Objective To analyze the composing principles of the prescriptions for Lijie disease by using The Prescription of Traditional Chinese Medicine Dictionary. Methods The prescriptions for Lijie disease in the book of The Prescription of Traditional Chinese Medicine Dictionary were collected. Database was established by using TCM Inheritance Support System (V1.1). Medicine using frequency, composing principles, and composition of new medicine were analyzed through association rules and entropy clustering method. Results Based on the analysis on 187 cases of prescriptions, 42 Chinese herbal medicines were with the frequency of 42 times and Angelicae Sinensis Radix was the core herbal medicine. 69 core combinations with three to four herbal medicines and 6 new prescriptions were obtained through evolution. Conclusion According to the analysis of composing principles and clustering new prescriptions of Lijie disease, the treatment for Lijie disease is toning liver and spleen, and nourishing liver and kidney.

Objective To analyze the protein expression profile of human glomerular mesangial cells (HMCs) under high glucose and to characterize molecular functions and biological processes. Methods HMCs were divided into high glucose cultured group (30 mmol/L) and normal glucose cultured group (5 mmol/L). The total proteins were extracted after culture for 48 hours. The total proteins of the two groups were separated using two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and analyzed using DeCyder 2-D difference analysis software. The differentially expressed proteins were further identified using in-gel digestion with trypsin, of which peptide extracts were prepared for MALDI-TOF-MS analysis. Protein identifications were searched in the NCBI protein database using the Mascot search engine. Results One hundred and forty-seven protein spots whose expression levels were significantly increased or decreased more than 1.5 folds under high glucose were identified. Ninety-six differentially expression protein spots were analyzed by peptide mass fingerprinting and 37 kinds of proteins were identified. The protein spots of phosphatidylethanolamine binding protein 1 (PEBP-1), granulysin,ATP synthase H + transporting mitochondrial FO complex subunit F2 were observed only in high glucose group. The expression of 24 proteins was up-regulated by high glucose, including eosinophil cationic protein, RGS membrane-interacting proteins 16 (MIR16), peptidyl-prolyl cis-trans isomerase, disks large homolog DLG2, breast cancer 2, early onset (BRCA2), Catechol-O-methyltransferase etc. The expression of 5 proteins was down-regulated by high glucose, including O-GlcNAc transferase-interacting protein 106 000 isoform 1, proteasome beta 6 subunit precursor,NEFA-interacting nuclear protein NIP30 etc. Conclusions Expression of 147 proteins in HMCs alters under high glucose. These proteins are involved in the regulation of cytoskeleton, glucose metabolism, cell division, gene transcription, signal transduction, phosphorylation, cell proliferation,apoptosis etc. In-depth analysis of these differentially expressed proteins' function and crosstalk is expected to provide an important experimental basis for clarifying the pathogenesis of diabetic nephropathy.

OBJECTIVETo observe the effects of Porphyromonas gingivalis (P. gingivalis) ATCC 33277 infection on expression of intercellular adhesion molecule-1 (ICAM-1) in rat vascular smooth muscle cells(VSMC).

METHODSAn infection model of rat VSMC invaded by P. gingivalis was established in vitro. The mRNA of ICAM-1 was measured through reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSCompared with the control group, an apparent and statistically significant increase in expression of ICAM-1 mRNA was observed after 8, 16, and 24 h in P. gingivals-infected rat VSMC (P<0.05). The expression reached its peak at 16 h. Statistically significant differences were observed in the 8 h group and in the other two experimental groups (P<0.05).

CONCLUSIONInfection of P. gingivals in rat VSMC can cause increased expression of ICAM-1, which may have an important function in the progression of atherosclerosis.

