Effects of total ginsenosides from Panax ginseng stems and leaves on gut microbiota and short-chain fatty acids metabolism in acute lung injury mice / 中国中药杂志

This study aimed to investigate the biological effects and underlying mechanisms of the total ginsenosides from Panax ginseng stems and leaves on lipopolysaccharide(LPS)-induced acute lung injury(ALI) in mice. Sixty male C57BL/6J mice were randomly divided into a control group, a model group, the total ginsenosides from P. ginseng stems and leaves normal administration group(61.65 mg·kg~(-1)), and low-, medium-, and high-dose total ginsenosides from P. ginseng stems and leaves groups(15.412 5, 30.825, and 61.65 mg·kg~(-1)). Mice were administered for seven continuous days before modeling. Twenty-four hours after modeling, mice were sacrificed to obtain lung tissues and calculate lung wet/dry ratio. The number of inflammatory cells in bronchoalveolar lavage fluid(BALF) was detected. The levels of interleukin-1β(IL-1β), interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α) in BALF were detected. The mRNA expression levels of IL-1β, IL-6, and TNF-α, and the levels of myeloperoxidase(MPO), glutathione peroxidase(GSH-Px), superoxide dismutase(SOD), and malondialdehyde(MDA) in lung tissues were determined. Hematoxylin-eosin(HE) staining was used to observe the pathological changes in lung tissues. The gut microbiota was detected by 16S rRNA sequencing, and gas chromatography-mass spectrometry(GC-MS) was applied to detect the content of short-chain fatty acids(SCFAs) in se-rum. The results showed that the total ginsenosides from P. ginseng stems and leaves could reduce lung index, lung wet/dry ratio, and lung damage in LPS-induced ALI mice, decrease the number of inflammatory cells and levels of inflammatory factors in BALF, inhibit the mRNA expression levels of inflammatory factors and levels of MPO and MDA in lung tissues, and potentiate the activity of GSH-Px and SOD in lung tissues. Furthermore, they could also reverse the gut microbiota disorder, restore the diversity of gut microbiota, increase the relative abundance of Lachnospiraceae and Muribaculaceae, decrease the relative abundance of Prevotellaceae, and enhance the content of SCFAs(acetic acid, propionic acid, and butyric acid) in serum. This study suggested that the total ginsenosides from P. ginseng stems and leaves could improve lung edema, inflammatory response, and oxidative stress in ALI mice by regulating gut microbiota and SCFAs metabolism.

Short-chain fatty acids ameliorate spinal cord injury recovery by regulating the balance of regulatory T cells and effector IL-17+ γδ T cells / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Spinal cord injury (SCI) causes motor, sensory, and autonomic dysfunctions. The gut microbiome has an important role in SCI, while short-chain fatty acids (SCFAs) are one of the main bioactive mediators of microbiota. In the present study, we explored the effects of oral administration of exogenous SCFAs on the recovery of locomotor function and tissue repair in SCI. Allen's method was utilized to establish an SCI model in Sprague-Dawley (SD) rats. The animals received water containing a mixture of 150 mmol/L SCFAs after SCI. After 21 d of treatment, the Basso, Beattie, and Bresnahan (BBB) score increased, the regularity index improved, and the base of support (BOS) value declined. Spinal cord tissue inflammatory infiltration was alleviated, the spinal cord necrosis cavity was reduced, and the numbers of motor neurons and Nissl bodies were elevated. Enzyme-linked immunosorbent assay (ELISA), real-time quantitative polymerase chain reaction (qPCR), and immunohistochemistry assay revealed that the expression of interleukin (IL)‍-10 increased and that of IL-17 decreased in the spinal cord. SCFAs promoted gut homeostasis, induced intestinal T cells to shift toward an anti-inflammatory phenotype, and promoted regulatory T (Treg) cells to secrete IL-10, affecting Treg cells and IL-17+ γδ T cells in the spinal cord. Furthermore, we observed that Treg cells migrated from the gut to the spinal cord region after SCI. The above findings confirm that SCFAs can regulate Treg cells in the gut and affect the balance of Treg and IL-17+ γδ T cells in the spinal cord, which inhibits the inflammatory response and promotes the motor function in SCI rats. Our findings suggest that there is a relationship among gut, spinal cord, and immune cells, and the "gut-spinal cord-immune" axis may be one of the mechanisms regulating neural repair after SCI.

Jujubae Fructus alleviates intestinal injury caused by toxic medicinals in Shizao Decoction based on correlation between intestinal flora and host metabolism / 中国中药杂志

