Technical guidelines for risk assessment of heavy metals in traditional Chinese medicines.

BACKGROUND: Heavy metals are considered a global concern because they can deteriorate human health. This guideline aims to scientifically evaluate health risk of heavy metals in TCM and to propose a reference for decision making in developing TCM-related health policies. METHODS: Using a multidisciplinary approach, a steering committee oversaw the development of the guideline. To obtain a reasonable and accurate risk assessment, key exposure assessment parameters for TCM, e.g., exposure frequency (EF), exposure duration (ED), and daily ingestion rate (IR) were obtained from surveys. In addition, transfer rates for heavy metals from Chinese medicinal materials (CMM) to decoctions or preparations were examined. RESULTS: Based on the scientific theory of risk control, the guideline was designed systematically, and principles and procedures for the risk assessment of heavy metals in TCM were identified. The guideline can be utilized to assess the risk of heavy metals in CMM and Chinese patent medicines (CPM). CONCLUSION: This guideline may help standardize the risk assessment of heavy metals in TCM, advance regulatory standards for heavy metals in TCM, and ultimately improve human health through scientific TCM usage in clinic.

A strategy for quality control of ginkgo biloba preparations based on UPLC fingerprint analysis and multi-component separation combined with quantitative analysis.

BACKGROUND: Many studies have assessed the fingerprint and quantitative analysis of Ginkgo biloba preparations, but the fingerprint mainly focuses on flavonoid glycosides. However, according to our previous study, the differences among diverse manufacturers mainly involve organic acids. METHODS: A novel reverse-phase liquid chromatography assay using diode array detection was developed for evaluating Ginkgo biloba preparations for quality based on a chromatographic fingerprint allowing the simultaneous assessment of eleven compounds, including four organic acids, six flavonol glycosides and one flavonoid aglycone. And the method was applied to 51 batches of Ginkgo biloba preparations from manufacturers in China. Chemometric approaches were performed for evaluating 51 batches of Ginkgo biloba preparations from various manufacturers. RESULTS: The similarity values among the chromatograms of 51 samples ranged from 0.45 to 1.00, showing that the quality of Ginkgo biloba preparations produced by different manufacturers varied greatly. Data analysis of the 51 batches of GBP samples suggested significant variations of the total contents of all 11 targets, also demonstrating the quality difference of GBP samples. There were significant differences in organic acids in particular. CONCLUSION: Combining the chemical fingerprint and quantitative assessment revealed significant variations in the examined commercial products with regard to organic acids. Thus, this study provided a more comprehensive tool for monitoring the quality consistency of Ginkgo biloba preparations.

Association of Fecal Microbiota with Irritable Bowel Syndrome-Diarrhea and Effect of Traditional Chinese Medicine for Its Management.

Changes in intestinal microbiota have been linked to the development of diarrhea predominant irritable bowel syndrome (IBS-D). In order to better elucidate the relationship between intestinal microbiota changes and IBS-D, we compared fecal microbiota of IBS-D rats and healthy control using pyrosequencing of bacterial 16S rRNA gene targeted. Furthermore, we explored the effects of different traditional Chinese medicine (TCM) on intestinal microbiota of IBS-D in dose-dependent manner. Our results showed that there was no significant difference in fecal microbial community diversity among the healthy control group, IBS-D rats and IBS-D rats treated with traditional Chinese medicine, but the fecal microbial composition at different taxonomic levels have changed among these groups. Interestingly, the weight of IBS-D rats treated with moderate doses (13.4 g/kg) of TCM increased significantly, and the diarrhea-related symptoms improved significantly, which may be related to the enrichment in Deferribacteres, Proteobacteria, Tenericutes, Lachnospiraceae, and Ruminococcaceae and the reduction in Lactobacillus in fecal samples.

[Simultaneous determination of six pyrrolizidine alkaloids in different parts of Emilia sonchifolia by UPLC-MS/MS].

