Objective To observe the clinical efficacy of Chuankezhi acupoint injection combined with nursing health care education for treatment of patients with chronic obstructive pulmonary disease (COPD) at stable stage complicated with spleen kidney deficiency syndrome.Methods A prospective study was conducted. 118 patients with stable COPD and spleen kidney yang deficiency syndrome admitted to Hangzhou Third People's Hospital and Zhejiang Xinhua Hospital from February to April 2014 were enrolled, and they were divided into an experimental group (60 cases) and a control group (58 cases) according to the random number table. Both groups were given conventional treatment [including oxygen therapy, bronchodilators, corticosteroids, rehabilitation therapy in accord with the Global Initiative COPD (GOLD)]. Simultaneously, in the experimental group, Chuankezhi acupoint injection was additionally given, once daily for 12 weeks, and appropriate corresponding health education was mentioned. The lung function, quality of life and prognoses of the patients were regularly followed up for 9 months and compared between the two groups.Results After 12 weeks, one second forced expiratory volume (FEV1), FEV1/forced vital capacity (FVC) and maximum voluntary ventilation (MVV) per minute in the experimental group were significantly higher than those in control group [FEV1 (L): 1.57±0.42 vs. 1.35±0.35, FEV1/FVC: (58.62±4.56)% vs. (50.36±4.35)%, MVV (L/min): 60.62±7.56 vs. 56.95±6.33, allP < 0.05]. The quality of life evaluation form for COPD (QLICD-COPD) was used to evaluate the physical function, psychological function, social function and specific modules scores, showing that the above items in the experimental group were significantly higher than those in control group (total score: 74.69±8.92 vs. 68.62±8.95, physical function score: 74.16±8.55 vs. 66.31±7.15, mental function score: 78.69±9.79 vs. 72.64±8.45, social function score: 75.45±6.75 vs. 70.36±6.12, specific modules score: 67.52±6.33 vs. 61.36±6.17, allP < 0.05); Barthel index score was also significantly higher than that in the control group (75.41±9.24 vs. 68.43±7.95,P < 0.05). Within 9 months of follow-up, the incidence of acute exacerbation of COPD in the experimental group was significantly lower than that in the control group [5.00% (3/60) vs. 17.24% (10/58),P < 0.05]. The re-admission rate was also significantly less in patients in experimental group compared with that of control group [1.67% (1/60) vs. 12.07% (7/58),P < 0.05]. Conclusion Chuankezhi acupoint injection with health education for treatment of patients with COPD at stable stage and spleen kidney yang deficiency syndrome has good clinical efficacy, worthy to be applied more widely.

Objective To explore the relationship of serum parathyriod hormone ( PTH) level with renal ane-mia in patients with chronic kidney disease (CKD).Methods Serum levels of PTH,Hb,Hct,BUN,Cr were observed in 126 patients with CKD at stages 3-5,the changes of 42 patients in maintenance hemodialysis(MHD) with recombi-nant human erythropoietin (rhEPO) and calcitriol-1.25(OH)2D3 treatment for three months were determined. Results The serum PTH level was significantly increased in CKD patients at stage 3 and more in CKD patients at stage 4 and 5.There were positive relationship between blood PTH and BUN ,Cr(P<0.01),but a negative relation-ship between PTH and Ccr,Hb(P <0.01).42 patients of MHD treated with rhEPO,1.25(OH)2D3 after three months,according to the changes of HCT ,the effective rate was 73.8% and the ineffective rate was 26.2%.The PTH,Hb,HCT of the effective group had statistically significant differences compared with the ineffective group ( all P<0.01).Hb and Hct significantly increased and PTH significantly decreased in the effective group .Conclusion CKD patients had decreased renal function ,elevated blood PTH .High serum PTH might aggravate renal anemia and fail to respond to rhEPO.The high serum PTH corrected by using 1.25(OH)2D3 might improve renal anemia,with high plasma PTH correcting .

