Effect of Dahuang Lingxian prescription on the scf/c-kit signaling pathway in gallbladder interstitial cells of Cajal in a guinea pig model of cholesterol gallstone / 临床肝胆病杂志

Objective To investigate the expression levels of scf and c-kit under the regulation of Dahuang Lingxian prescription and the possible mechanism of its effect on gallbladder dynamics, and to provide a theoretical basis for Dahuang Lingxian prescription in preventing the development and recurrence of cholesterol gallstone. Methods A total of 45 specific pathogen-free healthy male guinea pigs were randomly divided into normal group, model group, and traditional Chinese medicine (TCM) group. The guinea pigs in the normal group were fed with normal diet, and those in the model group and the TCM group were fed with high-fat lithogenic diet. After 8 weeks of feeding, 5 guinea pigs were randomly selected from each group, and successful modeling was determined if gallstone was observed with the naked eye in more than 4 guinea pigs. After successful modeling, the guinea pigs in the TCM group were given Dahuang Lingxian prescription by gavage, and those in the model group were given an equal volume of normal saline by gavage. After 8 consecutive weeks of administration by gavage, gallbladder tissue samples were collected, and HE staining was used to observe the pathological changes of gallbladder tissue; Western blot was used to measure the expression level of tumor necrosis factor-α (TNF-α) in gallbladder tissue; immunohistochemistry was used to measure the protein expression levels of scf and c-kit in gallbladder smooth muscle tissue. A one-way analysis of variance was used for comparison of normally distributed continuous data between multiple groups, and the least significant difference multiple comparison method was used for further comparison between two groups. Results HE staining showed marked inflammation of gallbladder tissue in the model group, and compared with the model group, the TCM group had a significantly lower degree of inflammation. Western blot showed that the model group had the highest expression level of TNF-α in gallbladder tissue, followed by the TCM group and the normal group ( P < 0.05); immunohistochemistry showed that compared with the model group, the normal group and the TCM group had significantly higher protein expression levels of scf and c-kit in gallbladder smooth muscle tissue ( P < 0.05). Conclusion Dahuang Lingxian prescription can enhance the dynamic function of the gallbladder, possibly by upregulating the scf/c-kit signaling pathway in interstitial cells of Cajal in gallbladder.

Analysis on mechanisms of Jindan Tablets, Xiaoyan Lidan Tablets and ursodeoxycholic acid in the treatment of gallstones and cholecystitis based on network pharmacology / 国际中医中药杂志

Objective:To analyze the mechanism of Jindan Tablets, Xiaoyan Lidan Tablets and ursodeoxycholic acid in the treatment of gallstone and cholecystitis based on network pharmacology; To conduct a comparative analysis.Methods:The chemical components of Jindan Tablets, Xiaoyan Lidan Tablets and ursodeoxycholic acid and their drug targets were collected from Traditional Chinese Medicine Database and Analysis Platform (TCMSP). DAVID 6.8 database was used to search for the associated diseases of the drug targets. The disease targets of gallstone and cholecystitis were collected from GeneCards and other databases. The protein-protein interactions network was established based on the intersecting targets of three drugs and two diseases. KEGG enrichment analysis was performed based on the DAVID 6.8 database. Cytoscape 3.7.1 software was used to construct a complex network and topology analysis of component- target- disease between three drugs and diseases.Results:222 chemical components and 3 133 drug targets were collected for Jindan Tablets. 104 chemical components and 1 425 action targets were collected for Xiaoyan Lidan Tablets. 1 chemical component and 119 action targets were collected for ursodeoxycholic acid. The three drugs were associated with 31 diseases. 1 382 disease targets for gallstones and cholecystitis were collected. There were 237, 163 and 33 targets for gallstones and cholecystitis in the three drugs, of which 17 were shared by the three drugs and 20 were shared by Jindan Tablets and Xiaoyan Lidan Tablets. Based on the DAVID database, 113, 74 and 10 significant KEGG enrichment pathways were obtained for the three drugs respectively.Conclusions:The three drugs shared many targets and pathways in the treatment of gallstones and cholecystitis, which all had the function of regulating metabolism and inhibiting inflammatory response, while participating in apoptosis, oxidative stress and cancer pathology process. However, they had their own special effects, with Jindan Tablets favoring involving in the cancer process and inhibition of inflammation, and promoting angiogenesis. Xiaoyan Lidan Tablets and ursodeoxycholic acid focused on regulating cholesterol metabolism, and Xiaoyan Lidan Tablets also regulated steroid metabolism and inhibit inflammation, while ursodeoxycholic acid regulated bile acid metabolism.