Genkwa Fols, Kansui Radix, and Euphorbiae Pekinensis Radix in Shizao Decoction(SZD) are toxic to intestinal tract. Jujubae Fructus in this prescription can alleviate the toxicity, but the mechanism is still unclear. Therefore, this study aims to explore the mechanism. To be specific, 40 normal Sprague-Dawley(SD) rats were classified into the normal group, high-dose and low-dose SZD groups, and high-dose and low-dose SZD without Jujubae Fructus(SZD-JF) groups. The SZD groups were given(ig) SZD, while SZD-JF groups received the decoction without Jujubae Fructus. The variation of body weight and spleen index were recorded. The patho-logical changes of intestinal tissue were observed based on hematoxylin and eosin(HE) staining. The content of malondialdehyde(MDA) and glutathione(GSH) and activity of superoxide dismutase(SOD) in intestinal tissue were measured to evaluate the intestinal injury. Fresh feces of rats were collected to detect intestinal flora structure by 16S ribosomal RNA gene(16S rDNA) sequencing technology. The content of fecal short chain fatty acids and fecal metabolites was determined by gas chromatography-mass spectrometer(GC-MS) and liquid chromatography-mass spectrometer ultra-fast liquid chromatography-quadrupole-time-of-flight mass spectrometer(UFLC-Q-TOF-MS), separately. Spearman's correlation analysis was employed to analyze the differential bacteria genera and differential metabolites. RESULTS:: showed that high-dose and low-dose SZD-JF groups had high content of MDA in intestinal tissue, low GSH content and SOD activity, short intestinal villi(P<0.05), low diversity and abundance of intestinal flora, variation in the intestinal flora structure, and low content of short chain fatty acids(P<0.05) compared with the normal group. Compared with high-dose and low-dose SZD-JF groups, high-dose and low-dose SZD groups displayed low content of MDA in intestinal tissue, high GSH content and SOD activity, recovery of the length of intestinal villi, increased abundance and diversity of intestinal flora, alleviation of dysbacteria, and recovery of the content of short chain fatty acids(P<0.05). According to the variation of intestinal flora and fecal metabolites after the addition of Jujubae Fructus, 6 differential bacterial genera(Lactobacillus, Butyricimonas, Clostridia＿UCG-014, Prevotella, Escherichia-Shigella, Alistipes),4 differential short chain fatty acids(such as acetic acid, propionic acid, butyric acid, valeric acid) and 18 differential metabolites(such as urolithin A, lithocholic acid, and creatinine) were screened out. Beneficial bacteria such as Lactobacillus were in positive correlation with butyric acid and urolithin A(P<0.05). The pathogenic bacteria such as Escherichia-Shigella were in negative correlation with propionic acid and urolithin A(P<0.05). In summary, SZD-JF caused obvious intestinal injury to normal rats, which could lead to intestinal flora disorder. The addition of Jujubae Fructus can alleviate the disorder and relieve the injury by regulating intestinal flora and the metabolites. This study discusses the effect of Jujubae Fructus in relieving the intestinal injury caused by SZD and the mechanism from the perspective of intestinal flora-host metabolism, which is expected to serve as a reference for clinical application of this prescription.

Effects of electroacupuncture at "Siguan" points on the expression of colonic 5-hydroxytryptamine and fecal short-chain fatty acids in rats with post-stroke depression / 中国针灸

OBJECTIVE@#To observe the effects of electroacupuncture at "Siguan" points on behavior, colonic 5-hydroxytryptamine (5-HT) and fecal short-chain fatty acids (SCFAs) in rats with post-stroke depression (PSD), and explore the effect mechanism of electroacupuncture at Siguan points on PSD.@*METHODS@#Fifty SD rats were randomly divided into a sham-operation group, a stroke group, a PSD group, a drug group and an electroacupuncture group, with 10 rats in each one. The stroke model was established by middle cerebral artery occlusion (MCAO) method in the stroke group; except for the sham-operation group, the rats in the other groups were intervened with MCAO combined with solitary and chronic unpredictable mild stress (CUMS) to establish PSD model. In the electroacupuncture group, electroacupuncture was delivered at "Hegu" (LI 4) and "Taichong" (LR 3), with disperse-dense wave, 2 Hz/10 Hz in frequency, for 30 min in each intervention, once daily, for consecutive 21 days. Simultaneously, distilled water (0.01 L•kg-1•d-1) was administrated intragastrically. Fluoxetine solution (2.33 mg•kg-1•d-1) was given by gavage , once a day and for 21 days in the drug group. The same procedure of fixation and gavage with distilled water were adopted in the sham-operation group, the stroke group and the PSD group. Separately, before stroke modeling, after PSD modeling and after 21-day intervention, the consumption of sugar water and the scores of horizontal movement and vertical movement in open-field test were observed. After 21-day intervention, the content of colonic 5-HT was detected by immunohistochemical method, and that of fecal SCFAs was determined by gas chromatography mass spectrometry.@*RESULTS@#After PSD modeling, compared with the stroke group, the sugar water consumption, the horizontal movement scores and vertical movement scores of the open-field test were all reduced in the PSD group, the drug group and the electroacupuncture group (P<0.05). After 21-day intervention, the sugar water consumption and the scores of horizontal movement and vertical movement of the open-field test were increased in the drug group and the electroacupuncture group (P<0.05) when compared with the PSD group; and the horizontal movement score in the electroacupuncture group was lower than that of the drug group (P<0.05). Compared with the sham-operation group, the contents of total fecal SCFAs and acetic acid were lower in the stroke group (P<0.05), and the contents of colonic 5-HT and total fecal SCFAs, acetic acid, propionic acid and butyric acid were reduced in the PSD group (P<0.05). In comparison with the PSD group, the contents of colonic 5-HT and total fecal SCFAs, acetic acid and propionic acid were increased in the drug group and the electroacupuncture group (P<0.05); and the content of colonic 5-HT in the electroacupuncture group was lower than that of the drug group (P<0.05). The level of colonic 5-HT was positively correlated with the contents of total fecal SCFAs and propionic acid (r=0.424, P=0.005; r=0.427, P=0.004).@*CONCLUSION@#Electroacupuncture at "Siguan" points can relieve the depression-like behavior of PSD rats, and its underlying mechanism may be related to the regulation of fecal SCFAs, which affects the release of colonic 5-HT.

