Transcriptome and Gut Microbiota Profiling Analysis of ANIT-Induced Cholestasis and the Effects of Da-Huang-Xiao-Shi Decoction Intervention.

Cholestasis is characterized by bile acid (BA) circulation disorders, which is usually related to damage of hepatocyte barrier function. Currently, patients with cholestasis face several obstacles in seeking diagnosis and therapy. Da-Huang-Xiao-Shi decoction (DHXSD) is an ancient classic formula that has been used clinically for cholestasis treatment. Nevertheless, the underlying biological activities and therapeutic mechanisms remain unclear. In this study, an alpha-naphthylisothiocyanate (ANIT)-induced cholestasis rat model was established to examine the anticholestatic effects of DHXSD using histopathological and molecular analyses. Transcriptomic analysis combined with 16S rRNA gene sequencing analysis was systematically applied to study the mechanism of action of DHXSD. Simultaneously, the effect of DHXSD on gut microbiota, short-chain fatty acids (SCFAs), and intestinal barrier function were evaluated based on the ANIT-induced cholestasis model in rats. The results showed that DHXSD effectively attenuated ANIT-induced cholestasis by reducing liver function indicators (alanine transaminase [ALT], P < 0.05; alkaline phosphatase [ALP], P < 0.05; total bile acid [TBA], P < 0.01; γ-glutamyl transpeptidase [GGT], P < 0.001) and levels of hepatotoxicity-related enzymes (P < 0.05), thus improving the recovery of histopathological injuries, and regulating levels of inflammatory cytokines (P < 0.05). In addition, 16S rRNA gene sequencing analysis combined with intestinal barrier function analysis revealed that the DHXSD significantly ameliorated ANIT-induced gut microbiota dysbiosis. Significantly altered genes in the model and treatment groups were screened using transcriptomic analysis. Sixty-eight genes and four microbial genera were simultaneously altered with opposing trends in variation after ANIT and DHXSD treatments. We built a framework for predicting targets and host-microbe interaction mechanisms, as well as identifying alternative treatment for cholestasis, which should be validated further for clinical application. In conclusion, DHXSD appears to be a promising agent for protection against liver injury. IMPORTANCE Cholestasis is a serious manifestation of liver diseases resulting in liver injury, fibrosis, and liver failure with limited therapies. To date, only ursodeoxycholic acid (UDCA) has been approved by the U.S. Food and Drug Administration for the treatment of cholestasis. However, approximately one-third of patients with cholestasis are unresponsive to UDCA. Therefore, it is urgent to search for appropriate therapeutic agents for restoring stoppage status of the bile components to treat cholestasis. In this study, we investigated how the microbiome and transcriptome data sets correlated with each other to clarify the role of microbiome alterations in host metabolism. In combination, this research offers potential molecular biomarkers that should be validated for more accurate diagnosis of cholestasis and the clinical utilisation of gut microbiota as a target for treatment.

The application of Tong-fu therapeutic method on ulcerative colitis: A systematic review and meta-analysis for efficacy and safety of rhubarb-based therapy.

Background: Tong-fu therapeutic method (TFTM) is a traditional Chinese medicine treatment method for ulcerative colitis, which is a novel treatment strategies and have purgative effect. As the most representative medicinal of TFTM, Rhubarb has been reported to have a therapeutic impact on ulcerative colitis by regulating intestinal flora, anti-inflammation, and improving intestinal microcirculation. Although rhubarb has been widely used in Chinese medicine for the treatment of ulcerative colitis, the appropriate protocol is still demanded to its rational use in clinic, which promoted to evaluate the efficacy and safety for rhubarb-based therapy on ulcerative colitis. Method: Clinical trials were searched through PubMed, Cochrane Library, Web of Science, Excerpta Medica Database, Chinese National Knowledge Infrastructure, WAN FANG Database, Chinese Scientific Journal Database, and Chinese Biomedical Literature Database. The subgroup analyses were performed with three groups: medication, course of treatment, and route of administration. The statistical analyses were performed on Review Manager software (version 5.4.1). Results: A total of 2, 475 patients in 30 original studies were analyzed in this article. It was found that rhubarb-based therapy could increase clinical efficacy and reduce the recurrence rate. Subgroup analyses showed that rhubarb-based therapy was more effective than 5-aminosalicylic acid or sulfasalazine alone. In addition, the hypercoagulable state of ulcerative colitis could be ameliorated by decreasing platelet (PLT) and fibrinogen (FIB), and increasing prothrombin time (PT) significantly. Moreover, C-reaction protein (CRP), tumor necrosis factor-α (TNF-α), interleukin (IL)-6, IL-8, and IL-1ß expression were significantly reduced, while IL-10 production was increased, which mediated the alleviation of intestinal inflammation stress. Conclusion: Rhubarb-based therapy could effectively improve ulcerative colitis. Of note, the rhubarb-based medicinal formulas combined with 5-ASA or SASP are more effective than the 5-ASA or SASP alone. In addition, although rhubarb has side effect, the results of our analysis showed that rhubarb-based therapy did not exhibit significant side effects. This means it has a high safety profile in clinical use. Moreover, the use of rhubarb-based therapy is recommend to use within 1-13 weeks or 3 months via administered orally or by enema, which is contributes to ensure the curative effect and avoid its toxic and side effects. As an important case of TFTM, rhubarb-based therapy provides evidence for the practical application of TFTM.

