Objective The modified rapid one-step extraction (ROSE) method was employed in the Mycoplasma pneumoniae (Mp)genomic DNA extraction,in order to establish an easy DNA extraction method,which is used in Mp Loop Mediated Isothermal Amplification(LAMP) and suitable for high throughput without any expensive instruments.Methods In line with the character of Mp,a one-step Mp acid extraction method was established on the basis of ROSE.The studies had been carried out on comparing the modified ROSE DNA extraction with the classical methods of Boiling,CTAB,Alkali solution and reagent kit in extraction quality,quantity,reagent kinds,operation steps and extraction time.The extracted DNA of the Mp culture solution by these five methods had been used in LAMP to test the extraction efficiency.20 cases of Throat swab specimens from children with mycoplasma pneumonia were selected from January to March of 2016,and the throat swab specimens were extracted by the improved ROSE method then applied to LAMP detection system.Results Purity,comparison of ROSE method and other four methods were P＜0.05,the difference was statistically significant and ROSE was slightly better than the alkaline lysis method and boiling method,but lower than the CTAB method and kit method;Yield,results were compared with ROSE method and other four methods:there was no significant difference in q=0.95 (P＞0.05) compared with ROSE method and boiling method.The ROSE method was significantly different from the other three methods (P＜0.05),so the yield of ROSE method was higher than that of alkaline lysis method,CTAB method and kit method.Instrument and equipment,extracting procedure,extracting time and quantity ROSE,Boiling and Alkali solution were obvious ly better than CTAB and reagent kit.The extracted DNA by these five methods could be used in LAMP reaction,and the results showed no significant difference.Conclusion This study established an original one-step DNA genomic extraction method of Mp,which had been successfully applied in the Mp LAMP withoutany expensive instruments.Offer a groundbreaking method detecting Mp for the hospitals at all levels including the primary medical organizations.

OBJECTIVE To analyze the correlation factors of nosocomial infection in patients after receiving craniocerebral operation.METHODS Totally 388 hospitalized patients after receiving craniocerebral operation from Jan 2006 to Dec 2006 were studied retrospectively.RESULTS From them 45 pateints were suffering from nosocomial infection and total incidence of nosocomial infection was 11.59%.Among them,60.0% was lower respiratory tract infection,11.11% was surgical wound infection and 8.89% was upper respiratory infection.Aging,length of stay and invasive operation(deep vein needle detaining,trachea incision,tracheal intubation,urinary canal and drainage tube dwelling) were the main correlation tactors.CONCLUSIONS Minimizing invasive operation and early removing various kinds of dwelling tubes,using antibiotics correctly,improving hospitalizd environment and strengthening wards management are the measures to reduce nosocomial infection according its correlation factors.

Objective To study the effect of relaxation therapy on sleeping quality of patients with post cran- iocerebral traumatic syndrome. Methods 90 cases with sleep disorders were randomly divided into the treatment group and the control group. The control group was given routine care and the treatment group was given relaxation therapy combined with routine care. Take pittsburgh sleep quality index (PSQI) as evaluation criteria. Results There was significant difference between the two groups( P < 0.01 ). Sleep quality was more improved by relaxation therapy in the experimental group than that of the control group. Conclusion Relaxation therapy can effectively im- prove sleep quality in patients with post craniocerebral traumatic syndrome.

Objective To explore the clinical effect of both lower extremities exercise therapy on the prevention of deep vein thrombosis.Methods According to the randomized principal,200 cases of patients with limb paralysis caused by cerebrovascular disease were divided into the experimental group and the control group by random number table,and each group contained 100 cases.The experimental group was given both lower extremities exercise therapy in addition to routine care of limbs paralysis caused by cerebrovascular diseases.The control group received routine care and simple postoperative activity health education by nurses.The occurrence of deep venous thrombosis before the intervention and one month after intervention was evaluated in two groups.Results The incidence of deep vein thrombosis in the experimental group after intervention one month was lowered more significantly than that in the control group.Conclusions Both lower extremities exercise therapy can improve lower extremity venous blood flow velocity,and safely and effectively prevent the occurrence of DVT of patients with limb paralysis caused by cerebrovascular disease.