Effect of Zuogui Jiangtang Tongmai Prescription on SCFAs/GPR43/GLP-1/GLP-1R Signaling Pathway in Rats with Diabetes Mellitus Complicated with Cerebral Infarction / 中国实验方剂学杂志

ObjectiveTo explore the potential mechanism of Zuogui Jiangtang Tongmai prescription （ZJT） in the treatment of diabetes mellitus complicated with cerebral infarction （DM-CI） in rats based on the short-chain fatty acids （SCFAs）/G protein-coupled receptor 43 （GPR43）/glucagon-like peptide-1 （GLP-1）/GLP-1 receptor （GLP-1R） signaling pathway. MethodSixty SD rats were randomly divided into sham operation group， model group， low- and high-dose ZJT groups （12， 24 g·kg-1）， western medicine group （140 mg·kg-1 pioglitazone metformin tablets + 27 mg·kg-1 enteric-coated aspirin tablets）. Except for the sham operation group， all other groups were fed a high-sugar high-fat diet for 4 weeks and then subjected to intraperitoneal injection of 1% streptozotocin at 35 mg·kg-1 combined with middle cerebral artery occlusion （MCAO） to establish a DM-CI rat model. The corresponding interventions were performed with distilled water， low-dose ZJT， high-dose ZJT， pioglitazone metformin tablets， and enteric-coated aspirin tablets. After surgery， National Institutes of Health Stroke Scale （NIHSS） scoring and triphenyltetrazolium chloride （TTC） staining to measure the rat's cerebral infarct volume were carried out. Random blood glucose levels were measured， and hematoxylin-eosin （HE） staining was used to observe histopathological changes in rat brain tissues. Gas chromatography was employed to detect the content of SCFAs in the cecum contents. Enzyme-linked immunosorbent assay （ELISA） was adopted to measure serum GLP-1 level. Western blot was used to detect the protein expression of GPR43 in rat ileal tissues and GLP-1R in the ischemic brain tissues. ResultCompared with the sham operation group， the model group showed significantly increased NIHSS scores， random blood glucose levels， and cerebral infarct volumes （P<0.01）， and significantly decreased SCFAs content， GLP-1 levels， and GPR43 and GLP-1R protein expression （P<0.01）. Compared with the model group， the high-dose ZJT group and the western medicine group exhibited significantly reduced NIHSS scores， random blood glucose levels， and cerebral infarct volumes （P<0.05， P<0.01）， and significantly increased SCFAs content， GLP-1 levels， and GPR43 and GLP-1R protein expression （P<0.01）. ConclusionZJT can improve glucose metabolism disorder and reduce neurological damage in DM-CI rats， and its mechanism may be related to the increase in SCFAs content and the upregulation of the GPR43/GLP-1/GLP-1R signaling pathway.

Protective effect of Clostridium butyricum on renal tissue in db/db mice and its mechanism / 中华内分泌代谢杂志

Objective:To investigate the effect of Clostridium butyricum on renal tissue of db/db mice and to explore its mechanism. Methods:Fourteen-week-old db/db mice were divided into db/db group( n=10) and db/db+ Cb group( n=7) according to random number table method. Age-matched db/m mice were selected as the normal control group. The db/m and db/db mice were administered 0.9% sodium chloride solution by gavage, while the db/db+ Cb mice were administered an equivalent amount of Clostridium butyricum solution by gavage for 8 weeks. Serum creatinine , fasting blood glucose, urinary albumin to creatinine ratio(ACR) and other biochemical indicators were also detected. HE staining was used to observe the pathological changes of kidney tissue. The expressions of peroxisome proliferators-activated receptor γ coactivator-1α(PGC-1α) mRNA were detected by realtime PCR, while the expressions of nuclear factor-κB(NF-κB), glucagon-like peptide 1 receptor(GLP-1R), and adenosine monophosphate-activated protein kinase(AMPK) in kidney tissue were determined by immunohistochemistry and Western blotting. The levels of intestinal flora, serum and fecal short-chain fatty acids(SCFAs) were measured by 16S rRNA, liquid chromatograph-mass spectrometer, and gas chromatograohy-mass spectrometry respectively. Results:Compared to db/db mice, db/db+ Cb mice showed improvement in general condition after supplementation with Clostridium butyricum. Fasting blood glucose, blood urea nitrogen, albumin-to-creatinine ratio(ACR), blood creatinine, and levels of interleukin-6(IL-6) in kidney tissue were reduced(all P<0.05). The pathology showed various degrees of amelioration of kidney tissue injury in mice. The expression of PGC-1α mRNA increased in kidney tissue( P<0.05). Decreased expression of NF-κB protein, as well as increased expression of GLP-1R and phosphorylated(p-)AMPK/AMPK protein(all P<0.05) were detected in kidney tissues. Clostridium butyricum modulated the composition of the gut microbiota with elevated total SCFAs in blood and feces. Conclusion:Clostridium butyricum increased the expression of GLP-1R in kidney tissue, promoted AMPK phosphorylation, and alleviated renal tissue damage in mice. This suggests that it may be associated with regulating the abundance of SCFA-producing bacterial populations.

Stoichiometric models for prediction of enteric methane production

This study aimed to empirically evaluate the adjustment quality of two stoichiometric models of methane production from diets that used different rumen fermentation modulators. We used the models proposed by Moss et al. (2000) and Blümmel et al. (1997). The data set consisted of 169 observations of in vitro methane production and volatile fatty acids (acetate, propionate, and butyrate) from dissertations, theses, and scientific articles. The model's adequacy evaluation was only possible through combination of several statistical analyzes. The models were unable to predict the enteric methane precisely and accurately. Despite this, the model proposed by Moss et al. (2000) showed a better fit based on the mean bias (% of observed, MB), MEF, RMSEP, Cb and CCC measures compared to the model proposed by Blümmel et al. (1997). However, both models presented strong evidence against the general hypothesis H0: a = 0 & b = 1. Thus, the model proposed by MOSS et al. (2000) was more efficient, whereas the model by Blümmel et al. (1997) showed a better fit for diets with high availability of H2.

