Expression, purification, and functional verification of recombinant human glycoprotein hormone beta 5/alpha 2 fusion protein in CHO-S cells / 中国药理学通报

Aim To express and purify recombinant hCGH-CTP fusion protein in high-density suspension culture of Chinese hamster ovary cells (CHO-S), and to verify the lipid accumulation effect of rhCGH-CTP on 3T3-L1 mature adipocytes. Methods The recombinant protein expression vector (pcDNA3. 1-rhCGH-CTP) was constructed, achieved by fusing the human glycoprotein hormone beta 5/alpha 2 cDNA with CTP Linker. The expression plasmid was transiently transfected into the suspended CHO-S to express rhCGH-CTP protein and then purified, and the protein biological activity was verified. Intervention with 3T3-L1 mature adipocyte cells for 24 h was performed to detect the changes of intracellular triglyceride (TG) level. Results Western blot results showed that rhCGH-CTP protein was successfully expressed in CHO-S cells, and the yield was up to 715. 4 mg • L~ . The secreted protein was purified by AKTA pure system with higher purity that was up to 90% as identified by SDS-PAGE. In addition, the intracellular cAMP content of mature adipocytes with high expression of TSHR gene significantly increased after intervention with different concentrations of rhCGH-CTP protein by ELISA kit, indicating that rhCGH-CTP protein had biological activity. Oil red 0 staining showed that compared with the control group, the lipid content of mature adipocytes in the intervention groups with different concentrations of rhCGH-CTP protein significantly decreased (P < 0. 05) . Conclusions The rhCGH-CTP protein has been successfully expressed and purified with biological activity, and effectively reduce TG. This research provides an important theoretical basis for further revealing the physiological role of CGH protein and its potential application in clinical practice.

Chemical constituents from the fruits of Alpinia oxyphylla and their neuroprotective effects / 药学学报

Fourteen compounds were isolated from the ethyl acetate fraction of 90% EtOH extracts of the dried fruits of Alpinia oxyphylla by silica gel, MCI, RP-18, Sephadex LH-20, TLC and semi-preparative HPLC column chromatography. Their structures were identified by HR-ESI-MS, UV, IR, NMR, ECD and X ray single crystal diffraction spectroscopic data as: (2R,5R,7R,10S)-2,7-dihydroxyl-eudesmane-3(4),11(12)-diene (1), α-rotunol (2), diketone I (3), (1S,4S,5R,7S)-1-hydroxyl-eremophilane-9(10),11(12)-diene-8-one (4), cyperusol A1 (5), (6R,9S,10S)-10-hydroxyl-11,12,13-trinor-cadinane-4(5)-ene-3-one (6), (2E,4E)-6-hydroxy-2,6-dimethylhepta-2,4-dienal (7), oxyphyllacinol (8), yakuchinone A (9), (5R)-5-hydroxy-1,7-diphenylhept-3-heptanone (10), (5S)-5-hydroxy-7-(4″-hydroxyphenyl)-1-phenylhept-3-heptanone (11), (5S)-5-hydroxy-7-(4″-hydroxyl-3″-methoxyphenyl)-1-phenyl-3-heptanone (12), 7-(4″-hydroxy-3″-methoxyphenyl)-1-phenyl-3,5-heptadione (13), bis-(2-ethylhexyl) terephthalate (14). Compounds 1-6 were sesquiterpenoids in which compound 1 is a new eudesmane sesquiterpenoid and compound 7 was a monoterpenoid. Compounds 8-13 were diarylheptanoids, and compounds 2-6 and 14 were isolated from A.oxyphylla for the first time. The experiments on H2O2 induced SH-SY5Y cells showed that compounds 2, 6, 7, 12 and 13 had neuroprotective effects at low and medium concentrations. In particular, compound 6 showed obvious neuroprotective effect at low, medium and high concentrations whose cell viability was higher than that of the positive control.