Curcumin Supplementation Ameliorates Bile Cholesterol Supersaturation in Hamsters by Modulating Gut Microbiota and Cholesterol Absorption.

Curcumin is a polyphenol that has been shown to have prebiotic and cholesterol-lowering properties. This study aimed to investigate the impact of curcumin on bile cholesterol supersaturation and the potential mechanistic role of intestinal microbiota and cholesterol absorption. Male hamsters (n = 8) were fed a high-fat diet (HFD) supplemented with or without curcumin for 12 weeks. Results showed that curcumin significantly decreased cholesterol levels in the serum (from 5.10 to 4.10 mmol/L) and liver (from 64.60 to 47.72 nmol/mg protein) in HFD-fed hamsters and reduced the bile cholesterol saturation index (CSI) from 1.64 to 1.08 due to the beneficial modifications in the concentration of total bile acids (BAs), phospholipids and cholesterol (p < 0.05). Gut microbiota analysis via 16S rRNA sequencing revealed that curcumin modulated gut microbiota, predominantly increasing microbiota associated with BA metabolism and short-chain fatty acid production, which subsequently up-regulated the expression of hepatic cholesterol 7-alpha hydroxylase and increased the synthesis of bile acids (p < 0.05). Furthermore, curcumin significantly down-regulated the expression of intestinal Niemann−Pick C1-like protein 1(NPC1L1) in hamsters and reduced cholesterol absorption in Caco-2 cells (p < 0.05). Our results demonstrate that dietary curcumin has the potential to prevent bile cholesterol supersaturation through modulating the gut microbiota and inhibiting intestinal cholesterol absorption.

Effects of Coffee on the Gastro-Intestinal Tract: A Narrative Review and Literature Update.

The objective of the present research was to review the state of the art on the consequences of drinking coffee at the different levels of the gastrointestinal tract. At some steps of the digestive process, the effects of coffee consumption seem rather clear. This is the case for the stimulation of gastric acid secretion, the stimulation of biliary and pancreatic secretion, the reduction of gallstone risk, the stimulation of colic motility, and changes in the composition of gut microbiota. Other aspects are still controversial, such as the possibility for coffee to affect gastro-esophageal reflux, peptic ulcers, and intestinal inflammatory diseases. This review also includes a brief summary on the lack of association between coffee consumption and cancer of the different digestive organs, and points to the powerful protective effect of coffee against the risk of hepatocellular carcinoma. This review reports the available evidence on different topics and identifies the areas that would most benefit from additional studies.

Analysis of pathogenesis of bile's tri-pa tumor or gallstones and the mechanism of some traditional drug action / Монголын Анагаах Ухаан

