Objective:To observe the effect of Changji'an prescription on intestinal permeability in diarrhea-predominant irritable bowel syndrome (IBS-D) rats and explore its mechanism for treatment of IBS-D. Method:Male SD neonatal rats were randomly divided into five groups:normal group, model group, pinaverium bromide group(0.018 g·kg^-1), high-dose(33.48 g·kg^-1) and low-dose (16.74 g·kg^-1)Changji'an prescription groups. Except for the normal group, the IBS-D model was established by the combination of maternal and infant separation+acetic acid stimulation+restraint stress. After drug treatment, the ultrastructure of rat intestinal mucosa was observed by using transmission electron microscopy and the plasma D-lactate level was detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of tight junction proteins Occludin and Claudin-1 were detected by Western blot. The mRNA expression levels of Occludin,Claudin-1 and zonula occluden(ZO)-1 were detected by real time polymerase chain reaction (Real-time PCR). Result:As compared with the normal group, the intestinal mucosal epithelial cells were damaged in IBS-D model group, and the microvilli arrangement was sparse and tight junction was widened, and some were not obvious,and the plasma D-lactate level in IBS-D rats was increased significantly (P<0.05),and the mRNA and protein expression levels of Occludin and Claudin-1 in the colon were decreased and the mRNA expression of ZO-1 was also decreased (P<0.05). After drug treatment, as compared with the model group, all drug-administered groups can repair intestinal mucosal cell damage. Plasma D-lactate level in pinaverium bromide group and high-dose Changji'an prescription group was significantly decreased (P<0.05) while D-lactate level in the low-dose group Changji'an prescription group had a tendency to decrease with no statistical difference. The mRNA and protein expression levels of Occludin and Claudin-1 and the mRNA expression of ZO-1 in the colon of rats in each administration group were higher than those in the model group (P<0.05). Conclusion:The therapeutic effect of Changji'an prescription on IBS-D may be achieved by improving the intestinal permeability.

OBJECTIVETo investigate the antibacterial activity of glycyrrhizic acid (GA) against Streptococcus mutans (S. mutans) under acidic environment in vitro.

METHODSWorking culture were prepared by inoculation of S. mutans into TPY broth followed by static incubation under anaerobic condition at 37 degrees C for 24 h. TPY broth was supplemented with three kinds density of GA (0.78, 1.57, 3.13 mg x mL(-1)), whose acidity was regulated to pH7.0, pH 5.5 and pH4.0. And the group of pH 7.0 was used as negative control. The growth of S. mutans was measured by A600 of bacteria suspension and counting colony forming unit (CFU). In addition, the survival rate of S. mutans was calculated.

RESULTSIn pH 5.5 groups, the survival rates of 0.78, 1.57 and 3.13 mg x mL(-1) GA groups were 60.96%, 60.27% and 45.58%, respectively, and in pH4.0 groups, the survival rates were 68.75%, 53.12% and 45.83%. In 0.78, 1.57 and 3.13 mg x mL(-1) GA groups, the survival rates of pH5.5 and pH4.0 were 52.25% and 39.05%, 74.39% and 43.11%, 86.38% and 55.30%, respectively.

CONCLUSIONGA could inhibit the growth of S. mutans under acidic environment, which the effect is improved as the acidity increased.

Anti-Bacterial Agents ; Bacteria ; Glycyrrhizic Acid ; In Vitro Techniques ; Streptococcus mutans