This study aims to develop a UPLC-MS/MS method for simultaneous determination of six pyrrolizidine alkaloids(PAs)--intermedine N-oxide(ImNO), lycopsamine N-oxide(LyNO), seneciphylline(Sp), seneciphylline N-oxide(SpNO), senecionine N-oxide(SnNO), and senkirkine(Sk) in different parts of Emilia sonchifolia. UPLC conditions are as follows: ACQUITY UPLC HSS T3 column(2.1 mm×100 mm, 1.8 µm), mobile phase consisting of 0.05% formic acid and 2.5 mmol·L~(-1) ammonium formate in water(A)-0.05% formic acid and 2.5 mmol·L~(-1) ammonium formate in acetonitrile(B) for gradient elution. MS conditions are as below: electrospray ionization(ESI) in the positive ion mode, multiple reaction monitoring(MRM), and the content of the six PAs was calculated with the external standard method. The results suggested the differences in the six PAs among different parts of E. sonchifolia. Sk was detected in all the four parts, with similar content. SnNO also existed in all the four parts, but the content in roots was significantly higher than that in other parts. Sp and SpNO were found in both roots and flowers, with the content higher in the former than in the later. ImNO and LyNO were only found in leaves, and the content was low. Among the six components detected, ImNO, LyNO, and SpNO were found and determined for the first time, which enriched the toxic components and laid a scientific basis for the quality and safety evaluation of E. sonchifolia.

Research on the mechanism of microwave-toughened starch on glucolipid metabolism in mice.

Potato resistant starch (RS) was prepared by microwave-toughening treatment (MTT). This study investigated the beneficial effects of RS on high-fat diet (HFD)-induced hyperlipidemia in C57BL/6J mice by evaluating changes in the gut microbiota. The mice were fed low-fat diet with corn starch, HFD with corn starch, HFD with potato starch (HFP), or HFD with RS (HFR) for 6 weeks. The results showed that the HFR group had lower body weight and total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels compared with the HFP group. Moreover, the brown adipose tissue levels of uncoupling protein 1 (UCP-1), ß3-adrenoceptor (ß3-AR), peroxisome proliferator-activated receptor-γ (PPAR-γ), and PPAR-γ coactivator-1α (PGC-1α) were increased. Our results showed that RS supplementation increased the Bacteroidetes/Firmicutes ratio and the abundance of short-chain fatty acid-producing Allobaculum, Ruminococcus, and Blautia. Our data suggest that RS prepared by MTT may be used as a prebiotic agent to prevent gut dysbiosis and obesity-related chronic diseases, such as hyperlipidemia, and obesity.

Association between circulating 25-hydroxyvitamin D and systemic lupus erythematosus: A systematic review and meta-analysis.

AIM: The indicators for measuring vitamin D are various, and 25-hydroxyvitamin D (25(OH)D) is considered as the optimal indicator of total vitamin D levels. In this study, we aim to deeply explore the 25(OH)D status in systemic lupus erythematosus (SLE) patients, and evaluate its relation to SLE risk and disease severity. METHODS: Literature about 25(OH)D status and its associations with SLE were searched in Pubmed, Embase and Cochrane Library databases. Standardized mean difference (SMD), odds ratio (OR) and corresponding 95% confidence interval (95% CI) were illustrated by forest plots, and correlation coefficients (r) were combined by generic inverse variance method. Heterogeneity and publication bias were quantified by I-squared (I2 ) test, funnel plot and Egger's test, respectively. Sensitivity analyses were further examined by leave-one-out method. RESULTS: Nineteen articles were included into our meta-analysis. The overall results showed that compared with the healthy controls, the circulating 25(OH)D levels were significantly lower in SLE patients (pooled SMD = -1.63, 95% CI: -2.51 to -0.76). Subgroup analysis revealed that compared with the healthy controls, SLE patients of Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) ≥ 10, Arab and European ethnicity, all 4 seasons, no vitamin D supplement, had significantly lower circulating 25(OH)D levels; no significant differences were observed in SLE patients of SLEDAI < 10, mixed ethnicity, spring, summer, vitamin D supplement, respectively; no matter the changes of age, disease duration, and the therapy of corticosteroid or immunosuppressive or neither, circulating 25(OH)D levels were significantly reduced in SLE patients. The deficiency, insufficiency and sufficiency of vitamin D could significantly elevate, slightly decrease (not significantly), significantly decrease SLE risk, respectively (pooled OR = 4.37, 95% CI: 1.49 to 12.84; pooled OR = 0.52, 95% CI: 0.22 to 1.26; pooled OR = 0.31, 95% CI: 0.15 to 0.63). Circulating 25(OH)D levels were inversely associated with SLEDAI (pooled correlation coefficient = -0.50, 95% CI: -0.8278 to -0.1689). CONCLUSIONS: Compared with healthy controls, 25(OH)D levels are significantly lower in SLE patients, which is influenced by disease activity, ethnicity, seasons and vitamin D supplement; no matter the change of age, diseases duration and therapy of corticosteroid or immunosuppressive or neither, 25(OH)D levels are significantly decreased in SLE patients; the deficiency, insufficiency and sufficiency of vitamin D could significantly elevate, slightly decrease, and significantly decrease SLE risk, respectively; and 25(OH)D levels inversely correlate with SLEDAI.