Determination of multiple active constituents in Da-Huang-Xiao-Shi decoction using HPLC-LTQ-Orbitrap mass spectrometry: Application in comparing the differences in the formula and its constituent herbs.

Da-Huang-Xiao-Shi decoction (DHXSD) is a traditional Chinese medicine formula and is used to treat cholestasis. In this study, we developed a reliable and comprehensive HPLC coupled with a linear ion trap-Orbitrap mass spectrometry method for the separation and determination of 21 components including six alkaloids, five anthraquinones, three tannins, three terpenes, two iridoid glycosides, one organic acid and one flavonoid in DHXSD. A C18 column was eluted using a gradient mobile phase at a flow rate of 1 ml/min. Detection was operated with an electrospray ionization source in positive and negative ion modes using selective ion monitoring. The calibration curves for all analytes showed good linearity (r > 0.9901), and the inter- and intra-day precision did not exceed 4.98%. The recovery, repeatability and stability were also within the acceptable limits. The method was successfully applied to determine multiple active constituents in DHXSD and its constituent herbs. Compared with Da Huang, the total contents of the five anthraquinones were significantly higher in DHXSD. However, the changes in the components from Zhi Zi/Huang Bo were complicated in DHXSD. The study could serve as a fundamental reference for establishing comprehensive DHXSD quality control measures and be helpful to understand some compatibility laws of DHXSD.

Da-Huang-Xiao-Shi decoction protects against3, 5-diethoxycarbonyl-1,4-dihydroxychollidine-induced chronic cholestasis by upregulating bile acid metabolic enzymes and efflux transporters.

ETHNOPHARMACOLOGICAL RELEVANCE: Chronic cholestasis is a usual clinical pathological process in hepatopathy and has few treatment options; it is classified under the category of jaundice in Chinese medicine. Da-Huang-Xiao-Shi decoction (DHXSD) is a classic Chinese prescription which is used to treat jaundice. AIM OF THE STUDY: We aimed to examine the protective effect of DHXSD on liver and its potential mechanism of action against chronic cholestasis. MATERIALS AND METHODS: Chronic cholestasis was induced using 3, 5-diethoxycarbonyl-1,4-dihydroxychollidine (DDC) in mice. Mice were then administered DHXSD intragastrically at doses of 3.68, 7.35, and 14.70 g/kg for four weeks followed by further analyses. Serum biochemical indices and liver pathology were explored. Eighteen individual bile acids (BAs) in mice serum and liver were quantified using ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). The expression of BA related metabolic enzymes, transporters, along with nuclear receptor farnesoid X receptor (FXR) was detected by real-time qPCR and Western blot. RESULTS: DHXSD treatment reduced the serum biochemical indices, ameliorated pathological injury, and improved the disordered BA homeostasis. Mice treated with DHXSD showed significantly upregulated expression of the metabolic enzymes, cytochrome P450 2b10 (Cyp2b10), Cyp3a11, and UDP-glucuronosyltransferase 1a1 (Ugt1a1); and the bile acid transporters, multidrug resistance protein 2 (Mdr2), bile salt export pump (Bsep), and multidrug resistance-associated protein 3 (Mrp3). DHXSD treatment also significantly upregulated FXR expression in mice with DDC-induced chronic cholestasis. CONCLUSIONS: DHXSD exerted protective effects on chronic cholestasis in DDC-treated mice by alleviating the disordered homeostasis of BAs through increased expression of BA related metabolic enzymes and efflux transporters.

Study protocol for daiobotanpito combined with antibiotic therapy for treatment of acute diverticulitis: a study protocol for a randomized controlled trial.

