Objective To explore the effects of Angiotensin (ANG)-( 1-7 ) on postangioplasty fibrotic remodeling and the involvement of TGF-β/Smad signaling pathway in this process. Methods Thirty two healthy New Zealand white rabbits were randomly divided into 4 groups: sham group, control group,ANG-(1-7) group and ANG-( 1-7 ) + A-779 group. Rabbits underwent angioplasty in the abdominal aorta or sham surgery. Subsequently, an osmotic minipump was implanted for saline, ANG-(1-7) (576 μg· kg~(-1) ·d~(-1)) or ANG-(1-7) + A-779 (576 μg · kg~(-1) · d~(-1)) delivery. Before and after 4 weeks treatment, the levels of ANG Ⅱ in plasma were measured by ELISA. At week 4, angiography and histomorphometric analysis were performed, mRNA levels of collagen Ⅰ and Ⅲ were assayed by RT-PCR and protein levels of TGF-β_1 and Stand2 in local vessel were assayed by Western blot. Results Following 4 weeks treatment,ANG-( 1-7 ) and ANG-( 1-7 ) + A-779 group displayed a significant elevation in lumen diameter [ (4. 11±0.10) mm and (3.34 ±0. 11) mm vs. (2.88 ±0.08) mm,P <0.05, respectively] and reduction in neointimal thickness [ (208 ± 17) μm and (407±25) μm vs. (448 ± 15) μm, P <0. 05, respectively],neointimal area [(0.27 ±0.09) mm~2 and (0.38 ±0.01) mm~2 vs. (0.41 ±0.02) mm~2, P<0.05,respectively] and restenosis rate [(28. 1 ±2.7)% and (36.8 ±2.2)% vs. (40. 1 ±2.7)% ,P<0.05,respectively] compared with control group. Collagen Ⅰ , Ⅲ mRNA and TGF-β_1 , Smad2 protein levels were significantly elevated in control group,ANG-(1-7) group and ANG-(1-7) + A-779 group compared to sham group ( P < 0. 01 or P < 0. 05 ) and reduced in ANG- (1-7) group compared to control group ( all P < 0. 05).Co-treatment with A-779 reversed the inhibitory action of ANG-( 1-7 ) . Plasma levels of ANG Ⅱ postangioplasty were similar in control and ANG-( 1-7 ) group and both were significantly higher than preoperation levels. Conclusion ANG-(1-7) attenuates postangioplasty collagen synthesis in rabbits possibly through down-regulating the expression of TGF-β_1 and inhibiting the activation of Smad2 pathway.

To develop an attenuated vaccine against the highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) virus, the HP-PRRS virus strain TJ was attenuated by serial passages and plaque cloned every 5 to 10 passages in Marc-145 cells. Genetic variation and pathogenicity of HP-PRRSV strain TJ in the course of attenuation were analyzed. The results showed that the strain TJ sustained various sequence changes during the course of attenuation. Fifty-eight amino acids changes and a new continuous 120 amino acids deletion after the discontinuous 30 amino acids deletion (sites 481 and 533-561) occurred in strain TJ passages 140, and the position of 120 amino acids deletion was between 628 to 747 according to VR-2332. Animal test showed that the pathogenicity of strain TJ passages 20 was attenuated obviously, so we presume that genetic variation in nonstructural protein nsp2-nsp5, nsp7 and structural protein GP5 during the attenuation provides the molecular bases for the observed attenuated phenotype.
Amino Acid Sequence ; Animals ; Genetic Variation ; Molecular Sequence Data ; Porcine Reproductive and Respiratory Syndrome ; virology ; Porcine respiratory and reproductive syndrome virus ; classification ; genetics ; isolation & purification ; pathogenicity ; Sequence Deletion ; Serial Passage ; Swine ; Vaccines, Attenuated ; genetics ; isolation & purification ; Virulence

BACKGROUNDChronic heart failure (CHF) is associated with calcium transients and calcium handling proteins. Angiotensin converting enzyme (ACE) inhibitor has been demonstrated to have beneficial effect on CHF. Yet studies addressed to the relationship between ACE inhibitor and calcium transients in CHF are rare. The aim of this study was to investigate the influence of ACE inhibitor (perindopril) on the contractility and calcium transients and calcium handling proteins in ventricular myocytes from rats with experimental heart failure.

