AIM: To study the effect of sesamin on expression of inducible nitric oxide synthase(iNOS)and nitrotyrosine(NT)in rat liver tissue with metabolic syndromic hepatic steatosis. METHODS: The rat model of metabolic syndromic hepatic steatosis was induced by operation of two kidneys with one clip(2K1C)and high-fat. The rats taken from that successful model were randomly divided into model group and sesamin(120, 60, and 30 mg·kg~(-1)·d~(-1))groups. In addition, the sham-operated group was set up. The rats in treated group were given sesamin intragastrically everyday for 8 weeks. The levels of blood lipids(TC, TG and FFA)in serum were detected. The activity of SOD and MDA level in the liver homogenate were determined. The expressions of iNOS and NT proteins were detected by Western blotting analysis. The histopathological changes were observed by HE staining in the liver tissues. RESULTS: Compared to model groups, sesamin(120, 60 mg·kg~(-1))significantly inhibited the elevation of serum TC, TG, FFA, and MDA in liver homogenate(P<0.05), and increased the activity of SOD(P<0.05). It also decreased the protein expression of iNOS and NT(P<0.05), and ameliorated the degree of hepatic steatosis. CONCLUSION: Sesamin prevents and cures the metabolic syndromic hepatic steatosis. The mechanism is probably mediated through decreasing the protein expression of iNOS and NT, and alleviating the oxidative stress in addition to regulating the lipid metabolism.

Aim To investigate the difference of aortic function between renal hypertension rat(2K1C) and renal hypertensive-hyperlipidemia rat(RHHR).Methods Animals were divided into 3 groups: the sham-operated group,RHHR and 2K1C model groups.The vascular function test was performed in vitro.The response of aortic ring to phenylephrine(PE),acetylcholine(ACh) and sodium nitroprusside(SNP) was measured.Then aortic ring was incubated with nitricoxide synthase inhibitor N-nitro-L-arginine-methyl-ester(L-NAME) and its response to Ach was observed.Results The contraction in response to PE was augmented in the two model groups.ACh-induced vasorelaxation was reduced in both model groups,and such relaxation was more depressed in RHHR than that in 2K1C.The ability of relaxation evoked by NO was impaired in two model groups.SNP elicited complete relaxation in three groups,but the sensitivity to SNP was more decreased in RHHR than those in sham-operated and 2K1C groups.Conclusion These findings suggest that the vasodilation function of aorta is more impaired in RHHR than that in 2K1C group.The decrease in utilization of vascular smooth muscle to NO induced by hyperlipidemia may contribute to the main cause.

Peptide cyclization, a pivotal approach to modifying linear precursors of proteins and pepticles, has been used to enhance their biological activities and serum stabilities. Recently, sortase A (SrtA) from Staphyloccus aureus becomes a promising new technology for efficiently incorporating site specific modifications into proteins, conjugating the cell surface and cyclizing the linear peptides. In this study, we constructed two recombinant expression systems, one with chitin binding domain and the other with six-histidine tag and chitin binding domain on the N-terminal of SrtA, separately. The results of enzymatic kinetics indicate that the two recombinant tags do not impair the transpeptidase activity of SrtA compared with the standard reaction reported under the same reaction condition. The two synthesized peptides with N-ternimal three glycines and C-terminal penta-amino acid motif, LPETG, were cyclized using immobilized and recycled SrtA. The SrtA-based cyclization promises to represent a simple method for easy and efficient enzymatic synthesis of large cyclic peptides.
Aminoacyltransferases ; metabolism ; Bacterial Proteins ; metabolism ; Cyclization ; Cysteine Endopeptidases ; metabolism ; Enzymes, Immobilized ; metabolism ; Kinetics ; Peptides ; metabolism ; Peptides, Cyclic ; biosynthesis ; Staphylococcus aureus ; enzymology

Pancreatic cancer is malignant with a poor prognosis,and its incidence is rising worldwide in recent years.Multiple slices spiral computed tomography and computed angiography are the first choice for the diagnosis of pancreatic cancer,while misdiagnosis of pancreatic cancer still exists.From August 2009 to October 2011,80 patients with pancreatic or periampullary cancer were diagnosed using the medical image three dimensional visualization system (MI-3DVS).The threedimensional models of the liver,pancreas,vascular system and tumors were reconstructed successfully based on the 64-slice spiral computed tomography data.According to the analysis of the three dimensional models,4 patients underwent standard pancreaticoduodenectomy,1 received palliative surgery.MI3DVS plays an important role in the diagnosis and assessement of resectability of pancreatic and periampullary cancer.