Background@#Some scholars believe that the study ancient medical works to gain insight into their scientific concepts and correctly understand the profound techniques of doctors are very important to development of traditional medicine in the 21st century. In “The WHO Traditional Medicine Strategy 2014-2023”, strengthen the study of traditional medical books is mentioned. According to epidemiological surveys, gallstones are a common disease worldwide. In the “Oral Instruction Tantra”, pathogenesis and treatment of gallstones mentioned and further refined in the Interpretation books. Although in modern times, doctors have published many papers and books on traditional medical treatment of gallstones and their pathogenesis based on the contents of ancient books and their own clinical experience, but in-depth analysis of the literature information and explanations have not been mentioned in detail.@*Goal @#According to the traditional medical theory to analyze the pathogenesis of bile’s Tri-pa tumor or gallstones and the mechanism of some drug action.@*Materials and methods@#In our study, we used the book named “Oral Instruction Tantra” as main reference material and for research methods, we used the checklist method to list relevant information about the pathogenesis of gallstones and Nemert Yvuunii Uns-10 traditional drug from the literature, so as to provide preconditions for further analysis. The pathogenesis of gallstones and the action mechanism of Nemert Yvuunii Uns-10 were analyzed by theoretical analysis methods and the analysis-synthesis method was used to summarize the analysis results. </br>The study was carried out in accordance with “Academic Staff Code of Ethics” in the appendix of Ministerial Decree No. 454 in 2007.12.03 by Ministry of Education, Culture and Science,Mongolia. @*Results @#Affected by constitution, unaccustomed diet, inharmonious diet and naturally difficult to digest food, stomach fire reduced leads to food can not be corroded by decomposing phlegm, can not be melting by digestive bile and can not be separated by fire-accompanying wind. This causes food waste enter the nutritional essence which is transported to the liver. The pathological blood is produced when color-producing bile in the liver processes nutritional essence and eventually, bile’s Tri-pa tumor or gallstones are caused by the accumulation of bile deposits from the pathological blood under the influence of wind. </br>We using Mongolian medicine theory to analyze the mechanism of action of Nemert Yvuunii Uns-10 in treating gallstones. In the prescription, Manufactured Monetaria moneta (Linnaeus), Calcitum and Boar dung are main ingredients, Have a crushing Tri-pa tumor or gallstones effect. The function of the Aucklandia lappa Decne supplement the efficacy of the main ingredients. Trogopterori Faeces, Lagotis Integrifolia and Carthamus tinctorius L. treat pathologic blood in the liver. Gentiana Barbata and Herpetospermum caudigerum Wall hava a reducing bile heat effect. Punica granatum L. has the effect of increasing stomach fire and promoting digestive function. Terminalia chebula Retz. homogenize the three humorus.@*Conclusions@#Reduced stomach fire leads to the production of pathological blood in the liver and eventually accumulation of deposits in bile is the cause of bile’s Tri-pa tumor or gallstones. The mechanism of action of Nemert Yvuunii Uns-10 traditional drug for treat bile’s Tri-pa tumor or gallstones is by breaking stones, treating pathological blood, reducing bile heat, increasing stomach fire to promote digestion and homogenizing the three elements

Morphological and elemental mapping of gallstones using synchrotron microtomography and synchrotron X-ray fluorescence spectroscopy.

BACKGROUND AND AIM: Regional differences in gallstone (GS) composition are well documented in the Indian subcontinent. The reasons for the same are unknown. Etiopathogenesis of GS remains elusive despite advances in instrumentation. This was an in-depth analysis of the chemical, structural, and elemental composition of GS with special reference to synchroton studies. METHODS: We used high-end sensitive analytical complementary microscopic and spectroscopic methods techniques, such as X-ray diffraction, scanning electron microscopy, Fourier transform infrared, synchrotron X-ray fluorescence spectroscopy (SR-XRF), and 2D and 3D synchrotron microtomography (SR-µCT), to study the ultra structure and trace element composition of three major types of GS (cholesterol, mixed, and pigment). SR-XRF quantified the trace elements in GS. RESULTS: The cholesterol GS (monohydrate and anhydrate) were crystalline, with high calcium content. The pigment GS were amorphous, featureless, black, and fragile, with high calcium bilirubinate and carbonate salts. They had the highest concentration of iron (average 31.50 ppm) and copper (average 92.73 ppm), with bacterial inclusion. The mixed stones had features of both cholesterol and pigment GS with intermediate levels of copper (average 20.8 ppm) and iron (average 17.78 ppm). CONCLUSION: SR-µCT has, for the first time, provided cross-sectional computed imaging delineating the framework of GS and mineral distribution. It provided excellent mapping of cholesterol GS. SR-XRF confirmed that pigment GS had high concentrations of copper and iron with bacterial inclusions, the latter possibly serving as a nidus to the formation of these stones.

Efficacy and safety of a novel topical agent for gallstone dissolution: 2-methoxy-6-methylpyridine.

