Phenols from Potentilla anserina L. Improve Insulin Sensitivity and Inhibit Differentiation in 3T3-L1 Adipocytes in Vitro.

Potentilla anserina L., a well-known perennial herb, is widely used in traditional Tibetan medicine and used as a delicious food in humans. The present investigation reports on the activity of P. anserina phenols (PAP) in regulating glycolipid metabolism in 3T3-L1 adipocytes. Insulin sensitivity tests showed that PAP improved insulin-stimulated glucose uptake by promoting the phosphorylation of serine/threonine kinase Akt. Moreover, an assay involving the differentiation of 3T3-L1 preadipocytes demonstrated that PAP also decreased the accumulation of lipid droplets by suppressing the expression of adipokines during the differentiation process. In addition, the underlying mechanism from the aspects of energy metabolism and oxidative stress is also discussed. The improvement in energy metabolism was supported by an increase in mitochondrial membrane potential (MMP) and intracellular ATP. Amelioration of oxidative stress was supported by decreased levels of intracellular reactive oxygen species (ROS). In summary, our findings suggest that PAP can ameliorate the disorder of glycolipid metabolism in insulin resistant 3T3-L1 adipocytes by improving energy metabolism and oxidative stress and might be an attractive candidate for the treatment of diabetes.

In Situ Deposition of Gold Nanoparticles and L-Cysteine on Screen-Printed Carbon Electrode for Rapid Electrochemical Determination of As(III) in Water and Tea.

In this study, a screen-printed carbon electrode (SPCE) based on in situ deposition modification was developed for the sensitive, rapid, easy and convenient determination of As(III) in water and tea by linear sweep anodic stripping voltammetry (LSASV). The screen-printed carbon electrodes were placed in a solution consisting of As(III) solution, chlorauric acid and L-cysteine. Under certain electrical potential, the chloroauric acid was reduced to gold nanoparticles (AuNPs) on the SPCE. L-cysteine was self-assembled onto AuNPs and promoted the enrichment of As(III), thus enhancing the determination specificity and sensitivity of As(III). The method achieved a limit of determination (LOD) of 0.91 ppb (µg L-1), a linear range of 1~200 µg L-1, an inter-assay coefficient of variation of 5.3% and good specificity. The developed method was successfully applied to the determination of As(III) in tap water and tea samples, with a recovery rate of 93.8%~105.4%, and further validated by inductively coupled plasma mass spectrometry (ICP-MS). The developed method is rapid, convenient and accurate, holding great promise in the on-site determination of As(III) in tap water and tea leaves, and it can be extended to the detection of other samples.

Mechanism of Traditional Tibetan Medicine Grubthobrildkr Alleviated Gastric Ulcer Induced by Acute Systemic Hypoxia in Rats.

Objective: This study was aimed at investigating the potential mechanism of Grubthobrildkr (GTB) on systemic hypoxia-induced gastric ulcers in rats and at detecting the chemical profile of GTB. Methods: Male Sprague-Dawley rats were separated into control, hypoxia, hypoxia+omeprazole, and hypoxia+GTBs (0.25, 0.5, and 1.0 g·kg-1·d-1) groups. Systemic hypoxia was created in a hypobaric chamber to simulate 5000 m high altitude by adjusting the inner pressure and oxygen content for 6 days. After that, the ulcer index, pH, and volume of gastric juice were assessed. The levels of endothelin-1 (ET-1), gastrin (GAS), motilin (MTL), phospholipase A2 (PLA2), and prostaglandin E2 (PGE2) were detected by ELISA. The expression level of hydrogen potassium ATPase (H+-K+-ATPase), cyclooxygenase-1 (COX-1), and cyclooxygenase-2 (COX-2) was tested by western blotting. Chemical profile of GTB was revealed by UHPLC-Q-exactive hybrid quadrupole-orbitrap mass (UHPLC-Q-Orbitrap MS). Results: GTB decreased the ulcer index in rats under hypoxia for six days, which was related to increased pH and volume of gastric juice, enhanced MTL and PGE2 levels, and decreased ET-1 and PLA2 levels of gastric mucosa. Furthermore, GTB decreased the level of H+-K+-ATPase and COX-2 while increased COX-1 levels in gastric mucosal tissue. 44 constituents were identified by UHPLC-Q-Orbitrap MS in GTB. Conclusion: GTB exerted a gastroprotective effect to alleviate gastric ulceration induced by acute systemic hypoxia in rats. The effect of GTB increasing the volume and pH of gastric juice in rats under acute systemic hypoxia could be regulated by gastrointestinal hormones, including MTL and ET-1. Mechanically, gastrointestinal protection of GTB was based on inhibition of the protons pumping H+-K+-ATPase and regulation of prostaglandin family in rats.