Effect of a novel phosphodiesterase 5 inhibitor, CPD1, on renal interstitial fibrosis after unilateral renal ischemia-reperfusion injury / 生理学报

This study was designed to evaluate the protective effect of CPD1, a novel phosphodiesterase 5 inhibitor, on renal interstitial fibrosis after unilateral renal ischemia-reperfusion injury (UIRI). Male BALB/c mice were subjected to UIRI, and treated with CPD1 once daily (i.g, 5 mg/kg). Contralateral nephrectomy was performed on day 10 after UIRI, and the UIRI kidneys were harvested on day 11. Hematoxylin-eosin (HE), Masson trichrome and Sirius Red staining methods were used to observe the renal tissue structural lesions and fibrosis. Immunohistochemical staining and Western blot were used to detect the expression of proteins related to fibrosis. HE, Sirius Red and Masson trichrome staining showed that CPD1-treated UIRI mice had lower extent of tubular epithelial cell injury and deposition of extracellular matrix (ECM) in renal interstitium compared with those in the fibrotic mouse kidneys. The results from immunohistochemistry and Western blot assay indicated significantly decreased protein expressions of type I collagen, fibronectin, plasminogen activator inhibitor-1 (PAI-1) and α-smooth muscle actin (α-SMA) after CPD1 treatment. In addition, CPD1 dose-dependently inhibited the expression of ECM-related proteins induced by transforming growth factor β1 (TGF-β1) in normal rat kidney interstitial fibroblasts (NRK-49F) and human renal tubular epithelial cell line (HK-2). In summary, the novel PDE inhibitor, CPD1, displays strong protective effects against UIRI and fibrosis by suppressing TGF-β signaling pathway and regulating the balance between ECM synthesis and degradation through PAI-1.

Effect of a novel phosphodiesterase type 5 inhibitor, CPD1, on paraquat-induced lung fibrosis in rats / 中国药理学通报

Aim To investigate the effects of CPD1, a novel phosphodiesterase 5 inhibitor, on lung pathological phenotype and epithelial-mesenchymal transition of alveolar epithelial cells in lung fibrosis model rats caused by paraquat (PQ). Methods Lung fibrosis model was constructed by a single intraperitoneal injection of PQ (30 mg·kg

Effect of a novel phosphodiesterase type 5 inhibitor,CPD1,on unilateral renal interstitial fibrosis caused by ureteric obstruction in mice / 中国药理学通报

Aim To investigate the effects of CPD1,a novel phosphodiesterase 5 inhibitor,on renal pathological phenotype and fibrotic protein expression in renal fibrosis model mice. Methods Male C57BL/6 J mice were divided into three groups randomly(sham group,UUO group and UUO+CPD1 group). Unilateral ureteric obstruction model was constructed by surgery,and CPD1(5 mg·kg-1·d-1)was administered by intragastric administration two hours after the modeling for seven days. HE and Sirius Red staining were used to observe the distribution of tissue structural lesions and fibrosis. Immunohistochemical staining and Western blot were used to detect the expression of fibronectin(FN),α-SMA,collagen-I and kidney injury molecule-1(Kim-1). Results Compared with sham operation group,the renal tubules of mice were dilated and accompanied by a large amount of inflammatory infiltration. Moreover,the expressions of FN,α-SMA,collagen-I and Kim-1 proteins increased significantly(P<0.05)in UUO group. CPD1 treatment improved the kidney structure and decreased the expression of collagen fibers. Furthermore,CPD1 inhibited the expression of FN,α-SMA,collagen-I and Kim-1 markedly(P<0.05). Conclusions Phosphodiesterase 5 inhibitor CPD1 alleviates the progression of renal fibrosis induced by unilateral ureteral obstruction through down-regulating ECM deposition in the extracellular matrix and expression of Kim-1. The specific mechanism remains to be further studied.