AIM:To investigate the effects of Xinkang recipe on myocardial miR-25-3p expression and sarco-plasmic reticulum calcium ATPase 2a ( SERCA2a) activity in heart failure rats .METHODS:Male SD rats were randomly divided into normal group , sham group , model group , Xinkang recipe group ( Xinkang group ) , and captopril group .The heart failure rat model was induced by intraperitoneal injection of doxorubicin .Distilled water , Xinkang recipe and capto-pril were administrated by gastric gavage for 35 d, respectively .The indexes of cardiac function and plasma level of brain natriuretic peptide (BNP) were measured.The SERCA2a activity was determined by the inorganic phosphorus method . The myocardial protein expression of SERCA 2a and phospholamban ( PLB) was detected by Western blot .The myocardial expression of miR-25-3p was detected by stem-loop RT-qPCR.RESULTS:Cardiac output (CO), left ventricular fraction-al shortening ( LVFS) and left ventricular ejection fraction ( LVEF) in Xinkang group and captopril group were significantly higher while the plasma levels of BNP were significantly lower than those in model group (P<0.01).The myocardial ex-pression levels of miR-25-3p in Xinkang group and captopril group were significantly lower while the myocardial protein le -vels of SERCA2a and PLB were significantly higher than those in model group (P<0.01).The SERCA2a/PLB ratio and SERCA2a activity in Xinkang group were significantly higher than those in model group (P<0.05), and no significant change was observed between captopril group and model group .CONCLUSION:Xinkang recipe therapy may improve car-diac function in heart failure rats , which may be related to inhibiting the expression of miR-25-3p, increasing the SER-CA2a/PLB ratio and enhancing SERCA 2a activity in the myocardium .

Objective: To observe the effect of bisoprolol on cardiac function in heart failure (HF) rats and to explore the mechanism. Methods: The experimental rats were divided into 6 groups: Control group, with normal healthy rats, Sham group, the rats received intraperitoneal injection of normal saline; chronic heart failure (CHF) model was successfully established in 40 rats and divided into 4 groups: CHF group, CHF+bisoprolol (Bis) group, CHF+captopril (Cap) group and CHF+Bis and Cap group.n=10 in each group. The cardiac function was observed among different groups; plasma BNP level was measured by ELISA, myocardial miR-25-3p expression was examined by RT-PCR, protein expressions of SERCA2a and phospholamban (PLB) were detected by Western blot analysis and SERCA2a activity was determined by inorganic phosphorus method. Results: Compared with Control group, CHF group showed decreased cardiac output (CO), left ventricular fractional shortening (LVFS), left ventricular ejection fraction (LVEF), reduced expression of cardiac SERCA2a, PLB, the ratio of SERCA2a/PLB and SERCA2a activity; while increased plasma BNP and miR-25-3p expression, allP<0.01. Compared with CHF group, CHF+Bis, CHF+Cap and CHF+Bis and Cap groups had increased CO, LVFS, LVEF, elevated expression of cardiac SERCA2a, PLB, the ratio of SERCA2a/PLB and SERCA2a activity; while decreased plasma BNP and miR-25-3p expression, allP<0.05.Conclusion: Bisoprolol could improve cardiac function in HF rats, which might be related to down regulating myocardial miR-25-3p expression, up regulating myocardial protein expressions of SERCA2a, PLB and enhancing SERCA2a activity.

Objective To investigate the effects of bisoprolol on myocardial SERCA2a activity in rats with heart fail-ure.Methods Male SD rats were randomly divided into normal control group (control group), sham operation group ( sham group ) , model group , bisoprolol group ( Bis group ) , captopril group ( Cap group ) and bisoprolol plus captopril group[(Bis+Cap)group], heart failure rat model was induced by intraperitoneal injections of doxorubicin .Distilled water, bisoprolol, captopril or bisoprolol plus captopril were administrated by gastrogavage for 35 days, respectively. Indices of cardiac function and plasma levels of B-type natriuretic peptide ( BNP) were measured , myocardial expres-sion of miR-25-3p was detected by Stem-loop RT-qPCR, myocardial levels of SERCA2a and phospholamban (PLB) were detected by Western blot , myocardial SERCA2a activity was determined by the inorganic phosphorus method . Results Cardiac function in model group decreased significantly while plasma levels of BNP were significantly higher than those of control group ( P<0.01 ) .Myocardial expression of miR-25-3p in model group was significantly higher while myocardial levels of SERCA 2a and PLB,SERCA2a activity were significantly lower than those of con-trol group(P<0.01).Cardiac function in Bis group , Cap group and Bis +Cap group improved significantly while plasma levels of BNP were significantly lower than those of model group ( P<0.01 ) .Myocardial expression of miR-25-3p in Bis group, Cap group and Bis +Cap group were significantly lower while myocardial levels of SERCA2a and PLB were significantly higher than those in model group (P<0.01).The SERCA2a/PLB ratio and SERCA2a activity in Bis group and Bis +Cap group were significantly higher than those of model group ( P<0.05 ) .Conclu-sions Bisoprolol therapy improves cardiac function in rats with heart failure , which may be related to inhibition of myocardial miR-25-3p, increasing myocardial SERCA2a and PLB levels, enhancing SERCA2a activity.