Acupuncture plus Chinese Herbal Medicine for Irritable Bowel Syndrome with Diarrhea: A Systematic Review and Meta-Analysis.

PURPOSE: To comprehensively evaluate the efficacy and safety of acupuncture combined with Chinese herbal medicine (CHM) in treating irritable bowel syndrome with diarrhea (IBS-D). METHODS: Relevant randomized controlled trials (RCTs) were systemically retrieved from electronic databases from inception to March 2018, including the Cochrane Central Register of Controlled Trials (CENTRAL), PubMed, EMBASE, China National Knowledge Infrastructure (CNKI), Chinese Biological Medical Database (CBM, SinoMed), China Science and Technology Journal Database (VIP), and Wan Fang Data. Meanwhile, pooled estimates, including the 95% confidence interval (CI), were calculated for primary and secondary outcomes of IBS-D patients. Besides, quality of relevant articles was evaluated using the Cochrane Collaboration's risk of bias tool, and the Review Manager 5.3 and Stata12.0 softwares were employed for analyses. RESULTS: A total of 21 RCTs related to IBS-D were included into this meta-analysis. Specifically, the pooled results indicated that (1) acupuncture combined with CHM might result in more favorable improvements compared with the control group (relative risk [RR] 1.29; 95% CI 1.24-1.35; P =0.03); (2) the combined method could markedly enhance the clinical efficacy in the meantime of remarkably reducing the scores of abdominal pain (standardized mean difference [SMD] -0.45; 95% CI -0.72, -0.17; P = 0.002), abdominal distention/discomfort (SMD -0.36; 95% CI -0.71, -0.01; P = 0.04), diarrhea (SMD -0.97; 95% CI -1.18, -0.75; P < 0.00001), diet condition (SMD -0.73; 95% CI -0.93, -0.52; P<0.00001), physical strength (SMD -1.25; 95% CI -2.32, -0.19; P = 0.02), and sleep quality (SMD -1.02; 95% CI -1.26, -0.77; P < 0.00001) compared with those in the matched groups treated with western medicine, or western medicine combined with CHM. Additionally, a metaregression analysis was constructed according to the name of prescription, acupuncture type, treatment course and publication year, and subgroup analyses stratified based on the names of prescriptions and acupoints location were also carried out, so as to explore the potential heterogeneities; and (3) IBS-D patients treated with the combined method only developed inconspicuous adverse events; more importantly, the combined treatment had displayed promising long-term efficacy. CONCLUSIONS: Findings in this study indicate that acupuncture combined with CHM is suggestive of an effective and safe treatment approach for IBS-D patients, which may serve as a promising method to treat IBS-D in practical application. However, more large-scale, multicenter, long-term, and high-quality RCTs are required in the future, given the small size, low quality, and high risk of the studies identified in this meta-analysis.

[Study on application of salvianolic acids reference extract in quality control of Salvia miltiorrhiza and salvianolic acids for injection].