BACKGROUND: Colonic diverticular disease has been increasing in prevalence due to the rapidly aging global population, but standard treatment has not changed dramatically in recent years. Daiobotanpito (DBT; Da Huang Mu Dan Tang in Chinese) has been used in medical treatment of acute abdominal abscesses, such as appendicitis or diverticulitis in traditional Japanese (Kampo) medicine for many years, based on more than 3000 years of experience. Prior to this study, a retrospective open-label trial was conducted to compare patients with acute diverticulitis who received oral DBT combined with intravenous antibiotics with those who received intravenous antibiotic alone; it showed a positive effect of DBT on acute diverticulitis. We aim to investigate whether moderate to severe acute diverticulitis shows greater improvement with intravenous antibiotics plus orally administered DBT compared with intravenous antibiotics plus placebo. METHODS: This is a two-group, randomized, double-blind, placebo-controlled, multi-center trial, which is designed to evaluate the efficacy and safety of DBT in patients with moderate to severe diverticulitis treated with intravenous antibiotics. Eligible participants will be randomized to either a treatment group receiving a 10-day oral DBT regimen plus conventional therapy or a control group receiving a 10-day placebo regimen plus conventional therapy. The primary outcome will be success in treating diverticulitis: the success rate will be defined as elimination of abdominal pain within 4 days in all patients, and in patients with fever (body temperature ≧ 37.5 °C) on inclusion into this study, fever relief with reduction in body temperature to < 37.5 °C within 3 days. Secondary endpoints will include the number of hospitalization days, changes in inflammatory response (C-reactive protein (CRP), white blood cell (WBC) and neutrophil counts), fever type, number of days before beginning food intake, recurrence rate (observation for 1 year after registration), and adverse event expression rate. Assessments will be performed at baseline and on the day of discharge. The recurrence rate will be recorded at 1 year after registration. DISCUSSION: This study is expected to provide evidence to support the clinical benefits of DBT in the treatment of acute diverticulitis. It may also provide evidence on the efficacy and safety of DBT in the recurrence of acute diverticulitis. TRIAL REGISTRATION: UMIN-CTR: UMIN000027381. Registered on 27 April 2017. https://upload.umin.ac.jp/cgi-bin/ctr/ctr_view_reg.cgi?recptno=R000031377, and changed to jRCTs041180063, registered on 30 July 2019; as a result of the revision of the domestic law in 2018 in Japan.

Rheum officinale extract promotes the innate immunity of orange-spotted grouper (Epinephelus coioides) and exerts strong bactericidal activity against six aquatic pathogens.

We investigated the antimicrobial properties and the effects of Rheum officinale extract (ROE) on nonspecific immune parameters of orange-spotted grouper (Epinephelus coioides) in vitro and in vivo. The in vitro analysis was conducted by treating grouper primary head kidney leukocytes with various concentrations of ROE. The phagocytic rate of the leukocytes was elevated in a dose-dependent manner from 0.01 to 0.1 mg/ml, but decreased with higher concentrations of ROE (0.5 and 1.0 mg/ml). The production of reactive oxygen species (ROS) was strongly enhanced in a dose-dependent manner by treatment with ROE doses of 0.1-10.0 mg/ml. However, morphological changes (e.g., rounding and shrinkage of cells, chromatin condensation, fragmentation, and appearance of apoptotic bodies) were observed in the leukocytes after incubation with higher concentrations of ROE (1.0 and 10.0 mg/ml). A 28-day feeding trial was performed to assess the impact of dietary administration of ROE on grouper innate immunity parameters. Fish were fed with feed supplemented with 0, 0.1, 1.0, or 5.0 g ROE per kg of feed. The phagocytic activity of the animals' leukocytes was significantly elevated in all ROE-fed groups on day 1 and in groups fed with ROE at 0.1 or 1.0 g/kg on day 14. Production of ROS was substantially increased on day 1 in fish fed with ROE at 1.0 and 5.0 g/kg, but decreased steadily later on. The ability to generate ROS increased steadily until day 7 in fish fed the lowest concentration of ROE (0.1 mg/ml), but decreased thereafter. ROE showed excellent antibacterial activity against six pathogens of aquatic animals: Vibrio parahaemolyticus, V. vulnificus, V. alginolyticus, V. carchariae, Aeromonas hydrophila, and Edwardsiella tarda. The minimum inhibitory and bactericidal concentrations of measured ROE-derived anthraquinones were 10.57-84.53 µg/ml and 10.57-169.05 µg/ml, respectively.

Simultaneous determination of multiple compounds of Da-Huang-Xiao-Shi decoction in rat plasma by LC-MS/MS and its application in a pharmacokinetic study.

