Polypharmacology-based approach for screening TCM against coinfection of Mycoplasma gallisepticum and Escherichia coli.

Natural products and their unique polypharmacology offer significant advantages for finding novel therapeutics particularly for the treatment of complex diseases. Meanwhile, Traditional Chinese Medicine exerts overall clinical benefits through a multi-component and multi-target approach. In this study, we used the previously established co-infection model of Mycoplasma gallisepticum and Escherichia coli as a representative of complex diseases. A new combination consisting of 6 herbs were obtained by using network pharmacology combined with transcriptomic analysis to reverse screen TCMs from the Chinese medicine database, containing Isatdis Radix, Forsythia Fructus, Ginkgo Folium, Mori Cortex, Licorice, and Radix Salviae. The results of therapeutic trials showed that the Chinese herbal compounds screened by the target network played a good therapeutic effect in the case of co-infection. In summary, these data suggested a new method to validate target combinations of natural products that can be used to optimize their multiple structure-activity relationships to obtain drug-like natural product derivatives.

Ammonia exposure causes the disruption of the solute carrier family gene network in pigs.

Ammonia is the main harmful gas in livestock houses. However, the toxic mechanism of ammonia is still unclear. Therefore, we examined the effects of ammonia exposure on different tissues of fattening pigs by histological analysis and transcriptome techniques in this study. The results showed that there were varying degrees of pathological changes in liver, kidney, hypothalamus, jejunum, lungs, spleen, heart and trachea of fattening pigs under ammonia exposure. Notably, the extent of damage in liver, kidney, jejunum, lungs, hypothalamus and trachea was more severe than that in heart and spleen. Transcriptome results showed that ammonia exposure caused changes in 349, 335, 340, 229, 120, 578, 407 and 115 differentially expressed genes in liver, kidney, spleen, lung, trachea, hypothalamus, jejunum and heart, respectively. Interestingly, the changes in solute vector (SLC) family genes were found in all 8 tissues, and the verified gene results (SLC11A1, SLC17A7, SLC17A6, SLC6A4, SLC22A7, SLC25A3, SLC28A3, SLC7A2, SLC6A6, SLC38A5, SLC22A12, SLC34A1, SLC26A1, SLC26A6, SLC27A5, SLC22A8 and SLC44A4) were consistent with qRT-PCR results. In conclusion, ammonia exposure can cause pathological changes in many tissues and organs of fattening pigs and changes in the SCL family gene network. Importantly, the SCL family is involved in the toxic mechanism of ammonia. Our findings will provide a new insight for better assessing the mechanism of ammonia toxicity.

In vitro and in vivo antioxidant activities of the flavonoid-rich extract from Flos populus.

Flos populi is a well-known traditional Chinese medicine, obtained from the male inflorescence of Populus tomentosa Carr. or Populus canadensis Moench. In this study, we aimed to evaluate the antioxidant activities of Flos populi extract by various in-vitro and in-vivo methods. In-vitro results showed that Flos populi extract had strong antioxidant potential in terms of Superoxide radical and ABTS radical scavenging capacity, nitrogen dioxide radical inhibition and the ability to inhibit lipid peroxidation. Mice were given with Flos populi extract via gavage for 1 month to scrutinize the in-vivo antioxidant effects of Flos populi. The results revealed that Flos populi extract had markedly enhanced the levels of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD) and catalase (CAT), while decreased the level of malondialdehvde (MDA) in serums and livers of mice compared to CCl4 treated group. Additionally, the flavonoids content were determined by the colorimetric method was 553.23 ± 23.45mg/g. The extract was further examined by HPLC and showed the presence of flavonoids-including quercetin (16.26±0.58mg/g), luteolin (9.97±1.07mg/g), apigenin (8.57±0.46mg/g), pinocembrin (119.71±1.05mg/g) and chrysin (16.12±0.53mg/g). The study revealed that Flos populi extract had antioxidant activity and could be utilized as a potential natural antioxidant.