Interactions between gut microbiota and berberine,a necessary procedure to understand the mechanisms of berberine / 药物分析学报

Berberine(BBR),an isoquinoline alkaloid,has been found in many plants,such as Coptis chinensis Franch and Phellodendron chinense Schneid.Although BBR has a wide spectrum of pharmacological effects,its oral bioavailability is extremely low.In recent years,gut microbiota has emerged as a cynosure to un-derstand the mechanisms of action of herbal compounds.Numerous studies have demonstrated that due to its low bioavailability,BBR can interact with the gut microbiota,thereby exhibiting altered pharma-cological effects.However,no systematic and comprehensive review has summarized these interactions and their corresponding influences on pharmacological effects.Here,we describe the direct interactive relationships between BBR and gut microbiota,including regulation of gut microbiota composition and metabolism by BBR and metabolization of BBR by gut microbiota.In addition,the complex interactions between gut microbiota and BBR as well as the side effects and personalized use of BBR are discussed.Furthermore,we provide our viewpoint on future research directions regarding BBR and gut microbiota.This review not only helps to explain the mechanisms underlying BBR activity but also provides support for the rational use of BBR in clinical practice.

Wang's Yijing Decoction Combined with Conventional Therapy： Performance in Treating Polycystic Ovary Syndrome with Kidney Deficiency and Phlegm-dampness Syndrome and Regulating Effect on Intestine-brain Axis / 中国实验方剂学杂志

ObjectiveTo observe the therapeutic effect of Wang's Yijing decoction combined with conventional therapy on the treatment of polycystic ovary syndrome （PCOS） with kidney deficiency and phlegm-dampness syndrome and its regulating effect on the intestine-brain axis. MethodThe 132 patients from the gynecological clinic of Shanxi Bethune Hospital were randomly assigned into a control group and an observation group， with 66 patients in each group. The patients in the control group were treated with drospirenone and ethinylestradiol tablets and metformin tablets， and those in the observation group were administrated with Wang's Yijing decoction on the basis of the therapy in the control group. The treatment in both groups lasted for 3 menstrual cycles. The changes in menstrual blood volume， color， and texture， as well as that in the menstrual cycle before and after treatment were scored. Further， the Rosenfield， hirsutism， and kidney deficiency and phlegm-dampness syndrome were scored before and after treatment. Pelvic ultrasound was performed before and after treatment to compare the bilateral ovarian volume. The levels of anti-mullerian hormone （AMH）， estradiol （E2）， luteinizing hormone （LH）， follicle-stimulating hormone （FSH）， testosterone （T）， dihydrotestosterone （DHT）， and sex hormone-binding globulin （SHBG） were measured before and after treatment， and the free androgen index （FAI） was calculated. After the fasting insulin and fasting blood glucose were examined before and after treatment， the homeostatic model assessment of insulin resistance （HOMA-IR） was calculated. The levels of ghrelin， glucagon-like peptide-1 （GLP-1）， peptide tyrosine-tyrosine （PYY） in serum and short-chain fatty acids （SCFAs） in fresh stool were measured before and after treatment. ResultThe treatments in both groups lowered the scores of irregular menstruation， Rosenfield， hirsutism， and kidney deficiency and phlegm-dampness syndrome （P<0.01）， and the scores in the observation group were lower than those in the control group （P<0.01）. After treatment， the T， DHT， FAI， AMH， E2， LH， and FSH levels lowered （P<0.01）， while the SHBG level elevated （P<0.01）. In addition， the AMH， E2， LH， T， DHT， and FAI in the observation group were lower than those in control group， while the SHBG level presented an opposite trend （P<0.01）. After treatment， the bilateral ovarian volume and HOMA-IR of the two groups decreased （P<0.01）， and were lower in the observation group than in the control group （P<0.05， P<0.01）. The treatments in both groups elevated the levels of ghrelin， GLP-1， PYY， and SCFAs （P<0.01）， which were higher in the observation group than in the control group （P<0.01）. After treatment， the recovery rate in the observation group was 65.57% （40/61）， which was higher than 44.83% （26/58） in the control group （χ2=5.180， P<0.05）. ConclusionWang's Yijing decoction combined with conventional western medicine treatment can regulate the intestine-brain axis， endocrine and metabolic disorders， and sex hormones， adjust the menstrual cycle， and alleviate clinical symptoms， demonstrating good clinical effect for the PCOS patients with kidney deficiency and phlegm dampness syndrome.

The effect and mechanism of short-chain fatty acid regulate tacrolimus-related hyperglycemia in mice / 中华泌尿外科杂志