Da-Huang-Xiao-Shi decoction (DHXSD), a traditional Chinese medicinal formula, has been used mainly to treat jaundice for more than 1700 years in China. In this study, we developed a rapid, sensitive, and accurate LC-MS/MS method to simultaneously determine multiple, potentially bioactive compounds of DHXSD, including five alkaloids (berberine, phellodendrine, palmatine, jatrorrhizine, and magnoflorine), five anthraquinones (rhein, aloe-emodin, emodin, chrysophanol, and physcion), two iridoid glycosides (geniposide and genipin 1-gentiobioside), and one iridoid aglycone (genipin) in rat plasma. Plasma samples collected from rats were treated immediately with 5% acetic acid to avoid the degradation of genipin. After protein precipitation with acetonitrile containing 5% acetic acid, the compounds were reconstituted in acetonitrile-water (50:50, v/v) solution containing 6.5% formic acid and separated on the ACQUITY™ UPLC BEH C18 column (2.1 × 100 mm; 1.7 µm) using a mobile phase composed of 2 mM ammonium formate in water (solvent A) and acetonitrile (solvent B) at a flow rate of 0.3 mL/min. Quantitation was performed on a Triple Quand 5500 tandem mass spectrometer coupled with an electrospray ionization (ESI) source. Multiple reaction monitoring (MRM) was used to quantify compounds in positive and negative ion modes. The method validation results showed that the specificity, linearity, precision and accuracy, recovery, matrix effect, and stability of the 13 compounds met the requirements for their quantitation in biological samples. This newly established method was successfully used in a pharmacokinetic study on rats orally treated with DHXSD. Besides, glucuronide and sulfate metabolites were also determined in rat plasma after hydrolysis. This is the first method developed for the simultaneous quantification of multiple compounds of DHXSD in vivo. Our study provides relevant information on the pharmacokinetics of DHXSD and the relationship between the compounds of DHXSD and their therapeutic effects.

UPLC-MS-based metabonomic analysis of intervention effects of Da-Huang-Xiao-Shi decoction on ANIT-induced cholestasis.

ETHNOPHARMACOLOGICAL RELEVANCE: Cholestasis, caused by hepatic accumulation of bile acids, is a serious manifestation of liver diseases resulting in liver injury, fibrosis, and liver failure with limited therapies. Da-Huang-Xiao-Shi decoction (DHXSD) is a representative formula for treating jaundice and displays bright prospects in liver protective effect. AIM OF THE STUDY: This study was designed to assess the effects and possible mechanisms of DHXSD against alpha-naphthylisothiocyanate-induced liver injury based on ultra-high performance liquid chromatography-hybrid quadrupole-Orbitrap mass spectrometry (UHPLC-Q-Orbitrap MS) metabonomic approach. MATERIALS AND METHODS: The effects of DHXSD on serum indices (TBIL, DBIL, AST, ALT, ALP, TBA, and γ-GT) and the histopathology of the liver were analyzed. Moreover, UHPLC-Q-Orbitrap MS was performed to identify the possible effect of DHXSD on metabolites. The pathway analysis was conducted to illustrate the pathways and network by which DHXSD treats cholestasis. RESULTS: The results demonstrated that DHXSD could significantly regulate serum biochemical indices and alleviate histological damage to the liver. Twelve endogenous components, such as glycocholic acid, taurocholic acid and indoleacetaldehyde, were identified as potential biomarkers of the therapeutic effect of DHXSD. A systematic network analysis of their corresponding pathways indicates that the anti-cholestatic effect of DHXSD on alpha-naphthylisothiocyanate-induced cholestasis rats occurs mainly through regulating primary bile acid biosynthesis, arginine and proline metabolism, and arachidonic acid metabolism. CONCLUSIONS: DHXSD has exhibited favorable pharmacological effect on serum biochemical indices and pathological observation on cholestatic model by partially regulating the perturbed pathways. Moreover, these findings may help better understand the mechanisms of disease and provide a potential therapy for cholestasis.

Metabolic profile elucidation of Zhi-Zi-Da-Huang decoction in rat intestinal bacteria using high-resolution mass spectrometry combined with multiple analytical perspectives.

1. Zhi-Zi-Da-Huang decoction (ZZDHD) has been widely used for the treatment of alcoholic jaundice, alcoholic liver disease, and acute hepatitis in China for thousands of years. Conventionally decoctions are administered orally, after which the metabolism caused by the enzymes in intestinal bacteria may influence significantly on the curative effects or toxicity. 2. In this work, the comprehensive metabolic process of ZZDHD in intestinal bacteria was investigated reliably using high-resolution HPLC-DAD-ESI-TOF/MS. Besides, a novel strategy for major-to-trace metabolites identification which integrated information derived from diagnostic fragment ions, mass spectral similarity filter strategy, dynamic metabolic change of target compounds and relevant behavior in LC-MS was adopted. 3. As a result, 45 compounds, including 26 bio-converted prototypes and 19 newly generated metabolites were detected and tentatively identified. The metabolic profile of ZZDHD in gastro-intestinal was subsequently elucidated. Deglycosylation, oxidation, reduction, acetylation, and ring cleavage were all observed in the biotransformation of the decoction. Among the rest, deglycosylation was found to be the predominant metabolic pathway. 4. The results obtained herein provided a practical strategy for metabolic profile elucidation of traditional herbal medicines. Moreover, it would be helpful to unravel how the oral decoctions play the therapeutic role in vivo.

