Effect of Yupingfeng Granules （玉屏风颗粒） on Metabonomics of Alveolar Lavage Fluid and Gut Microflora in Chronic Obstructive Pulmonary Disease Model Rats / 中医杂志

ObjectiveTo explore the possible mechanism of Yupingfeng Granules （玉屏风散） in preventing and treating chronic obstructive pulmonary disease （COPD） from the perspective of “lung-gut axis”. MethodsThirty-two male Wistar rats were randomly divided into normal group，model group， roxithromycin group and Yupingfeng Granules group， with 8 rats in each group. Except for the normal group， the rat model of COPD was prepared by intratracheal instillation of lipopolysaccharide （LPS） combined with smoking for 12 weeks. Since the fifth week of modeling，the roxithromycin group and the Yupingfeng Granules group were given 31.5 mg/（kg·d） and 1.575 g/（kg·d） of corresponding drugs respectively by gavage，and normal group and model group were given 10 ml/（kg·d） physiolo-gical saline. Sample was collected 24 hours after the last administration. The pathological changes of lung tissue were observed using HE staining； Ultrahigh performance liquid chromatography quadrupole time of flight mass spectrometry （UHPLC-QTOFMS） was used to detect the differential metabolites in alveolar lavage fluid （BALF） in all groups but roxithromycin group；16S rDNA sequencing technology was used to detect the changes of intestinal flora， and the association analysis was conducted between the differential metabolites and the differential flora. ResultsCompared with the normal group， the model group showed an increase in goblet cells in the small bronchial wall， disappearance of the smooth muscle layer of the bronchial wall， and infiltration of inflammatory cells； compared with the model group， roxithromycin group showed slight alveolar interstital edema， and obviously reduced inflammatory cell， while no obvious alveolar interstital edema was observed in the Yupingfeng Granules group， showing a small amout of inflammatory cell infiltration. The results of the BALF differential metabolite screening showed that compared with the normal group， 12 substances were upregulated and 19 substances were downregulated in the model group； compared with the model group， 37 substances in the Yupingfeng Granules group were upregulated and 43 substances were downregulated KEGG analysis yielded a total of 2 metabolic pathways， glycerophospholipid metabolism， and unsaturated fatty acid biosynthetic metabolism； compared with the model group， choline， acetylcholine， glycerol-3-phosphate， glycerophosphate choline， palmitic acid， and arachidonic acid showed an upward trend， while stearic acid and docosahexaenoic acid showed a downward trend in Yupingfeng Granules group （P＜0.05）. The results of the intestinal flora showed that， there are 80 different species between the normal group and the model group， and 65 different species between the model group and Yupingfeng Granules group. Among the top 5 species with relative abundance levels，compared with the model group， the level of Prevotella_9，Ruminococcaceae_UCG-005，Ruminiclostridium_6 increase，and Lactobacillus，Bacteroides decrease（P＜0.05）.The results of the correlation analysis showed that， in the normal and model groups， arachidonic acid was negatively correlated with Oribacterium（r=-0.753，P＜0.01）； in the Yupingfeng Granules group and model group， stearic acid and Bacteroides（r=0.788）， Mycobacterium（r=0.826），［Eubacterium］_Ruminantium_Group（r=0.770） was positively correlated（P＜0.01）， Arachidic acid was negatively correlated with Roseiarcus（r=-0.779）， glycerol-3-phosphate was negatively correlated with Desulfovibrio（r=-0.758）， Arachidonic acid was negatively correlated with Oribacterium（r=-0.753）， and Palmitic acid was negatively correlated with Pseudolabs（r=-0.750，P＜0.01）. ConclusionYupingfeng Granules can affect the metabolism of BALF and the flora structure of intestinal microorganisms， and regulating the balance of “lung-gut axis” may be one of the mechanisms of Yupingfeng Granules in treatment of COPD.

Effect of early intervention with electroacupuncture on behavior and gut microbiota in mice with post-traumatic stress disorder / 中华行为医学与脑科学杂志