BACKGROUND: Although methyl-tertiary butyl ether (MTBE) is the only clinical topical agent for gallstone dissolution, its use is limited by its side effects mostly arising from a relatively low boiling point (55 °C). In this study, we developed the gallstone-dissolving compound containing an aromatic moiety, named 2-methoxy-6-methylpyridine (MMP) with higher boiling point (156 °C), and compared its effectiveness and toxicities with MTBE. METHODS: The dissolubility of MTBE and MMP in vitro was determined by placing human gallstones in glass containers with either solvent and, then, measuring their dry weights. Their dissolubility in vivo was determined by comparing the weights of solvent-treated gallstones and control (dimethyl sulfoxide)-treated gallstones, after directly injecting each solvent into the gallbladder in hamster models with cholesterol and pigmented gallstones. RESULTS: In the in vitro dissolution test, MMP demonstrated statistically higher dissolubility than did MTBE for cholesterol and pigmented gallstones (88.2% vs. 65.7%, 50.8% vs. 29.0%, respectively; P < 0.05). In the in vivo experiments, MMP exhibited 59.0% and 54.3% dissolubility for cholesterol and pigmented gallstones, respectively, which were significantly higher than those of MTBE (50.0% and 32.0%, respectively; P < 0.05). The immunohistochemical stains of gallbladder specimens obtained from the MMP-treated hamsters demonstrated that MMP did not significantly increase the expression of cleaved caspase 9 or significantly decrease the expression of proliferation cell nuclear antigen. CONCLUSIONS: This study demonstrated that MMP has better potential than does MTBE in dissolving gallstones, especially pigmented gallstones, while resulting in lesser toxicities.

Diet After Cholecystectomy.

Patients with gallstones are often affected by alimentary disorders contributing to the onset of gallstones disease. Cholecystectomy can have nutritional and metabolic consequences in the short-term (diarrhea, abdominal pain and bloating) and in the long-term (increased Body Mass Index with metabolic syndrome, gastritis, liposoluble vitamin deficiency). Pathogenic mechanisms behind these disturbances are reviewed and the need for an early post-operative nutritional intervention based on low-lipid, high-fibers diet, is highlighted.

"Aged" autopsy gallstones simulating dry bone context: A morphological, histological and SEM-EDS analysis.

OBJECTIVE: The aim of this paper is to provide information on the morphology and composition of gallstones based on clinical samples in order to assist paleopathologists and bioarchaeologists in recognizing their presence in archaeological contexts. MATERIALS AND METHODS: 270 gallstones were extracted and macerated from autopsies conducted at the Istituto di Medicina Legale in Milan (Italy) in order to simulate a dry bone recovered from archaeological contexts. Morphological, histological, and elemental variation was documented. RESULTS: Gallstones vary in size, shape, color and texture. The cross-sectional surface correlates with chemical composition and is a valuable tool for classification into subcategories of stones. Histological analysis can confirm the classification. Elemental analysis yielded a higher frequency of carbon, calcium and phosphorus. CONCLUSIONS: Although identification of gallstones in archaeological contexts can be challenging, familiarity with morphological, histological, and elemental variation can assist researchers in the field and laboratory. SIGNIFICANCE: Identifying gallstones in archaeological populations will assist researchers in estimating their frequency in the past and the environmental, cultural, and biological conditions leading to their presence. LIMITATIONS: Small sample size derived from a modern and limited autopsy population may minimize the types and degree of variation present in the past. Effects of climate, soil, and taphonomy were not evaluated. SUGGESTIONS FOR FURTHER RESEARCH: Examination of larger samples derived from diverse populations may reveal greater variation or more diagnostic aspects of stones.

Effect of Chaishao Shugan Lidan Paishitang on Inflammation, Stress Response and Gastrointestinal Function in Gallstones Patients with Chronic Cholecystitis / 中国实验方剂学杂志