METHODSMale Wistar rats were randomized to heart failure group treated with perindopril [CHF-T, 3 mg.kg(-1).d(-1)], heart failure group without treatment (CHF-C) and sham-operated group (PS). Heart failure was induced by abdominal aortic constriction. All groups were further followed up for 12 weeks. Left ventricular myocytes were then isolated. Single cell shortening fraction and [Ca(2+)]i were simultaneously measured by laser scanning confocal microscope under the field stimulation (1.0 Hz). Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were performed to evaluate the changes of mRNA and protein of Na(+)-Ca(2+) exchanger (NCX1), sarcoplasmic reticulum Ca(2+)-ATPase (SERCA2) and phospholamban (PLB).

RESULTSThe fraction of cell shortening (FS%) and [Ca(2+)]imax (nmol/L) were significantly reduced in group CHF-C compared with group PS (FS%: 7.51 +/- 1.15 vs 13.21 +/- 1.49; [Ca(2+)]i max: 330.85 +/- 50.05 vs 498.16 +/- 14.07; both P < 0.01), and restored at least partially in CHF-T group. In CHF-C group, the left ventricular mRNA of NCX1 and PLB were significantly upregulated in comparing with PS group (RNCX1/beta-Actin: 0.51 +/- 0.12 vs 0.19 +/- 0.06, P < 0.01; RPLB/beta-Actin: 0.26 +/- 0.12 vs 0.20 +/- 0.08, P < 0.05), while SERCA2 mRNA was downregulated (0.48 +/- 0.10 vs 0.80 +/- 0.11, P < 0.01). The mRNA levels of NCX1 and SERCA2 in CHF-T group were between the CHF-C and PS group, and the differences of the latter two groups were significant (all P < 0.05). In CHF-C and CHF-T groups, the protein expression of NCX1 were 1.141 +/- 0.047 and 1.074 +/- 0.081 times of that in PS group respectively (both P < 0.05), and SERCA2 protein levels were 0.803 +/- 0.100 and 0.893 +/- 0.084 times of that in PS group respectively (both P < 0.05). The protein expression of NCX1 and SERCA2 in the CHF-C and CHF-T groups is significantly different (both P < 0.05).

CONCLUSIONACE inhibitor could improve cardiac function of failing heart through directly enhancing the contractility of single cardiomyocyte, and these effects are probably mediated by its roles in preventing the deleterious changes of calcium transients and calcium handling proteins in CHF.

Angiotensin-Converting Enzyme Inhibitors ; pharmacology ; Animals ; Calcium ; metabolism ; Calcium-Binding Proteins ; genetics ; Calcium-Transporting ATPases ; genetics ; Heart Failure ; drug therapy ; metabolism ; Heart Ventricles ; drug effects ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; Perindopril ; pharmacology ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Sarcoplasmic Reticulum Calcium-Transporting ATPases ; Sodium-Calcium Exchanger ; genetics

OBJECTIVETo investigate the influence of ACE inhibitor (perindopril) on the contractility and calcium transient and calcium handling proteins in ventricular myocytes from rats with experimental heart failure.

METHODSMale Wistar rats were randomized to heart failure group treated with perindopril (CHF-T, 3 mg.kg(-1).d(-1)), heart failure group without treatment (CHF-C) and sham-operated group (PS) after heart failure was induced by constricting abdominal aorta for 16 weeks. All groups were further followed up for 12 weeks. Left ventricular myocytes were isolated, and single cell shortening fraction and [Ca(2+)](i) were simultaneously measured through laser scanning confocal microscope under the field stimulation (1.0 Hz). RT-PCR and Western blot were performed to evaluate the level of mRNA and protein of Na(+)-Ca(2+) exchanger (NCX(1)), sarcoplasmic Ca(2+)-ATPase (SERCA(2)) and phospholamban (PLB).

RESULTSThe fraction of cell shortening (FS%) and [Ca(2+)](i max) (nmol/L) were significantly smaller in group CHF-C than group PS (FS%: 7.51 +/- 1.15 vs 13.21 +/- 1.49; [Ca(2+)](i max): 330.85 +/- 50.05 vs 498.16 +/- 14.07; both P < 0.01). And in CHF-T group, FS and [Ca(2+)](i max) were greater than those in CHF-C group. In CHF-C group, the left ventricular mRNA of NCX(1) and PLB were significantly higher than those in PS group (R(NCX)(1)/beta-Actin: 0.51 +/- 0.12 vs 0.19 +/- 0.06, P < 0.01; R(PLB)/beta-Actin: 0.26 +/- 0.12 vs 0.20 +/- 0.08, P = 0.045), yet SERCA(2) mRNA was lower than PS group (0.48 +/- 0.10 vs 0.80 +/- 0.11, P < 0.01). In CHF-T group, the mRNA levels of NCX(1) and SERCA(2) were just in the midst of the CHF-C and PS group, and had statistical significance respectively (all P < 0.05). In CHF - C and CHF - T group, the protein levels of NCX(1) were 1.141 +/- 0.047 and 1.074 +/- 0.081 times PS group, respectively (both P < 0.05), and SERCA(2) protein levels were respectively 0.803 +/- 0.100 and 0.893 +/- 0.084 times as high as in PS group (both P < 0.05). The protein expression of NCX(1) and SERCA(2) were also different between CHF-C and CHF-T groups (both P < 0.05).