Objective:To investigate the effect of early intervention with electroacupuncture (EA) on the gut microbiota in a mouse model of post-traumatic stress disorder(PTSD).Methods:Totally 32 C57BL/6 mice were randomly assigned to the following 4 groups ( n=8 for each group): Control group, EA group, PTSD group and PTSD+ EA group.After 7 days acclimation, mice in the PTSD group and PTSD+ EA group were subjected to modified single prolonged stress (mSPS). Mice in the EA group and PTSD+ EA group received EA (2/15 Hz, 1 mA, dilatational wave, 30 min/d) on "Baihui" for 7 days. Mice in the Control group and PTSD group received false stimulation (stimulated the same acupiont without electricity) for 7 days. Seven days after the last stimulation, elevated plus maze test and fear conditioning test were conducted to observe the effect of EA on PTSD-like behavior of mice. At the same time, feces of the mice were collected for gut microbiota detection by 16S rRNA sequencing.SPSS 19.0 was used for statistical analysis.One-way ANOVA was used for multiple group comparison and Bonferrani test was done for further pairwise comparision. Results:(1) There were statistically differences in the open arm activity time of the elevated plus maze test and the immobility time in contextual and cued fear conditioning test among the four groups ( F=6.93, 5.26, 14.51, all P<0.01). In the elevated plus maze test, mice in PTSD group ((60.17±15.52) s) showed significant less time in the open arms than mice in Control group((96.37±14.62) s) and PTSD+ EA group ((86.89±15.02) s) (both P<0.05). In the fear conditioning test, mice in PTSD group ((121.99±29.67) s, (130.82±29.11) s) showed significant increased immobility time both in contextual and cued fear conditioning tests than mice in Control group((74.50±26.65) s, (39.50±23.52) s) and PTSD+ EA group ((76.77±22.60) s, (102.17±3.39) s)(both P<0.05). (2) There were no significant differences among the four groups in the alpha diversity of gut microbiota ( F=0.79-2.45, all P>0.05). (3)Correlation analysis showed that 13 gut microbiotas were negatively correlated with the immobility time in contextual fear conditioning test, 2 gut microbiotas were positively correlated with it; 7 gut microbiotas were negatively correlated with the immobility time in cued fear conditioning test, 1 gut microbiota was positively correlated with it; 3 gut microbiotas were positively correlated with time spent in open arms of elevated plus maze test. Conclusion:Early intervention with EA can improve anxiety-fear like behaviors and gut microflora disorder in PTSD model mice.

Significance of fermented rice beverage on management of antibiotic associated diarrhea (AAD) on Wistar rats

Antibiotic associated diarrhoea (AAD) is caused mostly by disruption of the physiological gut microflora. One potential strategy to prevent this is the concurrent use of probiotic bacteria or yeast. Tribal populations of West Garo Hill region of Meghalaya use locally available unique microflora to prepare fermented food products whichserve as a potential source of novel probiotic organisms and bioactive compounds. In this context, here, we have evaluated the potentiality of laboratory made functional fermented rice beverage in mitigating AAD on animal models viz. Wistar rats. The animal models were administered with fermented rice beverage (test group A1), combination of indigenous Lactobacillus isolates (test group A2), a yeast isolate (test group A3), loperamide (test group STD), normal control (test group NC) and disease control (test group DC). Furthermore, various diarrhea assessment parameters were checked from each group followed by analysis of fecal microbiome, haematological parameters, histopathology of colon, liver and cecal short chain fatty acids (SCFAs) determination. NC and A1 was least affected by AAD induction with a faecal consistency score of 1 on the final day of the study. After day 10, a significant reduction (P <0.05) in the faecal water content was observed in A1, A2 and STD till day 15. Compared to the NC, a slight decrease in body weight was found in the rest of five test groups at day 5, 10 and 15. Except NC, the remaining five test groups showed a significant decrease of lactobacilli and yeast counts in faecal microbiota at 5th day. An increase in the enterococci and coliform counts indicated severe diarrheal condition but A1 reported with significant increase (P <0.05) in the population of Lactobacillus at day 15. An increase in red blood corpuscles, haemoglobin, packed cell volume, mean cell haemoglobin, mean corpuscular volume, mean corpuscular hemoglobin concentration was observed. During the histopathology analysis of colon and liver, normal layers of mucosa, submucosa, muscularis and serous with absence of any abnormal changes or lesions was seen in A1. The cecal concentrations of lactate, acetate and propionate were significantly higher in A1 as compared to the other test groups. Therefore, fermented rice beverage possesses potential to be used in inhibition of antibiotic associated diarrhea with further clinical investigations.

Research progress of prebiotics in drug delivery system / 药学学报

With the in-depth study on the gut microflora, the impact of intestinal bacteria on human health has attracted more and more attention. It has become a research hotspot in life science and medicine, and is considered as an important target of disease control. Prebiotics can regulate the composition and function of intestinal flora and then improve host health. Carbohydrate is the most basic prebiotic. Its unique physiochemical characteristics and gut microbiota-regulating ability make it a promising ingredient for achieving drug target delivery and intestinal health promotion. In this paper, different kinds of prebiotics and their regulation mechanism of intestinal bacteria were illuminated. Moreover, the research progress of carbohydrate prebiotics in drug delivery system was elucidated, and its application prospect is prospected, so as to provide reference for related research.