Objective:To investigate the effect and mechanism of short-chain fatty acids (SCFAs) on the side-effect of tacrolimus on blood glucose.Methods:The C57BL/6 mice were treated with tacrolimus orally (10 mg/kg, tacrolimus group), tacrolimus plus 150 mmol/L sodium butyrate and isovalerate mixed solution (SCFAs group), broad-spectrum antibiotics (antibiotic group), and tacrolimus plus broad-spectrum antibiotics (tac&abx group). After 8 weeks intervention, the fasting blood glucose (FBG), oral glucose tolerance test (OGTT), hemoglobin A1C (HbA1c) were tested as indicators of glucose metabolism, and the gut microbiota, SCFAs concentration in the ileocecal, serum glucagon-like peptide-1 (GLP-1), fasting serum insulin, and GLP-1 expression in intestinal mucosa were performed for intestinal-glucose metabolism mechanism.Results:The FBG and HbA1c were significantly increased in tacrolimus group[(7.31±0.97)mmol/L, (8.34±1.12)%] than control group [(5.23±0.30)mmol/L, (4.32±0.80)%, all P<0.05], which remained normal in antibiotic group [(4.92±0.31)mmol/L, (5.61±0.98)%)], tac&abx group[(5.95±0.37)mmol/L, (4.56±0.26)%] and SCFAs groups [(5.87±0.68)mmol/L, (5.07±1.79)%]. The OGTT in the tacrolimus group showed glucose tolerance impairment, while other groups remained normal. The ileocecal butyric acid and isovaleric acid concentrations in the tacrolimus group were (722.3±262.2) μg/g and (10.0±5.1)μg/g, lower than the control group[ (1 321.3±165.5) μg/g, (19.7±3.6)μg/g, P<0.05]. The above acids in the SCFAs group remained normal as in the control group [(1 375.7±451.6) μg/g, (24.5±11.5)μg/g)]. The fasting serum insulin in the tacrolimus group decreased significantly to (3.2 ± 0.6)mIU/L, compared with control[ (4.4±0.9) mIU/L]and SCFAs groups [(7.0±1.1) mIU/L]. The GLP-1 test indicated a significant decrease in the tacrolimus group[ (4.7±2.9)pg/ml, P<0.05] compared with the SCFAs group and control group [(42.5±19.9) pg/ml, (33.1±9.1) pg/ml]. Conclusions:Tacrolimus affects glucose metabolism through the SCFAs-associated GLP-1 pathway in the intestine, and oral supplementation with mixed SCFAs provides a new insight for the prevention and treatment of tacrolimus-induced hyperglycemia in transplant recipients.

Regulatory Mechanism of Total Glucosides of Paeony on Gut Microenvironment in Non-obese Diabetic Mice with Sjogren's Syndrome / 中国实验方剂学杂志

Objective:To investigate the effects of total glucosides of paeony （TGPs） on intestinal motility， barrier function， and gut microbiota in non-obese diabetic （NOD） mice with Sjogren's syndrome （SS）. Method:Thirty NOD mice were randomly assigned into the model group （deionized water）， prebiotic fructo-oligosaccharide （FOS） group （700 mg∙kg^-1）， and the low- （160 mg∙kg^-1）， medium- （320 mg∙kg^-1）， and high-dose （640 mg∙kg^-1） TGP groups， with six mice in each group. Moreover， the BALB/c mice were employed as the normal control and administered with deionized water. The food and water intakes， number of discharged fecal particles， and fecal moisture content were observed to evaluate the effect of TGPs on intestinal motility in SS mice. The levels of <italic>D</italic>-lactate （<italic>D</italic>-Lac） content， diamine oxidase （DAO）， and junction-associated protein zonula occludens-1 （ZO-1） in mouse serum were detected by enzyme linked immunosorbent assay （ELISA）. The fecal samples collected at different time points were determined by spread plate method and gas chromatography for uncovering the intestinal microbial communities and the content of short-chain fatty acids. Result:Compared with the normal group， the model group exhibited decreased food and water intakes （<italic>P</italic><0.01）， weakened intestinal propulsion （<italic>P</italic><0.01）， elevated <italic>D</italic>-Lac and DAO （<italic>P</italic><0.05，<italic>P</italic><0.01）， lowered ZO-1 and SCFAs （<italic>P</italic><0.05，<italic>P</italic><0.01）， and reduced number of intestinal bacteria （<italic>P</italic><0.01）. The comparison with the model group revealed that TGPs significantly increased the number of discharged fecal particles and fecal moisture content （<italic>P</italic><0.05，<italic>P</italic><0.01）， enhanced intestinal propulsion （<italic>P</italic><0.05， <italic>P</italic><0.01）， decreased serum <italic>D</italic>-Lac and DAO levels （<italic>P</italic><0.05，<italic>P</italic><0.01）， and up-regulated ZO-1 expression （<italic>P</italic><0.01）. Apart from increasing the proportions of <italic>Bifidobacterium</italic> and <italic>Lactobacillus</italic> and decreasing the proportion of<italic> Enterobacter </italic>in intestinal flora （<italic>P</italic><0.05，<italic>P</italic><0.01）， TGPs also accelerated the production of acetic acid and butyric acid （<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:TGPs attenuate SS-mediated constipation and restore the impaired intestinal barrier function in mice by increasing fecal moisture content， boosting intestinal motility， regulating intestinal microbial communities， elevating acetic acid and butyric acid levels， and up-regulating tight junction protein expression.

Research progress of microbe gut brain axis / 中国综合临床

The emergence of gut microbes has become one of the key regulatory factors of gut brain function, and has made people realize the importance of microbe gut brain axis.The gut microbiota and brain are interconnected through immune system, tryptophan metabolism, vagus nerve and endocrine system.Many factors can affect the composition of gut microbiota, including infection, mode of delivery, use of antibiotics, stress, and host genetics.Recent studies have shown that gut microbiota may be associated with many diseases, including autism, irritable bowel syndrome and so on.Future research will focus on understanding the underlying mechanisms of the microbial gut brain axis and attempt to elucidate microbial based intervention and treatment strategies for neuropsychiatric diseases.

