[Effect of electroacupuncture at "Zusanli" (ST 36) on duodenal mast cells, NGF and NTRK1 in rats with functional dyspepsia].

OBJECTIVE: To observe the effect of electroacupuncture (EA) at "Zusanli" (ST 36) on duodenal mast cells, nerve growth factor (NGF) and neurotrophic tyrosine kinase receptor type 1 (NTRK1), and to explore the mechanism of electroacupuncture at Zusanli (ST 36) on functional dyspepsia (FD). METHODS: Sixty SPF-grade 10-day-old SD rats were randomly divided into a normal group, a model group, a ketotifen group and an EA group, 15 rats in each group. The FD model was prepared by iodoacetamide combined with rat tail clamping method in the model group, the ketotifen group and the EA group. The rats in the ketotifen group were injected intraperitoneally with ketotifen (1 mg•kg-1•d-1) for 7 days; the rats in the EA group were treated with EA at bilateral "Zusanli" (ST 36), with disperse-dense wave, frequency of 2 Hz/50 Hz and intensity of 0.5 mA, 20 min each time, once a day for 14 days. The gastric emptying rate and small intestinal propulsion rate in each group were observed; the morphology of duodenal mucosa was observed by HE staining; the toluidine blue staining was used to observe the number and degranulation of mast cells in duodenal mucosa; the protein and mRNA expressions of NGF, NTRK1 in duodenum were detected by Western blot and real-time PCR; the level of interleukin-1ß (IL-1ß) in duodenum was measured by ELISA. RESULTS: Compared with the normal group, the gastric emptying rate and small intestinal propulsion rate in the model group were decreased (P<0.01); compared with the model group, the gastric emptying rate and small intestinal propulsion rate in the ketotifen group and the EA group were increased (P<0.01); the small intestinal propulsion rate in the EA group was higher than that in the ketotifen group (P<0.01). In the model group, local defects in duodenal mucosa were observed with a small amount of inflammatory cell infiltration; no obvious abnormality was found in duodenal mucosa of the other groups. Compared with the normal group, the mast cells of duodenal mucosa in the model group were increased significantly with significant degranulation; compared with the model group, the mast cells of duodenal mucosa in the ketotifen group and the EA group were decreased significantly, and the degranulation was not obvious. Compared with the normal group, the protein and mRNA expressions of NGF, NTRK1 as well as the level of IL-1ß in duodenum in the model group were increased (P<0.01); compared with the model group, the protein and mRNA expressions of NGF, NTRK1 as well as the levels of IL-1ß in duodenum in the ketotifen group and the EA group were decreased (P<0.01, P<0.05); compared with the ketotifen group, the mRNA expression of NGF, as well as the protein and mRNA expressions of NTRK1 in duodenum in the EA group were decreased (P<0.05, P<0.01). CONCLUSION: EA at "Zusanli" (ST 36) could inhibit the activation of duodenal mast cells and regulate the expressions of NGF and its receptor to improve the low-grade inflammatory response of duodenum, resulting in treatment effect on FD.

Apolipoprotein AI-CIII-AIV gene cluster polymorphisms in relation to cholesterol gallstone.

OBJECTIVE: To investigate the frequency of variants at Xmn I, Msp I sites of apolipoprotein (Apo), A I-CIII-AIV gene cluster, and its relation to cholesterol gallstones in Chinese patients. METHODS: Restriction fragment length polymorphisms (RFLP) at Xmn I, Msp I sites of ApoAI-CIII-AIV gene cluster were studied using a polymerase chain reaction (PCR) in 161 patients with cholesterol gallstones and 94 healthy subjects from a Chinese population in Sichuan Province. RESULTS: In both the cholesterol gallstone group and the healthy control group, X1 and M1 alleles were the major alleles and homozygous X1X1 and M1M1 genotypes were the most frequent. However, the frequency of X2 allele mutation in female patients of the cholesterol gallstones group was significantly higher than that in women in the healthy control group (P<0.05), but no difference was found in the frequency of M2 alleles mutation (P>0.05). CONCLUSION: The data showed that Xmn I RFLP of ApoAI-CIII-AIV gene cluster is associated to some extent with cholesterol gallstones in female Chinese patients.

Effect of apolipoprotein E gene Hha I restricting fragment length polymorphism on serum lipids in cholecystolithiasis.