The purpose of this study was to investigate the feasibility of the salvianolic acids reference extract for quality control for Salvia miltiorrhiza and salvianolic acids for injection. An Agilent ZORBAX SB-C18( 4. 6 mm×250 mm,5 µm) column was used with mobile phase consisting of 0. 1% formic acid-water and 0. 1% formic acid-acetonitrile in gradient elution procedure. The column temperature was 30 ℃; the flow rate was 1 m L·min-1; and the detection wavelength was 288 nm. The content of rosmarinic acid,lithospermic acid and salvianolic acid B in S. miltiorrhiza was determined by using the salvianolic acids reference extract as control substance. The content of caffeic acid,salvianolic acid E,rosmarinic acid,lithospermic acid,salvianolic acid B,and salvianolic acid Y in the salvianolic acids for injection was also determined. The linear relationship between chemicals was good( r>0. 998 9),and the injection precision RSD was 0. 30%-0. 90%. The sexual RSD is between 1. 4% and 3. 0%,and the RSD of the reproducibility of the extract is between 2. 1% and 5. 2%. The recovery rate of the three components in S. miltiorrhiza was 96. 80%-99. 20%,and the recovery rate of the six components in salvianolic acids for injection was 88. 90%-107. 5%. The solution of S. miltiorrhiza and salvianolic acids for injection were stable within 48 h. A total of 8 batches of S. miltiorrhiza and injection were determined by the reference extract,and the difference was smaller than that measured by the monomer control. This study preliminarily verified that the salvianolic acids reference extract can be used as a substitute for the monomer control for the quality control of S. miltiorrhiza and salvianolic acids for injection.

The essential oil from the twigs of Cinnamomum cassia Presl inhibits oxytocin-induced uterine contraction in vitro and in vivo.

ETHNOPHARMACOLOGICAL RELEVANCE: The twigs and bark of Cinnamomum cassia Presl (Lauraceae) are widely used in traditional Chinese medicine in the treatment of tumor, abdominal pain, dysmenorrhea, digestive system disease and inflammatory diseases. The aim of this study was to determine the inhibitory effect of the essential oil from the twigs of Cinnamomum cassia Presl (EOCC) on uterine contraction in vitro and in vivo. MATERIALS AND METHODS: The Institute of Cancer Research (ICR) mouse uterine contraction was induced by oxytocin (OT) exposure following estradiol benzoate pretreatment. Mice were given the EOCC (60, 30, and 15mg/kg) by gavage. The level of prostaglandin F2α (PGF2α) in uterine tissue were determined according to specification of enzyme linked immunosorbent assay (ELISA) kit. Uterine tissue was collected for histopathological analysis (H&E). Myosin light chain 20 (MLC20), phosphorylation of myosin light chain 20 (p-MLC20) and cyclooxygenase-2 (COX-2) proteins in uterine tissue were assessed by Western Blot. Mouse isolated uterus strips were mounted in tissue organ baths containing Locke's solution. The contractile responses were recorded with Power Lab recording system. The effect of the EOCC on uterine contraction induced by OT, PGF2α, and acetylcholine (Ach) was observed. Myometrial cells were exposed to OT (7µM) to induce Ca2+ release, and the effect of the EOCC (100, 50, and 25µg/ml) on intracellular Ca2+ was analysed with fluorometry imaging. RESULTS: In vivo study demonstrated that the EOCC significantly reduced OT-induced writhing responses with a maximal inhibition of 66.5%. It also decreased the level of PGF2α in OT-induced mice uterine tissue. Moreover, Western blot analysis showed that COX-2 and p-MLC20 expressions in uterine tissue of dysmenorrhea mice were significantly reduced. EOCC inhibited spontaneous uterus contractions in a dose-dependent manner, and the concentration of the EOCC giving 50% of maximal contraction (IC50) value was 61.3µg/ml. The IC50 values of the EOCC on OT, PGF2α, and Ach-induced contractions were 113.0µg/ml, 94.7µg/ml, and 61.5µg/ml, respectively. Further in vitro studies indicated that the EOCC could restrain intracellular Ca2+ levels in favour of uterine relaxation. CONCLUSION: Both in vivo and in vitro results suggest that the EOCC possesses significant spasmolytic effect on uterine contraction. Thus, the EOCC yields a possible therapeutic choice for the prevention and treatment of primary dysmenorrhea.