Objective To evaluate the relationship between the mechanism underlying propofol-induced inhibition of migration of human breast cancer cells and glyc olysis.Methods Human breast cancer cell line MDA-MB-231 cells were inoculated in 12-well culture plates at a density of 3× 105 cells/well.After being cultured for 24 h,the cells were divided into 2 groups (n=12 each) by using a random number table:control group (group C) and propofol group (group P).Propofol at the final concentration of 5 μg/ml was added to the culture medium in group P,and the equal volume of phosphate buffer solution was added to the culture medium in group C.At 6 h of incubation,the culture media were changed to the common culture media containing no drugs,and the cells were then incubated for another 24 h.The culture media were collected for determination of glucose concentrations (by oxidase mnethod) and lactate concentrations (by chemical colorimetry).Glucose consumption and lactate production were calculated according to glucose and lactate concentrations.Cells were collected,the expression of lactate dehydrogenase A was detected by Western blot,and the migration of cells was determined by cell scratch test.Results Compared with group C,the consumption of glucose and production of lactate were significantly decreased,the expression of lactate dehydrogenase A was down-regulated,and the migration rate was decreased in group P (P＜0.05).Conclusion The mechanism by which propofol inhibits migration of human breast cancer cells may be associated with inhibition of glycolysis.

OBJECTIVE: To study the generality of the structure-activity relationships (SARs) of 3-deoxylation and 6-deoxylation of the sugar moiety in SGLT2 inhibitors. METHODS: Based on the earlier study, 3-deoxycanagliflozin (compound 5), 6-deoxyipra-gliflozin (compound 6), 6-deoxyempagliflozin (compound 7) and 3-deoxyempagliflozin(compound 8) were synthesized and evaluated by in vitro hSGLT2 and hSGLT1 inhibitory assay. RESULTS: The deoxylated SGLT2 inhibitors were synthesized from their corresponding aryl halides 9a-9c and 3-/6-deoxylated perbenzylated gluconolactones 11a-11b. In vitro hSGLT2 and hSGLT1 inhibitory assay showed that compounds 5 and 8 almost lost SGLT2 inhibitory activity, while compounds 6 and 7 exhibited similar activities to their corresponding parent compounds. CONCLUSION: It seems a general rule that 6-deoxylation of the sugar moiety in SGLT2 inhibitors has no effect on the SGLT2 inhibitory activity, whereas the effect of 3-deoxylation on SGLT2 inhibitory activity depends on the structures of the specific SGLT2 inhibitors, which does not show a universal rule.

Objective To investigate the effects of angiotensin receptor blocker (ARB)losartan on the glomerular protein expression profile of spontaneous type 2 diabetic KKAy mice by two-dimensional differential gel eleetrophoresis and MALDI-TOF mass spectrometry.Methods 8-week-old spontaneous type 2 diabetic KKAy mice were randomly divided into losartan (10 mg·kg-1·d-1 given in drinking water) treatment group and non-treatment group.Eight-week-old C57BL/6 mice were used as normal control.The glomeruli were separated by magnetic bead perfusion through thoracic aorta at age of 20 weeks,then glomerular protein was extracted.The glomerular protein expression profile was investigated by CyDyes minimal fluorescence labelling,two-dimensional differential gel electrophoresis and MALDI-TOF mass spectrometry.Results KKAy mice developed higher body weight and blood glucose,higher urinary microalbumin creatinine ratio at age of 20 weeks than C57BL/6 mice at the same age (all P＜0.05).Losartan treatment markedly reduced urinary microalbumin creatinine ratio [(539.71 ±100.23)mg/g vs (728±177.19) mg/g],attenuated mesangial expansion and the thickening of glomerular basement membrane,but had no effect on the blood glucose.By DeCyder 2-D differential analysis software,62 protein spots of differential expression were found in glomeruli between losartan treatment and non-treatment KKAy mice at age of 20 weeks.Among them,41 proteins were identified by peptide mass fingerprinting.The expressions of 28 proteins were up-regulated by losartan treatment,including glycerokinase,sulfite oxidase,glycine amidinotransferase,adenosylhomocysteinase,etc.The expressions of 13proteins were down-regulaled by losartan treatment,including 3-mercaptopyruvate sulfurtransferase,ATP synthase subunit d,60 000 heat shock protein,stress-70 protein (alternative name 75 000glucose-regulated protein,GRP75),etc.Six differcntially expressed proteins were found in glomeruli between non-treatment KKAy mice and C57BL/6 mice,and the differential expressions were suppressed by losartan treatment,including dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex,succinyl-CoA ligase (GDP-forming) subunit beta,mitochondrial,ATP synthase subunit d,GRP75,nucleoside diphosphate-linked moiety X motif 19 and seleniumbinding protein 1.Conclusions Losartan significantly reduces the urinary protein excretion rate and renal pathological lesion of spontaneous type 2 diabetic KKAy mice,and suppresses the differential expression of mitochondrial ATP synthase subunit d,GRP75,selenium-binding protein 1,etc in glomeruli.Losartan may play a renoproteetive role by reducing glomerular mitochondrial reactive oxygen species genesis and inhibiting oxidative stress.