Objective To explore the effect of Xinkang Tablets on myocardial apoptosis index,collagen volume fraction and sarcoplasmic reticulum Ca2+-ATPase activity of rats with adriamycin-induced heart failure.Methods The chronic heart failure (CHF) SD rat model was established by intraperitoneal injection of doxorubicin.After successful modeling,the rats with CHF were randomly divided into 5 groups,namely model group,western medicine group,and low-,middle-and high-dose of Chinese medicine groups,10 rats in each group.The rats in the above groups were given intragastric administration of distilled water,22.5 μg/kg of Digoxin mixed suspension,9,18,36 g/kg of XinkangTablets,respectively,in the volume of 10 mL/kg of distilled water dilution,once a day,for 5 continuous weeks.Another the same batch of 10 SD rats were randomly allocated to the sham operation group,and were treated with intragastric administration of the same volume of distilled water.And then the apoptotic rate of myocardial cells was measured by TUNEL method,the collagen volume fraction (CVF) was measured after Masson staining,and the sarcoplasmic reticulum Ca2+-ATPase activity was determined by inorganic phosphate assay.Results Compared with the sham operation group,the apoptotic rate of myocardial cells and CVF in the model group were increased(P ＜ 0.01),indicating that the myocardial remodeling occurred in rats with CHF.Compared with the model Group,the apoptotic rate of western medicine group and three Chinese medicine groups was significantly decreased(P ＜ 0.01),suggesting that Digoxin and Xinkang Tablets can relieve apoptosis to certain extent.The CVF in Digoxin group and middle-and high-dose of Chinese medicine groups were lower than those in the model Group (P＜ 0.05 or P＜ 0.01),indicating that Digoxin and Xinkang Tablets can delay the myocardial fibrosis.Last but not least,the SERCA2a activities in the middle-and high-dose of Chinese medicine groups were higher than those in the model group (P ＜ 0.05 or P ＜ 0.01),suggesting that Xinkang Tablets may relieve myocardial remodeling and improve cardiac function through the regulation of SERCA2a activity.Conclusion Xinkang Tablets decrease the apoptotic rate and myocardial cell volume fraction probably through the regulation of SERCA2a activity,which may play a role in counteracting apoptosis and myocardial fibrosis,and ultimately delay the remodeling of the myocardium.

ABSTRACT:Objective To investigate the effects of Xiefei Lishui recipe on left ventricle remodeling in rats with heart failure.Methods Heart failure rat model was induced by intraperitoneal injections of doxorubicin. Rats were randomly divided into sham operation group (sham group),model group,traditional Chinese medicine group (TCM group),captopril group,and digoxin group.Distilled water,TCM [22 g/(kg · d)],captopril [19 mg/(kg·d)],and digoxin [32μg/(kg·d)]were administered by gastrogavage in rats in different groups for 35 days,respectively.Indices of ventricle remodeling and cardiac function,plasma levels of B-type natriuretic peptide (BNP),rennin (REN),angiotensin Ⅱ(AngⅡ)and aldosterone (ALD)were measured.Cardiomyocyte apoptosis index and collagen volume fraction (CVF)were analyzed.We also assayed myocardial mRNA expressions of MMP-2/9 and TIMP-1/2,and their tissue inhibiting factors TIMP-1 and TIMP-2.Results Compared with those in sham group,in model group cardiac function was significantly decreased,which could be significantly increased by TCM or captopril or digoxin,indices of cardiac remodeling were significantly increased,which could be significantly decreased by TCM or captopril (P<0.01 or P<0.05).Plasma levels of BNP,REN,AngⅡ and ALD,cardiomyocyte apoptosis index and CVF in model group were significantly increased,could be significantly decreased by TCM or captopril (P<0.01 or P<0.05).Myocardial mRNA expressions of MMP-2,MMP-9,TIMP-1 and TIMP-2 in model group were significantly upregulated compared with those in sham group, which could be significantly downregulated by TCM (P<0.01 or P<0.05).Conclusion Xiefei Lishui recipe can attenuate left ventricle remodeling and improve cardiac function in rats with heart failure, which may be related to downregulating myocardial mRNA expressions of MMP-2 ,MMP-9 ,TIMP-1 and TIMP-2 in the left ventricle as well as inhibiting cardiomyocyte apoptosis and myocardial fibrosis.