Effect of a novel phosphodiesterase type 5 inhibitor, CPD1, on carbon tetrachloride-induced liver fibrosis in mice / 中国药理学通报

Aim To investigate the effects of CPD1, a novel phosphodiesterase 5 inhibitor, on liver pathological phenotype and hepatic stellate cells (HSCs) activation in hepatic fibrosis model mice caused by carbon tetrachloride ( CCl

Expression, purification and functional verification of recombinant human α-galactosidase A in suspension CHO-S / 中国药理学通报

Aim To express and purify rhα-Gal A with a 6 X His tag via using a serum-free expression system in high-density suspension culture of Chinese hamster ovary cells ( CHO-S) , and to verify the scavenging effect of rhα-Gal A on globular trisaccharide ceramide (Gb3 or GL3) . Methods The construction of recombinant protein expression vector, pcDNA4-GLA, was achieved by fusing the human α-galactosidase cDNA, gla, with 6 X His tag and artificial DNA synthesis. The expression plasmid was transfected into the suspended CHO-S to express rhα-Gal A and then purified. Following this procedure, we determined rhα-Gal A's expression, the enzymatic activity, and the glycosylation of the recombinant enzyme. Co-incubation with cultured cells was performed to examine whether rhα-Gal A could be taken up into the cells and effectively remove Gb3 substrates. Results rhα-Gal A was successfully expressed and purified after transiently transfecting pcDNA4-GLA into the suspended CHO-S, and the yield was up to (100 ±20. 6) mg • L

Effect of menthol, a transient receptor potential melastatin-8 channel activator, on pulmonary arterial hypertensive rats / 中国药理学通报

Aim To investigate the effects of menthol, a transient receptor potential melastatin-8 channel activator, on treating pulmonary arterial hypertension (PAH) in PAH model rats caused by monocrotaline (MCT). Methods Male Sprague-Dawley rats were divided into six groups randomly (control group, MCT group, MCT + menthol 1 mg • kg

Effect of FKBP38 protein on endometrial precancerous lesions and mechanism / 中国药理学通报

Aim To build the model of the gene FKBP38(FK506 binding protein 38)conditional knock out in uterus and then investigate the effect on endometrial precancerous lesions and the underlying mechanism.Methods Transgenic mice whose FKBP38 gene was flanked with loxP were constructed by embryo microinjection. The conditional knockout of FKBP38 was obtained by breeding mice harboring two loxP sites in FKBP38(FKBP38

Amygdalin Ameliorates Liver Fibrosis through Inhibiting Activation of TGF-β/Smad Signaling / 中国结合医学杂志

OBJECTIVE@#To observe the effect of amygdalin on liver fibrosis in a liver fibrosis mouse model, and the underlying mechanisms were partly dissected in vivo and in vitro.@*METHODS@#Thirty-two male mice were randomly divided into 4 groups, including control, model, low- and high-dose amygdalin-treated groups, 8 mice in each group. Except the control group, mice in the other groups were injected intraperitoneally with 10% carbon tetrachloride (CCl4)-olive oil solution 3 times a week for 6 weeks to induce liver fibrosis. At the first 3 weeks, amygdalin (1.35 and 2.7 mg/kg body weight) were administered by gavage once a day. Mice in the control group received equal quantities of subcutaneous olive oil and intragastric water from the fourth week. At the end of 6 weeks, liver tissue samples were harvested to detect the content of hydroxyproline (Hyp). Hematoxylin and eosin and Sirius red staining were used to observe the inflammation and fibrosis of liver tissue. The expressions of collagen I (Col-I), alpha-smooth muscle actin (α-SMA), CD31 and transforming growth factor β (TGF-β)/Smad signaling pathway were observed by immunohistochemistry, quantitative real-time polymerase chain reaction and Western blot, respectively. The activation models of hepatic stellate cells, JS-1 and LX-2 cells induced by TGF-β1 were used in vitro with or without different concentrations of amygdalin (0.1, 1, 10 µmol/L). LSECs. The effect of different concentrations of amygdalin on the expressions of liver sinusoidal endothelial cells (LSECs) dedifferentiation markers CD31 and CD44 were observed.@*RESULTS@#High-dose of amygdalin significantly reduced the Hyp content and percentage of collagen positive area, and decreased the mRNA and protein expressions of Col-I, α-SMA, CD31 and p-Smad2/3 in liver tissues of mice compared to the model group (P<0.01). Amygdalin down-regulated the expressions of Col-I and α-SMA in JS-1 and LX-2 cells, and TGFβ R1, TGFβ R2 and p-Smad2/3 in LX-2 cells compared to the model group (P<0.05 or P<0.01). Moreover, 1 and 10 µmol/L amygdalin inhibited the mRNA and protein expressions of CD31 in LSECs and increased CD44 expression compared to the model group (P<0.05 or P<0.01).@*CONCLUSIONS@#Amygdalin can dramatically alleviate liver fibrosis induced by CCl4 in mice and inhibit TGF-β/Smad signaling pathway, consequently suppressing HSCs activation and LSECs dedifferentiation to improve angiogenesis.