The essential oil from the twigs of Cinnamomum cassia Presl alleviates pain and inflammation in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Cinnamomum cassia Presl (Lauraceae) can be found southern China and its bark is commonly used for centuries as ingredient in food and cosmetic industry. The twigs of Cinnamomum cassia Presl is popularly used in China to treat inflammatory processes, pain, menstrual disorders, hypertension, fever etc. The aim of this study is to evaluate the antinociceptive and anti-inflammatory properties of the essential oil (EO) from the twigs of Cinnamomum cassia Presl. MATERIAL AND METHODS: The chemical characterization of the EO was performed by gas chromatography coupled with mass spectrometry (GC-MS). The EO doses of 15, 30, and 60mg/kg were employed in the biological assays. The antinociceptive effects of the EO were evaluated using the models of acetic acid-induced writhing, oxytocin-induced writhing, and formalin and complete Freund's adjuvant (CFA) -induced overt pain tests. we also investigated the effect of the EO in pain intensity to a mechanical stimulus (mechanical hyperalgesia) after carrageenan by using an electronic version of von Frey filaments. Evaluation of anti-inflammatory activity was based on paw edema induced by carrageenan (300µg/25µL/paw) in mice. The levels of cytokines, NO, and PGE2 in paw skin tissue were determined according to instructions. COX-2 and iNOS proteins in paw skin tissue were assessed by Western Blot. RESULTS: The EO (15, 30, and 60mg/kg) reduced the number of abdominal writhings induced by acetic acid with inhibition of 38.0%, 55.4% and 58.7%, respectively. The EO (15, 30, and 60mg/kg) also reduced the number of abdominal writhings induced by oxytocin with inhibition of 27.3%, 51.7% and 69.0%, respectively. The EO significant inhibited the inflammatory (second phase: 10-30min) phase of the formalin-induced paw flinching and licking at the doses of 15, 30, and 60mg/kg. The EO at the tested doses of 15, 30, and 60mg/kg showed inhibited CFA-induced paw flinching and licking. The EO (15, 30, and 60mg/kg) also inhibited carrageenan-induced mechanical hyperalgesia and paw edema. It also decreased the levels of cytokines (TNF-α, and IL-1ß), NO, and PGE2 in carrageenan-induced mice paw skin tissue. Moreover, Western blot analysis showed that COX-2 and iNOS expressions in paw skin tissue of mice were significantly reduced. CONCLUSIONS: These results demonstrate that the antinociceptive and anti-inflammatory properties of the EO from the twigs of Cinnamomum cassia Presl, corroborating its use in folk medicine.

Application of immunoaffinity purification technology as the pretreatment technology for traditional Chinese medicine: Its application to analysis of hesperidin and narirutin in traditional Chinese medicine preparations containing Citri reticulatae Pericarpium.

In the present study, the feasibility of immunoaffinity chromatography (IAC) as a purification technology for the analysis of bioactive components in Traditional Chinese Medicine (TCM) was evaluated. IAC was used to analyze hesperidin (HP) and narirutin (NR) in TCM preparations containing Citri reticulatae Pericarpium (CRP, Chenpi in Chinese). An IAC column for the specific extraction and enrichment of HP and NR from TCM preparations containing CRP was developed and characterized. After HP reacted with carbonyl diimidazole and coupled to protein, it was used to immune mice for the generation of antibody. Through cell fusion, cloning and screening, monoclonal antibody was obtained. The IAC column was constructed by covalently coupling specific monoclonal antibody against HP and NR to CNBr-activated Sepharose 4B and packed into a common solid phase extraction cartridge. The extraction conditions including loading, washing and eluting, as well as flow rate for the extraction of HP and NR were optimized. Under optimal conditions, the maximum capacity, extraction recovery rate and stability of IAC column was also characterized. Results revealed that the maximum capacity of IAC column for HP and NR was approximately 16µg and the relative binding capacity per 1mL of the column volume was 27µg. The extraction recovery rate of IAC column for HP and NR at three spiked levels was in the range of 94.05-109.15%. After the repeated application for 5 times, no significant loss of specific recognition was observed. Using high performance liquid chromatography (HPLC) as an effective analytic tool, HP and NR could be successfully separated via IAC column without the inference from impurities, suggesting that the extraction of HP and NR using the prepared IAC column is feasible. The application of IAC can solve the problem of quantitative analysis due to severe interference or low content. Furthermore, pretreatment methods in different matrixes can be unified. The IAC purification procedure can be used as an alternative effective analytical method for the pretreatment of bioactive components in TCM.