Objective To evaluate the value of DNA quantitative analysis in diagnosis of breast tumor.Methods The data of 128 cases diagnosed with breast disease from Feb.2005 to Jan.2006 in Department of Pathology in Wuhan Center Hospital were collected,among whom 72 cases had benign breast lesion,10 cases had boundary breast lesion and 46 cases had malignant breast disease.Automated DNA imaging cytometry was used to identify DNA indexes of the 128 cases.Results The positive rate was 1.39％ (1/72),30.00％ (3/10) and 78.26％ (36/46) respectively in benign breast lesion,boundary breast lesion and malignant breast disease.The difference had statistical significance (P ＜ 0.05).In invasive ductal breast cancer,the positive rate was 57.14％ (4/7),76.47％ (13/17) and 100％ (18/18) respectively for breast cancer of grade Ⅰ,Ⅱ and Ⅲ.Conclusion Automated DNA imaging cytometry is of some value in analyzing the malignant grade of breast cancer and predicting the prognosis of breast diseases.

Intracranial atherosclerosis is one of the leading causes of ischemic stroke and occurs more commonly in patients of Asian, African or Hispanic origin than in Caucasians. Although the histopathology of intracranial atherosclerotic disease resembles extracranial atherosclerosis, there are some notable differences in the onset and severity of atherosclerosis. Current understanding of intracranial atherosclerotic disease has been advanced by the high-resolution magnetic resonance imaging (HRMRI), a novel emerging imaging technique that can directly visualize the vessel wall pathology. However, the pathological validation of HRMRI signal characteristics remains a key step to depict the plaque components and vulnerability in intracranial atherosclerotic lesions. The purpose of this review is to describe the histological features of intracranial atherosclerosis and to state current evidences regarding the validation of MR vessel wall imaging with histopathology.
Asian Continental Ancestry Group ; Atherosclerosis ; Autopsy ; Hispanic Americans ; Humans ; Intracranial Arteriosclerosis* ; Magnetic Resonance Imaging* ; Microscopy* ; Pathology ; Stroke

AIMTo investigate the changes of large-conductance calcium-activated potassium channels (BKCa, MaxiK) during aging and relations between the changes and blood pressure.

METHODSMale spontaneously hypertensive rats (SHR) aged 9, 15, 21, 27, 33 weeks (the number of each weeks SHR was 4) were selected as hypertension group rats, corresponding gender, weeks and number Wistar-Kyoto rats (WKY) as control group rats. Blood pressure of abdominalis aorta of each weeks SHR and WKY were measured by BL-420F experimental system of biological function. The arteria mesenteric minor (AMM) were isolated in blunt dissection method. The vascular smooth muscle cells (VSMCs) of AMM were isolated with prolease. The potassium current, the current after BKCa were blockaded by Tetraethylammonium (TEA) and the capacitance of membrane (Cm) of VSMCs of AMM were recorded with using whole cell patch clamp, and calculated the BKCa current and the BKCa current density. Probe the correlation of the changes of BKCa current density with MABP during aging.

RESULTSThe potassium current density and BKCa current density of VSMCs of AMM of SHR were decreasing during aging, however, the changes of WKY had no statistically significance (P > 0.05). The BKCa current density was extremely correlative with MABP in SH R (the values of r were -0.7174), in WKY, the BKCa current density was correlative with MAB P r = -0.4832.

CONCLUSIONBKCa current and current density attenuate with aging, the level of blood pressure is response of the attenuated degree. The BKCa current density is extremely correlative with the blood pressure.