Effect of FKBP38 expression on occurrence and development of nonalcoholic fatty liver disease in mice / 中国药理学通报

Aim To investigate the effects of FKBP38 gene on nonalcoholic fatty liver disease ( NAFLD ) model induced by methionine and choline deficiency j J diet (MCD) in mice.Methods The mutant model of hepatocellular specific deletion of FKBP38 gene was successfully established.The mice were divided into wild-type group ( WT) and homologous knockout group (L-FKBP38 ).Mice were fed with MCD for four weeks to construct NAFLD model.Liver injury was e- valuated by the contents of alanine transaminase (ALT), aspartate transaminase (AST) in the serum samples.We also performed HE staining, examined lipid accumulation by triglyceride (TG) and total cholesterol (CHO) and oil red staining, as well as macrophage infiltration by F4/80 immunohistochemical stai-ning of the liver sections.Fatty acid metabolism-relat ed genes were quantifier] by Quantitative Real-time PCR assays.Results Comparer] with WT group, the levels of ALT, AST, TG and CHO in serum signifi- eantly inereased ( P < 0.05 ) ; liver damage , lipid ac- eumulation, and maerophage infiltration were markedly more severe, and the expressions of fatty aeirl oxidation related genes PPARa, ACOX-1 , CPT-la and SIRT3 markedly rleereaserl ( P < 0.05) in the liver samples of L-FKBP38 group.Conclusions Hepatocellular speeifie deletion of FKBP38 intensifies lipid accumula- tion by inhibiting fatty aeid oxidation in the liver, thus exaeerbating nonaleoholie fatty liver disease.

Effect of astragaloside IV on the expression of NOD-like receptor protein 3 inflammasome in neonatal rats with hypoxic-ischemic brain damage / 中国当代儿科杂志

OBJECTIVE@#To study the effect of astragaloside IV (AS-IV) on NOD-like receptor protein 3 (NLRP3) inflammasome in neonatal rats with hypoxic-ischemic brain damage (HIBD).@*METHODS@#A total of 24 Sprague-Dawley rats, aged 7 days, were randomly divided into a sham-operation group, an HIBD group, and an AS-IV treatment group, with 8 rats in each group. After 24 hours of modeling, brain tissue was collected for hematoxylin-eosin staining, yo-PRO-1 staining, and EthD-2 immunofluorescent staining in order to observe the cerebral protection effect of AS-IV in vivo. HT22 cells were used to prepare a model of oxygen-glycogen deprivation (OGD), and a concentration gradient (50-400 μmol/L) was established for AS-IV. CCK-8 assay was used to measure the viability of HT22 cells. RT-PCR and Western blot were used to observe the effect of different concentrations of AS-IV on the mRNA and protein expression of NLRP3, gasdermin D (GSDMD), caspase-1, and interleukin-1β (IL-1β).@*RESULTS@#Yo-Pro-1 and EthD-2 staining showed that compared with the sham-operation group, the HIBD group had an increase in pyroptotic cells with a small number of necrotic cells, and the AS-IV group had reductions in both pyroptotic and necrotic cells. Compared with the sham-operation group, the HIBD group had significantly higher protein expression levels of NLRP3, IL-1β, caspase-1, and GSDMD (@*CONCLUSIONS@#AS-IV may alleviate HIBD in neonatal rats by inhibiting the expression of NLRP3, GSDMD, caspase-1, and IL-1β.