Occurrence of Powdery Mildew Caused by Erysiphe buhrii on Dianthus chinensis in Inner Mongolia, China.

Dianthus chinensis is widely cultivated for ornamental and medicinal use in China (Guo et al. 2017). The plant has been used in traditional Chinese medicine for the treatment of urinary problems such as strangury and diuresis (Han et al. 2015). In June and July 2020, powdery mildew-like signs and symptoms were seen on leaves of D. chinensis cultivated on the campus of Inner Mongolia Agricultural University, Hohhot city, Inner Mongolia Province, China. White powder-like masses occurred in irregular shaped lesions on both leaf surfaces and covered up to 50% of leaf area. Some infected leaves were deformed on their edges and some leaf senescence occurred. More than 40 % of plants (n = 180) exhibited these signs and symptoms. Conidiophores (n = 50) of the suspect fungus were unbranched and measured 70 to 140 µm long × 6 to 10 µm wide and had foot cells that were 25 to 48 µm long. Conidia (n = 50) were produced singly, elliptical to cylindrical shaped, 30 to 45 µm long × 12 to 19 µm wide, with length/width ratio of 2.0 to 3.2, and lacked fibrosin bodies. No chasmothecia were found. Based on these morphological characteristics, the fungus was tentatively identified as an Erysiphe sp. (Braun and Cook 2012). Fungal structures were isolated from diseased leaves and genomic DNA of the pathogen extracted utilizing the method described by Zhu et al. (2019). The internal transcribed spacer (ITS) region was amplified by PCR employing the primers PMITS1/PMITS2 (Cunnington et al. 2003) and the amplicon sequenced by Invitrogen (Shanghai, China). The sequence for the powdery mildew fungus (deposited into GenBank under Accession No. MW144997) showed 100 % identity (558/558 bp) with E. buhrii (Accession No. LC009898) that was reported on Dianthus sp. in Japan (Takamatsu et al. 2015). Pathogenicity tests were done by collecting fungal conidia from infected D. chinensis leaves and brushing them onto leaves of four healthy plants. Four uninoculated plants served as controls. Inoculated and uninoculated plants were placed in separate growth chambers maintained at 19 ℃, 65 % humidity, with a 16 h/8 h light/dark period. Nine-days post-inoculation, powdery mildew disease signs appeared on inoculated plants, whereas control plants remained asymptomatic. The same results were obtained for two repeated pathogenicity experiments. The powdery mildew fungus was identified and confirmed as E. buhrii based on morphological and molecular analysis. An Oidium sp. causing powdery mildew on D. chinensis previously was reported in Xinjiang Province, China (Zheng and Yu 1987). This, to the best of our knowledge, is the first report of powdery mildew caused by E. buhrii on D. chinensis in China (Farr and Rossman 2020). The sudden occurrence of this destructive powdery mildew disease on D. chinensis may adversely affect the health, ornamental value and medicinal uses of the plant in China. Identifying the cause of the disease will support efforts for its future control and management.

Phased secondary small interfering RNAs in Panaxnotoginseng.

BACKGROUND: Recent results demonstrated that either non-coding or coding genes generate phased secondary small interfering RNAs (phasiRNAs) guided by specific miRNAs. Till now, there is no studies for phasiRNAs in Panax notoginseng (Burk.) F.H. Chen (P. notoginseng), an important traditional Chinese herbal medicinal plant species. METHODS: Here we performed a genome-wide discovery of phasiRNAs and its host PHAS loci in P. notoginseng by analyzing small RNA sequencing profiles. Degradome sequencing profile was used to identify the trigger miRNAs of these phasiRNAs and potential targets of phasiRNAs. We also used RLM 5'-RACE to validate some of the identified phasiRNA targets. RESULTS: After analyzing 24 small RNA sequencing profiles of P. notoginseng, 204 and 90 PHAS loci that encoded 21 and 24 nucleotide (nt) phasiRNAs, respectively, were identified. Furthermore, we found that phasiRNAs produced from some pentatricopeptide repeat-contain (PPR) genes target another layer of PPR genes as validated by both the degradome sequencing profile and RLM 5'-RACE analysis. We also found that miR171 with 21 nt triggers the generations of 21 nt phasiRNAs from its conserved targets. CONCLUSIONS: We validated that some phasiRNAs generated from PPRs and TASL genes are functional by targeting other PPRs in trans. These results provide the first set of PHAS loci and phasiRNAs in P. notoginseng, and enhance our understanding of PHAS in plants.