Safe extensive tumescent liposuction with segmental infiltration of lower concentration lidocaine under monitored anesthesia care.

Tumescent anesthesia makes it feasible to perform liposuction in an office setting. There are often patients who desire extensive liposuction on approximately 30% of total body surface area, which means the lidocaine total dose might be over the dosing recommendation. So the segmental infiltration is applied, although the concentration of lidocaine in tumescent fluid is gradually reduced to 0.0252%. Moreover, supplemental intravenous (IV) sedation using monitored anesthesia care is usually applied concurrently to help alleviate discomfort and pain of the patients during tumescent anesthetic infusion and fat extraction which in turn increases the risks of potential lidocaine toxicity due to possible drug interactions. This study was to demonstrate the safety of segmental infiltration of tumescent fluid with lower lidocaine concentration combined with IV sedation in extensive liposuction and determine whether the risk of lidocaine toxicity is increased in this protocol. Ten female patients who requested the extensive liposuction participated in the study. The targeted areas were divided into 2 segments and treated in turn in 1 session. Lidocaine (1600 mg) was infiltrated into the first segment, and approximately 928 mg lidocaine was subsequently infiltrated after accomplishment of the first segment operation. Serum levels of lidocaine were taken every 4 hours during the first 24 hours after the second infiltration. The average time of the procedure is 222 (33) minutes. The dose and total amount of lidocaine injected are 40.7 (5.8) mg/kg and 2528.2 (155.2) mg, respectively. The total volume of the infusates and aspirates are 9918.1 (494) and 6325 (1461.6) mL, respectively, the ratio of total infusates to total aspirates is 1.66 (0.45). The total aspirated fat and fluids are 3280 (1051.8) and 3045 (824.1) mL, respectively. The peak lidocaine levels [2.18 (0.63) µg/mL] occurred after 12 to 20 hours [16.4 (2.27) hours]. No significant correlation between dose per kilogram body weight or total dose of lidocaine infiltrated and its peak levels or time existed. The extensive liposuction covering the 30% total body surface areas was well tolerated by the patients under tumescent anesthesia in combination with the supplemental IV sedation. Our previous study on the fluid management has demonstrated the risk of hypovolemia or fluid overload is very low with this technique, although the patients who received only maintenance fluid (500 mL) in the operating room and could discharge and resume oral intake after 6 hours of recovery room stay. The adequate anesthesia support is available in our office-based setting with adequate recovery facilities in place. It has a high margin of safety, without increasing of lidocaine toxicity or adverse cardiopulmonary sequelae while using a segmental tumescent infiltration with lower concentration of lidocaine.

[Study on UIPLC-DAD fingerprint of Disporum cantoniense].

OBJECTIVE: To establish an UHPLC-DAD fingerprint of 30 batches of Miao medicine Disporum cantoniense from Guizhou, and to provide a theoretical evidence to evaluate its quality. METHODS: The analysis was carried out on an Agilent Eclipse ZOR-BAX Plus C18 (100 mm x 2. 1 mm, 1. 8 µm) column with a mobile phase consisting of acetonitrile-water(containing 0. 1% formic acid) with gradient elution, and the flow rate was 0. 3 mL/min. The column temperature was set at 40 °C and UV detection wavelength was set at 290 nm. The sample injection volume was 3 µL. The similarity evaluation and principal component analysis(PCA) of these fingerprints were carried out. RESULTS: The UHPLC-DAD fingerprint was established and compared by 30 batches of samples similarity with 12 common peaks and 3 peaks were identified. The similarities of 30 batches of Disporum cantoniense were between 0. 766 and 0. 994. The principal component analysis showed that compounds 1 and 12 were representative for the fingerprint. CONCLUSION: The method is accurate and credible,which can be used for identification and quality control of Disporum cantoniense.

[Effect of Fagopyrum cymosum (Trev.) Meisn alcohol extract on defecation and isolated colon of diarrhea-IBS rats and its mechanism].