Aging ; physiology ; Animals ; Blood Pressure ; physiology ; Cell Membrane ; physiology ; Hypertension ; metabolism ; physiopathology ; Large-Conductance Calcium-Activated Potassium Channel alpha Subunits ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; physiology ; Male ; Membrane Potentials ; physiology ; Mesenteric Arteries ; cytology ; Muscle, Smooth, Vascular ; cytology ; metabolism ; physiology ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; physiology ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY

Objective To establish the enzyme-linked immunosorbent assay (ELISA) for the detection of serum alcohol dehydrogenase (ADH) antibody and evaluate its role in its diagnosis of autoimmune hepatitis( AIH ). Methods The reactivity between yeast ADH and human anti-ADH serum antibody was tested by Western blot analysis. ELISA was established using yeast ADH. The method was applied in serums of 67 AIH patients,94 primary biliary cirrhosis(PBC) patients, 199 chronic hepatitis B (CHB) patients, 132 chronic hepatitis(CHC) patients, 24 alcohol hepatitis disease(ALD) patients, 99 connective tissue disease(CTD) patients and 31 healthy individuals. The positive rate of ADH antibody in the patients and healthy individuals was measured. The χ2 test was used to compare the positive rates. Results The ELISA method for detecting human anti-ADH serum antibody was established successfully and the optimum reaction conditions were defined. Western blot showed that yeast ADH has cross reactivity with human anti-ADH antibody. The positive rate of anti-ADH antibody in the AIH group [59. 7% ,40/67 ] was higher than that in the normal control group(0,χ2 = 31. 271 ,P <0. 05), PBC group (6. 4% ,χ2 =54. 492,P <0. 05), CHB group( 14. 1% ,χ2 =54. 848,P <0. 05) ,CHC group(21.2% ,χ2 = 29.269,P<0.05), ALl) group ( 25. 0% ,χ2 =8.512,P <0.05)and CTD group ( 43. 4% ,χ2 =4.229, P <0. 05). Conclusions Compared with the PBC, CHB, CHC, ALD and CTD group, the anti-ADH antibody positive rate in the serums of AIH was significantly increased. The antibody may be helpful to the diagnosis of autoimmune hepatitis.

Objective:To stablish of chemiluminescent Southern blot detection system for examining HBV DNA replication intermediates in HepG2.2.15,and analyse the inhibition of HBV replication with three kind of drugs with different targets.Methods:The HBV DNA replication intermediates were extracted and analyzed by Southern blot with HBV probe,which(pTHBV1047) was labelling with digoxigenin.The results of the hybridization were detected by chemiluminescent,and the condition of hybridization was optimized.After treated with lamivudine,Bay41-4109,?-Galcer in different concentration,the HBV DNA from the HepG2215 cells were detected with the system.Results:the sensitivity of the system was 1pg of pTHBV1047,and HBV specific positive signals was detected with the DNA from HepG2.2.15.The three kinds of drugs can inhibit the HBV replication obviously with chemiluminescent Southern blot detection system,the IC50 were 1.53?mol/L,0.41?mol/L,0.01?mol/L.Conclusion:The HBV replication intermediates from the cell of HepG2.2.15 can reflect the antiviral effect accurately with different targets drugs and this mothod would be used in the study of Chinese-midicine.

BACKGROUND:Hypertrophic differentiation of chondrocytes is the sign of starting endochondral ossification, and it is also an essential step in endochondral ossification, which is a cascade reaction and difficult to be blocked once started. The end result is the formation of bone structure. RNA interference is a post-transcriptional gene silencing. Relevant studies have shown that the use of RNA interference to block the expression of core binding factorα1 (Cbfα1) can effectively inhibit the formation of heterotopic ossification. OBJECTIVE:To use RNA intereference technology to suppress Cbfα1 expression so as to achieve the purpose of blocking the hypertrophic diferentiation of chondrocytes. METHODs: We constructed an adenovirus containing siRNA against Cbfα1 (Ad-Cbfα1-siRNA). Retinoic acid and interleukin-1α were used to induce hypertrophic differetiation of chondrocytes, and then Ad-Cbfα1-siRNA was utilized to inhibit the hypertrophic differentiation of chondrocytes. Immunohistochemistry method was used to analyze the expression of Cbfα1. RESULTS AND CONCLUSION:After induction with retinoic acid and interleukin-1α, the chondrocytes in the negative control virus group appeared to have hypertrophy and the expression of Cbfα1 was positive. In the Ad-Cbα1-siRNA group, the expression of Cbfα1 was negative. These findings suggest that the inhibition of Cbfα1 by RNA interference can be a powerful way to prevent the hypertrophic differentiation of chondrocytes .