Small RNA profiles from Panax notoginseng roots differing in sizes reveal correlation between miR156 abundances and root biomass levels.

Plant genomes encode several classes of small regulatory RNAs (sRNAs) that play critical roles in both development and stress responses. Panax notoginseng (Burk.) F.H. Chen (P. notoginseng) is an important traditional Chinese herbal medicinal plant species for its haemostatic effects. Therefore, the root yield of P. notoginseng is a major economically important trait since the roots of P. notoginseng are the parts used to produce medicine. To identify sRNAs that are critical for the root biomass of P. notoginseng, we performed a comprehensive study of miRNA transcriptomes from P. notoginseng roots of different biomasses. We identified 675 conserved miRNAs, of which 180 pre-miRNAs are also identified, and three TAS3 loci in P. notoginseng. By using degradome sequencing, we identified 79 conserved miRNA:target or tasiRNA:target interactions, of which eight were further confirmed with the RLM 5'-RACE experiments. More importantly, our results revealed that a member of miR156 family and one of its SPL target genes have inverse expression levels, which is tightly correlated with greater root biomass contents. These results not only contributes to overall understanding of post-transcriptional gene regulation in roots of P. notoginseng but also could serve as markers for breeding P. notoginseng with greater root yield.

Uric acid lowering effect of Tibetan Medicine RuPeng15 powder in animal models of hyperuricemia.

OBJECTIVE: To evaluate the influence of the Tibetan medicine RuPeng15 powder (RPP15) on uric acid levels, and explore its possible mechanisms of action in hyperuricemic animal models. METHODS: Hyperuricemic mice were generated by orally administering yeast extract paste twice daily (30 g/kg) for 8 days, to mimic human hyperuricemia induced by high-protein diets. Hyperuricemic rats were generated by intraperitoneal injection of 250 mg/kg potassium oxonate to each animal 1 h before the last oral administration of test compounds, which raised the serum uric acid level by inhibiting the decomposition of uric acid. Levels of uric acid and creatinine in serum and urine were detected by the phosphotungstic acid and picric acid methods respectively, and the activity of xanthine oxidase (XOD) was assayed using a commercial test kit. RESULTS: RPP15 (0.4, 0.8, 1.2 g/kg) significantly decreased the level of serum uric acid in healthy rats (P < 0.05). Furthermore, hyperuricemic rats treated with RPP15 (0.4, 0.8, 1.2 g/kg) had lower serum uric acid levels (P < 0.05), accompanied by lower urine uric acid (P < 0.05). For the hyperuricemic mice, the levels of uric acid in the serum decreased significantly (P < 0.05) and the activity of XOD in the liver was restored to normal levels after treatment with RPP15 (P < 0.05). CONCLUSION: RPP15 (0.4, 0.8, 1.2 g/kg) demonstrated an anti-hyperuricemic effect on both healthy and hyperuricemic animals, and the mechanism is most likely associated with inhibiting the activity of XOD.