Interactions between Cannabis Fructus and human gut microflora / 中草药

Objective To explore the characteristics of the interactions between traditional laxative medicine Cannabis Fructus and human gut microflora. Methods HPLC method was used to determine the content of the main unsaturated fatty acids linoleic acid and linolenic acid in Cannabis Fructus Decoction. At the same time, solid culture and liquid culture in vitro anaerobic culture method were combined with 16 S rRNA technology to analyze the interactions between Cannabis Fructus Decotion and human gut microflora. Moreover, the metabolits of linoleic acid and linolenic acid in Cannabis Fructus Decoction by human intestinal microflora were determined using HPLC method. At the same time, the possible conjugated linolenic acid and linoleic acid were determined. Results Cannabis Fructus Decoction promoted the growth of Proteobacteria significantly, which showed that Escherichia-shigella was significantly increased (P < 0.01), but the growth of Bacteroidetes was decreased (P < 0.01), and the content of unsaturated fatty acids linoleic acid and linolenic acid in Cannabis Fructus Decoction were reduced after being incubated with human intestinal bacteria, and the metabolites were conjugated linoleic acid and conjugated linolenic acid. Conclusion The interactions between Cannabis Fructus Decoction and human intestinal microflora are obvious. The Chinese medicine can change the structure of the gut microflora, and the gut microflora can metabolize the drug components. This analysis method partially restores the pharmacokinetics process of the oral administration drug in the human intestinal tract. It could provide a new insight of the mechanism research of Cannabis Fructus.

Human microecology, especially gut microflora provides unprecedented opportunities and challenges for new drug research and development / 中国药理学与毒理学杂志

Since entering the 21st century,the trend of high incidence of chronic diseases in the in?ternational community has not been effectively controlled and the pharmaceutical and medical fields are put forward new challenges. Both the development and evaluation of chemical drugs,biological drugs or traditional Chinese medicine are facing with how to significantly improve the efficacy and reduce the side effect of the drugs. With the coming of the accurate medical treatment as well as the big data era,new opportunities and challenges are needed either in the development of new drugs or new use of old drugs. However,as known that the human body consists of the symbiotic microbial groups of human microecological system especially the gut microecology system is more and more important in human health and disease,and even part diseases such as obesity and diabetes have a certain degree of causality with the gut microflora,so we need to reanalyze the core ideas about drug development or even the current medical theory system. Therefore,it is possible to step out of the old way of current drug development in the new period,to make new contributions on the real effective implementation of prevention and control of chronic diseases. According to international human body microecological especially the research progress of gut microecological field,systematic analysis of the problems existing in the current research and development of new drugs,will not only focus on the human body itself,but also the human microecology as well as the balance between human and micro?ecological,which will promote rapid development of research and development of new drugs and prevention and control of chronic disease in order to improve the physical and mental health services in our country.

Microbial diversity and digestive enzyme activities in the gut of earthworms found in sawmill industries in Abeokuta, Nigeria / Diversidad microbiana y actividad de enzimas digestivas en el intestino de las lombrices de tierra en los aserraderos de Abeokuta, Nigeria

The growing demand for wood has resulted in large volumes of wood wastes that are daily released to the soil from the activities of sawmills in South-Western Nigeria. In an attempt to setup a bioremediation model for sawdust, this study therefore aimed at evaluating microbial diversity, and the level of digestive enzymes in the gut of earthworms (Eudrilus eugeniae, Libyodrilus violaceous and Hyperiodrilus africanus) of sawmill origin. Four major sawmills located in Abeokuta (7o9’12” N - 3o19’35” E), namely Lafenwa, Sapon, Isale-Ake and Kotopo sawmills were used for this study. The arboretum of the Federal University of Agriculture, Abeokuta was used as control. Gut microbial analysis was carried out using the pour-plate method while digestive enzyme activities in the earthworm guts were done by the spectrophotometric method. Higher microbial counts (28.5±0.1x10³-97.0±0.1x10³cfu for bacteria and 7.0±0.1x10³-96.0±0.1x10³cfu for fungi) and microbial diversity were recorded in the gut of earthworms of the sawmill locations than those of the control site (17.5±0.1x10³cfu for bacteria and 4.5±0.1x10³cfu for fungi). Streptococcus mutans and Proteus spp.were common in the gut of E. eugeniae, and L. violaceous from the study sawmills, while Streptococcus mutans were also identified in H. africanus, but absent in the gut of E. eugeniae from the control site. Cellulase (48.67±0.02mg/g) and lipase (1.81±0.01mg/g) activities were significantly higher (p<0.05) in the gut of earthworms from the control site than those of the study sawmills. Furthermore, amylase (α and β) activity was highest in the gut of earthworms from the sawmills. Variations observed in the gut microbial and digestive enzyme activities of earthworms from the study sawmills as compared to the control site suggests that earthworms, especially E. eugeniae, could be a better organism for use as bioremediator of wood wastes. Rev. Biol. Trop. 62 (3): 1241-1249. Epub 2014 September 01.