Short-chain fatty acids production by Bifidobacterium species in the presence of salep

BACKGROUND: Salep is obtained by grinding dried orchid tubers and used as a valuable ingredient in the food industry. Because of the glucomannan content of salep, it is thought to have prebiotic potential. However, there is little information in studies concerning the fermentation characteristics and potential prebiotic properties of salep. The objective of this study was to investigate the effect of salep on bifidobacterial growth by measuring the highest optical density (OD), calculating the specific growth rates, and determining the production of lactic acid and short-chain fatty acids (acetic, propionic, and butyric acid) as a result of bacterial fermentation. RESULT: The OD and pH values obtained in this study showed that salep was utilized as a source of assimilable carbon and energy by the Bifidobacterium species (BS). All Bifidobacterium strains produced lactic, acetic, propionic, and butyric acid, indicating that salep is readily fermented by these bacteria. Salep at 1% (w/v) showed a similar effect on bifidobacterial growth as that promoted by 1% (w/v) glucose used as a traditional carbon source. CONCLUSIONS: Bifidobacterium species can develop in media containing salep as well as in glucose and exhibit the potential to be used as new sources of prebiotics.

Regulatory effects of Cannabis Fructus oil on gut microecology in D-galactose- induced aging mice / 中草药

Objective: To investigate the effects of Cannabis Fructus oil on the regulation of gut microecology in D-galactose-induced aging mice. Methods: The Kunming mice were divided into control group, model group, positive control group (Jin Shuangqi group), Cannabis Fructus oil (12.0, 6.0, 3.0 mL/kg) group with six males and six females in each group. In the control group, normal saline was injected subcutaneously into the back of the neck every day. The other five groups were injected subcutaneously with 1% D-galactose aqueous solution, and the volume was 10 mL/kg. After 1 h, the control group was given normal saline by intragastric administration. The other groups were intragastrically administered with different doses of Cannabis Fructus oil for 42 d, and the dosage volume was 20 mL/kg. After the end of the administration, the change in body weight was analyzed; The proximal intestinal tissue of the ileocecal area and the feces in the cecum and colon were retained. Gram staining was used for the detection of Gram-positive bacilli (G+b), Gram-negative bacilli (G-b), Gram-positive cocci (G+c) and Gram-negative cocci (G-c); The ileal mucosa changes were observed by HE staining; The pH value of the colon feces was determined by pH meter; And the content of short-chain fatty acids (SCFA) in feces was determined by gas chromatography. Results: The results showed that Cannabis Fructus oil increased the ratio of bacillus, reduced the ratio of cocci and the cecal coefficient, decreased the pH value of the colon, significantly improved the colon pathological changes of the model animals with unbroken membrane skin, regular glands and rich cup cells and fluff rich, increased the content of SCFAs in the intestine of mice, and significantly increased (P < 0.01) the content of acetic acid, propionic acid, butyric acid, isobutyric acid, and significantly reduced (P < 0.01) the content of isovaleric acid. Conclusion: It could conclude that Cannabis Fructus oil can up-regulate the ratio of D-galactose-induced mice intestinal bacteria structure, improve the intestinal microecology, so as to provide theoretical support for clinical application and product development of Cannabis Fructus oil.

Effect of Wumeiwan on Intestinal Microflora, Inflammatory Factor and Short Chain Fatty Acids in Type 2 Diabetic Rats / 中国实验方剂学杂志

Objective::To study the effects of Wumeiwan on blood glucose, intestinal microflora, tumor necrosis factor-α (TNF-α), interleukin-10 (IL-10) and dietary fiber fermented by intestinal microflora in type 2 diabetic (T2DM) rats. Method::Totally 80 SD clean rats were selected as experimental subjects, and 10 of them were randomly selected as the normal group. The remaining 70 rats were given high-sugar and high-fat emulsion for 8 weeks and intraperitoneally injected with streptozotocin (STZ, 35 mg·kg-1) to establish the rat model of T2DM. The fasting blood glucose higher than 11.10 mmol·L-1 was considered as a successful model. The rats that were not successfully modeled were removed, and the remaining rats that were successfully modeled were randomly divided into model group, metformin group, high-dose Wumeiwan group, medium-dose Wumeiwan group and low-dose Wumeiwan group, with 10 rats in each group. Normal group and model group received (ig) normal saline (20 mL·kg-1·d-1), while metformin group (ig metformin 200 mg·kg-1·d-1), Wumeiwan high, medium and low dose groups (ig Wumeiwan 20, 10, and 5 g·kg-1·d-1) received corresponding drugs respectively. Blood glucose and body weight of rats were monitored regularly before and after administration of the drugs. Blood and feces were collected after four weeks of administration.16S-rDNA high-throughput sequencing technology was used for gene sequencing of intestinal flora. Enzyme linked immunoassay (ELISA) was used to detect the serum levels of inflammatory factors TNF-α and IL-10 in rats, and the contents of acetic acid, propionic acid and butyric acid in feces were detected by gas chromatography. Result::As compared with the normal group, the body weight decreased significantly (P<0.01). Bacteroidetes, Actinobacteria, Bacteroides, Clostridium increased, Firmicutes, Deltaproteobacteria, and Lactobacillus decreased, fasting blood glucose and serum TNF-α levels increased significantly (P<0.01), IL-10 level decreased (P<0.01), and the contents of acetic acid, propionic acid and n-butyric acid of short-chain fatty acids decreased(P<0.05, P<0.01) in model group. As compared with the model group, the body weight decreased; Bacteroidetes, Actinobacteria, Bacteroides and Clostridium decreased. Firmicutes, DeltaProteobacteria and Lactobacillus increased; fasting blood glucose and serum TNF-α decreased (P<0.01), and IL-10 increased (P<0.01), contents of acetic acid, propionic acid and n-butyric acid increased in Wumeiwan high-dose, medium-dose and low-dose groups and metformin group (P<0.05, P<0.01). Conclusion::Wumeiwan may prevent and treat T2DM by regulating intestinal flora, improving inflammatory response, increasing SCFAs content and reducing blood glucose.

