Objective To prepare and identify anti‐CD47 monoclonal antibodies .Methods The gene fragment of CD47 was am‐plified by polymerase chain reaction and cloned into prokaryotic expressing vector pET‐32a(+ ) .Purified reconstructed protein was used to immunize BALB/c mice .The immunized spleen cells were isolated and fused with Sp2/0 cells .After screened ,hybridomas secreting anti‐CD47 monoclonal antibody were acquired .Biological activities of antibodies were investigated by Western blot and flow cytometry .Results The recombinant CD47 extracellular domain protein was successfully expressed in BL21 ,and certificated by sodium dodecyl sulfate polyacrylamide gel electropheresis and Western blot .Data of flow cytometry detection demonstrated that the antiserum had high affinity to CD47 protein .Conclusion Recombinant CD47 and its monoclonal antibody ,with high affinity , were successfully prepared ,which could provide reliable tools for the future study of CD47 .

ObjectiveTo study the prevalence of chronic kidney disease(CKD) among population with rheumatoid arthritis(RA) in Luohe city.Methods 3072 residents (older than 45 years) with eligible data from Luohe city were randomly selected using a stratified,multistage sampling.All residents were interviewed and given physical examination,tested for sample of uria and blood,and also given special examination about RA.Results The prevalence of RA was 6.90％ in 3072 subjects.The albuminuria was detected in 10.42％ of subjects,hematuria in 8.59％,reduced renal function in 1.82％,Howerer,which was higher in patients with RA,was 14.62％,10.85％ and 4.72％ respectively.Especially the prevalence of albuminuria and reduced renal function in RA patients was significantly higher compared to that in population without RA (14.62％ vs 10.10％,P＜0.05 and 4.72％ vs 1.61％,P＜0.01),but no significant differences in hematuria (10.85％ vs 8.43％,P＞0.05).The prevalence of CKD was 16.93％ in tougher,and RA patients had higher prevalence of CKD than those population without RA (22.17％ vs 16.54％,P＜0.05).ConclusionThe prevalence of CKD among population older than 45 years in Luohe city is high,and the patients with RA have special characteristic in epidemiology of CKD.

Objective To explore the effect of serum uric acid (SUA) on the clinicopathological manifestation and prognosis of IgA nephropathy(IgAN)patients. Methods A total of 348 patients with renal biopsy-proven IgAN in our hospital were enrolled in this study.The data were retrospectively analyzed to examine the association of SUA level with clinicopathological manifestation and prognosis of IgA nephropathy(IgAN)patients. Results There were no significant differences of 24 hour proteinuria,BUN and Scr between patients of high SUA level with various GFR and those of normal SUA level.While differences of glomerular sclerosis,tubulointerstitial scores and vascular injury between these two groups were significant (P＜0.05).At the end of follow-up,prevalence of GFR decline and ESRD was significantly higher in patients with high SUA as compared to those with normal SUA(40.82%vs 15.70%,64.71% vs 35.00%,respectively,P＜0.05). Conclusions Patients with different SUA levels have similar clinical manifestations,but different pathological findings and prognosis.It is important to pay attention to the follow-up of SUA level in IgAN patients.

OBJECTIVE:To optimize the ultrasonic extraction and purification technology of total saponins from Marsdenia te-nacissima,and to prepare purified total saponins from M. tenacissima. METHODS:Using the extraction rate of total saponins from M. tenacissima as index,ethanol volume fraction,solvent-solid ratio and extraction temperature as factors,ultrasonic extraction technology of total saponins from M. tenacissima was optimized by single factor test and Box-Behnken response surface design. Us-ing the content of total saponins in elution,the effects of ethanol elution volume fraction on purification technology of total sapo-nins by D101 type macroporous adsorption resin was investigated. RESULTS:The optimal ultrasonic extraction technology was as follows as ethanol volume fraction of 85%,solvent-solid ratio of 18.5∶1 (ml/g),extraction temperature of 80 ℃. In verification test,the average extraction rate of total saponins from M. tenacissima was 4.80%(RSD=1.06%,n=3);it was close to predicted value 4.89%,and the deviation was 1.84%. The optimal purification technology was as follows as 45% ethanol as eluant,90%ethanol for purification. Verification test showed that the average content of total saponins from M. tenacissima was 62.45%(RSD=0.88%,n=3). CONCLUSIONS:The optimized technology can be used for the extraction and purification of total saponins from M. tenacissima. The technology is stable and reliable.