Optimization of baicalin water extraction process from Scutellaria baicalensis (a traditional Chinese medicine) by using orthogonal test and HPLC

ABSTRACT In this study, we optimized the baicalin water extraction process from Scutellaria baicalensis Georgi, Lamiaceae (a traditional Chinese medicine). Orthogonal test design L9(3)4 was used to analyze the optimization of water extraction process of baicalin from S. baicalensis. The effect of solid-liquid ratio, extraction time and soaking time on the yield of baicalin were investigated and optimized by orthogonal test. High-performance liquid chromatography was employed for the determination of extraction yield of baicalin. Analysis of variance was carried out to study the effects of the above three factors. The results showed that solid-liquid ratio plays a significant role in attaining maximum extraction yields of baicalin. However, the other two factors had some effect (not statistically significant) on the extraction yield of baicalin. Conclusively, the optimum experimental conditions such as the solid-liquid ratio (1:12), extraction time (30 min) and soaking time (1 h) for the water extraction of baicalin were proposed which can provide the maximum extraction yield of baicalin. In addition, the score based on the content of baicalin and total solid residues yield were used as evaluation indexes for baicalin uterus suppositories evaluation.

Effects of Traditional Chinese Medicine Qinbai Qingfei Concentrated Pellet on Cellular Infectivity of Mycoplasma pneumoniae.

Aim. To study the effect and mechanism of traditional Chinese medicine Qinbai Qingfei concentrated pellet (QQCP) against Mycoplasma pneumoniae (MP). Methods. Rat airway smooth muscle (ASM) cells were used to examine the antimycoplasmal activity of QQCP via four drug-adding modes: pre- and postadding drugs, simultaneous-adding after drug and MP mixed, and simultaneous-adding drug and MP; taking roxithromycin dispersive tablets (RDT) as positive control, the cellular A 570 values were determined by MTT method. Results. All of A 570 values in QQCP group were significantly higher than those of the corresponding MP control group (P < 0.01) in four drug-adding modes; there was no significant difference in A 570 values between the QQCP group and that of the positive control group (P > 0.05), confirming that QQCP could significantly inhibit the infectivity of MP to ASM cells. Conclusion. QQCP had significant activity in preventing and treating MP infection, killing MP, and antiabsorption.

Baicalin inhibits colistin sulfate-induced apoptosis of PC12 cells.

Baicalin, a type of flavonoid extracted from the dried root of Scutellaria baicalensis georgi, has been shown to effectively inhibit cell apoptosis. Therefore, we assumed that baicalin would suppress co-listin sulfate-induced neuronal apoptosis. PC12 cells exposed to colistin sulfate (62.5-500 µg/mL) for 24 hours resulted in PC12 cell apoptosis. In addition, caspase-3 activity, lactate dehydrogenase level and free radical content increased in a dose-dependent manner. Subsequently, PC12 cells were pretreated with baicalin (25, 50 and 100 µg/mL), and exposed to 125 µg/mL colistin sulfate. Cell morphology markedly changed, and cell viability increased. Moreover, caspase-3 activity, tate dehydrogenase level and free radical content decreased. Results indicated that baicalin inhi-bited colistin sulfate-induced PC12 cell apoptosis by suppressing free radical injury, and reducing caspase-3 activity and lactate dehydrogenase activity.

Extraction and purification of a lectin from red kidney bean and preliminary immune function studies of the lectin and four Chinese herbal polysaccharides.

Reversed micelles were used to extract lectin from red kidney beans and factors affecting reverse micellar systems (pH value, ionic strength and extraction time) were studied. The optimal conditions were extraction at pH 4-6, back extraction at pH 9-11, ion strength at 0.15 M NaCl, extraction for 4-6 minutes and back extraction for 8 minutes. The reverse micellar system was compared with traditional extraction methods and demonstrated to be a time-saving method for the extraction of red kidney bean lectin. Mitogenic activity of the lectin was reasonably good compared with commercial phytohemagglutinin (extracted from Phaseolus vulgaris) Mitogenic properties of the lectin were enhanced when four Chinese herbal polysaccharides were applied concurrently, among which 50 µg/mL Astragalus mongholicus polysaccharides (APS) with 12.5 µg/mL red kidney bean lectin yielded the highest mitogenic activity and 100 mg/kg/bw APS with 12.5 mg/kg/bw red kidney bean lectin elevated mouse nonspecific immunity.

[Studies on the alkaloids from roots of Corydalis impatiens].