Objective The effects of candesartan,an angiotensin Ⅱ type 1 receptor blocker (ARB) were investigated on advanced glycation end-products accumulation and the receptor for AGE (RAGE) expression in type 2 diabetic KK/Ta mouse kidneys.MethodsKK/Ta mice(n=72)were random divided into three groups(n=24) and it was treated with candesartan [4 mg/(kg·d)] or vehicle from 6 or 12 to 28 weeks of age.BALB/c mice(n=24) treated with vehicle were used as controls.Body weight,blood pressure,blood glucose,urinary microalbumin,urinary creatinine and serum creatinine were measured every four weeks.At 28 weeks,renal expressions of carboxymethyllysine and RAGE were evaluated by immunohistochemistry and/or competitive RT-PCR.Results KK/Ta mice developed high body weight,high blood glucose,and high urinary microalbumin/creatinine ratio in KK/Ta mice at 28 weeks of age,and it was significantly higher than that of BALB/c mice [(427.49±89.37)mg/g vs (9.54±3.25)mg/g,P＜0.01 ].Protein and mRNA expressions of RAGE were upregulated in KK/Ta kidneys with increased immunostaining intensities of carboxymethyllysine.Candesartan treatment has markedly reduced urinary microalbumin/creatinine ratio [Early treatment group (32.18±9.41)mg/g,Late treatment group (53.20±7.26)mg/g,P＜0.01 ].Treatment with candesartan down-regulated the protein and mRNA expressions of RAGE and reduced the accumulation of carboxymethyllysine.There were no significant differences between the two treatment groups (from 6 or 12 weeks).ConclusionsThe results suggest that candesartan,an ARB,reduces advanced glycation end-products accumulation and subsequent albuminuria by down-regulating RAGE expression in type 2 diabetic KK/Ta mouse kidneys.

Objective To study the differences in body composition and blood lipids between the pre-and post-menopausal women of Maonan ethnicity , and to explore the correlations between body fat content , fat distribution and blood lipids.Methods Totally 200 Maonan pre-and post-menopausal women were randomly selected from Huanjiang county in Guangxi.Body composition were measured by bioelectrical impedance analysis (BIA), and blood lipids were tested from blood samples .Results Compared with the pre-menopausal women , the visceral fat level (/area ) , waist-hip ratio (WHR), left (/right) lower limbs fat, total cholesterol (TC), triglyceride (TG) and low-density lipoprotein( LDL-C) in post-menopausal women were significantly higher ( P <0.01 ) , and the detection rate of hypercholesterolemia , mixed hyperlipidemia and dyslipidemia in postmenopausal group was also significantly higher ( P<0.01 ) .All the blood lipids were closely related to WHR and visceral fat content (P<0.05 or P<0.01).In addition, TG, high-density lipoprotein ( HDL-C) and LDL-C except TC were significantly correlated to %BF, BMI and subcutaneous fat content ( P<0.05 or P<0.01).Conclusion The accumulation of visceral and abdominal fat in Maonan postmenopausal women is significantly correlated to dyslipidemia .The results may provide references for making preventive program for the Maonan women .

