Objective:To study the feasibility,safety and effectiveness of intrapericardial injection with a trans-diaphragmatic approach in rats and its value in the study of heart diseases. Methods:Blue-black ink,recombinant adeno-associated virus carrying enhanced green fluorescent protein((eGFP)) gene,0.9% sodium chloride solution were respectively injected into pericardium of rats in three different groups(Group ink,Group eGFP and Group sodium chloride) by intrapericardial injection with a trans-diaphragmatic approach.Autopsy was done and hemodynamic parameters were measured and cryosection was analyzed by fluorescence microscopy. Results:The injection was proved successful in all rats in Group ink.Green fluorescence was detected in cryosection of all hearts in Group eGFP and the expression of green fluorescent protein was ubiquitously,but not homongeneous.No rat died during and after the operation among the operated rats(rats in Group eGFP and Group sodium chloride).There was no difference between hemodynamic parameters of the operated rats and those of the controls.Conclusion:The intrapericardial injection with a trans-diaphragmatic approach is a simple,feasible,safe and valid operation and suitable for small experimental animals.And it suggests an easy, safe,efficient and cheap technique in the study of heart diseases.

Objective To study the short-term therapeutic effects of adeno-associated viral gene transfer of sarcoplasmic reticulum Ca2+-ATPase (SRCA2a) on chronic heart failure (CHF). Methods The rats were divided into four groups: control group, CHF group, CHF+eGFP group and CHF+SRCA2a group. CHF rats were obtained by creating descending aortic constriction. 0.9% sodium chloride solution, recombinant adeno-associated virus carrying enhanced green fluorescent protein (eGFP) gene, and recombinant adeno-associated virus carrying SRCA2a gene, were respectively injected into pericardium of heart failure rats in different groups by intrapericardial injection via trans-diaphragmatic approach. 10 days after gene transfer, hemodynamic parameters, the protein expression of SRCA2a and the SRCA2a activity were measured and analyzed respectively. Results 10 days after gene transfer, the protein expression and activity of SRCA2a were increased by 80.7% and 89.9% respectively, but still lower than those in the control rats. The heart function was also improved along with the increase of SRCA2a expression and activity. The left ventricular systolic pressure and the maximal rate of decline of left-ventricular pressure in CHF+SRCA2a group were similar with those of the controls. The left ventricular end-diastolic pressure and the maximal rate of rise of left-ventricular pressure were much improved, but did not reach the level of the controls. Conclusions SRCA2a protein levels and activity are significantly decreased in CHF rats. Adeno-associated viral gene transfer of SRCA2a can improve the function of the heart in short term.

Clinical data of 27 children with renal calculi (11 cases on the right side and 16 on left)who were treated with extracorporeal shockwave lithotripsy (ESWL) using HB-ESWL-VG lithotripter from April 2006 to October 2008 were retrospectively reviewed.The size of stones ranged from 5 mm to 17 mm in diameter (mean 11 mm).In 22 out of 27 cases (82%) stones were crushed completely after the first course.Eleven of 27 ESWL cases were stone-free in 7 days,8 patients in a month after treatment;the stonefree rate was 96% when patients were followed up for 1 year.The results suggest that ESWL is a safe and effective method for treatment of renal calculi in children.

AIM: To study the feasibility, security a nd validity of the intrapericardial injection with a trans-diaphragmatic approach in chronic heart failure rats and its value in the study of gene therapy for hea rt diseases, and further investigate whether adeno-associated virual gene transf er of sarcoplasmic reticulum Ca 2+-ATPase (SERCA2a) gene can improve ven tricular function in chronic heart failure (HF) rats. METHODS: An animal model of heart failure was obtained by creati ng descending aortic constriction in rats. Recombinant adeno-associated virus, c arrying enhanced green fluorescent protein (EGFP) gene and recombinant adeno-ass ociated virus carrying SERCA2a gene, were respectively injected into pericardium of heart failure rats in different groups (group HF+EGFP and group HF+SERCA2a) by intrapericardial injection with a trans-diaphragmatic approach. After 30 days , hemodynamic parameters were measured and analyzed. Cryosection was analyzed by fluorescence microscopy to examine the expression of green fluorescent protein, and Western blotting was performed to detect the expression of SERCA2a. RESULTS: Green fluorescence was detected in cryosection of the h earts in all rats in group HF+EGFP and the expression of green fluorescent prote in was ubiquitously. The expression of SERCA2a in all rats in group HF+SERCA2a w as more than those in group HF and group HF+EGFP. And overexpression of SERCA2a improved the systolic and diastolic function of heart failure rats significantly and the hemodynamic parameters were similar with those of the controls. CONCLUSION: The intrapericardial injection with a trans-diaphrag matic approach suggests a simple, safe, efficient and cheap technique for the ge ne therapy of chronic heart failure. Gene thransfer of SERCA2a may be a new appr oach for the treatment of chronic heart failure.