Objective To investigate the binding of the chemosynthetic analogue of N-acetyl-seryl-aspartyl-lysyl-proline (Ac-SDKP) FAM-Aca-SDKP to hepatic stellate cell-T6 (HSC-T6) cells and basic physical characteristics.Methods The Ac-SDKP analogue short-peptide FAM-Aca-SDKP carrying green fluorescence was synthesized chemically.Quantitative real-time PCR was used to evaluate its effect on the secretion of HSC collagen and verify the consistency in the biological effect between FAM-Aca-SDKP and Ac-SDKP.A fluorescence microscope was used to observe the binding between FAM-Aca-SDKP and HSC-T6,and flow cytometry was used to evaluate the time-concentration effect of the binding between FAM-Aca-SDKP and HSC-T6.The t-test or rank sum test was used for the statistical analysis of different types ofdata.Results After HSC-T6 was incubated with Ac-SDKP or FAM-Aca-SDKP for 24 hours,the expression of type Ⅰ collagen in HSC-T6 was increased,when the action time was 0.5 hour,Ac-SDKP and FAM-Aca-SDKP caused a 30％-50％ reduction in the expression of type Ⅰ collagen.After HSC-T6 was incubated with FAM-Aca-SDKP,strong green fluorescence was observed on cell surface under a fluorescence microscope,and after Ac-SDKP was added,Ac-SDKP significantly reduced the fluorescence intensity on cell surface due to competitive inhibition.Flow cytometry showed that when the concentration of FAM-Aca-SDKP was 0-50 μmol/L,the rate of fluorescence-positive cells rapidly increased from 0 to 12％;when the concentration was 50-100 μmol/L,the rate of fluorescence-positive cells only increased from 12％ to 14％;co-incubation with Ac-SDKP significantly reduced the rate of fluorescence-positive cells.The number of positive cells reached the peak at the 45-minute point of the incubation and then decreased gradually.Conclusions FAM-Aca-SDKP can bind to the surface of HSC-T6 cells,and this process has ligand-receptor binding characteristics such as competitive inhibition,saturability,and time-concentration effect.

Six month old Cinnamomum cassia seedlings were used to simulate drought stress with polyethylene glycol(PEG 6000). The physiological indicators(osmotic substances, antioxidant enzymes, etc.) and chemical components of seedlings under different drought levels and the correlation between the two were studied. The results showed that the chlorophyll content and relative water content decreased gradually with the increase of PGE 6000(0, 5%, 10%, 15%) concentration and time(3, 5, 7 d), while the soluble protein content, soluble sugar content and catalase(CAT) activity increased, but the rising rate slowed down with the time. The activities of peroxidase(POD), superoxide dismutase(SOD), malondialdehyde(MDA) and proline content increased at first and then decreased. The content of coumarin, cinnamaldehyde, cinnamic acid and dimethoxycinnamaldehyde decreased, while the content of cinnamyl alcohol continued to increase.Under drought stress, the fluorescence signals of reactive oxygen species and no contents in roots of C. cassia seedlings were significantly stronger than those of the control.Further correlation analysis showed that coumarin content, di-methoxycinnamaldehyde content and osmoregulation substance content were significantly negatively correlated(P<0.05), cinnamic acid content was significantly negatively correlated with POD and SOD activities(P<0.01).It was found that C. cassia seedlings showed a certain degree of drought tolerance under short-term or mild drought stress, but if the drought exceeded a certain degree, the physiological metabolism of the seedlings would be unbalanced.
Catalase ; Cinnamomum aromaticum ; Droughts ; Malondialdehyde ; Seedlings ; Stress, Physiological ; Superoxide Dismutase

To study the SSR loci information and develop molecular markers, a total of 43 683 Unigenes in transcriptome of Andrographis paniculata were used to explore SSR. The distribution frequency of SSR and the basic characteristics of repeat motifs were analyzed using MicroSAtellite software, SSR primers were designed by Primer 3.0 software and then validated by PCR. Moreover, the gene function analysis of SSR Unigene was obtained by Blast. The results showed that 14 135 SSR loci were found in the transcriptome of A. paniculata, which distributed in 9 973 Unigenes with a distribution frequency of 32.36%. Di-nucleotide and Tri-nucleotide repeat were the main types, accounted for 75.54% of all SSRs. The repeat motifs of AT/AT and CCG/CGG were the predominant repeat types of Di-nucleotide and Tri-nucleotide, respectively. A total of 4 740 pairs of SSR primers with the potential to produce polymorphism were designed for maker development. Ten pairs of primers in 20 pairs of randomly picked primers produced fragments with expected molecular size. The gene function of Unigenes containing SSR were mostly related to the basic metabolism function of A. paniculata. The SSR markers in transcriptome of A. paniculata show rich type, strong specificity and high potential of polymorphism, which will benefit the candidate gene mining and marker-assisted breeding.