Deciphering the absorption profile and interaction of multi-components of Zhi-Zi-Da-Huang decoction based on in vitro-in silico-in vivo integrated strategy.

Zhi-Zi-Da-Huang decoction (ZZDHD) has been acknowledged with striking therapeutic effects for hepatobiliary disorders in the history of China. As decoctions are usually administrated orally, intestinal absorption, the prerequisite task of exerting therapeutic effects, is of utmost significance for screening potential active compounds and understanding the mechanism of drug action. In this work, an in vitro-in silico-in vivo strategy based on HPLC-DAD-ESI-TOF/MS was adopted for precisely profiling the intestinal absorption of ZZDHD, which integrated information obtained from rat everted gut sac model, octanol-water partition model, in silico prediction and in vivo experimental data. Besides, 34 main absorbed ingredients were selected as chemical markers to investigate the compatible interaction of the decoction on absorption level using rat everted gut sac experiment. In total, 106 compounds of ZZDHD were speculated as potential absorptive. Among them, 90 constituents predicted absorbable in at least two experimental models were finally recognized as intestinal absorbable ingredients. In addition, the absorption level of iridoids, terpenoids and flavonoid glycosides were found improved and the absorption of catechins and anthraquinones were inhibited after prescription compatibility. Taken together, this study presents a reliable strategy for evaluating intestinal absorption of herbal medicines and offers a reference for the rationality of herbal compatibility and the modernization of traditional Chinese medicine (TCM).

The activity of Hou-Po-Da-Huang-Tang is improved through intestinal bacterial metabolism and Hou-Po-Da-Huang-Tang selectively stimulate the growth of intestinal bacteria associated with health.

Hou-Po-Da-Huang Tang (HPDHT) was used for the treatment of intestinal tract diseases in China. However, the underlying mechanisms via the intestinal bacteria remain largely unclear. Therefore, the aim of this study was to evaluate the metabolism of HPDHT by the human intestinal bacteria and its modulating effect on the intestinal bacteria. As a result, a total of 34 compounds were identified in HPDHT and transformed HPDHT (T-HPDHT). Among them, 12 metabolites were proved to be transformed by human intestinal bacteria. In vitro assays showed that T-HPDHT exhibited more significant elevation of free radical scavenging activity and suppression on the production of nitric oxide (NO) and TNF-α when comparing to HPDHT. Additionally, in vivo experiment confirmed that HPDHT significantly increased activity of superoxide dismutase (SOD), attenuated the malondialdehyde (MDA) and TNF-α levels in the conventional rats compared with that of pseudo germ-free (PGF) rats. In addition, HPDHT could significantly enhance the mean counts of Bifidobacterium and Lactobacillus and inhibit the growth of Clostridium, and Enterobacteriaceae, relative to controls. Due to the transformation of HPDHT being dependent on the bacterial strain, the effect of HPDHT on the selective growth of Bifidobacterium bifidum 29521 and Lactobacillus plantarum 8014 was evaluated. The kinetic parameters of microbial growth and prebiotic activity scores indicated that HPDHT could selectively stimulate the growth of the strains Bifidobacterium bifidum 29521 and Lactobacillus plantarum 8014. Taken together, metabolism of HPDHT by intestinal bacteria is a critical step towards the emergence of their anti-oxidation, anti-inflammation and prebiotic activities. This study provided valuable information for further pharmacological research on HPDHT.

Treatment effect of Da Huang Mu Dan Tang in rats with pancreatic carcinoma and protective effects on liver and kindey function / 吉林大学学报(医学版)