Objective To evaluate the role of cadiocyte connexin 43 (Cx43) in mito-KATP channel mediated cardioprotection against ischemia-reperfusion(I/R) injury induced by sevoflurane preconditioning in isolated rat hearts.Methods Forty hearts from male adult SD rats weghing 200-250 g were excised and perfused in a Langendorff apparatus with K-H solution with 95 ％ O2-5 ％ CO2 at 36.5-37.5 ℃.Their hearts were randomly divided into 5 groups (n =8 each): control group (group C),group I/R,sevoflurane preconditioning group (group S),sevoflurane preconditioning + 5-HD group (group SH) and 5-HD group (group H).Myocardial I/R was induced by occlusion of the anterior descending branch of left coronary artery (LAD) for 30 min followed by 120 min reperfusion.After 10 min of equilibration,group C received continuous perfusion and LAD was exposed but not occluded.Group I/R received continuous perfusion for 30 min and LAD was occluded.In group S,SH and H,3％ sevoflurane,3 ％ sevoflurane mixed with 100μmmol/L 5-HD and 100 tmmol/L 5-HD was added into K-H solution to perfuse for 15 min respectively.After that,the hearts were washed by K-H solution for 15 min.HR,left ventricular systolic pressure ( LVSP),left ventricular diastolic pressure (LVDP) and ± dp/dtmax were recorded before administration (T0),immediately after administration (T1),immediately before ischemia ( T2 ),at 30 min of ischemia (T3) and 120 min of reperfusion (T4).At the end of reperfusion,left ventricular tissue was removed for determination of myocardial infarct size and expression of Cx43 and phosphor Cx43 (p-Cx43) by immunohistochemistry and Western blot respectively.Results Compared with group C,HR,LVSP and ± dp/dtmax were significantly decreased,LVDP was increased and the expression of Cx43 and p-Cx43 were down-regulated in groups I/R,SH and H (P ＜ 0.05).Compared with group I/R,HR,LVSP and ± dp/dtmax were significantly increased,LVDP and myocardial infarct size decreased,and the expression of Cx43 and p-Cx43 up-regulated in group S ( P ＜ 0.05),no significant difference was found in groups S + H and H( P ＞ 0.05 ).Conclusion Sevoflurane preconditioning can open mito-KATP channel,promote cadiocyte Cx43 phosphorylation,attenuate I/R injury in isolated rat hearts.

Objective To explore the prevalence of chronic kidney disease(CKD) and its related risk factors in the people aged 60 years or over in Zhengzhou.Methods One thousand two hundred and ninety-one residents aged 60 years or over were selected with stratified-multi-steps-cluster sampling method in 4 communities from 4 different districts in Zhengzhou city.The indicators of renal damage and its related factors were collected with questionnaires.Results Eligible data of 1247 subjects were collected.After adjustment for age and gender, the prevalence of albuminuria,hematuria and reduced eGFR were 13.3% ,6.0% and 2.0% respectively, and the prevalence of CKD was 17.5%.Multiple logistic regression analysis showed that hypertension, diabetes and overweight were independent risk factors for CKD.Conclusions The prevalence of chronic kidney disease in the urban people aged 60 years or over in Zhengzhou is higher than in the non-elderly people and preventive measures should be adopted.

Objective To study the prevalence of chronic kidney disease (CKD) among patients with bronchial asthma in Zhengzhou.Methods A total of 655 patients (older than 14years) were interviewed and received physical,uria and blood examination.Bronchial asthma and CKD were diagnosed according to related definition.Results The prevalence of albuminuria,hematuria,reduced eGFR and CKD was 9.41%,10.37%,3.03% and 17.38% respectively in the above special patients,which was higher than that of the general population.Women had higher prevalence of CKD than men (21.47% vs 13.99%,χ2=6.060,P=0.014).Prevalence of 1 to 5 stage of CKD was 8.61%,5.10%,3.03%,0.48% and 0.16% respectively.In the acute phase of asthma,prevalence of CKD (24.42%) was significantly higher than that in chronic duration and clinical remission period (χ2=12.445,P=0.002) and the same significant differences were found in albuminuria and reduced eGFR (χ2=19.619,P＜0.01 and χ2=9.305,P=0.010).Conclusion The bronchial asthma patients have higher incidence of renal damage and should be estimated,especially in the acute phase.

Objective To study the prevalence and risk factors of chronic kidney disease (CKD) among adults in Zhengzhou. Methods One thousand eight hundred and fifty five residents (≥ 20 years) from 4 communities in 4 districts of Zhengzhou city were randomly selected by using a stratified,multistage sampling. They were interviewed, and received physical examination and measurements of urine and blood for renal damage as well as risk factors. Results Eligible data of 1752 subjects were included in the study. After the adjustment of age and gender component, albuminuria was found in 5.78% of the subjects, hematuria in 8.19%, and reduced renal function in 1.58%. Male had lower prevalence of albuminuria and hematuria (4.37% vs 7.29%, X2=6.252, P=0.012; 5.08% vs 11.51%, X2=24.499, P＜0.01), but higher prevalence of reduced eGFR(2.26% vs 0.86%, X2=5.830, P=0.016) as compared with female. The prevalence of albuminuria and reduced eGFR increased with age. The crude prevalence of CKD was 14.50%, while the standardized rate was 13.57%. The prevalence of female was higher than that of male (17.83% vs 9.59%, X2=23.132, P＜0.O1), which also increased with age. The most common manifestations of CKD were hematuria and albuminuria. Gender, age, smoking, hypertension,diabetes mellitus, obesity and hyperuricaemia were independently associated with CKD. The awareness rate of CKD was 8.27% and only 7.09% of the subjects received treatment. Conclusions The prevalence of CKD is 13.57% and the recognition is 8.27% in urban adult population of Zhengzhou.lndependent risk factors associated with kidney damage are gender, age, smoking, hypertension, diabetes mellitus, obesity and hyperuricaemia.