Advances in Pharmacological Research of Traditional Chinese Medicine Based on "Butyric Acid Producing Bacteria-Polysaccharides of Traditional Chinese Medicine" / 中国实验方剂学杂志

The latest research shows that the intestinal flora is closely related to the pharmacology of traditional Chinese medicine（TCM）, and the study of microecological mechanism is a new direction of pharmacology of TCM. Among many functional groups of intestinal flora, butyric acid producing bacteria is an important functional group of intestinal flora. It can ferment dietary fiber, carbohydrate, endogenous protein and so on to produce metabolites. The imbalance of its flora is also related to the occurrence of a variety of diseases. The reason is that butyric acid is an important secondary metabolite of butyrobacteria. As an important short chain fatty acid, butyric acid can maintain intestinal health, regulate immune system and inflammatory response, regulate energy metabolism, and affect cell fraction Chemokines and apoptosis play an important role. Polysaccharide of TCM has the characteristics of high content, not easy to be digested and absorbed by the host, but can be decomposed and utilized by intestinal flora. It can be used as the carbon source of bacteria to regulate the intestinal flora, including butyric acid producing bacteria, and improve the structure of intestinal flora to achieve the purpose of disease treatment. Therefore, based on "butyric acid producing bacteria-polysaccharide of TCM", it is a new field in the study of microecological mechanism of TCM to study the pharmacology of TCM from the function of intestinal bacteria and the polysaccharide component of flora carbon source. Based on the latest literature and microecological pharmacology of TCM, this paper reviews the role of butyric acid, the molecular mechanism of butyric acid producing bacteria using polysaccharide of TCM, the relationship between polysaccharide of TCM and butyric acid producing bacteria, and discusses the relationship between butyric acid producing bacteria and polysaccharide of TCM. It also looks forward to the research of TCM pharmacology based on "butyric acid producing bacteria-polysaccharide of TCM", in order to provide a reference for the research progress of TCM pharmacology based on "butyric acid producing bacteria-polysaccharide of TCM".

Effects of Guizhi Fuling Capsules and Its Main Components on Intestinal Flora of Primary Dysmenorrhea Model Rats / 中国药房

OBJECTIVE：To investigate the effects of Guizhi ful ing capsules and its principal components （paeoniflorin， paeonol and amygdalin ）on the intestinal flora of primary dysmenorrhea model rats. METHODS ：Female SD rats were randomly divided into normal group ，model group ，capsule group（Guizhi fuling capsule ，1 000 mg/kg），paeoniflorin group （15.0 mg/kg）， paeonol group （10.3 mg/kg）and amygdalin group （12.1 mg/kg），with 6 rats in each group. Except for normal group ，other groups were given estradiol benzoate subcutaneously on the back of rats and oxytocin intraperitoneally to induce primary dysmenorrhea model. From the 4th day after subcutaneous injection of estradiol benzoate ，normal group was given constant volume of normal saline intragastrically ；model group was given constant volume of 0.5％CMC-Na solution intragastrically ；administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 7 days. The writhing times and the contents of and MDA in uterus tissue of rats were determined ，and then com the contents of short-chain fatty acids （SCFAs）such as acetic acid，propionic acid ，butyric acid in colonic contents were detected by GC method. Using diver sity index as index ， Rep-PCR and Eric-PCR were used to evaluate the d iversity of intestinal flora in feces of rats. RESULTS ：Compared with normal group，the writhing times of rats were increased significantly in model group ；the contents of NO and MDA in uterus were increased significantly ，while the contents of acetic acid ，propionic acid and butyric acid in colonic contents and total content of SCFAs were decreased significantly （P＜0.05 or P＜0.01）；the number of DNA electrophoresis bands of intestinal flora was significantly reduced ，the brightness of most bands was significantly reduced ，and the diversity indexes （by Rep-PCR and Eric-PCR method ，hereinafter）1 h after administration were significantly reduced （P＜0.05 or P＜0.01）. Compared with model group，writhing times of rats were decreased significantly in capsule group ，paeoniflorin group and paeonol group ；the contents of NO in uterus of rats in capsule group and paeoniflorin group as well as the contents of MDA in capsule group ，paeoniflorin group and paeonol group were decreased significantly （P＜0.05 or P＜0.01）；the propionic acid content and total content of SCFAs in colon of rats in capsule group ，the contents of acetic acid ，propionic acid and butyric acid ，total content of SCFAs in paeoniflorin group as well as the contents of propionic acid and butyric acid ，total content of SCFAs in paeonol group were increased significantly；the content of isovaleric acid was decreased significantly in paeoniflorin group （P＜0.05 or P＜0.01）；DNA electrophoresis bands and its brightness of intestinal flora changed to different extents in administration groups ，and the diversity indexes of intestinal flora 1 h after administration were increased significantly in capsule group and paeoniflorin group ，while those indexes were decreased significantly in paeonol group and amygdalin group （P＜0.05 or P＜0.01）. CONCLUSIONS ：Guizhi fuling capsules can significantly reduce writhing times and the contents of NO and MDA in uterus of primary dysmenorrhea model rats. At the same time ，the capsules also can regulate SCFAs content in colonic contents and intestinal flora diversity of rats. The above effects may be related to paeoniflorin and paeonol in the capsules.

