Objective To study the effect of carbachol(CCH) on membrane potential of smooth muscle cells of gastric antrum of diabetic rats to verify the mechanism of gastric motility disturbance.Methods Fifty healthy Wistar rats of 2-month old,weighing 100-160 g,were divided into the control group(n=20) and diabetes mellitus group(n=30).Three months after the establishment of the rat model of diabetes by intraperitoneal injection of 60 mg/kg streptozotocin,gastric emptying time and gastric electrical activity were measured,and the resting membrane potentials of smooth muscle cells in antrum were measured by confocal laser scanning microscopy,then treated with carbachol of 10~(-9) mmol/L,10~(-8) mmol/L,10~(-7) mmol/L,the membrane potentials were measured.Results As compared with the normal rats,gastric emptying time in diabetic rats was significantly longer and abnormal gastric electric rhythm was significantly increased,the abnormal rhythm index(ARI) and the coefficient of variation(CV) of slow wave frequency in diabetic rats were significantly higher,but the resting membrane potentials remained unchanged and the sensitivity of CCH-induced membrane depolarization was increased.Conclusion The increase of sensitivity of CCH-induced membrane depolarization may be involved in the diabetes-induced gastric motor disorders.

Objective To clarify the changes of ultrastructure of interstitial cells of Cajal in the small intestine of diabetic rats. Methods Rats were randomly divided into diabetic group and control group. The rate of the small intestinal transit was measured and the tissues of the small intestine were observed by transmission electron microscopy at 3 months after the establishment of rat model of diabetics. Results In the diabetic rats, the rate of small intestinal transit was delayed as compared with that in the control group. The number of the gap junction of interstitial cells of Cajal in the small intestine of diabetic rats decreased significantly, and the rest structures were damaged. Damaged organelles and formation of vacuoles were also found. Conclusion The changes of ultrastructure of interstitial cells of Cajal in the small intestine may be one of the mechanisms resulting in slow rate of the small intestinal transit in diabetic rats.

Objective In the present study,we use GC/MS-based metabolomics approaches to make analysis of serum metabolic profiles of healthy people and the hyperactivity of liver yang type of constitution in patients with essential hypertension.Try to establish the discriminant model,to discover biomarkers (group) of the hyperactivity of liver yang type of essential hypertension,and to explore the essential material basis of Traditional Chinese medicine syndrome theory.Methods Classified according to the guiding principles of Chinese medicine research,the hyperactivity of liver yang type of constitution in male patients with essential hypertension (n=18),as well as health volunteers (n=15) were randomly selected from Guang An Men Hospital clinic,wards and medical center in the first half of 2010,selected patients with essential hypertension requirements are not taking any drugs or Chinese herbs,or stop taking the various drugs more than one week.Extracted Venous Blood of subjects fasting for 12 hours,and serum was separated through centrifugation.Serum samples are stored and at-86℃ refrigerator.Survey and evaluate endogenous metabolism in serum samples of health control group and types of syndrome mentioned above by gas chromatograph mass spectrometry (GCMS)analysis.Then,analyze the metabolites with Partial Least Squares-Discriminant Analysis.Further use PCA to analyze the principal component factor loadings matrix analysis,and for variable scatter plot (Loading plot),significant increase or decrease the variables can be found from the figure.The combination of these variables is the lesion biomarkers group.Results Compared with the health control group,13 differentially expressed metabolites in the essential hypertension hyperactivity of liver yang type group can be identified (P＜0.05).8 metabolites were up-regulated expression:Uric acid,citrate,Octadecanoic acid,Hexadecanoic acid,Octadecadienoic acid,Leucine,Cholesterol,Norvaline,and 5 metabolites were down-regulated expression:arachidonate,Oleate,Alanine,Aspartic acid,glycine.Conclusion We are incline to regard that the 13 of EH patient serum differentially expressed metabolites are EH hyperactivity of liver yang syndrome metabolic biomarkers:Uric acid,citrate,Octadecanoic acid,Hexadecanoic acid,Octadecadienoic acid,Leucine,Cholesterol,Norvaline,Arachidonate,Oleate,Alanine,Aspartic acid,glycine.

This study was aimed to set up the fingerprint chromatograms of chemical principles of Dendrobii Caulis by HPLC-DAD-ELSD. This analysis was performed at 30°C on a Dikma Cl8 column (250 mm × 4.6 mm, 5 μm) with a mobile phase consisting of acetonitrile-0.5% glacial acetic acid, gradient elution, at the flow rate of 1.0 mL?min-1 with DAD and ELSD detection. The HPLC fingerprint of Dendrobii Caulis was established. And 9 fingerprint peaks were marked. The similarity among different species was low. This method was stable, reliable and repeatable. It is useful for the study on the relationship between chemical components and species of Dendrobii Cauli.

To investigate the transgenic expressing efficacy of helper-dependent adenoviral vector (HDAd) in vitro, we constructed a HDAd encoding enhanced green fluorescent protein (EGFP), denominated as HDAd/EGFP, performed large scale preparation and purification, and then identified the purified HDAd/EGFP under fluorescent microscope and electron microscope. After the concentration of HDAd/EGFP was determined by spectrophotometer, the transgenic expression efficiency of HDAd/EGFP was compared with first generation adenoviral vector encoding EGFP (FGAd/EGFP) in vitro. Therefore, we infected A549 cells with 2000 virus particles (vp) per cell by HDAd/EGFP and FGAd/EGFP respectively and analyzed EGFP expressing level by flow cytometry. Consequently, the fluorescent expression rate and fluorescent intensity of EGFP were higher in early infected A549 cells by HDAd/EGFP than by FGAd/EGFP. HDAd, capable of expressing transgene instantly and efficiently in vitro, is a potential vaccine vector.
Adenoviridae ; genetics ; metabolism ; Cell Line, Tumor ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Helper Viruses ; genetics ; metabolism ; Humans ; Transgenes ; Viral Fusion Proteins ; genetics ; metabolism

Objective To explore the specific plasma exosomal protein expression in Coronary Heart Disease(CHD)with phlegm-turbidity syndrome(PTS),so as to provide biological reference for the diagnosis of CHD with PTS.Methods A total of 20 CHD patients with PTS and 20 healthy subjects at the same period were included.Plasma exosomes were extracted from all subjects using the qEV size exclusion method.First,10 CHD cases with PTS and 10 healthy subjects were randomly selected,and differentially expressed proteins between the two groups were screened using label free non-targeted protein quantification analysis.Then,MRM targeting protein labeling technique was applied to verify the differentially expressed proteins in the remaining subjects.Results Compared with healthy controls,214 differentially expressed plasma exosomal proteins(61 up-regulated and 153 down-regulated)were found in CHD patients with PTS,mainly related to cholesterol metabolism,complement and coagulation cascade,and immune effects.ANXA6,C4BPB,F8,CFB,APOE,C9,and CLU proteins were further validated by MRM targeting protein.Conclusion CHD patients with PTS had differences in plasma exosomal protein expression from healthy controls,and the differential proteins are mainly related to cholesterol metabolism,complement,and the coagulation cascade.