AIM:To investigate the role of apolipoprotein E (apoE) polymorphism in the lithogenesis of gallstone and the hereditary pathogenesis of the disease.METHODS: Polymerase chain reaction (PCR) was used to study apoE phenotypes and allele frequencies in patients with gallstones and control, and the fasting serum lipids of subjects were also measured by enzymatic methods.RESULTS:The levels of triglyceride (TG) and very low density lipoprotein cholesterol (VLDL-C) were much higher in E(2/3) patients than that in E(2/3) control. E(3/3) patients were accompanied with remarkably low levels of high density lipoprotein cholesterol (HDL-C) and its subforms.But in E(3/4) patients there were only slight changes in levels of VLDL-C and low density lipoprotein cholesterol (LDL-C).CONCLUSION:Different apoE phenotype patients with gallstones have different cheracteristics of dyslipidemia and the average level of serum lipids in patients with gallstones are higher than subjects without gallstones in the same apoE gene phenotype.epsilon2 allele is possibly one of the dangerous factors in the lithogenesis of cholecystolithiasis.

Detection of bacterial DNA from cholesterol gallstones by nested primers polymerase chain reaction.

AIM:To search for bacterial DNA sequences in cholesterol gallstones with negative bacterial culture.METHODS:DNA was extracted from cholesterol gallstones in gallbladders and nested primers polymerase chain reaction (NP-PCR) was used to amplify bacterial gene fragments for identifying the existence of bacteria. The samples of bacterial DNA extracted from potentially causative or unrelated living bacteria were amplified in vitro as the standard markers and comparative 16S ribosomal RNA sequence analysis was made for bacterial identification.RESULTS:The gallbladder gallstones of 30 patients were analyzed and bacterial DNA was found in 26 patients. Among them, gallstones with cholesterol content between 30%-69% were seen in 5 (5/5) patients, 70%-90% in 11(11/14) patients, and more than 90% in 10(10/11) patients. There was no difference either in cholesterol and water content of gallstones or in harboring bacterial DNA of gallstones.E.coli-related DNA fragments appeared in the stones of 8 (26.67%) patients; propionibacteria type DNA in 7 (23.33%); and harbored bacterial gene fragments in 2 patients, similar to Streptococcus pyogenes. A more heterogenous sequence collection was found in 7 (23.33%) patients, which could belong to multiple bacterial infections.Two (6.67%) patients had bacterial DNA with low molecular weight which might be related to some unidentified bacteria.CONCLUSION:Most cholesterol gallstones harbor bacterial DNA.It is important to determine whether these microorganisms are innocent bystanders or active participants in cholesterol gallstone formation.

Changes of lipid metabolism in plasma, liver and bile during cholesterol gallstone formation in rabbit model.

AIM:To find out the relationship between the disturbances of lipid metabolism and the formation of cholesterol gallstones by studying the changes of lipid metabolism in plasma, liver tissue and the bile.METHODS:Male and female white Japanese rabbits were divided randomly into a control group (Con) and four experimental groups of 10 rabbits each fed with a diet containing 1.2% cholesterol for one, two, three and four weeks (1wk, 2wk, 3wk and 4wk group). The measurement of plasma triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C) and its subfractions (HDL(2)-C, HDL(3)-C), very low and low density lipoprotein cholesterol (VLDL-C, LDL-C) was taken with standard enzymatic techniques. Apolipoprotein (apo) concentrations in plasma were measured by radial immunodiffusion assay for apoA1, apoB100, aopCII and apoC . Total cholesterol of liver was measured by the enzymatic procedure for each animal.Bile acids, mainly glycocholate (GCA) and glycodeoxycholate (GDCA) were detected by dual wavelength thin layer scanner.RESULTS:In all the experimental groups fed with dietary cholesterol, cholesterol crystal was found in the gallbladder in 2/10 cases of the 1wk group, 4/10 of the 2wk group,6/10 of the 3wk group and 7/10 of the 4wk group respectively. The concentration of plasma total cholesterol (TC),triglyceride (TG), phospholipid (pl), VLDL-C, LDL-C, apoB100, apoCII, apoC gradually increased (P < 0.05)with the prolonged feeding time of dietary cholesterol. High density lipoprotein cholesterol and its subfractions (HDL(2)-C, HDL(3)-C) showed a tendency to decrease, but without statistical significance (P > 0.05). ApoA1 was reduced with increased feeding time of dietary cholesterol (P < 0.05).The hepatic and biliary cholesterol increased 1-1.5 times as compared with the control group (t =5.221 and 3.445, P < 0.05).The GCA gradually decreased beginning from the control group to the 4wk group (P <0.05).CONCLUSION:Owing to the high cholesterol diet, the increased concentrations of plasma TC, TG, VLDL-C, LDL-C, hepatic TC and TG, apoB100, apoCII and apoC possibly enhanced the secretion of biliary cholesterol into bile; the decreased plasma apoA1 level might reduce the secretion of antinucleating factor into bile.All those factors mentioned above probably contribute to the formation of cholesterol gallstones.