Objective To investigate the effects of hydrogen gas on the myocardial injury in rats with endotoxemia.Methods Forty-eight healthy male Sprague-Dawley rats,weighing 200-220 g,were randomly divided into 4 groups (n =12 each ):control group (group C ),hydrogen gas control group (group HC ),endotoxemia group (group E) and hydrogen gas + endotoxemia group (group LH).Endotoxemia was induced by intraperitoneal lipopolysaccharide (LPS) 20 mg/kg in groups E and LH,while the equal volume of normal saline was given in groups C and HC.After LPS administration,the rats were exposed to the air containing 2％ hydrogen gas for 6 h in groups HC and LH,and the rats were exposed to the air for 6 h in groups C and E.Blood samples were taken from the abdominal aorta after 6 h inhalation of hydrogen gas to determine the serum level of cTnI.The hearts were then removed to determine the content of TNF-α and IL-6 and expression of phosphorylated nuclear factor κB ( NF-κB) inhibitory protein (p-IκB-α) and p38 mitogen-activated protein kinase (p-p38MAPK) in the myocardial tissues.Results Compared with group C,no significant change was found in the levels of cTnI,TNF-α and IL-6 and expression of p-IκB-α and p-p38MAPK in group HC ( P ＞ 0.05),and the levels of cTnI,TNF-α and IL-6 were significantly increased and the expression of p-IκB-α and p-p38MAPK was up-regulated in groups E and LH ( P ＜ 0.05).Compared with group E,the levels of cTnI,TNF-α and IL-6 were significantly decreased and the expression of p-IκB-α and p-p38MAPK was down-regulated in group LH ( P ＜ 0.05).Conclusion Hydrogen gas can reduce the myocardial injury in rats with endotoxemia,and inhibition of the p38MAPK and NF-κB pro-inflammatory pathway and reduction of the inflammatory response of myocardial tissues are involved in the mechanism.

Objective To evaluate the diagnostic accuracy of ultrasound bladder detrusor wall thickness (DWT) measurement for female bladder outlet obstruction (FBOO) and investigate the application of this non-invasive method for diagnosis of FBOO.Methods DWT was measured by linear ultrasound (7.5 MHz) either at a filling volume of 50％ of cystometric capacity or at 250 ml filling in 93 women undergoing pressure flow study (PFS) for lower urinary tract symptoms (LUTS).FBOO was defined as maximal flow rate (Qmax) of less than 12 ml/s combined with a detrusor pressure at maximal flow rate (Pdet Qmax)greater than 25 cm H2O.All the patients were divided into 2 groups according to FBOO and non-FBOO.The age,urodynamic parameters and DWT of the 2 groups were compared.Meanwhile DWT was evaluated for diagnosing BOO through receiver operating characteristic (ROC) curve.ResultsThe age (61.2 ± 8.3 vs 59.9 ± 7.7 years,P =0.44) and maximal cystometric capacity (292.2 ± 82.3 vs 308.1 ± 87.5 ml,P =0.37 ) between the 2 groups had no significant difference.DWT was significantly higher ( P =0.00 ) in FBOO group (42 cases,DWT 1.8 ±0.3 mm) compared to non-FBOO group (51 cases,1.4 ±0.2 mm).Furthermore maximal detrusor pressure (43.1 ± 11.2 vs 16.2 ± 7.1 cm H2O,P =0.00),Pdet Qmax (34.3 ±8.2 vs 13.1 ±7.8 cm n2O,P =0.00),Qmax(7.4 ±3.2 vs 17.4 ±4.1 ml/s,P =0.00),voided volume (157.1 ±63.7 vs 251.2 ±77.4 ml,P=0.00) and post-void residual volume (117.5 ±71.3 vs 37.7 ± 18.1 ml,P =0.00) had significant differences between the 2 groups.For a diagnosis of FBOO,DWT of 1.9 mm or greater had a positive predictive value of 100％,a negative predictive value of 62％,specificity of 100％ and sensitivity of 38％.ROC analysis revealed that DWT had a high predictive value for FBOO with an area under the curve of 0.88 ± 0.06.Conclusions Ultrasound DWT measurement for FBOO is non-invasive,convenient and reliable.DWT 1.9 mm or greater assessed by ultrasound has a high predictive value and specificity for FBOO and can replace PFS in some extent.However,this cutoff value needs to be validated in multiple center and larger population study.