Objective: To explore the effect of Chaishao Shugan Lidan Paishitang on inflammation, stress response and gastrointestinal function in gallstones patients with chronic cholecystitis. Method: Gallstones patients with chronic cholecystitis treated in our hospital from March 2017 to May 2018 were randomly divided into two groups, with 65 cases in each group. The control group was orally given ursodeoxycholic acid combined with metronidazole. In addition to the therapy of the control group, the Observation group was also given Chaishao Shugan Lidan Paishitang. Traditional Chinese medicine symptom scores, interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), hypersensitive C-reactive protein (hs-CRP), superoxide disproportionation alcohol (SOD), propylene glycol (MDA) and carbohydrate antigen 19-9 (CA19-9), carcinoembryonic antigen (CEA), gastric dynamic element (MOT), gastrin-releasing (GAS) and somatostatin (SS), alanine aminotransferase (ALT), aspertate aminotransferase (AST), total bilirubin (TBIL) and total bile acid (associates), and total cholesterol (TC) levels before and after treatment were observed in two groups, and the curative effect, reactions and relapse were also observed. Result: The inflammatory effect, stone ablation effect and clinical effect of observation group were significantly better than those of control group (Z=2.329, PZ=2.686, PZ=2.940, PPα, hs-CRP, IL-6, CEA, CA19-9, SS and MDA in observation group were lower than those in control group (PPχ2=5.140, PConclusion: The treatment of gallstones patients with chronic cholecystitis by Chaishao Shugan Lidan Paishitang has a good curative effect, and can alleviate clinical symptoms, inhibit oxidative stress and inflammation, improve liver and bile functions and gastrointestinal function, and reduce the recurrence rate.

Prospective randomized controlled trial comparing standard analgesia with combined intra-operative cystic plate and port-site local anesthesia for post-operative pain management in elective laparoscopic cholecystectomy.

BACKGROUND: Various mechanisms, including somatic and visceral nociceptive stimulation, have been suggested as a cause for pain after laparoscopic cholecystectomy (LC). We therefore conducted a prospective randomized controlled trial (PRCT) to evaluate whether somatovisceral pain blockade reduces pain after LC. HYPOTHESIS: Analgesic efficacy of multimodal analgesia is superior to standard analgesia for patients undergoing elective LC for symptomatic cholelithiasis. Specifically, topical cystic plate and port-site injection with 0.25 % bupivacaine significantly reduces pain after LC. DESIGN: This study was designed as single-blinded PRCT. SETTING: This study was conducted in an academic medical center. PATIENTS AND METHODS: Between February and May 2010 we randomly assigned 63 patients with symptomatic cholelithiasis in a 1:1 ratio to non-opioid/opioid analgesic combinations (Control Group, n = 32) and non-opioid/opioid analgesic combinations plus topical 0.25 % bupivacaine onto the cystic plate and local 0.25 % bupivacaine port-site injection, post-LC (Study Group, n = 31). Primary endpoint was patient-reported pain 1, 4, 6, 12, 24 h and 1 week post-LC using the Visual Analog Scale (VAS 0-10). RESULTS: Study groups were comparable clinicopathologically. There were no adverse events. A statistically significant reduction in mean pain score was apparent in Study Group patients in comparison with Control Group (mean VAS 4.83 ± 2.33 vs. 6.80 ± 1.87; p < 0.001) at all early (1-6 h) post-operative time points following LC. CONCLUSION: This PRCT shows significantly improved pain control with somatovisceral pain blockade over non-opioid/opioid analgesic combinations following LC for symptomatic cholelithiasis. For centers not utilizing adjunctive local anesthetic for LC, this topical use of bupivacaine may improve patient comfort during recovery. This trial was registered on www.ClinicalTrials.gov NCT# 01972620.

Study on relationship between gallbladder calculi with hyperlipidemia and Chinese medicine constitution / 重庆医学

Objective To investigate the relationship between gallbladder stone with blood lipid levels ,body weight index (BMI) and distribution of TCM constitution .Methods A total of 698 cases of gallstone in our two hospital from September 2013 to February 2015 were selected as the experiment group ,and contemporaneous 253 cases of abdominal hernia ,gallbladder polyp and other diseases were included into the control group .The TCM constitution identification investigation was performed and the TCM constitution distribution was observed .The blood lipid levels and BMI were compared between the two groups .The risk of blood lipid level and BMI to gallstone occurrence was evaluated .Results The normal constitution in the experimental group accounted for 12 .2% ,the top three places of the biased constitution were phlegm‐damp nature ,Qi deficiency nature and damp heat nature ,which accounted for 29 .4% ,17 .0% and 11 .0% respectively .The TC and VLDL‐C level and BMI in the male and female patients of the experiment group were higher than those in the control group ,moreover the TG level in the female patients of the experiment group was significantly higher than that in the control group ,the difference was statistically significant (P< 0 .05) .The increase of TC (OR=2 .87) ,VLDL‐C(OR=6 .32) and BMI (male:OR=2 .43 ,female:OR=2 .01) were the risk factors of gallstone occurrence . Conclusion Increase of TC ,VLDL‐C and BMI are the risk factors of gallstone occurrence ,the TCM constitution of gallstone pa‐tients is dominated by the phlegm‐damp nature .