BACKGROUND:In chronic congestive heart failure (CHF),both protein expression and activity of sarcoplasmic reticulum (SR) Ca2+ ATPase (SERCA2a) are decreased, which results in abnormal regulation of calcium ion and systole and diastole dysfunction.OBJECTIVE:To observe the short-and middle-term therapeutic effects of adeno-associated viral gene transfer of SERCA2a in treatment of CHF in rats,and to investigate the possible mechanisms.DESIGN:Randomized-controlled design.SETTING:Department of Geriatric Cardiology and Department of Pathophysiology,General Hospital of Chinese PLA.METHODS:Male Sprague-Dawiey rats,were randomized into 4 groups:sham-operation group,CHF group,CHF+ green fluorescent protein (GFP) group and CHF+SERCA2a group. Gene transfer 10 and 30 days two time points were set in each group.CHF rat models were established by coarctation of abdominal aorta.At postoperative 18 to 20 weeks,the successful rats in CHF group were intraperitoneally injected with 500 μL aseptic normal saline,and those in the CHF+ GFP group and CHF+ SERCA2a group were injected with the same dose of Adeno-associated viral gene with GFP and SERCA2a gene respectively.MAIN OUTCOME MEASURES:①At 10 and 30 days after gene transfer,hemodynamic parameters,the SERCA2a protein expression was analyzed by Western Blot,and SERCA2a activity was carded out by modified Jones method.②The difference of proteome of hearts was detected by expression proteomics in between CHF+ SERCA2a group and CHF group at 30 days.③Electrophoretic separation and quantitation of cardiac myosin heavy chain(MHC) isoforrns were done at 30 days.RESULTS: ①In CHF group and CHF+GFP group, SERCA2a expression and activity were lower than those in sham-operation group.At 10 days after gene transfer.SERCA2a expression was increased by 80.7%and still lower than that of sham-operation group,and activity was also increased by 89.9%,but still lower than that of sham-operation group.And at 30 days,the SERCA2a protein expression and activity in the CHF+SERCA2a group reached to levels of sham-operation group. ②At day 10,LVSP and LV +dp/dtmax and LV-dp/dtmax were increased and LVEDP was decreased dramatically,but they still did not reach to the levels of sham-operation group.At day 30,the parameters were normalized back to control levels.There were no significant differences in each index between CHF+SERCA2a group and sham-operation group.③Overexpression of SERCA2a increased some energy metabolic enzymes of hearts at 30 days.④There were a downregulation of α-MHC and an upregulation of 3-MHC in failing hearts.Overexpression of SERCA2a increased the expression of α-MHC and decreased the expression of β-MHC to the levels of sham-operation group at 30 days.CONCLUSION: Adeno-associated viral gene transfer of SERCA2a can enhance SERCA2a functions, maintain calcium homeostasis,improve cardiac energy metabolism,and normalize the expression of MHC isoforms in CHF rats.As a result,the heart functions can be improved. So adeno-associated viral gene transfer of SERCA2a has good therapy effects on CHF rats in short and relatively long periods.

Objective To investigate the G protein-coupled receptor kinase 2 (GRK 2) level in peripheral blood lymphocytes with cardiac func-tion in elderly patients with acute myocardial infarction. Methods This study enrolled 40 patients with acute ST-segment elevation myo-cardial infarction (STEMI) and 40 patients with unstable angina. All patients were 65 years or older. Cardiac function was evaluated by echocardiography, and the GRK 2 level in peripheral blood lymphocytes was measured. Patients with STEMI were followed up for 2 years. Results The GRK 2 level in peripheral blood lymphocytes was significantly higher in patients with STEMI than in patients with unstable angina, and was negatively correlated with left ventricular ejection fraction, cardiac output, stroke volume, and left ventricular fractional shortening. The GRK 2 level was significantly elevated in some patients with acute STEMI and poor cardiac function. Conclusions In-creased GRK 2 level in patients with acute STEMI may contribute to poor myocardial systolic function and myocardial remodeling. Meas-urement of the GRK 2 level in peripheral blood lymphocytes may assist in the evaluation of cardiac function and myocardial remodeling in elderly patients with acute STEMI.

OBJECTIVETo investigate the dynamic changes in the expression of clara cell protein (CC16) and surfactant protein D (SP-D) in the lung tissues and bronchoalveolar lavage fluid (BALF) of silica-treated rats.

METHODSEighty-four Wistar rats were randomly divided into control group (n = 42) and silica group (n = 42). The silica group was subsequently divided into 3, 7, 14, 21, 28, and 60 d subgroups. The silicotic model was made by instilling silica suspension directly through the trachea into rat lungs. At 3, 7, 14, 21, 28, and 60 d after silica instillation, 8 rats in each group were sacrificed and their lung tissues and BALF were collected. The expression of SP-D and CC16 in lung tissues was detected by immunohistochemistry. Enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of SP-D and CCl6 in BALF.