Effect of a novel phosphodiesterase type 5 inhibitor, CPD1, on vasoconstriction in rats with pulmonary arterial hypertension / 中国药理学通报

Aim To investigate the effect of CPD1 , a novel phosphodiesterase 5 inhibitor, on contractile ten- sion of pulmonary artery and aorta in rats with pulmonary arterial hypertension ( PAH ) .Methods MCT- induced PAH was generated by a single intraperitoneal injection of MCT(50 mg • kg"1) in rats.Seven days after MCT injection, the rats were treated with CPD1 ( 10 mg • kg-1 • d"1) for 14 days.The tension of vascular rings was examined in MCT-induced PAH rats.Results MCT treated rats exhibited profound PAH when examined 3 weeks after injection.In contrast, gavage administration of CPD1 led to significant decrease in the right ventricle systolic pressure ( RVSP) and right ventricular mass index (RVMI), as well as MCT-induced pulmonary arterial wall thinning and enlarged lumen, indicating that CPD1 inhibited the de- velopment of PAH.Cavage administration of CPD1 also reduced phenylephrine and endothelin-1-induced pulmonary artery contraction and aorta contraction in MCT-treated rats.Conclusions Treatment with CPD1 attenuates vascular reactivity, lessens vascular smooth muscle cell proliferation and remodeling, and inhibits PAH via inhibition of non-voltage dependent Ca2∗ channels in normal and PAH rats.

Impact of autophagy gene Atg5 deficiency on sperm maturation / 中国药理学通报

Aim To investigate the role of autophagy core protein Atg5 in maintaining epididymal physiological functions and sperm maturation. Methods The Atg5 conditional knockout mouse model of principle cells in the 4 - 5 segments of the epididymal caput was constructed by Cre/LoxP system. The mice were divided into three groups according to genotype: Atg5v(, Atg5u~ and Atg5~/~ . The pregnancy rate and litter sizes were recorded by mating experiments at the age of 8 weeks-old. Sperm motility, sperm counts were evaluated with computer-aided sperm analysis ( CASA) after 14 weeks of feeding. HE staining was conducted to observe the morphology of the epididymal caput. Western blot and immunofluorescence technologies were applied to verify the expression level of Atg5 and the impediment of autophagy after Atg5 conditional knock out. Results After tissue specific knockout of Atg5 in the 4-5 epididymal principle cells, the autophagy marker proteins, LC3- I and p62, were accumulated and autophagy was successfully blocked. There was no significant difference in the morphology of epididymal tissues of the three genotypes of mice, nor any statistically significant difference in sperm motility, sperm count, litter size and pregnancy rate. Conclusions Under normal conditions without external challenge, Atg5 conditional knockout led to autophagy blocking of epididymal caput, but had no effect on mouse epididymal function and sperm maturation process.

Research Progress on Mechanism of Yiguanjian Intervention in Liver Cirrhosis / 中国实验方剂学杂志

Liver cirrhosis caused by the repeated action of one or more causes is a pathological stage characterized by diffuse fibrosis of the liver parenchyma， formation of false lobules and regenerative nodules， portal hypertension which caused by abnormal blood vessels inside and outside the liver. The progression of cirrhosis to decompensation is characterized by severe liver damage， with ascites， gastroesophageal varices bleeding， hepatic encephalopathy and other complications， and most of the treatments are symptomatic， with high mortality and poor prognosis. At present， the traditional Chinese medicine treatment of decompensated cirrhosis can not only effectively improve liver function， but also significantly improve the 5-year survival rate of patients， which suggests that Chinese medicine has potential advantages in preventing and treating end-stage liver disease， and promoting liver cirrhosis tissue reconstruction. Modern Chinese medicine doctors believe that liver cirrhosis is mainly caused by Qi Yin deficiency （liver， spleen， kidney），internal invasion of damp-heat epidemic toxin， and collateral stasis. Deficiency of liver and kidney Yin is a common symptom of decompensated cirrhosis. Yiguanjian， one of Kidney-Nourishing and Liver-Replenishing decoction in traditional Chinese medicine ， is the representative prescription for modern clinical treatment of chronic liver disease with "deficiency of liver and kidney Yin" syndrome. Yiguanjian， created by WEI Yu-zhen （WEI Zhi-xiu）in the Qing Dynasty， Contained in "Xu Ming Yi Lei An ". Clinical studies show that Yiguanjian can effectively improve liver function in patients with liver cirrhosis， promote ascites resolution， and reduce the occurrence of hepatic encephalopathy and other related complications. Experimental research has suggested that Yiguanjian has the characteristics of multi-path， multi-level， and multi-target comprehensive regulation. The mechanism of prevention and treatment of liver cirrhosis may be mainly related to anti-oxidative stress， improving liver inflammation， improving liver cell biosynthesis， inhibiting hepatic stellate cell activation， reducing collagen deposition， improving sinusoidal vascularization and promoting liver cell regeneration. This paper reviews the progress of clinical and experimental research of Yiguanjian in the treatment of liver cirrhosis in the past 5 years， to provide some references for the clinical application and in-depth study of Yiguanjian.