Objective To analyze the risk factors and correlation between clinical indicators and the four main pathological lesions of IgA ne?phropathy in the Oxford classification:mesangial hypercellularity(M0/1),endocapillary proliferation(E0/1),segmental sclerosis or adhesion(S0/1), and tubular atrophy/interstitial fibrosis(T0/1/2). Methods Clinical and pathological data were collected from 514 patients with biopsy?proven IgA nephropathy admitted in our hospital from February 17,2006 to October 11,2011. These patients were all above 18 years old. Cases with sec?ondary causes of mesangial IgA deposition were excluded,such as Henoch?chonlein purpura,ankylosing spondylitis and psoriasis et al. The inde?pendent risk factors affecting the pathological classification were analyzed by Spearman rank correlation analysis and two?category and multi?classi?fication logistic regression using SPSS 17.0 statistical software. Results In 514 IgAN patients,the ratio of males to females was 1.06:1. The aver?age age was 35.70±11.99 years,and the average disease duration was 18.31±30.42 months. M0E0S0T0 was the major pathologic classification of isolated hematuria. Chronic kidney disease(CKD)stage,24 hours proteinuria,albuminuria,urine transferrin and IgG levels were positively corre? lated with M lesion;serum albumin,C3 and PLT showed a negative correlation with M lesion. Twenty four hours proteinuria and blood platelet count were the independent risk factors for M lesion. As shown by stratified analysis ,the proportion of M1 in cases with 24 hours proteinuria≥3.5 g/d is much higher than that in cases with non?nephrotic range proteinuria. Age,systolic blood pressure,uRBC,24 hours proteinuria,albuminuria urine transferrin and IgG levels were positively correlated with E lesion,Duration,serum albumin showed a negative correlation with E lesion. Age and duration of nephritis were independent risk factors for E lesion. 73.3%of patients that above 60 years old showed endothelial proliferation. CKD stage,24 hours proteinuria were positively correlated with S lesion. Age,CKD stage,systolic blood pressure,diastolic blood pressure,C4,TC, LDL?C,CRP,Fib,UA,Cys?C and 24 hours proteinuria,urineβ2?microglobulin,albumin,transferrin and IgG levels were positively associated with T lesion;hemoglobin,serum albumin,serum IgG showed a negative correlation with T lesion. Infection history,high CRP levels,DBP more than 90 mmHg,hypoalbuminemia,high low density lipoproteinemia,and anemia were independent risk factors for T lesion. Conclusion Twenty four hours proteinuria,blood platelet count,age,duration of nephritis,hypoalbuminemia,anemia,hyperlipidemia,DBP≥90 mmHg and high CRP lev?els were risk factors for the Oxford classification of IgA nephropathy. Renal biopsy should be carried out in time to make clear the pathological clas?sification and individual treatment,so as to improve the prognosis.

Objective To evaluate the role of conventional protein kinase Cγ (cPKCγ)/growthassociated protein-43 (GAP-43) signaling pathway in ketamine-induced apoptosis in hippocampal neurons of developing rats in an in vitro experiment.Methods Primarily cultured hippocampal neurons were seeded in culture plates at a density of 1×10.6 cells/ml and divided into 2 groups (n=10 each) using a random number table:control group (C group) and ketamine group (K group).Group C received no treatment.Ketamine was added with the final concentration of 300 μmol/L in group K.At 12 h of culture or incubation,the apoptosis in hippocampal neurons was detected by flow cytometry.The apoptotic rate was calculated.The expression of cPKCγ,GAP-43 and phosphorylated GAP-43 in hippocampal neurons was measured by Western blot.Results Compared with group C,the apoptotic rates of hippocampal neurons were significantly increased,and the expression of cPKCγ,GAP-43 and phosphorylated GAP-43 was down-regulated in group K (P<0.01).Conclusion The mechanism by which ketamine induces apoptosis in hippocampal neurons of developing rats may be related to inhibition of cPKCγ/GAP-43 signaling pathway activation in an in vitro experiment.