Vitamin D Deficiency and Gallbladder Stasis.

Cholelithiasis or presence of calculi/gallstones in the gallbladder occurs more frequently in pregnant women. Vitamin D deficiency is suggested to be associated with gallbladder stasis, and a role for vitamin D supplementation is thought to have potential to prevent gallstones in this special population.

New Evidences of Key Factors Involved in "Silent Stones" Etiopathogenesis and Trace Elements: Microscopic, Spectroscopic, and Biochemical Approach.

The knowledge of the key factors involved in etiopathogenesis of the gallstone disease requires chemical, structural, and elemental composition analysis. The application of different complementary analytical techniques, both microscopic and spectroscopic, are aimed to provide a more comprehensive determination of the gallbladder calculi ultrastructure and trace element identification. High sensitivity techniques such as electron microscopy (SEM), Fourier transform infrared (FTIR), electron paramagnetic resonance (EPR) spectroscopy, and X-ray diffraction (XRD) along with biochemical analysis are used in a new attempt to investigate various factors which play a regulatory role in the pathogenesis of gallstones. The microstructure of different types of gallbladder stones has specific characteristics which are related to the elemental composition. The binding of metal ions with bile salts and bilirubin plays important roles in gallstone formation as revealed by FTIR spectrum of calcium bilirubinate complex in pigment gallstones. The EPR results demonstrated the generation of bilirubin free radicals and variation of its electronic structure and conjugation system in the skeleton of bilirubin molecule during complex formation. EPR spectra of pigment gallstones demonstrate the coexistence of four paramagnetic centers including stable bilirubin free radical, Mn2+, Cu2+, and Fe3+ with distinct magnetic parameters and well-resolved hyperfine structure in the case of Mn2+ ions. The result confirms a macromolecular network structure with proteins and the formation of bilirubin-coordinated polymer. Bilirubin and bilirubinate free radical complexes may play an important role in pigment gallstone formation.

Influence of dietary tender cluster beans (Cyamopsis tetragonoloba) on biliary proteins, bile acid synthesis and cholesterol crystal growth in rat bile.

Tender cluster beans (CBs; Cyamopsis tetragonoloba) are observed to possess anti-lithogenic potential in experimental mice. Formation of cholesterol gallstones in gallbladder is controlled by procrystallizing and anticrystallizing factors present in bile in addition to supersaturation of cholesterol. This study aimed at evaluating the influence of CB on biliary glycoproteins, low molecular weight (LMW) and high molecular weight (HMW) proteins, cholesterol nucleation time, and cholesterol crystal growth in rat hepatic bile. Groups of rats were fed for 10 weeks with 0.5% cholesterol to render the bile lithogenic. Experimental dietary interventions were: 10% freeze-dried CB, 1% garlic powder or their combination. Incorporation of CB into HCD decreased the cholesterol saturation index in bile, increased bile flow and biliary glycoproteins. Dietary CB prolonged cholesterol nucleation time in bile. Electrophoresis of biliary proteins showed the presence of high concentration of 27 kDa protein which might be responsible for the prolongation of cholesterol nucleation time in the CB fed group. Proteins of 20 kDa and 18 kDa were higher in CB treated animals, while the same were less expressed in HCD group. Biliary proteins from CB fed animals reduced cholesterol crystal growth index which was elevated in the presence of proteins from HCD group. Cholesterol-7α-hydroxylase and cholesterol-27-hydroxylase mRNA expression was increased in CB treated animals contributing to the bile acid synthesis. Thus, the beneficial anti-lithogenic effect of dietary CB which primarily is due to reduced cholesterol saturation index was additionally affected through a modulation of the nucleating and anti-nucleating proteins that affect cholesterol crystallization.

Determination of neuroprotective oxysterols in Calculus bovis, human gallstones, and traditional Chinese medicine preparations by liquid chromatography with mass spectrometry.