OBJECTIVE: To explore the intervention of Fagopyrum cymosum (Trev.) Meisn alcohol extract (FAE) on defecation function and motor functions of isolated colons of diarrhea-predominant irritable bowel syndrome (D-IBS) rats and to study its underlying mechanism. METHODS: The D-IBS rat model was established by neonatal pups maternal separation (NMS) combined with intracolonic infusion of acetic acid (AA). Adult IBS rats were randomly divided into the pre-intervention control group (n = 10, with no gastrogavage), the normal saline control group (n = 10, administered with normal saline by gastrogavage), the pre-treatment model group (n = 8,with no gastrogavage),the normal saline model group (n = 8, administered with normal saline by gastrogavage), the low dose FAE group (n = 8, administered with 6 g/kg FAE by gastrogavage), the high dose FAE group (n = 8, administered with 24 g/kg FAE by gastrogavage), and the Pinaverium Bromide group (n = 8, administered with 0.02 g/kg Pinaverium Bromide by gastrogavage). All medication was performed once daily for 2 weeks. The abdominal withdrawal reflex (AWR) was employed to evaluate the visceral hypersensitivity; their loose and watery stool grade was assessed by Bristol scores for stool consistency; and their fresh feces weight was calculated. In vitro effect of different concentrations of FAE and Pinaverium Bromide (0.02 µg/mL) on spontaneous contraction and spasmodic contraction induced by acetylcholine (Ach) in rats' isolated colon were observed and the influence on the intestinal calcium channel was evaluated. RESULTS: Compared with the pre-intervention control group, the pain pressure threshold and the maximum tolerance pressure decreased significantly in the pre-intervention model group (P < 0.05), and the loose and watery stool grade and fresh feces weight increased drastically (P < 0.01). Compared with the normal saline control group, the pain pressure threshold and the maximum tolerance pressure decreased significantly in the normal saline model group (P < 0.05), and the loose and watery stool grade and fresh feces weight increased markedly (P < 0.01). Compared with the normal saline model group, the pain pressure threshold of 24 g/kg FAE and Pinaverium Bromide group significantly increased (P < 0.05). The loose and watery stool grade and fresh feces weight decreased obviously in the low dose FAE group, the high dose FAE group, and the Pinaverium Bromide group (P < 0.05). FAE (30, 100, 300, 1,000, and 3,000 µg/mL) and Pinaverium Bromide could significantly inhibit spontaneous contraction of isolated intestines (P < 0.05, P < 0.01), and FAE (30, 100, and 300 x 10(-6) g/mL) could remarkably inhibit their spasmodic contraction and contractile tension induced by Ach and Ca2+ respectively (P < 0.05, P < 0.01) in a concentration-dependent manner. Pinaverium Bromide also could significantly inhibit Ach and Ca2+ induced contraction. CONCLUSION: Effective components of FAE improved the defecation function and inhibited enterospasm induced intestinal hyperactivity in IBS model rats via antagonizing calcium channel competitively and inhibiting colonic motility dose-dependently.

[Study of cajal interstitial cells in stomach and small intestine of rats with crude, honey-stir-baked radix polygalae and its saponins].

OBJECTIVE: To observe the changes of interstitial cells of Cajal (ICC) in stomach and small intestine of rats with honey-stir-baked Radix Polygalae, crude Radix Polygalae and its saponins, so as to study mechanism of crude Radix Polygalae reducing the motility disorder in gastrointestinal tract. METHODS: Immunohistochemical staining was used to investigate the distribution of c-kit positive ICC in stomach and small intestine. RESULTS: Compared with control group, the c-kit positive ICC in stomach and small intestine of crude Radix Polygalae and its saponins groups were both markedly decreased in model group (both P < 0.01), while honey-stir-baked Radix Polygalae group can not (P > 0.05). CONCLUSION: The motility disorder in gastrointestinal tract caused by crude Radix Polygalae and its saponins may be associated with the changes of ICC number in stomach and small intestine. Honey-stir-baked Radix Polygalae can protect ICC in some extent.

[Changes of gastrointestinal myoelectric activity and bile acid pool during cholesterol gallstone formation in guinea pigs].

