Objective To investigate the effects of Hui-hui Gan-song Yin(HGY) on the accumulation of extracellular matrix of rat glomerular mesangial cells(MCs) induced by high glucose.Methods The 40 SD rats were randomly divided into normal control group(distilled water), glurenorm group(10 mg/kg), HGY high-dose group(10 g/kg) and HGY low-dose group(5 g/kg), 10 rats in each group.The rats in each group were treated with corresponding drugs, twice a day.After 3 days, the serum containing each drug were prepared to culture rat MCs in vitro.The MCs were divided into the normal control group( 10% serum of rats in normal control group ) , high glucose group ( 30 mmol/L glucose +10% serum of rats in normal control group), glurenorm group(30 mmol/L glucose+10% serum of rats in glurenorm group), HGY high-dose group(30 mmol/L glucose+10% serum of rats in HGY high-dose group) and HGY low-dose group(30 mmol/L glucose+10% serum of rats in HGY low-dose group).The fibronectin(FN), ColⅠand ColⅣ levels were detected by Western blot.Results Compared with normal control group, the expression of FN, ColⅠand ColⅣ in high glucose group increased(P<0.01).The HCY suppressed the protein expression of FN, ColⅠand ColⅣ significantly(P<0.05).Conclusion The serum containing HGY could suppressed protein expression of FN , ColⅠand ColⅣ and inhibit the accumulation of extracellular matrix of MCs induced by high glucose, which could protect glomerulus and delay the development of diabetic nephropathy.

Aim To observe the effects of theaflavin on intracellular calcium concentration(i) level in rat ventricular myocytes and discuss the possible mechanisms.Methods The effects of theaflavin oni were investigated in rat ventricular myocytes.i was detected by laser confocal microscopy and represented by relative fluorescent intensity(FI-FI0)/FI0,%;FI0:control;FI:administration of drugs).Results ① Theaflavin(10,20,40 ?mol?L-1) had no effect on the i of ventricular myocytes in normal Tyrode′s solution.However,it reduced the i of ventricular myocytes in simulated ischemia solution in a concentration-dependent manner.② Pretreatment with Bay k8644(0.5 ?mol?L-1) mostly abolished the effects of theaflavin(20 ?mol?L-1) in simulated ischemia solution.③ Theaflavin(20 ?mol?L-1) markedly inhibited the low concentration of ryanodine-induced i increase in Ca2+-free Tyrode′s solution.④ When the propagating waves of elevated i(Ca2+ waves) were produced by increasing extracellular Ca2+ concentration from 1 mmol?L-1to 10 mmol?L-1,theaflavin(20 ?mol?L-1) could block the propagating waves of elevated i(Ca2+ waves),reduce the frequency and duration of propagating waves,and reduce i as well.Conclusion Theaflavin may reduce the i in isolated rat ventricular myocytes via inhibiting Ca2+ influx by voltage-dependent Ca2+ channel and alleviating Ca2+ release from sarcoplasmic reticulum(SR).

Acoustic Stimulation ; Animals ; Corpus Striatum ; physiology ; Estrogens ; metabolism ; Estrus ; physiology ; Evoked Potentials, Auditory ; physiology ; Female ; Rats ; Rats, Sprague-Dawley

Adult ; Chemical and Drug Induced Liver Injury ; etiology ; Dinitrobenzenes ; poisoning ; Humans ; Male ; Occupational Diseases ; chemically induced

The intracellular hepatitis B surface antigen (HBsAg) content per cell was increased by 7.2-fold in the culture with 1.5% dimethyl sulfoxide (DMSO) compared with that in the control without DMSO, while the extracellular HBsAg production and specific productivity were only improved by 70% and 3.2-fold, respectively. Electron microscope has been employed to reveal large dilated structures within recombinant CHO cells in the presence DMSO. The dilated structures have a distribution within whole cytoplasm, and some dilated areas were engulfed in the nucleus. These large, dilated structures were not observed in the control. Immunogold labeling was used to discover the accumulated HBsAg was localized within these dilated areas, and some HBsAg-specific labels were detected in the nucleus membrane, owing to the encroachment of the dilated areas upon nucleus. The result could help to reveal the mechanism of intracellular HBsAg accumulation in the presence of DMSO.