So far, the components responsible for the neuroprotective effects of Calculus bovis are unclear. Cholesterol, one of the major components in Calculus bovis, is easily oxidized into oxysterols, which possess direct or indirect neuroprotective effects proved by our and others' previous studies. Therefore, a liquid chromatography with mass spectrometry method coupled with ultrasonic extraction and solid-phase extraction was developed for the determination of neuroprotective oxysterols in Calculus bovis, human gallstones, and traditional Chinese medicine preparations. Chromatographic separation was achieved on a C18 column with isocratic elution at a flow rate of 1 mL/min. The established method showed good linearity (R(2) > 0.998), sensitivity with low limits of detection (0.06-0.39 µg/g), acceptable precisions (relative standard deviations ≤ 7.4%), stability (relative standard deviations ≤ 5.9%), and satisfactory accuracy (92.4-102.9%) for all analytes identified by different retention times, which could be applied for the determination of oxysterols. Five kinds of oxysterols proved to function as neuroprotectants were detected at different concentrations. Among them, 7ß-hydroxycholesterol and cholestane-3ß,5α,6ß-triol were rather abundant in the samples. It could be concluded that the potential neuroprotective components in Calculus bovis may be these oxysterols.

Dietary fat and protein intake are not associated with incident biliary sludge and stones during pregnancy.

BACKGROUND: Dietary composition can cause insulin resistance, elevated serum lipid levels, and obesity, all of which predispose to gallstone formation. The effects of dietary fat (including individual fatty acids) and protein on gallstone formation are controversial. The aim of this study was to examine the effects of dietary fat and protein intake on incident gallstone disease during pregnancy, a high-risk time for stone formation. METHODS: We prospectively studied 3070 pregnant women who underwent serial gallbladder ultrasound examinations during pregnancy and at 4-6 weeks postpartum. All women had at least 2 study ultrasounds for comparison. A semi-quantitative food frequency questionnaire was completed by subjects in the early third trimester. Multivariate logistic regression was performed to assess the risk of incident gallbladder disease across quartiles of intake of total fat, individual fatty acids (polyunsaturated, monounsaturated, saturated, and total trans-fatty acids, as well as cholesterol), protein, and protein subtype (animal or vegetable based). RESULTS: The cumulative incidence of new biliary sludge/stones or progression of baseline sludge to stones was 10.2% by 4-6 weeks postpartum. There was no association between total dietary fat (odds ratio [OR], 1.18; 95% confidence interval [CI], 0.64-2.18 comparing lowest and highest quartiles) or protein intake (OR, 0.83; 95% CI, 0.44-1.22 comparing lowest and highest quartiles) and incident gallbladder disease. There was also no association between individual fatty acids or protein subtype and gallbladder disease. CONCLUSIONS: Neither total nor subtype of dietary fat or protein was associated with incident biliary stone or sludge formation in this cohort of pregnant women.

Therapeutic uses of animal biles in traditional Chinese medicine: an ethnopharmacological, biophysical chemical and medicinal review.

Forty-four different animal biles obtained from both invertebrates and vertebrates (including human bile) have been used for centuries for a host of maladies in traditional Chinese medicine (TCM) beginning with dog, ox and common carp biles approximately in the Zhou dynasty (c. 1046-256 BCE). Overall, different animal biles were prescribed principally for the treatment of liver, biliary, skin (including burns), gynecological and heart diseases, as well as diseases of the eyes, ears, nose, mouth and throat. We present an informed opinion of the clinical efficacy of the medicinal uses of the different animal biles based on their presently known principal chemical components which are mostly steroidal detergent-like molecules and the membrane lipids such as unesterified cholesterol and mixed phosphatidylcholines and sometimes sphingomyelin, as well as containing lipopigments derived from heme principally bilirubin glucuronides. All of the available information on the ethnopharmacological uses of biles in TCM were collated from the rich collection of ancient Chinese books on materia medica held in libraries in China and United States and the composition of various animal biles was based on rigorous separatory and advanced chemical identification techniques published since the mid-20(th) century collected via library (Harvard's Countway Library) and electronic searches (PubMed and Google Scholar). Our analysis of ethnomedical data and information on biliary chemistry shows that specific bile salts, as well as the common bile pigment bilirubin and its glucuronides plus the minor components of bile such as vitamins A, D, E, K, as well as melatonin (N-acetyl-5-methoxytryptamine) are salutary in improving liver function, dissolving gallstones, inhibiting bacterial and viral multiplication, promoting cardiac chronotropsim, as well as exhibiting anti-inflammatory, anti-pyretic, anti-oxidant, sedative, anti-convulsive, anti-allergic, anti-congestive, anti-diabetic and anti-spasmodic effects. Pig, wild boar and human biles diluted with alcohol were shown to form an artificial skin for burns and wounds one thousand years ago in the Tang dynasty (618-907 CE). Although various animal biles exhibit several generic effects in common, a number of biles appear to be advantageous for specific therapeutic indications. We attempt to understand these effects based on the pharmacology of individual components of bile as well as attempting to identify a variety of future research needs.