Use status and metabolism of realgar in Chinese patent medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Realgar is widely used in combination with other herbs as Chinese patent medicine to treat a wide range of diseases in China. It is also a well known arsenical toxicant. Chronic arsenic poisoning events caused by long-term usage of realgar-containing medicines have been reported in literatures. Given to the paradoxical role of realgar, comprehensive outline of its usage status in Chinese patent medicine might provide basal data for evaluating its toxicology risks in populations. Unfortunately, the relevant information is limited. Also, a metabolic process after intake of realgar-containing medicine in humans is poorly understood. MATERIALS AND METHODS: The Traditional Chinese Patent Medicine Prescription Database was reviewed to get the information on the usage status of realgar. Realgar powder was dissolved in different pH-value solutions (1, 3, 5, 7, 9 and 11) to determine the soluble arsenic concentrations from realgar. Ten volunteers aged 24-26 years old were recruited to take four pills of Niu Huang Jie Du Pian (NHJDP), a very common Chinese patent medicine with realgar, to analyze the arsenic metabolism after exposure to realgar-containing medicine. The four pills were taken according to the medical instruction. Concentrations of soluble arsenic from realgar and urinary arsenic metabolites in humans were determined by hydride generation atomic absorption spectrometry. RESULTS: A total of 191 (2.25%) realgar-containing traditional Chinese patent medicines were obtained from the database, and almost 86.91% of them were for oral application. 73 (38.22%) medicines were found to be available for children. The mass fraction of arsenic in realgar-containing medicine ranged from 0.11% to 27.52%. According to medical instructions, the amount of average daily arsenic intake ranged from 0.47 to 2895.53mg. Nearly 86% medicines with daily intake of arsenic >10mg. Only inorganic arsenic (iAs) was detected from realgar in dissolution experiment and the levels of soluble iAs increased with pH values. After intake NHJDP, arsenic excretion in urine significantly increased, with a maximum excretion of iAs and monomethylarsonic acid at 6h post-ingestion and a peak excretion of dimethylarsinic acid at 9h post-ingestion. Arsenic methylation capacity was decreased after intake NHJDP. Females carried a more efficient arsenic methylation process than males. CONCLUSIONS: Realgar is widely used in traditional Chinese medicine. The arsenic solubility from realgar may be enhanced under alkaline conditions. The levels of urinary arsenic metabolites significantly increased while the arsenic methylation capacity significantly decreased after intaking realgar-containing medicine, which may suggest that a potential health hazard exists if people use arsenical medicines for long-term.

Polycyclic aromatic hydrocarbons in deposited bedroom dust collected from Xinxiang, a fast developing city in North China.

To investigate the contamination levels of polycyclic aromatic hydrocarbons (PAHs) in the indoor environment, 16 PAHs were measured in 20 deposited bedroom dust (DBD) samples collected from four residential areas in Xinxiang City. The total PAH concentrations (∑PAHs, defined as the sum of 16 PAHs) in 20 DBD samples ranged from 1.47 to 21.8 µg/g dry weight. PAH ratios indicated that main sources of PAHs in most DBD samples were coal, grasses, and wood combustion. Correlation analysis showed that there were no significant positive correlations between ∑PAH concentrations in DBD and the number of family members or duration of residence. Analysis using the t test revealed that tobacco smoke was a source of PAHs in DBD and that an unattached kitchen may be a significant influencing factor related to PAHs in DBD collected from residential areas in Xinxiang City. Our results showed that the kitchen exhaust fan or hood had no significant influence on total PAHs in DBD, and there was no significant difference in ∑PAH concentrations in DBD in homes which used liquefied petroleum gas and those which used natural gas.

[Effect of Tibetan medicine zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2].

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.

Redox signaling mediates the expression of a sulfate-deprivation-inducible microRNA395 in Arabidopsis.

MicroRNA395 (miR395) is a conserved miRNA that targets a low-affinity sulfate transporter (AST68) and three ATP sulfurylases (APS1, APS3 and APS4) in higher plants. In this study, At2g28780 was confirmed as another target of miR395 in Arabidopsis. Interestingly, several dicots contained genes homologous to At2g28780 and a cognate miR395 complementary site but possess a gradient of mismatches at the target site. It is well established that miR395 is induced during S deprivation in Arabidopsis; however, the signaling pathways that mediate this regulation are unknown. Several findings in the present study demonstrate that redox signaling plays an important role in induction of miR395 during S deprivation. These include the following results: (i) glutathione (GSH) supplementation suppressed miR395 induction in S-deprived plants (ii) miR395 is induced in Arabidopsis seedlings exposed to Arsenate or Cu(2+) , which induces oxidative stress (iii), S deprivation-induced oxidative stress, and (iv) compromised induction of miR395 during S deprivation in cad2 mutant (deficient in GSH biosynthesis) that is defective in glutaredoxin-dependent redox signaling and ntra/ntrb (defective in thioredoxin reductases a and b) double mutants that are defective in thioredoxin-dependent redox signaling. Collectively, these findings strongly support the involvement of redox signaling in inducing the expression of miR395 during S deprivation in Arabidopsis.

Electroacupuncture at Qiuxu (GB 40) for treatment of migraine--a clinical multicentral random controlled study.