Value of absolute counts of lymphocyte subsets in the early prediction of refractory Mycoplasma pneumoniae pneumonia in children / 中国当代儿科杂志

OBJECTIVE@#To study the value of absolute counts of lymphocyte subsets in the early prediction of refractory Mycoplasma pneumoniae pneumonia (RMPP) in children.@*METHODS@#A retrospective analysis was performed for the clinical data of 244 children with Mycoplasma pneumoniae pneumonia (MPP). Among these children, 166 had MPP, and 58 had RMPP. The two groups were compared in terms of clinical features and laboratory markers such as lymphocyte subsets, lactate dehydrogenase, C-reactive protein, procalcitonin and immunoglobulin E (IgE). The receiver operating characteristic (ROC) curve was used to evaluate the specific indices for predicting RMMP.@*RESULTS@#There were significant differences between the two groups in the absolute counts of CD3, CD4, CD19, and CD56 lymphocytes and the serum levels of lactate dehydrogenase, C-reactive protein, and IgE (P<0.05). The ROC curve analysis showed that the absolute counts of CD3, CD4 and CD19 lymphocytes had an area under the ROC curve (AUC) of 0.866, 0.900 and 0.842 respectively in the differential diagnosis of RMPP and MPP, with a sensitivity of 86%, 90% and 82% respectively and a specificity of 75%, 70% and 80% respectively.@*CONCLUSIONS@#The absolute counts of CD3, CD4 and CD19 lymphocytes can be used to predict RMPP in children.

Evaluation on safety of puerarin injection in clinical use / 中国中药杂志

Puerarin injection is commonly used in clinical treatment of coronary heart disease, angina pectoris, retinal artery, vein occlusion, sudden deafness and so on. This paper is aimed to evaluate the safety of puerarin injection in clinical use and explore the related factors that may cause its adverse reactions (ADRs), so as to find the warning signal of safety medication in time, put forward early warning, make early judgment and treatment, and ensure the safety of drug use. By strengthening surveillance, the best medication plan was established to prevent the occurrence of adverse reactions of puerarin injection and enhance people's awareness on the safety of puerarin injection. Database were searched to collect literature related to ADRs of puerarin injection. The data were extracted and analyzed by decision tree with treeage software and ² test was used to verify the data. A total of 62 papers involving 129 cases were included. The results showed that ADRs occurred mostly in patients aged 50-79 years, with the immune system and blood system accounting for the majority (88.3%), and ADRs occurred mostly 48 h after drug administration (61.1%). The severity of ADRs was not related to the dosage of puerarin, but it was related to the choice of the infusion solvent. In puerarin injection, most of the ADRs were moderate or severe (64.3%), 13 out of 129 cases were of death. Therefore, the indications and methods of use should be strictly controlled, and the allergic history of patients should be carefully questioned before medication to strengthen the monitoring of drug use.

The Clinical Evaluation of Danning Tablets on Regulating Gut Flora Displacement to Eliminate Biliary Inflammation and Prevent Gallstone Formation / 昆明医科大学学报