Castanea sativa Mill. extract contracts gallbladder and relaxes sphincter of Oddi in guinea pig: a natural approach to biliary tract motility disorders.

Impaired gallbladder motility is a contributing factor to gallstone formation. Since many drugs delaying intestinal motility inhibit gallbladder emptying, the aim of the present study was to evaluate the effect on gallbladder and sphincter of Oddi motility of a Natural Chestnut Wood Extract (NEC) that reduces intestinal motility. In order to evaluate the effect of the extract in normal- and high-risk gallstone conditions, the investigation was performed using tissues from animals fed normal and lithogenic diet. Fifty guinea pigs were administered either control or lithogenic diet. The spontaneous motility of the gallbladder and sphincter of Oddi were recorded on isolated gallbladder tissues; thereafter, the effect of NEC on motility was tested and compared with carbachol (CCh), potassium chloride (KCl), noradrenaline (NA), and A71623. Compared to controls, the lithogenic diet induced an irregular and disordered motor pattern in both the gallbladder and sphincter of Oddi. NEC increased gallbladder and decreased sphincter of Oddi spontaneous motility independently of cholinergic, adrenergic, and CCK-1 receptor-mediated pathways both in controls and in lithogenic diet-fed animals, although the effect was lower in the latter group. The effect was reversible and mediated by calcium channels. The natural extract of chestnut increasing gallbladder contraction and inducing the relaxation of the sphincter of Oddi can be of benefit in pathological conditions associated with increased transit time at risk of gallstones.

Two glucosinolates and their effects related to the prevention of cholesterol gallstones: a review / Dos glucosinolatos y sus efectos asociados con la prevención de cálculos biliares de colesterol: una revisión

Two glucosinolates (glucoraphasatin and glucoraphanin) and their degradation products (raphasatin and sulforaphane) are secondary metabolites which have shown antioxidant properties and inhibitory properties against the hepatic cholesterol; these effects are very important for the prevention of cholesterol gallstones because in their pathophysiology there is an imbalance in the transport and secretion of cholesterol. These effects produce oxygen reactive species formation, which damages the hepatic and biliary tissues. Cholesterol gallstones are a public health problem; their pharmacological treatment is very limited and the invasive surgical treatment for symptomatic gallstones is the cholecystectomy. Current research focuses on the search for preventive treatments, as there are many risk factors associated with the development of gallstones; therefore, a natural therapeutic alternative may be the use of these glucosinolates and their degradation products.

Dos glucosinolatos (glucorafasatina y glucorafanina) y sus productos de degradación (rafasatina y sulforafano) son metabolitos secundarios que han demostrado propiedades antioxidantes y propiedades inhibidoras contra el colesterol hepático; estos efectos son muy importantes para la prevención de cálculos biliares de colesterol porque en su fisiopatología existe un desajuste en el transporte y secreción del colesterol. Estos efectos producen la formación de especies reactivas de oxígeno, que dañan los tejidos hepático y biliar. Los cálculos biliares de colesterol son un problema de salud pública, su terapia farmacológica es muy limitada y el tratamiento quirúrgico invasivo para cálculos biliares sintomáticos es la colecistectomía. Las investigaciones actuales están orientadas a la búsqueda de tratamientos preventivos, porque hay muchos factores de riesgo asociados al desarrollo de cálculos biliares; por lo tanto, una alternativa terapéutica natural podría ser el uso de estos glucosinolatos, así como sus productos de degradación.