Objective: To establish the pancreatic carcinoma models of rats with dimethybenzanthracene (DMBA)embedding method,to discuss the treatment effect of Da Huang Mu Dan Tang (DHMDT)in the rats with pancreatic carcinoma and the protective effects on the liver and kidney function,and to clarify the mechanisms. Methods:Sixty-five SD male rats were divided into blank control (n=10)and modeling group (n=55).DMBA was embedded in the capsule of pancreas to establish the pancreatic carcinoma models of rats. After 3 months, 50 successfully modeling rats were randomly divided into model,5-FU (intravennous injection,12 mg· kg-1 ), low,middle and high doses of DHMDT (intragastric administration,20,40 and 80 mg· kg-1 )groups.After 10 weeks of administration,the levels of alanine aminotransferase (ALT),aspartate transaminase (AST),urea nitrogen (BUN),creatinine (Cr)and total bilirubin (TBiL)in serum of the rats in various groups were measured;the weights and the inhibitory rates of the pancreatic tumor were detected;TUNEL method was used to analyze the apoptotic rates of tumor tissue;the expression levels of SST2R,Bax and Bcl-2 genes in pancreatic tumor tissue of the rats were detected by Q-PCR method.Results:Compared with model group,after treated for 10 weeks,the levels of ALT,AST,BUN,Cr and TBiL in serum of the rats in low,middle and high doses of DHMDT groups were significantly decreased (P <0.05 or P <0.01);the weights of tumor of the rats in middle and high doses of DHMDT groups were decreased (P <0.05 or P <0.01);the apoptotic rates of the cells in pancreatic tumor tissue of the rats in low,middle and high doses of DHMDT groups were increased (P <0.05 or P <0.01);the expression levels of SST2R gene in pancreatic tumor tissue of the rats in low,middle and high doses of DHMDT groups were significantly increased (P <0.05 or P <0.01);the expression levels of Bax gene in pancreatic tumor tissue of the rats in middle and high doses of DHMDT groups were significantly increased (P < 0.05 or P < 0.01),while the expression levels of Bcl-2 in pancreatic tumor tissue of the rats were significantly decreased (P < 0.05 ). Conclusion:DHMDT has the treatment effect in the rats with pancreatic carcinoma and can promote the apoptosis of tumor cells;DHMDT can protect the liver and kidney function of the rats with pancreatic carcinoma.

Therapeutic Observation of Acupoint Sticking plus Hot Compress by Moxa-salt Bag for Constipation After Acute Cerebral Stroke / 上海针灸杂志

Objective To observe the clinical efficacy of acupoint sticking plus hot compress by moxa-salt bag in treating constipation after acute cerebral stroke.Method Eighty patients with constipation after cerebral stroke were randomly allocated to a treatment group and a control group by using random number table method according to registration order, 40 cases each. The control group was intervened by acupoint sticking at umbilicus with rawDa Huang (Rheum Officinale), and the treatment group was additionally given hot compress using moxa-salt bag. The treatment was given once a day, for consecutive 3 days in total. The first defecation time, stool shape and defecation sensation were observed in the two groups. The clinical efficacies were compared between the two groups.Result The total effective rate was 90.0％ in the treatment group versus 72.5％ in the control group, and the difference was statistically significant (P<0.05). After the treatment, there was statistically significant difference in comparing the first defecation time between the two groups (P<0.05). After the treatment, the incidence rate of soft stool was 85.0％ and that of hard stool was 15.0％ in the treatment group versus 40.0％ and 60.0％ in the control group, and the differences were statistically significant (P<0.05). The incidence rate of pleasant defecation sensation was 80.0％ and that of difficult defecation was 20.0％ in the treatment group versus 35.0％ and 65.0％ in the control group after the treatment, and the differences were statistically significant (P<0.05).Conclusion Acupoint sticking plus hot compress by moxa-salt bag is an effective method in treating constipation after acute cerebral stroke.

Therapeutic Observation of Acupoint Sticking plus Hot Compress by Moxa-salt Bag for Constipation After Acute Cerebral Stroke / 上海针灸杂志

Objective To observe the clinical efficacy of acupoint sticking plus hot compress by moxa-salt bag in treating constipation after acute cerebral stroke.Method Eighty patients with constipation after cerebral stroke were randomly allocated to a treatment group and a control group by using random number table method according to registration order, 40 cases each. The control group was intervened by acupoint sticking at umbilicus with rawDa Huang (Rheum Officinale), and the treatment group was additionally given hot compress using moxa-salt bag. The treatment was given once a day, for consecutive 3 days in total. The first defecation time, stool shape and defecation sensation were observed in the two groups. The clinical efficacies were compared between the two groups.Result The total effective rate was 90.0％ in the treatment group versus 72.5％ in the control group, and the difference was statistically significant (P<0.05). After the treatment, there was statistically significant difference in comparing the first defecation time between the two groups (P<0.05). After the treatment, the incidence rate of soft stool was 85.0％ and that of hard stool was 15.0％ in the treatment group versus 40.0％ and 60.0％ in the control group, and the differences were statistically significant (P<0.05). The incidence rate of pleasant defecation sensation was 80.0％ and that of difficult defecation was 20.0％ in the treatment group versus 35.0％ and 65.0％ in the control group after the treatment, and the differences were statistically significant (P<0.05).Conclusion Acupoint sticking plus hot compress by moxa-salt bag is an effective method in treating constipation after acute cerebral stroke.