La creciente demanda de madera ha resultado en grandes volúmenes de residuos de madera que se lanzan diariamente a la tierra, procedentes de las actividades de los aserraderos en el suroeste de Nigeria. Se realizó este estudio en un intento de configurar un modelo de biorremediación de aserrín, con el objetivo de evaluar la diversidad microbiana y el nivel de las enzimas digestivas en el intestino de las lombrices de tierra (Eudrilus eugeniae, Libyodrilus violáceo y Hyperiodrilus africanus) de origen aserradero. Para este estudio se utilizaron cuatro grandes aserraderos ubicados en Abeokuta (7°9’12 ‘’ N - 3°19’35 ‘’ E), a saber: Lafenwa, Sapon, Isale-Ake y Aserraderos Kotopo. El arboreto de la Universidad Federal de Agricultura, Abeokuta se utilizó como control. El análisis microbiano se llevó a cabo utilizando el método de vertido de placa, mientras que las actividades de enzimas digestivas en los intestinos de las lombrices se realizaron por el método espectrofotométrico. Los recuentos microbianos más altos fueron 28.5±97.0±0.1x10³-0.1x10³cfu para las bacterias y 7.0±0.1x10³-96.0±0.1x10³cfu para los hongos y la diversidad microbiana registrada en el intestino de las lombrices de tierra de los lugares de aserraderos y las del sitio de control fueron: 17.5±0.1x10³cfu para bacterias y 4.5±0.1x10³cfu para hongos. Streptococcus mutans y Proteus spp. eran comunes en el intestino de E. eugeniae, y L. violaceous de los aserraderos de estudio, mientras que también se identificaron Streptococcus mutans en H. africanus, que estuvieron ausentes en el intestino de E. eugeniae en el sitio de control. Actividades de celulasa (48.67±0.02mg/g) y lipasa (1.81±0.01mg/g) fueron significativamente mayores (p<0.05) en el intestino de las lombrices de tierra en el sitio de control que las de los aserraderos de estudio. Por otra parte, la actividad de la amilasa (α y β) fue mayor en el intestino de las lombrices de tierra de los aserraderos. Las variaciones observadas en la microbiota intestinal y la actividad de las enzimas digestivas de las lombrices de tierra de los aserraderos de estudio, en comparación con el sitio de control, sugieren que las lombrices de tierra, especialmente E. eugeniae, podrían ser un mejor organismo para su uso como biorremediador de residuos de madera.

Correlation of gut microflora and intestinal mucosal immunity / 医学研究生学报

Gut microflora , an important part in maintaining the intestinal homeostasis , can participate in the development of intestinal mucosal immune system , promote the synthesis of secreted IgA ( sIgA) and interact with intestinal immune cells .Gut micro-flora also plays a significant role in the development of inflammatory bowel diseases , irritable bowel syndrome , pediatric allergic disea-ses and other disorders .This paper reviews the advances about the correlation of gut microflora and intestinal mucosal immunity .

In vivo intestinal absorption characteristics of α-hederin in rats / 中草药

Objective: To investigate the intestinal absorption characteristics of α-hederin and to explore the causes of poor bioavailability. Methods: In vivo single-pass perfusion model was used and the concentration of α-hederin was determined by HPLC. The effects of intestinal segment, drug concentration, pH value, gut microflora, and P-gp inhibitor on the intestinal absorption of the drug were investigated. Results: The absorption rate constant (Ka) of α-hederin decreased following the sequence of ileum > colon > jejunum > duodenum. Absorption parameters of α-hederin had no significant difference at different concentration of 75, 150, and 300 μg/mL and those increased with the increase of pH value. The intestinal flora which were disrupted may affect the absorption of α-hederin. There was no significant difference in Ka and Peff values between P-gp inhibitor and no P-gp inhibitor groups. Conclusion: α-Hederin can be absorbed in whole intestine, but better in lower intestine. The saturate phenomena was not observed under the test range of drug concentration, and the absorption mechanism may be the passive diffusion transport. The absorption can be better under basic condition. The absorption is significantly affected by the intestinal flora and α-hederin is not the substrate of P-gp.