[ ABSTRACT] AIM:To explore the effect of dexmedetomidine ( DEX) on the CCAAT/enhancer-binding protein-homologous protein ( CHOP) pathway during lung ischemia-reperfusion ( I/R) in mice.METHODS:C57BL/6J male mice were randomly divided into sham operation group ( sham group) , lung ischemia/reperfusion group ( I/R group) , ischemia/reperfusion +normal saline group ( I/R+NS group ) and ischemia/reperfusion+dexmedetomidine group ( I/R+DEX group) .Dexmedetomidine was infused intraperitoneally with 25 μg/kg for 30 min prior to the ischemia period in I/R+DEX group, the normal saline was administrated with the same volume of dexmedetomidine in I/R+NS group.After fini-shed the 3 h-reperfusion period , the left lung tissues were harvested to determine lung wet/dry weight ( W/D) , the total lung water content ( TLW) , and index of quantitative evaluation for alveolar damage ( IQA) .Morphological observation and terminal-deoxynucleotidyl transferase mediated nick end labeling ( TUNEL) were applied to evaluate the structure changes and the apoptosis index (AI) of the lung tissues.The expression of CHOP and glucose-regulated protein 78 (GRP78) at mRNA and protein levels in the lung tissues was detected by Western blot and RT-PCR.RESULTS:Compared with sham group, the W/D, TLW, IQA, AI, the mRNA and protein expression of CHOP and GRP78 obviously increased, and the left lung tissues structure were damaged more obviously both in I/R group and I/R+NS group.Compared with I/R group, the W/D, TLW, IQA, AI and the protein and mRNA expression of CHOP in I/R+DEX group decreased, the injury of the left lung tissue structures induced by I/R in I/R+DEX group were also alleviated .CONCLUSION:DEX alleviates the
lung I/R injury, which may be related to inhibition of apoptosis mediated by CHOP pathway.

Objective To investigate the effects of Xinkang recipe on myocardial collagen metabolism in rats with doxorubi-cin-induced heart failure.Methods Male SD rats were randomly divided into sham operation group(sham group),model group, Xinkang group and captopril group.The rat model of heart failure was established by intraperitoneal injection of doxorubi-cin.Distilled water,Xinkang decoction and captopril were respectively administrated to rats by gavage for 35 d.The indexes of ventricular remodeling and cardiac function were measured.Myocardial fibrosis was assessed by Masson's trichrome stai-ning.The expression levels of collagen Ⅰ,collagen Ⅲ,TGF-β1,IκB and p56 were detected in myocardial tissues by Western blotting.Myocardial protein and mRNA levels of matrix metalloproteinase-2(MMP-2),MMP-9,and tissue inhibitor of matrix metalloproteinase-1(TIMP-1)and TIMP-2 were detected by Western blotting and RT-qPCR,respectively.Results The expres-sion levels of collagenⅠand collagen Ⅲ protein,the collagen volume fraction(CVF),and the expression levels of TGF-β1 and p56 protein were significantly increased(P< 0.01),and the expression level of IκB protein was significantly decreased in the model group as compared with the sham group(P<0.05 for all).The myocardial protein and mRNA levels of MMP-2,MMP-9, TIMP-1,and TIMP-2 were significantly higher in the model group than in sham group(P<0.05 or P<0.01).The cardiac out-put(CO),left ventricular fractional shortening(FS),and left ventricular ejection fraction(LVEF)were significantly lower while the left ventricular end diastolic volume(LVEDV),left ventricular end systolic volume(LVESV),and left ventricular mass index (LVMI)were significantly higher in model group than in sham group(P<0.01).Compared with the model group,the expres-sion levels of collagen Ⅰand collagen Ⅲ protein,the CVF and the TGF-β1 and p56 expression levels were significantly decreased and the IκB protein expression was significantly increased in Xinkang and the captopril groups(P<0.05).The myocardial pro-tein and mRNA levels of MMP-2,MMP-9,TIMP-1,and TIMP-2 were significantly lower in Xinkang and captopril groups than in control group(P<0.05 or P<0.01).CO,FS,and LVEF were significantly higher while LVEDV,LVESV,and LVMI were significantly lower in Xinkang and captopril groups than in model group(P<0.01).Conclusion Xinkang recipe can reduce col-lagen deposition in the myocardia of rats with doxorubicin-induced heart failure,attenuate left ventricular remodeling,and im-prove cardiac function,which is associated with the inhibition of the NF-κB-mediated upregulation of TGF-β1.