A study on the microbial transformation of glycyrrhetinic acid (GA) was conducted by a fungus, Cunninghamella blakesleeana CGMCC 3.970 systematically. After incubation with the cell cultures of C. blakesleeana CGMCC 3.970 at 25 degrees C for 7 days on a rotary shaker operating at 135 r x min(-1), GA was converted into one major product and five minor products. The products were extracted and purified by solvent extraction, macroporous adsorbent resin, silica gel column chromatography, and semi-preparative RP-HPLC chromatography. Their structures were identified as 3-oxo-15α-hydroxy-18β-glycyrrhetinic acid(1), 3-oxo-15β-hydroxy-18β-glycyrrhetinic acid (2), 7β,15α-dihydroxy-18β-glycyrrhetinic acid (3), 3-oxo-7β, 15α-dihydroxy-18β-glycyrrhetinic acid (4), 7β-hydroxy-18β-glycyrrhetinic acid(5) and 15α-hydroxy-18β-glycyrrhetinic acid(6) by the analyses of MS, 1H-NMR and 13C-NMR spectroscopic data respectively. Among them, 2 was a new compound. These results suggest that C. blakesleeana CGMCC 3.970 has the capability of selective ketonization and hydroxylation for GA. [Key words] glycyrrhetinic acid; Cunninghamella blakesleeana CGMCC 3. 970; microbial transformation
Biotransformation ; Cunninghamella ; metabolism ; Glycyrrhetinic Acid ; analogs & derivatives ; chemistry ; metabolism ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization

Objective: To explore aspirin resistance (AR) phenomenon in patients with coronary artery disease (CAD) for secondary prevention and to study the relationships between AR and COX1, COX2, TBXA2R gene polymorphisms. Methods: A total of 2881 CAD patients taken aspirin (100 mg/day) in 7 consecutive days were enrolled. Among them, 2 groups were established as AR group, n=166 and Control group, n=200 aspirin sensitive patients. Platelet aggregation function was induced by arachidonic acid (AA), COX1, COX2 and TBXA2R gene polymorphisms were examined by polymerase chain reaction-restricted fragment length polymorphisms (PCR-RFLP) method. Results: The occurrence rate of AR was 5.76% (166/2881). There were 8 tagSNPs locus in 3 genes as in COX1:(rs3842788), (rs4273915), (rs7866582); in: COX2 (rs3218625); in TBXA2R: (rs2238630), (rs2238631), (rs2238633), (rs3786989). The frequencies of wild type, heterozygous genotype and homozygous genotype were similar between 2 groups. Conclusion: The incidence rate of AR is not high in CHD patients with regular aspirin medication; single nucleotide gene polymorphisms of COX1, COX2 and TBXA2R have no obvious correlation to AR.

Objective To investigate the neural circuit of inhibitory control in late-onset depressed patients(LOD) by functional magnetic resonance imaging(fMRI). Methods Fourteen late-onset depressed patients (LOD group) and thirteen elderly healthy subjects( control group) were recruited. The two groups were age, gender, and education matched. All the subjects performed a visual Go/Nogo task during the fMRI scan. Erect or inverted isosceles triangular figures were used for stimuli. The two groups were instructed to press a button as quickly and correctly as possible when the erect triangular figures(Go) were presented, but not to response when the inverted triangular figures(Nogo) were presented. The differences of brain activation between the two groups were compared. Results ( 1 ) During Go trials, there were no significant differences in reaction time and hit rate between the two groups (P ＞ 0.05 ). During Nogo trials, however, the late-onset depressed patients showed much higher false alarm rate(0.09 ±0.06) compared with control group(0.04 ±0.02) (P＜0.05＝. (2) During Go trials , LOD group showed significantly greater activity in left postcentral gyrus, left inferior parietal lobule, right precentral gyrus, left paracentral lobule, right inferior parietal lobule, right anterior cingulate cortex, left middle frontal gyrus, right middle frontal gyrus, right superior frontal gyrus compared with the control group. Whereas during Nogo trials, LOD group exhibited greater activity in left inferior parietal lobule and left middle frontal gyrus compared with the control group. Conclusion This study suggests that inhibitory control dysfunction in late-onset depressed patients may be closely related to frontostriatal circuit impairment. Over activation in left middle frontal gyrus, right middle frontal gyrus and right anterior cingulate cortex may contribute to the pathogenesis of late-onset depression.