Protective effect of Zhi Zi Da Huang Tang on rats with alcoholic fatty liver / 吉林大学学报(医学版)

Objective:To establish the alcoholic fatty liver(AFL) animal models,and to explore the protective effect of Zhi Zi Da Huang Tang (ZZDHT) on the rats with AFL and its dosage.Methods:A total of 54 SD rats were divided into normal control group (n=10) and model control group (n=44).The rats in model control group were given alcohol by lavage (50% ethanol solution 6.0 mL·kg-1) combined with high-fat feed to establish the rat models of AFL.After 4 weeks,the rats in model control group were randomly divided into model group(treated with water),simvastatin group (10 mg·kg-1),low and high doses (5.0 and 10.0 g·kg-1) of ZZDHT groups,and there were 10 rats in each group.Every 2 weeks during the process,the body weights and levels of serum total cholesterol (TC),triglyceride (TG),aspertate aminotransferase (AST),and alanine aminotransferase (ALT) were detected.After 4 weeks,the body weights and liver weights of the rats were detected;the levels of TC,TG,AST,ALT,alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in serum and liver tissue of the rats of were detected;the levels of tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6) and interleukin-1β(IL-1β) in serum of the rats were detected;the morphology of liver tissue was observed by HE staining and the pathological examination was performed.Results:Compared with normal control group,the levels of serum TC,AST,ALT,TG,TNF-α,IL-6,and IL-1β of the rats in model group 4 weeks after administration were significantly increased (P<0.05 or P<0.01).Compared with model group,the levels of serum TC,TG,AST,and ALT of the rats in low and high doses of ZZDHT groups 4 weeks after administration were decreased (P<0.05 or P<0.01);the levels of serum TNF-α and IL-6,IL-1β of the rats in high dose of ZZDHT group were decreased (P<0.01);the levels of serum TNF-α and IL-1β of the rats in low dose of ZZDHT group were decreased (P<0.01);the levels of serum ADH and ALDH in liver tissue of the rats in low and high doses of ZZDHT groups and simvastatin group were decreased(P<0.05 or P<0.01).The HE staining result showed that compared with model group,the pathological conditions of the liver tissue of the AFL rats in ZZDHT groups were significantly improved.Conclusion:ZZDHT can significantly improve the liver injury caused by high fat diet combined with alcohol and fat liver lesions.

Dynamic metabolic profile of Zhi-Zi-Da-Huang decoction in rat urine based on hybrid liquid chromatography-mass spectrometry coupled with solid phase extraction.

Zhi-Zi-Da-Huang decoction (ZZDHD) has been used for treatment of alcoholic liver disease in China for thousands of years. In order to reveal the dynamic biotransformation of the decoction in vivo, a high-throughout, sensitive and special method based on high performance liquid chromatography coupled with diode array detection and time-of-flight mass spectrometry (HPLC-DAD-TOF/MS) and high performance liquid chromatography coupled with triple quadrupole mass spectrometry (HPLC-QqQ/MS) was developed and validated. 25 parent compounds and 28 metabolites were characterized, among which, two metabolites were found for the first time and tentatively identified by neutral-loss scan and product-ion scan. All the compounds were assigned to iridoids, flavones, anthraquinones, coumarin or p-coumaric acid, and their biotransformation pathways were found to involve glucuronidation, sulfation, reduction and ring cleavage. Glucuronidation occurred as a major metabolic pathway of genipin and flavanone and the conjugates could be detected almost during the whole sampling duration. To compounds such as anthraquinones, coumarin and p-coumaric acid, sulfation is the only transformation pathway and the metabolites were found at 0-12, 4-18, 4-48h respectively after administration. Reduction and/or ring cleavage of genipin glucuronide and naringin were also observed obviously. The phenomena that parts of parent compounds and metabolites were able to be detected even 48h after administration implied that the accumulating effect of these constituents in vivo would happen and the potential toxicity of the decoction might appear if multiple dosing is adopted. The strategy used in this paper was proved helpful to offer important information for the clinical safe use of ZZDHD.

Identification and screening of chemical constituents with hepatoprotective effects from three traditional Chinese medicines for treating jaundice.

The constituents with hepatoprotective activity were investigated in three traditional Chinese medicine formulae for treating jaundice, namely, Zhi-Zi-Da-Huang-Tang, Yin-Chen-Hao-Tang, and Da-Huang-Xiao-Shi-Tang. By using liquid chromatography coupled with quadrupole time-of-flight mass spectrometry and liquid chromatography coupled with ion trap mass spectrometry, 79 chemical constituents were identified unambiguously or tentatively in three formulae based on the accurate molecular weight, mass spectrometric fragmentation behavior, and comparison with reference standards. Then the hepatoprotective activities of 27 constituents were evaluated on tert-butylhydroperoxide-injured BRL-3A cells. The results indicated that 11 constituents, including protocatechic acid (19), epijasminoside A (56), rutin (71), tetrahydropalmatine (76), rhaponticin (80), 3,4-dicaffeoylquinic acid (82), 3,5-dicaffeoylquinic acid (85), diosmetin-7-O-glucoside (90), jatrorrhizine (93), berberine (100), and daidzein (101) exerted hepatoprotective activities. Interestingly, most of the crude drugs in three formulae contained hepatoprotective active constituents, and the combinations of constituents from different crude drugs exhibited greater effects. This result provided evidence to the traditional Chinese medicine theory of combining several drugs together to exert synergistic efficacy.