OBJECTIVE: To observe the therapeutic effects of electroacupuncture (EA) at Qiuxu (GB 40) for treatment of migraine so as to provide clinical evidence for compilation of the Acupoints' Dictionary of the People's Republic of China. METHODS: 275 migraine patients admitted in 3 hospitals were randomly divided into a treatment group treated by EA at Qiuxu (GB 40), and a control group treated by EA at Tianshu (ST 25). The indexes of the migraine symptoms and the 5-HT level were observed in both the groups before and after treatment. RESULTS: There was a significant difference in VAS score between the two groups of the 3 clinical centers (P<0.01). The therapeutic effects of a 4-week treatment were much better in the treatment group than that of the control group. The 3-month follow-up survey showed that the long-term effects were in favor of the treatment group of the first and third clinical centers, though no significant difference was found in the treatment group of the second clinical center as compared with the control group. The results from the 6-month follow-up survey showed better effects in the treatment group of all the 3 clinical centers. CONCLUSION: EA at Qiuxu (GB 40) may show effect for migraine.

[The antioxidative effect of Sanwei Tanxiang powder on rats' hearts against myocardial ischemia and reperfusion injury].

OBJECTIVE: To study the effect of Sanwei Tanxiang powder on myocardial pathologic change, myocardium lipid peroxidation and antioxidation on anesthetized rats' hearts against myocardial ischemia and reperfusion injury. METHODS: A model of regional myocardial ischemia-reperfusion injury (IR) was established by 30 min ligation of the left anterior descending coronary artery followed by 40 min reperfusion in hearts of anesthetized rats'. The experimental animals were randomly divided into the sham operation, IR model group, positive control group and two dose Sanwei Tanxiang powder groups. The changes of ECG, LDH, CK, SOD, GSH-Px and MDA in the heart were measured and ultrastructural pathological changes were observed. RESULTS: In Sanwei Tanxiang groups the displacement of ST segment reduced (P < 0.05), the activity of LDH, CK decreased (P < 0.05), the activities of SOD, GSH-Px significantly increased and the contents of MDA reduced (P < 0.05). Moreover, the development of ultrastructral pathological changes obviously improved compared with the IR model group. CONCLUSION: The protective effects of Sanwei Tanxiang powder on anesthetized rats' hearts against myocardial ischemia and reperfusion injury may be related to protecting the antioxidants and inhibiting the lipid peroxidation in the myocardium.

[RAPD analysis on genetic relationships of Erigeron breviscapus germplasm resources].

OBJECTIVE: To investigate the genetic relationships of Erigeron breviscapus at the molecular biology level. METHOD: Thirty seven germplasm resources of E. breviscapus which collected from Yunnan, Sichuang and Guizhou province in 2005 were analyzed by Random amplified polymorphic DNA (RAPD) and cluster analysis based on NTSYS2. RESULT: A total of 10 primers were screened, and 107 bands were amplified, among which 94 (87.85%) bands were found to be polymorphic. Thirty seven germplasm resources of E. breviscapus were clustered into 3 groups at genetic distance 0.36, the I group include in 9 germplasm resources collected from Mile, Qiubei, Luxi, Gejiu, and Yanshan of south east of Yunnan province; the II group included 8 germplasm resources collected from Gucheng, and shangrila of north west of Yunnan province, and Mile and Qiubei of south east of Yunnan province; the III group included in 20 germplasm resources collected from the center of Yunnan province, north east of Yunnan province, Sichuan province, and Guizhou province. CONCLUSION: There were abundant genetic diversity in the germplasm resources of E. breviscapus, and the genetic relationships are closely related to geographical distance where they were collected.

[Study on the therapeutic effect of electroacupuncture on migraine and influencing factors].

OBJECTIVE: To compare therapeutic effects of electrical acupuncture (EA) at Qiuxu (GB 40) and Tianshu (ST 25), and to probe the factors of influencing the therapeutic effect. METHODS: Two hundred and seventy-five cases were randomly divided into the test group of 138 cases treated with EA at Qiuxu (GB 40) and the control group of 137 cases treated with EA at Tianshu (ST 25). Their therapeutic effects were observed and the factors influencing the therapeutic effect were analyzed. RESULTS: The transient effective rate was 70.3% in the test group and 58.4% in the control group with a significant difference between the two groups (P<0.05), and there were very significant differences between the two groups in the therapeutic effects at treatment of 4 weeks, and following-up 3 and 6 months later (P<0.001), the transient analgesic effect of EA at Qiuxu (GB 40) being better. EA had better transient effect on migraine for the mental workers, or the patient with high pain score at treatment, or more frequent attack. The long-term analgesic effect of EA at Qiuxu (GB 40) was better than Tianshu (ST 25). CONCLUSION: The analgesic effect of EA at Qiuxu (GB 40) is better than Tianshu (ST 25) and the profession and conditions of the patient are the factors influencing the therapeutic effect.