CONCLUSIONACE inhibitor could improve cardiac function in CHF through directly enhancing the contractility of single myocardial cell, and these effects were probably mediated by its role in preventing the deleterious changes of calcium transient and calcium handling proteins in CHF.

Animals ; Calcium ; metabolism ; Calmodulin ; metabolism ; Heart Failure ; drug therapy ; metabolism ; Male ; Myocytes, Cardiac ; drug effects ; metabolism ; Perindopril ; pharmacology ; therapeutic use ; Rats ; Rats, Wistar

OBJECTIVE To explore the efficacy of Sour Jujube Seed Decoction combined with Huanglian Wendan Decoction on adult chronic insomnia and its effect on hypnotic withdrawal.METHODS 187 patients with chronic insomnia were included for anal-ysis,including 102 in the traditional Chinese medicine(TCM)group and 85 in the western medicine group.The TCM group was trea-ted with Sour Jujube Seed Decoction combined with Huanglian Wendan Decoction,while the western medicine group was treated with benzodiazepine under the consideration of doctor.The intervention period was 1 month,with assessments using the Pittsburgh Sleep Quality Index(PSQI)conducted before and after the intervention.Follow-up evaluations were performed at 3 months and 6 months re-spectively after the intervention.RESULTS There was no significant difference between the two groups at baseline.After the inter-vention,the PSQI scores of patients in both groups were significantly improved(P<0.01).Among them,the TCM group was better than the western medicine group in the improvement of sleep quality and sleeping pills,total PSQI score reduction(P<0.01).The re-sults of linear regression analysis showed that after controlling for confounding factors,the regression coefficients of the TCM group in two different models were1.821 and 1.922 respectively,and the former was statistically significant(P<0.05).By screening patients who took hypnotics at baseline in the TCM group and comparing them with those in the western medicine group,the influencing factors of hypnotic withdrawal were analyzed.During the 3-month follow-up,25 out of 39 patients in the TCM group and 17 out of 80 patients in the western medicine group had hypnotic withdrawal(χ2= 19.25,P<0.001);during the 6-month follow-up,23 of the 39 patients in the TCM group and 18 of the 79 patients in the western medicine group had hypnotic withdrawal(χ2= 13.53,P<0.001),the with-drawal rate of patients in the TCM group was significantly higher than that in the western medicine group.Further regression analysis showed that after adjusting for confounding factors,the results showed that the western medicine group had a significantly higher rate of not withdrawal than the TCM group at 3 months(OR=5.50,95%CI:2.30～13.72)and 6 months(OR=6.43,95%CI:2.54～17.77),and the results were statistically different(P<0.05).CONCLUSION Sour Jujube Seed Decoction combined with Huangli-an Wendan Decoction is effective in treating adult chronic insomnia and assisting in hypnotic withdrawal.

The gut microbiota and its metabolites play a critical role on health maintenance, because they are involved in the absorption and metabolism of nutrients in the human bodies. This is also similar to traditional Chinese medicine (TCM) view that the ascending and descending of Qi movement affects Yin-Yang, Qi-blood, pneuma and body fluid, viscera and meridians of our bodies. More and more studies have demonstrated that gut microbiota is closely related to the development and progression of diabetes and its complications. Gut microbiota disorder could affect host metabolic signaling pathways, thereby promoting the formation and development of diabetes. The smooth ascending and descending of Qi movement is the basic form of maintaining host metabolic homeostasis, whose dysfunction however can lead to internal environment disturbance. Based on the theory of ascending and descending of Qi movement, this paper focuses on the pathogenesis of imbalanced intestinal flora in the process of the induction of diabetes mellitus from a dynamic perspective. It is assumed that the imbalance of Qi ascending and descending may act as a trigger for such symptoms as lung Qi impairment, spleen deficiency to dissipating essence, liver Qi stagnation and kidney Yang deficiency. Under this circumstance, gut microbiota will be out of balance, which will further lead to the nutrient substance metabolic disturbance in the body, and thus induce diabetes. Thus, it is significant to explore the regulatory mechanism of gut microbiota and its metabolites on diabetes based on the theory of ascending and descending of Qi movement, so as to reveal the scientific connotation of TCM in regulating substance metabolism homeostasis in the body.