OBJECTIVE: To study the alkaloids from Corydalis impatiens. METHODS: The alkaloids were isolated and purified by chromatography and their structures were identified by spectral data and others methods. RESULTS: Seven alkaloids were isolated and identified as bicuculline(1), ochotensine(2), ochotensimine(3), ochrobirine(4), tetrahydrothalifendine(5), norochotensimine(6), N-methylactinodaphnine(7). CONCLUSION: All these compounds are isolated from this plant for the first time.

[Study of huchang qingfei pellets on immune function in rats infected with Mycoplasma pneumoniae].

OBJECTIVE: To study the effects and mechanism of Huchang Qingfei pellets on immune function in rats infected with mycoplasma pneumoniae. METHOD: A rat model of mycoplasmal pneumonia (MP) was developed in repeated intranasal infectious routes and then the humoral and cellular immunocompetences were detected by radioimmunoassay, immune-turbidimetry and flow cytometry. RESULT: The levels of serum IgG,IgM and IL-2, IL-6 were enhinced obviously, the complement C3 and TNF-alpha were decreased and the ratio of CD4+ /CD8+ was improved significantly in the Huchang groups as compared with MP model group. CONCLUSION: Huchang Qingfei pellets can reinforce immune function via preventing both cellular and humoral immunity from depression in the rats with MP.

[Effect of Huchang Qingfei concentrated pellets on E-cadherin expression in lung tissue of mice infected with Mycoplasma pneumoniae].

OBJECTIVE: To investigate the effects of Huchang Qingfei concentrated pellets on the expression of E-cadherin (E-cd) in the lung tissue from mice infected with Mycoplasma pneumoniae (MP). METHOD: A mice model of Mycoplasmal pneumonia (MPP) was developed by repeatedly intranasal infectious route. Transmission electronic microscope (TEM) and immunohistochemistry stain were performed to observe the pathological changes and expression of E-cd in lung tissues. RESULT: Under TEM it was found that the cellular membrane was ruptured, mitochondria was denatured, crista was broken in the pulmonary cells of the model group; the all above parameters in Huchang medicated group were improved obviously. The immunohistochemistry test showed that strong positive brown stain of E-cd expression was found in the pulmonary epithelial cell membrane and bronchial periphery in the model group, however, in the medicated group, the E-cd expression level in the cellular membrane was decreased and the expression ratio was dropped significantly as compared with the model controls. CONCLUSION: Huchang Qingfei concentrated pellets can inhibit the overexpression of E-cd in the lung tissue of mice with MP-infection, which may be helpful for prevention and treatment of pulmonary injury caused by MPP.

Kinetic modulation of HERG potassium channels by the volatile anesthetic halothane.

BACKGROUND: (human ether-a-gogo related gene) encodes the cardiac rapidly activating delayed rectifier potassium currents (I(kr)), which play an important role in cardiac action potential repolarization. General anesthetics, like halothane, can prolong Q-T interval, suggesting that they act on myocellular repolarization, possibly involving HERG channels. Evidence for direct modulation of HERG channels by halothane is still lacking. To gain insight on HERG channel modulation by halothane the authors recorded macroscopic currents expressed in Xenopus oocytes and conducted non-stationary noise analysis to evaluate single channel parameters modified by the anesthetic. METHODS: Macroscopic currents were recorded in 120 mM K(+) internal-5 mM K(+) external solutions with the cut open oocyte technique. Macropatch recordings for non-stationary noise analysis of HERG tail currents were made in symmetrical 120 mM K(+) solutions. Pulse protocols designed for HERG current recording were elicited from a holding potential of -80 mV. Halothane was delivered via gravity-fed perfusion. RESULTS: Halothane (0.7%, 1.5%, and 3%) decreased macroscopic currents in a concentration-dependent manner (average reduction by 14%, 22%, and 35% in the range of -40 mV to 40 mV) irrespective of potential. HERG currents had slower activation and accelerated deactivation and inactivation. Non-stationary noise analysis revealed that halothane, 1.5%, decreased channel P(o) by 27%, whereas single-channel current amplitudes and number of channels in the patch remained unchanged. CONCLUSIONS: Halothane inhibits HERG currents expressed in oocytes in a concentration-dependent manner. It slowed down activation and accelerated deactivation and inactivation of HERG channels. The authors' results demonstrate that halothane decreased HERG currents by modulating kinetic properties of HERG channels, decreasing their open probability. Partial block of I(kr) currents could contribute to delayed myocellular repolarization and altered cardiac electrophysiology.