OBJECTIVE: To explore the rules of acupoint selection and drug use in treatment of hypertension with acupoint application therapy. METHODS: The articles of the clinical research of hypertension treated with acupoint application therapy were retrieved from Chinese journal full-text database (CNKI), VIP database (VIP) and Wanfang databases from the time of establishment to January 20, 2019. The database was set up with Microsoft Excel 2010. Using the cloud platform of the ancient and modern medicine record, the frequency statistical and clustering analyses were conducted. RESULTS: A total of 117 articles were collected, including 191 prescriptions, 60 aucpoints and 236 kinds of herbal drugs. It was found in the frequency statistical analysis that the top 6 acupoints in use frequency were Yongquan (KI 1), Quchi (LI 11), Taichong (LR 3), Shenque (CV 8), Sanyinjiao (SP 6) and Neiguan (PC 6). According to the correlation analysis, corresponding to these top 6 acupoints, the pairs of acupoints were Sanyinjiao (SP 6) and Yongquan (KI 1), Shenque (CV 8) and Yongquan (KI 1), Neiguan (PC 6) and Yongquan (KI 1), Zusanli (ST 36) and Sanyinjiao (SP 6), Sanyinjiao (SP 6) and Neiguan (PC 6) with Yongquan (KI 1), as well as Yongquan (KI 1) and Neiguan (PC 6) with Sanyinjiao (SP 6). The dominant meridians were the kidney meridian, the conception vessel and the bladder meridian. The special acupoints referred to -source point, -connecting point, back- point and front- point. The top 3 herbal drugs in use frequency included , and . The herbs used were mainly warm and slight cold in nature and neutral in property. The frequencies of the drug use were similar in the application for cold and heat purposes. The common flavors of the herbal medicines were pungent, sweat and bitter and the liver, kidney and spleen meridians were generally involved in meridian tropism. CONCLUSION In treatment of hypertension with acupoint application therapy, the commonly used single acupoint is Yongquan (KI 1), which is generally combined with Sanyinjiao (SP 6), Shenque (CV 8), Neiguan (PC 6) and Zusanli (ST 36). The correlation is emphasized on the application of special acupoints, meridian points and organs. The vesicatory herbal drugs are predominant in the drug use. In generally, this therapy embodies the treatment principles as tonifying for the deficiency and reducing for the excess, as well as balancing of cold and heat.
Acupuncture Points ; Acupuncture Therapy ; Humans ; Hypertension ; therapy ; Meridians

OBJECTIVETo isolate and identify active neuraminidase constituents of Polygonum cuspidatum against influenza A (H1N1) influenza virus.

METHODOn the basis of the bioassay-guided fractionation,such chromatographic methods as silica gel, sephadex LH-20 and HPLC were adopted to isolate active constituents of extracts from Polygonum cuspidatum, and their molecular structures were identifiied on the basis of their spectral data such as NMR and MS and physico-chemical properties.

RESULTSeven compounds were isolated from the ethyl acetate extract of P. cuspidatum and identified as 2-methoxystypandrone (1), emodin (2), resveratrol (3), polydatin (4), emodin-8-O-beta-D-glucopyranoside (5), (E)-3, 5, 12-trihydroxystilbene-3-O-beta-D-glucopyranoside-2'-(3", 4", 5"-trihydroxybenzoate) (6) and catechin-3-O-gallate (7), respectively. Among them, the NA test showed that compounds 3, 6 and 7 had inhibitory effect against NAs activity, with IC50 values of 129.8, 44.8 and 21.3 micromol x L(-1), respectively. Moreover, the further CPE test showed compounds 6 and 7 had significant inhibitory effect against H1N influenza virus (EC50 = 5.9, 0.9 micromol x L(-1), respectively), with very low cytotoxicity to the host cells, their therapeutic selective index(SI) in MDCK cells ranged from 56 to 269.

CONCLUSIONThe neuraminidase inhibitors against H1N1 anti-influenza virus isolated from extracts of P. cuspidatum on the basis of the bioassay-guided fractionation are significant in specifying their therapeutic material basis and drug R&D against influenza.

Cell Line ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Enzyme Inhibitors ; chemistry ; isolation & purification ; pharmacology ; Fallopia japonica ; chemistry ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; enzymology ; Influenza, Human ; virology ; Molecular Structure ; Neuraminidase ; antagonists & inhibitors