OBJECTIVE: To investigate the changes of gastrointestinal migrating myoelectric complex (MMC) and bile acid pool size during cholesterol gallstone formation in guinea pigs. METHODS: Seventy-five guinea pigs were randomized into 5 equal groups and given normal diet for 8 weeks, 1% cholesterol diet for 2, 4, and 8 weeks, and cholesterol diet with Vitamin C for 8 weeks, respectively. At the end of the feeding period, the gallbladder motility was assessed by deploying 4 bipolar silver electrodes on the antrum, duodenum, jejunum and ileum. Seven days later, MMC was recorded and the total bile acid pool size measured according to the isotope dilution method. RESULTS: After cholesterol feeding, the gallbladder motility declined in 4- and 8-week groups, which showed a reduction of bile acid pool size by 42.75% (P<0.05) and 57.24% (P<0.05), respectively. Compared to the control group, MMCs started mostly in the duodenum. MMC cycle duration was prolonged in the duodenum by 1.32 times in 4-week group (P<0.05) and 1.44 times in 8 week group (P<0.05) whereas MMC amplitude was lowered in the duodenum by 40.91% in 4-week group (P<0.05) and 60.17% in 8 week group (P<0.05). Supplementation of vitamin C caused changes in MMC similar to those observed in the guinea pigs of 2-week group. CONCLUSIONS: High-cholesterol diet inhibits MMC activity and reduces bile acid pool size in guinea pigs, which, along with impairment of the gallbladder motility, contributes to the formation of cholesterol stone, whereas vitamin C supplement in the diet may help prevent cholelithiasis.

[Study on RP-HPLC fingerprint of compound hongcao injection].

OBJECTIVE: To establish the fingerprint detecting standard of Compound Hongcao Injection by reversed phase high performance liquid chromatography. METHOD: The chromatographic conditions were as follows: an Inersil-ODS-3 column was used; the mobile phases (acetontrile and 0.1% phosphoric acid) with gradient elution; the flow rate 1.0 mL x min(-1) and the UV absorbance detection at 300 nm. RESULT: Under the same selected chromatographic conditions, the HPLC fingerprints of raw drugs and their intermediate, finished products were obtained with good separation and correlation according to the technical requirements of fingerprint on Injection of Chinese traditional medicine. CONCLUSION: 17 common peaks as well as their retention times and peak area ratios on HPLC fingerprint can be used as the important parameters of the quality control for Compound Hongcao Injection.

[Studies on chemical constituents in herb of Polygonum orientale].

OBJECTIVE: To study the chemical constituents of the essential substance from Polygonum oriental. METHOD: Compounds were isolated with silica gel and polyamide chromatography and their structures were determined by spectral analysis and chemical evidence. RESULT: Six compounds were obtained and identified as myricitrin, luteolin, gallic acid, catechin, protocatechuic acid, p-hydroxycinnamic acid. CONCLUSION: Six compounds were isolated from P. oriental for the first time.

[Determination of gallic acid in Melastoma dodecandrum by RP-HPLC].

OBJECTIVE: To establish a RP-HPLC method for the determination of gallic acid in Chinese herb Melastoma dodecandrum, and to evaluate the quality of the herb. METHOD: The RP-HPLC analysis was achieved by using a Polaris C18 column and tetrahydrofuran-methanol-0.2% phosphoric acid (0.5:0.5:99) as the mobile phase, with a flow rate of 1.0 mL x min(-1), and detected by UV at 274 nm. RESULT: The content of gallic acid was from 0.020% to 0.081%, in ten groups of M. dodecandrum collected from different locations. CONCLUSION: The method is a simple, convenient and rapid. It can be used for quality evaluation of M. dodecandrum.

[Qualitative and quantitative studies on qianliebeixi capsules].

OBJECTIVE: To establish a method of quality control on qianliebeixi capsules. METHOD: To clarify semen vaccarine by TLC, to determinate the contents of quertin and kaempferol in prepations, by HPLC. Chromatographic conditions: C18 column with METH-0.2%H3PO4(47:53) as mobile phase; the temperature of column was 40 degrees C, and quercetin was detected at 370 nm with flowing rate 1 ml/min. RESULT: The spots of semen vaccarine were clear in TLC, no interference in negative contrast. The average recovery rate of quertin was 98.9% and the RSD was 2.2%; the average recovery rate in kaempferol was 100.4% and the RSD was 1.7%. CONCLUSION: The resolution of spots were perfect in TLC, the spots were easy to distinguish, The method is simple, rapid as well as reproducible and can control the quality of qianliebeixi capsules effectively.