Objective To explore the mechanism of Danning Tablet on intestinal flora migration to eliminate biliary inflammation and prevent gallstone formation. Methods 104 cases of cholelithiasis admitted to the First Affiliated Hospital of Xi'an Jiaotong University School of Medicine were treated by choledocholithiasis but the master tube was cut and T tube drainage was performed from January 2014 to December 2016. Within a month did not take gallbladder drugs. Randomly divided into the Danning group and the control group, Danning group in addition to conventional treatment of biliary tract after surgery, in the first 2 days after starting to drink liquid according to the instruction manual dose of oral Danning tablets;the control group was given only biliary surgery After routine care, avoid taking gall bladder drugs. All patients were collected on the 7th day and the 14 th day after operation, about 100mL fasting T-tube was collected. Pay attention to the day before the collection of bile T tube pre-clamp to prevent the drainage of T tube affect the bile enterohepatic circulation. Detection of bile composition in the flora, observed before and after the flora differences, long-term follow-up of cholelithiasis recurrence. Results:The changes of the components before and after treatment:On the seventh day, the two groups of HS and UCB％were higher than the control group, the difference was statistically significant (<0.05) . On the 14th day in the treatment group, the three items of TBS, UCB％and HS were higher than the control group, but CH, LI and Z were lower than the control group, the difference was statistically significant ( <0.05) . The trend of stone formation before and after treatment:Compared with UCB％in control group, there was no significant change in LI and Z, but the HS value decreased, the difference was statistically significant ( <0.05) . There was no significant change of HS and UCB％in the treatment group, but the LI and Z values decreased, the difference was statistically significant (<0.05) . Conclusion Dan Ning Tablets can prevent cholesterol gallstone formation by regulating the intestinal flora, and affect the cholesterol content in bile , that plays a important role in prevention of cholesterol gallstone.

Alteration of tetraethylammonium-sensitive potassium current in pulmonary artery smooth muscle cells of pulmonary hypertension rats / 中国药理学通报

Aim To investigate the alteration of volt-age-depending potassium channel(KV) current in pul-monary arterial smooth muscle cells(PASMCs) of pul-monary hypertension (PH) rats, and the effect of tet-raethylammonium (TEA,a blocker of KV) on potassi-um channel current in different PH models. Methods The whole-cell patch clamp techniques were applied to record the KVcurrents from PASMCs cultured with Ham's F-12 (1% FBS). Furthermore, the effects of TEA on the KVcurrents were examined in different PH models. Results The whole-cell KVcurrents were ob-viously inhibited in PASMCs of chronic hypoxia (CH)and monocrotaline (MCT)-treated rats. TEA signifi-cantly decreased the whole-cell KVcurrents in PASMCs of control and PH rats,and the inhibitory effect of TEA was dramatically reduced in PH group. Conclusions The degree of the voltage-dependent potassium chan-nels opening is significantly inhibited in PASMCs of CH and MCT-treated rats,accordingly,the TEA-sen-sitive KVcurrents obviously decrease.

Comparison of the synchronous changes of vascular tension and intracellular Casignal in third-order branches of mesenteric arteries under the different objective amplification of confocal microscope / 生理学报

This study was aimed to establish an optimized method to observe the synchronous changes of vascular tension and intracellular Casignal in the third-order branches of mesenteric arteries (sMA, diameter: 100-300 μm). The vascular tension and intracellular Casignal changes in response to potassium chloride (KCl), endothelin-1 (ET-1) and Gdwere detected using confocal wire myograph system and confocal laser scanning microscopy imaging technique, respectively. The experimental results were analyzed to explore the optimal experimental conditions. The results showed that KCl caused contraction in sMA significantly, and the intracellular Calevel of vascular smooth muscle cells (VSMCs) was also increased under 20× and 40× objective lens. Compared with those under the 40× objective lens, the Casignal change was larger and the fluorescence value was more stable under the 20× objective lens, whereas the Casignal change was not obvious under the 10× objective lens. ET-1 (1-10 nmol/L) caused concentration dependent contraction in sMA significantly, and the intracellular Casignal of VSMCs was also enhanced in a concentration dependent manner. Additionally, Gdsignificantly reduced the contraction of sMA and the intracellular Casignal of VSMCs caused by ET-1. The results suggest that the intracellular Casignal of VSMCs changes with vascular contraction or relaxation caused by the agonists or antagonists of Cachannels. We successfully recorded both changes synchronously using confocal wire myograph system and confocal laser scanning microscopy imaging technique at the same time. Based on the analysis of the experimental results, we concluded that 20× objective lens provides the best experimental condition. Compared to combination of vascular tone detection method and real-time cellular fluorescence imaging technique, the present synchronous method is convenient and helpful to reduce experimental error.