of the present study is to investigate the role of Api in isolated rat heart model of ischemia/reperfusion (I/R). Langendorff-perfused isolated rat hearts were used in our study. Api was added to the perfusate before ischemia and during reperfusion in the isolated pulsed rat heart exposed to 30-min ischemia followed by 50-min reperfusion. The treatment with Api conferred a cardioprotective effect, and the treated hearts demonstrated an improved ischemic cardiac functional recovery, a decreased myocardial infarct size, a reduced activities of creatine kinase isoenzyme and lactate dehydrogenase in the coronary flow, a reduced number of apoptotic cardiomyocytes, a reduced activity of caspase-3, up-regulation of the anti-apoptotic protein Bcl-2 and down-regulation of the pro-apoptotic protein Bax. In addition, Api inhibited the phosphorylation of p38 MAPKS during I/R. In conclusion, these observations provide preliminary evidence that Api can protect cardiomyocytes from I-/R-induced injury, at least partially, through the inhibition of p38 MAPKS signaling pathway.

Therapeutic Observation of Comprehensive Acupuncture for Wet Age-related Macular Degeneration / 上海针灸杂志

ObjectiveTo systematically and objectively evaluate the therapeutic efficacy of comprehensive acupuncture treatment in managing wet age-related macular degeneration (ARMD), and to seek a standard acupuncture protocol for this disease, for promoting the application in clinic.MethodTotally 37 patients (67 eyes) were observed to compare the vision, Amsler grid and rating scale of quality of life for visual impairment before and after the 3-month treatment. The clinical efficacy was then evaluated. ResultThe vision and score of quality of life were significantly changed after intervention (P<0.05); according to Amsler grid, the total improved rate was 79.2% and the total effective rate was 77.6%; age and disease duration were negatively correlated with the therapeutic efficacy (P<0.05).ConclusionComprehensive acupuncture treatment can significantly improve the vision and quality of life in patients with wet ARMD, and the younger the patients and the shorter the disease duration, the higher the therapeutic efficacy.

[Protective effect and action mechanism of petroleum ether extracts from Saussurea involucrate on brain tissues of hypoxia rats].

OBJECTIVE: To investigate the protective effect and action mechanism of petroleum ether extracts from Saussurea involucrate on brain tissues of hypoxia rats under constant pressure and closed conditions. METHOD: The PESI dosage-dependent experiment for hypoxia rats was conducted under constant pressure and closed conditions by intraperitoneally injecting 125, 250, 500 mg x kg(-1) to finalize that the optimum dosage is the high dose of PESI. Afterwards, 90 Wistar rats were randomly divided into the hypoxic model group, the acetazolamide 250 mg x kg(-1) group and the PESI high dose group. Each group was further divided into three subgroups according to different hypoxia times, with 10 rats in each subgroup. Under the same hypoxia and administration conditions, the rats were sacrificed after 0, 3, 6 h respectively. Their brain samples were collected for common pathological observation and immunohistochemical staining of HIF-1alpha. Real-time RT-PCR was used to detect HIF-1alpha, EPO, HO-1 and Caspase-3 gene expressions. And the Western blot assay was adopted to detect HIF-1alpha protein expression. RESULT: The brain tissues of the hypoxia model group were severely damaged with the increase in the hypoxia time. The acetazolamide group and the PESI high does group were damaged in a much lower degree. According to the gene expression and the Western blot assay, high dose of PESI could inhibit HIF-1alpha expression. According to the pure gene expression test, high dose of PESI could increase EPO and HO-1 mRNA expressions, but inhibit Caspase-3 mRNA expression. CONCLUSION: PESI's protective mechanism for brain tissues of hypoxia rats under constant pressure and closed conditions may be related to its effects in inhibiting HIF-1alpha expression, increasing EPO expression and resisting cell apoptosis.

Suppression effects of betaine-enriched spinach on hyperhomocysteinemia induced by guanidinoacetic acid and choline deficiency in rats.

Betaine is an important natural component of rich food sources, especially spinach. Rats were fed diets with betaine or spinach powder at the same level of betaine for 10 days to investigate the dose-dependent effects of spinach powder supplementation on hyperhomocysteinemia induced by guanidinoacetic acid (GAA) addition and choline deprivation. The GAA-induced hyperhomocysteinemia in rats fed 25% casein diet (25 C) was significantly suppressed by supplementation with betaine or spinach, and it was completely suppressed by taking 11.0% spinach supplementation. The choline deprivation-induced enhancement of plasma homocysteine concentration in rats fed 25% soybean protein diet (25S) was markedly suppressed by 3.82% spinach. Supplementation with betaine or spinach partially prevented the effects of GAA on hepatic concentrations of methionine metabolites. The decrease in activity of betaine-homocysteine S-methyltransferase (BHMT) and cystathionine ß-synthase (CBS) in GAA-induced hyperhomocysteinemia was recovered by supplementation with betaine or spinach. Supplementation with betaine or spinach did not affect BHMT activity, whereas it partially restored CBS activity in choline-deprived 25S. The results indicated that betaine or spinach could completely suppress the hyperhomocysteinemia induced by choline deficiency resulting from stimulating the homocysteine removal by both remethylation and cystathionine formation.

[Isolation and identification of nine iridoid glycosides from Lamiophlomis rotata by HPLC combining PDA and MS].

OBJECTIVE: To establish a rapid chromatographic method to separate the iridoid glycosides from Lamioplomis rotata, and to identify the target compounds with PDA and MS. METHODS: Methanol-water gradient elution was used to separate and analyze the target compounds. The fluid fractions were gathered according to the chromatogram and dried with the nitrogen airflow. The mass fractions of the target compounds were determined with RP-HPLC and the structures were identified with PDA and MS. RESULTS: The purity of some compounds exceeded 90% and these 9 compounds were identified as iridoid glycosides, which were Phlorigidoside C (1), Schismoside (2), Sesamoside (3), Shanzhiside methylester (4), 6-O-Acetyl shanzhiside methylester (5), Phloyoside II (6), Penstemoside (7), Loganin (8) and 8-O-Acetyl shanzhiside methylester (9). CONCLUSION: The method is simple and practicable with high efficiency. It can be used to qualitative and quantitative analysis of the 9 iridoid glycosides in Lamiphlomis rotata and its preparations.

[Effect of ethanol extract from Saussurea involucrata on biochemical indicators of simulated high-altitude hypoxia induced mice].

OBJECTIVE: To estimate the effect of ethanol extract from Saussurea involucrata (EES) on biochemical indicators of simulated high-altitude hypoxia induced mice and its mechanism. METHODS: The oxidative stress indicator( MDA content, SOD activity) and metabolism parameters (LD content, LDH activity, ATP content, Na+ -K+ -ATPase and Ca2+ -Mg2+ -ATPase activity) in both brain and heart of the simulated high-altitude hypoxia induced mice were detected. RESULTS: Compared with the model group, the ESS group could significantly increase the activity of SOD and LDH and decrease the content of MDA and LD in both brain and heart, the content of ATP and the activity of Na+ -K -ATPase and Ca2+ -Mg2+ -ATPase were also elevated. CONCLUSION: The results demonstrate that EES can increase the antioxidant ability, decrease the injury of free radical and ease the disfunction of energy metabolism caused by hypoxia.