OBJECTIVETo compare the DNA methylation-related alteration induced by trichloroethylene (TCE) in human hepatic L-02 cells (L-02 cells) and SET deficient cells, and reveal the role of SET on the mechanisms in TCE-induced epigenetic pathway.

METHODSThe L-02 cells and pre-established SET deficient cells were treated with different TCE concentrations, and the changes of total cell viability, DNA methylation level and DNA methyltransferases (DNMTs) activity were measured, respectively. In addition, the TCE-induced alteration in the protein expression of DNMT1, DNMT3a and DNMT3b were analyzed by Western blotting.

RESULTSAfter treatment with TCE for 24 h, the cell proliferation level was significantly decreased in both cell lines. When concentrations of TCE were 0, 1.0, 2.0, 4.0 and 8.0 mmol/L, the proliferation levels of L-02 cells were 100.00±2.70, 83.34±2.38, 75.56±4.51, 71.67±2.77 and 66.67±1.63, respectively (F = 58.29, P < 0.001); the cell proliferation levels of SET deficient cells were 101.12±1.67, 85.01±2.33, 79.44±1.67, 78.337±3.89 and 76.11±3.33, respectively (F = 42.41, P < 0.001). When concentration of TCE reached 4.0 mmol/L, the difference of cell proliferation level between two groups was statistically significant (t = -3.51; P = 0.013). After treated by TCE for 24 h, the global DNA methylation significantly decreased in both cell lines (F value was 212.87 and 79.32, respectively, P < 0.001). The difference between two groups was not statistically significant. After treated by TCE for 24 h, the methyltransferases activities were significantly decreased in both cell cells (F values were 77.92 and 113.80, respectively, P-0.001). The SET deficiency could inhibit the decrease of methyltransferases activity under TCE treatment. When the concentration of TCE reached 8.0 mmol/L, the enzymatic activity of L-02 cells and SET deficient cells decreased to 67.61%±2.85% and 72.97%± 1.94%, respectively. The difference between two groups was statistically significant (t = -3.94, P = 0.008). After treated with TCE for 24 h, concentrations of TCE were 0, 1.0, 2.0, 4.0 and 8.0 mmol/L, and the relative protein levels of DNMT1 in normal L-02 cells increased significantly to 1.00±0.03, 1.28±0.04, 1.20±0.04, 1.62±0.05, 1.43±0.04 (F = 103.00, P < 0.001); In SET deficient cells, the expressions of DNMT1 were 1.00±0.04, 0.96±0.02, 1.19±0.05, 0.85±0.03, 0.83±0.03, which was significantly down-regulated under TCE treatment (F = 44.18, P < 0.001).

CONCLUSIONSET deficiency can significantly attenuate the TCE-induced decreases of cell viability and DNMTs activity, as well as alteration of protein expression of DNMT1 in L-02 cells, which indicated that SET was involved in the mechanism of TCE-induced cytotoxicity and epigenetic pathway in L-02 cells.