A phase 2 randomized controlled trial of oral resistant starch supplements in the prevention of acute radiation proctitis in patients treated for cervical cancer

Background: Radiation induced proctitis is frequently encountered during the radiation therapy of cervical and prostate cancers that causes pain and occasionally with bleeding and may affect the continuity of radiation therapy. Aims and Objectives: The purpose of the study is to look at the benefit of administration of an oral prebiotic amylase resistant starch in reducing the incidence of acute radiation proctitis, a distressing symptom in patients receiving radiation therapy for cancer of the cervix. Material and Methods: The study was conducted between 2011 and 2014 in 104 patients receiving radical chemo-radiotherapy for carcinoma cervix. Patients were randomized in to two arms, one receiving 30 gm of resistant starch and the other digestible starch on a daily basis throughout the course of the external radiotherapy. All patients received standard 4-field box radiation portals, 50 Gy in 25 fractions with 4 cycles of weekly concurrent Cisplatin. At completion of external beam radiotherapy, all patients underwent LDR/HDR brachytherapy. The study was double blinded and allocation was concealed from the investigators. The investigator recorded the radiotherapy related toxicity of the patients according to CTC V 3.0. The incidence and severity of grade 2-4 diarrhoea and proctitis were documented on a weekly basis and compared across the two groups and analyzed. Stool short chain fatty acid concentrations were measured at baseline at 2nd and 4th week and after 6 weeks of completion of radiotherapy in both study placebo arms and reported. The pattern of microbiota in the stool were also estimated in all patients at 4 time points. Two patients who progressed during therapy were not included in the analyses and two patients discontinued the intervention. A per protocol analyses was done. Results: At analysis there were 50 patients in each arm. The severity of clinical proctitis was found to be similar in both groups of patients with 12.2 % of patients experiencing toxicity of grade 2 and above in digestible starch group versus 14.6% in the resistant starch group. Functional proctitis was similarly graded and it was found that 16.3 % patients in digestible starch group experienced toxicity against 10.2 % patients in the resistant starch group. This difference was seen at 4th week and continued in the subsequent weeks till the end of radiation. Both groups had similar reported toxicity at 6 weeks post intervention and similar incidence of grade 2 and above diarrhea. The resistant starch group was found to have 8% incidence as compared to 2% in the other group at the 5th and 6th week. The short chain fatty acid concentrations were not significantly different in the groups at any point. Conclusion: The study did not demonstrate a significant benefit in administering resistant starch over and above normal diet to patients receiving pelvic radiotherapy. The reasons may be attributed to concurrent use of chemotherapy and decrease in intestinal probiotics. The use of digestible starch in the control arm may have contributed to lower incidence of the toxicity endpoints as well

Regulatory Effect of Sijunzi Tang and Its Single Herbs on Intestinal Flora in Rats with Spleen Deficiency / 中国实验方剂学杂志

Objective:To investigate the regulatory effect of Sijunzi Tang(SJZT) and its single herbs(Ginseng Radix et Rhizoma, Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, Atractylodis Macrocephalae Rhizoma and Poria) on intestinal flora in spleen-deficient rats. Method:Normal rats were randomly divided into the blank group, model group, Zhengchangsheng granules group, SJZT group and each single herb group, rats were orally administered Sennae Folium decoction to induce diarrhea for ten consecutive days to establish a spleen-deficient model(distilled water for the blank group), then treated with the corresponding drugs for seven consecutive days(distilled water for the blank group and the model group). Fresh feces were collected on pre-modeling(0th day), post-modeling(11th day), and post-treatment(18th day). Short-chain fatty acids(SCFAs) in feces were acidified by sulphuric acid and extracted by diethyl ether, then determined by gas chromatography. The structural change(diversity and similarity) of intestinal flora in feces was analyzed by 16S rDNA-polymerase chain reaction(PCR)-denaturing gel gradient electrophoresis(DGGE) technique. Result:Compared with blank group, the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of all administered groups were significantly decreased on the 11th day(PPth day, compared with model group, the contents of SCFAs as well as diversity and similarity indexes of intestinal flora in feces of Atractylodis Macrocephalae Rhizoma group were significantly increased(PPPPConclusion:Ginseng Radix et Rhizoma, Atractylodis Macrocephalae Rhizoma and Poria are the single herbs responsible for the regulatory effect of SJZT on intestinal flora in spleen-deficient rats, and Atractylodis Macrocephalae Rhizoma may play a major role.

Research Outlook of Short-chain Fatty Acids Detected by Ultra Performance Convergence Chromatography / 中国实验方剂学杂志

Gas chromatography (GC) is mainly used to detect the levels of short-chain fatty acids (SCFAs), but with the deepening of research,the drawbacks of GC have become more and more obvious in the fields of food,chemical engineering and clinical application. The analysis on existing research results showed that ultra performance convergence chromatography (UPC2) was appropriate for the analysis of lipid metabolism. The UPC2 is a new kind of chromatographic separation technology developed in recent five years and the level of SCFAs is associated with the research on multiple diseases. Therefore,application of UPC2 in the detection of SCFAs would be helpful for the scholars at home and abroad to carry out deeper researches,and also helpful to guide the treatment for various metabolic disorders. In this paper,the researches on SCFAs in recent ten years were reviewed; the shortcomings of GC and liquid chromatography (LC) in the detection of SCFAs were reviewed; the development process,basic characteristics and research status of UPC2 at home and abroad were introduced; feasibility and innovation of UPC2 in the detection of SCFAs were summarized. Pretreatment methods for UPC2 application to the detection of SCFAs in feces or serum were collected; the problems that should be noticed during the process of sample pretreatment were pointed out; meanwhile, an research outlook on methodology of UPC2 application in the detection of SCFAs was conducted. The effects of extracting solvent,mobile phase,and auxiliaryt solvent on chromatographic behavior as well as the physicochemical property, type and choice of UPC2 chromatographic column were mainly discussed in this paper. In addition, the choices of basic modifier,acid modifier,and salinity modifier were briefly outlined, in order to provide efficient,simple,environmental,and economic detection technologies for the research on SCFAs, and provide better reference solutions for the rapid detection of massive clinical samples.