Chemical profiling approach to evaluate the influence of traditional and simplified decoction methods on the holistic quality of Da-Huang-Xiao-Shi decoction using high-performance liquid chromatography coupled with diode-array detection and time-of-flight mass spectrometry.

Da-Huang-Xiao-Shi decoction, consisting of Rheum officinale Baill, Mirabilitum, Phellodendron amurense Rupr. and Gardenia jasminoides Ellis, is a traditional Chinese medicine used for the treatment of jaundice. As described in "Jin Kui Yao Lue", a traditional multistep decoction of Da-Huang-Xiao-Shi decoction was required while simplified one-step decoction was used in recent repsorts. To investigate the chemical difference between the decoctions obtained by the traditional and simplified preparations, a sensitive and reliable approach of high-performance liquid chromatography coupled with diode-array detection and electrospray ionization time-of-flight mass spectrometry was established. As a result, a total of 105 compounds were detected and identified. Analysis of the chromatogram profiles of the two decoctions showed that many compounds in the decoction of simplified preparation had changed obviously compared with those in traditional preparation. The changes of constituents would be bound to cause the differences in the therapeutic effects of the two decoctions. The present study demonstrated that certain preparation methods significantly affect the holistic quality of traditional Chinese medicines and the use of a suitable preparation method is crucial for these medicines to produce special clinical curative effect. This research results elucidated the scientific basis of traditional preparation methods in Chinese medicines.

Clinical Study onDa Huang (Rheum Officinale) Powder Application at Shenque (CV 8) for Incomplete Intestinal Obstruction / 上海针灸杂志

Objective To observe the clinical efficacy ofDa Huang (Rheum Officinale) powder application at Shenque (CV 8) in treating incomplete intestinal obstruction.Method Fifty-eight patients with incomplete intestinal obstruction were randomized into a treatment group and a control group, 29 cases in each group. The control group was by enema with laxative medicine, while the treatment group was byDa Huang (Rheum Officinale) application at Shenque (CV 8) in addition to enema. The time of gassing and defecation, and abdominal bloating intensity were observed.Result The time of gassing and defecation and abdominal bloating intensity in the treatment group were significantly different from that in the control group (P<0.05).ConclusionDa Huang (Rheum Officinale) application at Shenque (CV 8) is an effective approach in treating incomplete intestinal obstruction.

Clinical study of Dahuang-Mudan decoction by retention enema asan adjuvant treatment for the patients with acute pelvic inflammation disease and toxic heat flourishing type / 国际中医中药杂志

Objective To study the effect of Dahuang-Mudan decoctionby retention enemafor the thepatients with acute pelvic inflammation disease (APID) and toxic heat flourishing type. Methods Ninety patients with APID (toxic heat flourishing type) were randomly divided into treatment group and control group (45 case per group). The patients in both groups were given cefamandolenafate injection and tinidazole injection;and the Dahuang-Mudan decoction by retention enema was added to the treatment group.The C-reactive protein (CRP), tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6) and clinical symptoms scores in both groups before treatment and after 2 weeks of treatment were observed. Results After treatment, CRP (3.16 ± 1.25 mg/L vs. 8.68 ± 1.45 mg/L, t=19.126), TNF-α(4.02 ± 1.06 ng/L vs. 9.64 ± 2.08 ng/L, t=15.966), IL-6 (60.34 ± 14.35 ng/L vs. 88.24 ± 12.16 ng/L, t=9.839), temperature recovery time (3.22 ± 1.13 d vs. 4.73 ± 1.36 d, t=5.638), abdominal pain disappeared time (5.26 ± 1.43 d vs. 7.45 ± 1.32 d, t=7.418), leukocyte recovery time (7.34 ± 2.06 d vs. 9.58 ± 2.26 d, t=4.834), hospital stay time (10.24 ± 3.42 d vs. 13.43 ± 4.38 d, t=3.791) in the treatment group were significantly lower than those in the control group (P<0.01). The clinical effect rate (77.27% vs. 48.84%, χ2=7.562) in the treatment group were significantly higher than that in the control group (P<0.01). Conclusions The treatment of APID (toxic heat flourishing type) with Dahuang-Mudan decoction by retention enema can reduce the inflammatory response of pelvic, promote the absorption of exudate and adhesion in pelvic.