Loganin(LOG),a bioactive compound derived from Cornus officinalis Siebold & Zucc,has been understudied in the context of osteoarthritis(OA)treatment.In this study,we induced an inflammatory response in chondrocytes using lipopolysaccharide(LPS)and subsequently treated these cells with LOG.We employed fluorescence analysis to quantify reactive oxygen species(ROS)levels and measured the expression of NLRP3 and nuclear factor erythropoietin-2-related factor 2(NRF2)using real-time quantitative polymerase chain reaction(qRT-PCR),Western blotting,and immunofluorescence(IF)techniques.Additionally,we developed an OA mouse model by performing medial meniscus destabilization(DMM)surgery and monitored disease progression through micro-com-puted tomography(micro-CT),hematoxylin and eosin(H&E)staining,safranin O and fast green(S&F)staining,and immunohisto-chemical(IHC)analysis.Our results indicate that LOG significantly reduced LPS-induced ROS levels in chondrocytes,inhibited the activation of the NLRP3 inflammasome,and enhanced NRF2/heme oxygenase 1(HO-1)signaling.In vivo,LOG treatment mitigated cartilage degradation and osteophyte formation triggered by DMM surgery,decreased NLRP3 expression,and increased NRF2 expres-sion.These findings suggest that LOG has a protective effect against OA,potentially delaying disease progression by inhibiting the ROS-NLRP3-IL-1β axis and activating the NRF2/HO-1 pathway.

This study used nasal lavage fluid for metabolomics to explore its feasibility, and applied it to the clinical metabolomics study of Xiaoqinglong Decoction in the treatment of allergic rhinitis(AR), aiming to investigate the molecular mechanism of Xiaoqing-long Decoction in the treatment of AR through differential changes in local nasal metabolism. AR patients were selected as the research subjects, and nasal lavage fluid was collected as the sample. Metabolomics analysis using liquid chromatography-mass spectrometry was performed on normal group, AR group, and Xiaoqinglong Decoction group. The differences in metabolic profiles among the groups were compared using principal component analysis and partial least squares discriminant analysis, and differential metabolites were identified and subjected to corresponding metabolic pathway analysis. The results showed that Xiaoqinglong Decoction significantly improved the symptoms of AR patients. The metabolomics analysis revealed 20 differential metabolites between AR group and Xiaoqinglong Decoction group. The core metabolite with a trending return in comparison to normal group was trimethyladipic acid. The metabolites were involved in multiple pathways, including β-alanine metabolism, glutathione metabolism, and phenylalanine, tyrosine, and tryptophan biosynthesis. The feasibility of applying nasal lavage fluid in nasal metabolomics was preliminarily demonstrated. Differential metabolites and enriched pathways in the treatment of AR patients with Xiaoqinglong Decoction were identified, indicating that it may improve rhinitis symptoms through the regulation of various metabolites, including antioxidant effects and correction of Th1/Th2 imbalance.
Humans ; Nasal Lavage Fluid ; Rhinitis, Allergic/drug therapy* ; Metabolomics/methods* ; Metabolome

Objective: This article was to evaluate the applicability and feasibility of "Lingnanzhun" -an "Internet Plus-based HIV Self-testing Tool" targeting MSM in Guangzhou. Hopefully, the results could be used to improve the existing HIV testing services and to support the implementation and scale-up of HIV self-testing programs. Methods: Data were collected from a survey on HIV testing preferences among the Internet-using MSM in April to June, 2015. Univariate and multivariate logistic regression analysis were applied to identify factors associated with the use of HIV self-testing service provided by Lingnanzhun. Information related to the users of Lingnanzhun during September 2014 and December 2018 was also collected. Results: 769 MSM were recruited as participants. Of them, age distribution was 16-77(28.6±6.8) years old, 88.3%(679/769) were unmarried, 42.2%(325/769) were registered residents of Guangzhou, 82.1%(631/769) had university or college education. Among them, 195 (25.4%) used the HIV self- testing program of Lingnanzhun while 574 (74.6%) using the clinic service. Compared with the clinic service users, the Lingnanzhun users showed the following characteristics: longer experience in the MSM community ≥10 years (32.8% vs. 20.9%, 64/195 vs. 120/574); having male casual sexual partners ≥2 (42.1% vs. 29.6%, 82/195 vs. 170/574); having group sex (6.2% vs. 2.6%, 12/195 vs. 15/574) and having commercial sex with men (13.8% vs. 3.0%, 27/195 vs. 17/574). Data from the Lingnanzhun users showed that a total of 3 000 users had practised 5 038 times of self HIV-testings. 11.4% (343/3 000) of the Lingnanzhun users had never been tested. Conclusions It was applicable and feasible to provide Internet-based HIV self-testing service to MSM as a significant complement to the traditional facility-based HIV testing services. It was also useful in accessing those who were having higher risk or had never received HIV testing so as to increase the testing uptake and the frequency.