Objective To investigate the effect of transient receptor potential M4 (TRPM4) on autonomous regulation disorder of cerebral blood flow following subarachnoid hemorrhage (SAH) in rats.Methods A total of 120 clean grade male SD rats were selected.They were divided into sham operation,SAH,negative control,and treatment groups according to the random number table.The dead rats were excluded.A SAH model was induced by using the suprasellar cistern injection method with a stereotaxic apparatus.Isotonic saline 0.2 ml was injected into the rats of the sham operation group and negative control group respectively,and autologous tail arterial blood 0.2 ml was injected into the rats of the SAH group and the treatment group respectively.The isotonic saline solution was continuously pumped into lateral ventricle of rats via implantable micro-pump in the sham operation group and the SAH group respectively,and the concentration of 0.03 mol/L of TRPM4 blocking agent was continuously pumped into the lateral ventricles of rats in the control group and the treatment group respectively.The 4 groups of rats received the regional cerebral blood flow and whole cerebral blood flow detection on day 3,5,and 7,respectively.Results One hundred and six (88.3%) of the 120 SD rats survived to the time point of study,data analyses were performed in the 4 groups (with 21 rats in each group) respectively (n=7 in each time point).There were significant differences in cerebral cortex local and whole cerebral blood flow at day 3,5,and 7 in the sham operation,SAH and negative control groups (all P<0.05).Cerebral cortex local cerebral blood flow (141±18,148±24,and 168±19 PU,respectively at day 3,5,and 7) and whole cerebral blood flow (93±5,85±5,and 85±6 ml/[100 g·min],respectively at day 3,5,and 7 in the SAH group) were decreased significantly compared with the sham operation group (cortex local cerebral blood flow:235±17,220±24,and 224±20 PU),whole cerebral blood flow (141±10,147±8,and 143±8 ml/[100 g·min]),all P<0.05).Cerebral cortex local and whole cerebral blood flow (cortical local cerebral blood flow:183±26,173±26,and 187±15 PU,whole brain:114±10,104±9,and 119±5 ml/(100 g·min) in the treatment group were significantly increased compared with the SAH group (all P<0.05).Conclusion TRPM4 has an obvious effect on improving the autonomous regulation disorder of cerebral blood flow after SAH.

Aim To construct a eukaryotic expression vector of human MCHR2 and to analyze the stable expression and signal transduction pathways of MCHR2 in CHO cells. Methods The full-length MCHR2 cDNA fragment was amplified by PCR from the human fetal brain cDNA library and inserted into eukaryotic expres-sion vector pcDNA3.1(+), resulted in the recombinant expression vector pcDNA3.1-MCHR2. The recombinant plasmid was confirmed by the restriction enzymatic digestion and DNA sequencing analysis, and then transfected into CHO cells by lipofectamine. A stably-transfected cell line was obtained by the dominant G418 selection, and the expression of the MCHR2 gene on transcription and translation levels were identified by RT-PCR and Western blot. Signal transduction pathways mediated by MCHR2 were analyzed by measurement of intracellular cAMP and calcium. Results The eukaryotic expression vector pcDNA3.1-MCHR2 was successfully constructed and the MCHR2 gene was stably transfected into CHO cells. A stably-transfected cell line was established and the MCHR2 gene was efficiently transcribed and translated. MCH stimulation had no effect on the production of cAMP, however, it could induce a clear and transient increase of intracellular calcium, suggesting that MCHR2 was only coupled to Gq protein. Conclusion The stable expression of MCHR2 and analysis of its signal transduction pathways in CHO cell line provided a solid experimental foundation for further studies on the function of the MCHR2 gene in vitro.