RESULTSThe immunohistochemical assay indicated that CCl6 and SP-D were expressed in lung cells. The ELISA found that in 7, 14, 21, 28, and 60 d silica subgroups, the content of CCl6 in rat BALF was 8.14±0.70, 7.15±0.66, 7.00±0.69, 6.34 ± 0.59, and 5.27±0.49 ng/L, respectively; CCl6 expression decreased gradually with the silica exposure time prolonged, indicating a negative correlation (ra = -0.953, P < 0.01). Compared with the control group, all silica subgroups had significantly decreased CCl6 levels (P < 0.05). The content of SP-D in BALF was 12.20 ± 1.57, 14.41 ± 0.65, and 12.18 ± 0.74 ng/L, respectively, in the 7, 14, and 21 d silica subgroups, significantly higher than that in the control group (P < 0.05).

CONCLUSIONThe dynamic changes in SP-D and CCl6 protein levels in the lung tissues and BALF of rats could be induced by silica exposure and are related to silica exposure time. With the extension of silica exposure, CCl6 levels are gradually reduced, while the SP-D levels first increase and then fall.

Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Epithelial Cells ; metabolism ; Lung ; metabolism ; Pulmonary Surfactant-Associated Protein D ; metabolism ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity ; Uteroglobin ; metabolism

OBJECTIVETo evaluate the protective effect of amlodipine against contrast agent-induced renal injury in elderly patients with coronary heart disease.

METHODSA total of 189 elderly patients (>60 years) with coronary heart disease undergoing coronary artery angiography were randomly assigned into amlodipine group and control group to receive amlodipine or placebo, respectively, before and after administration of the contrast agent. At 24 h, 48 h and 5 days after contrast agent administration, the parameters of renal function were measured including serum cystatin C, urea nitrogen, creatinine, creatinine clearance rate, urine β2-microglobulin, and urine N-acetyl-β-glucosaminidase.

RESULTSIn both groups, the contrast agents obviously affected the renal functions of the patients (P<0.05). At 24 h after contrast administration, the levels of serum cystatin C, urine β2-microglobulin and urine NAG were significantly lower in amlodipine group than in the control group, but the other functional parameters showed no significant difference. At 48 h after contrast administration, the glomerular and tubular functional parameters were all superior in amlodipine group (P<0.05). At 5 days, the two groups showed significant differences in such glomerular and tubular functional parameters as urea nitrogen, creatinine, creatinine clearance rate, urine β2-microglobulin, and urine NAG (P<0.05), but not in serum cystatin C level. The incidence of contrast agent-induced nephropathy was significantly lower in amlodipine group than in the control group (5/95 vs 10/94, P<0.05).

CONCLUSIONSAmlodipine offers protection against radiographic contrast agent-induced renal injury in elderly patients with coronary heart disease.

Aged ; Amlodipine ; pharmacology ; therapeutic use ; Contrast Media ; adverse effects ; pharmacology ; Coronary Angiography ; Coronary Disease ; diagnostic imaging ; Female ; Humans ; Kidney Diseases ; chemically induced ; drug therapy ; Kidney Function Tests ; Male ; Middle Aged

Objective To investigate the predictive value of serum albumin level in patients with severe sepsis .Methods One hundred and twenty cases of patients with severe sepsis admitted to Qilu Hospital ,Shandong University from April 2014 to October 2014 were prospectively enrolled .The serum albumin levels were measured and the laboratory and clinical data were collected at the onset of severe sepsis .Acute Physiology and Chronic Health Evaluation (APACHE ) Ⅱ score and Sequential Organ Failure Assessment (SOFA) score were calculated .Patients were grouped according to the prognosis by day 28 or stratified by albumin level . Prognostic factors were analyzed by multivariable Logistic regression .Results A total of 120 patients were enrolled with mean age of (57 .6 ± 18 .3) years ,among which 75 were male .The mean duration of hospitalization was (20 .1 ± 17 .8) days .The 28‐day mortality was 25 .8% (31/120) .The most common infection sources were respiratory tract (56 .7% ) ,abdominal/pelvis (19 .2% ) and bloodstream (9 .2% ) .Serum albumin level in survival group was significantly higher than that in death group ([32 .1 ± 6 .4] g/L vs [27 .5 ± 5 .5] g/L ,t=3 .562 ,P=0 .001) .Compared with survival group ,the patients in death group had higher APACHE Ⅱ and SOFA scores (22 .0 ± 9 .1 vs 13 .4 ± 7 .2;7 .1 ± 3 .7 vs 4 .3 ± 3 .5 ;t= —5 .372 and —3 .690 ,both P<0 .05) .Along with the decrease of serum albumin level ,the incidence of bloodstream infection ,solid tumor ,septic shock ,acute kidney injury and liver injury significantly increased .Patients with lower albumin level had significantly higher SOFA scores and 28‐day mortality (all P<0 .05) .Multivariable regression analysis showed that albumin level lower than 28 g/L and higher APACHE Ⅱ score were independent risk factors for mortality (OR=4 .156 ,95% CI:1 .198—14 .415 ;OR=1 .121 ,95% CI:1 .039—1 .210;both P<0 .05) .Conclusions A significantly lowered serum albumin level would increase the risk of mortality in patients with severe sepsis .The combination of albumin level and APAHCE Ⅱ score might be beneficial to evaluate the prognosis .