Objective To investigate the significance of neutrophil gelatinase associated lipocalin (NGAL) and kidney injury molecule-1 (KIM-1) in the patients of primary nephmpathy syndrome (PNS) complicated with acute kidney injury (AKI) by detecting urinary level of NGAL and KIM-1.Methods Senventy-two patients of PNS were selected as our subjects including 34 case with minimal change disease (MCD),23 cases with membranous nephropathy(MN),15 cases with mesangial proliferative glomerulonephritis (MsPGN).Fifteen cases of healthy physical examination were selected as control group.Meanwhile subjects were also divided into PNS with ATN group (15 cases) and PNS without ATN group (57 cases) according to their pathology check.ELISA was applied to detect the urinary level of NGAL and KIM-1,whose correlation with pathological type and clinical index were analyzed.Results The urinary level of NGAL and KIM-1 in patients with PNS were (42.37 ± 28.24) μg/L,(2.76 ± 1.11) μg/L respectively,higher than that of control group (P ＜0.01).The urinary level of NGAL in the MCD group,MN group,MsPGN group were higher than that of control group (46.81 ± 15.75) μg/L,(22.09 ± 7.69) μg/L,(15.31 ± 3.74) μg/L,(8.03 ± 0.35) μg/L respectively,P ＜ 0.05).The urinary level of NGAL in MCD group was significantly highest than that in the other groups (P ＜ 0.05).The urinary level of KIM-1 in the MCD group,MN group,MsPGN group were higher than that of control group(2.41 ±0.58) μg/L,(2.54 ±0.67) μg/L,(2.87 ±0.50) μg/L,(0.73 ±0.35) μg/L respectively,P ＜0.05).But there was no significant difference among MCD,MN,MsPGN groups(P ＞ 0.05).The urinary level of NGAL and KIM-1 in patients PNS were (42.37 ± 28.34) μg/L and (2.76 ± 1.11) μg/L,significantly higher than that of control group (t =4.668,12.665,P ＜ 0.05).The urinary level of NGAL and KIM-1 in patients PNS with ATN were significantly higher than patient without ATN (NGAL:(74.98 ±9.52) μg/L vs.(31.31 ±2.34) μg/L;KIM-1:(3.60 ±0.92) μg/L vs.(2.54 ±0.81) μg/L,P ＜0.05).The correlation analysis showed that the urinary level of NGAL and KIM-1 was positively correlated with serum β2-MG,serum creatinine,serum blood urea nitrogen and 24-hour urine protein.(r =0.432,0.299,0.234,0.254,0.434,0.650,0.276,0.301 respectively,P ＜ 0.05).Conclusion The urinary level of NGAL and KIM-1 could be considered as the early,non-invasive biologic factors to reflect in patients of PNS with ATN.

Objective To analyze the impac factors of serum N?terminal pro?brain natriuretic peptide (NT?proBNP) in patients with renal failure in non?dialysis phase, and to determine the cut?off point of as a diagnostic values in these patients with heart failure (HF). Methods Cross?sectional study was applied. Clinical data of 145 patients (37 cases of CKD4, 89 cases of CKD5, and 19 cases of acute renal injury (AKI) with renal failure in non?dialysis phase were collected. Comparison between groups and lineal regression analysis were utilized to investigate the impact factors of NT?proBNP, and the receiver operating characteristic curve (ROC curve) to select a better cut?off point of diagnosis in these patients with HF. Results (1) Compared with patients without HF, patients with HF had significantly higher edema, cardiac troponin I, serum phosphorus concentration, and left atrial diameter (LA), while ALB and left ventricular ejection fraction (LVEF) were decreased (P<0.05). (2) The NT?proBNP was divided into 4 groups with four points: First groups of 36 cases, NT?proBNP 1 ?862 ng/L, second groups 37 cases, 866?2670 ng/L, third groups 37 cases, 2790?20 000 ng/L, fourth groups 35 cases, 20 900?35 000 ng/L. With the increase of NT?proBNP levels, the occurrence of AKI and CKD4 decreased gradually while the occurrence of CKD and edema were significantly increased (P<0.01). Systolic blood pressure, troponin I, uric acid, serum phosphorus, parathyroid hormone, 24 hours urine protein, LA, interventricular septum thickness (IVS), left ventricular posterior wall thickness (LVPW) level gradually increased. Hb, ALB, calcium, CO2, eGFR, LVEF significantly decreased (P<0.01). The serum NT?proBNP of patients with HF was significantly higher than that of patients without HF (19 150 ng/L vs 1530 ng/L, P<0.01). The serum NT?proBNP of patients with edema was significantly higher than that in patients without edema (5460 ng/L vs 1630 ng/L, P<0.01). (3) Single factor linear regression analysis indicated that higher NT?proBNP was positive correlated with HF, edema, cardiac troponin I, uric acid, serum phosphorus, LA, IVS and LVPW (P<0.05), while negative correlated with Hb, eGFR, ALB, serum calcium, CO2, LVEF (P<0.05), and not correlated with eGFR, uric acid, serum calcium (P>0.05). (4) The best cut?off point of NT?proBNP predicting HF in patients with renal failure in non?dialysis phase was 3805 ng/L, AUC=0.848, 95%CI 0.786?0.910. Sensitivity was 82.4%, specificity 74.5%, positive predictive value 62.1%, negative predictive value 87.3%, positive likelihood ratio 3.2, negative likelihood ratio 0.24. Conclusions The level of NT?proBNP>20 000 ng/L is mainly found in end?stage renal disease patients with HF. HF is a main factor for the increase of NT?proBNP in patients with renal failure in non?dialysis phase. High phosphorus viremia, anemia, and hypoalbuminemia are closely related to NT?proBNP. Therefore NT?proBNP predicting HF should take into account the effects of these confounding factors in these patients.

Objective To detect the M-type phospholipase A2 receptor (PLA2R),and thrombospondin type-1 domain-containing 7A (THSD7A) expression in renal tissue and the levels of their antibodies in adult idiopathic membranous nephropathy (IMN).Also to determine the value of the two markers in the diagnosis of IMN.Methods One hundred and sixteen patients with biopsyproven MN at the Second Hospital of Hebei Medical University from December 2014 to August 2015 were enrolled,including 86 patients with IMN,10 patients with HBV-MN and 10 patients with stage Ⅴ lupus nephritis (LN-Ⅴ).Twenty patients with minimal change disease (MCD) were regarded as control group.We conducted immunohistochemical analysis of the presence of THSD7A and PLA2R the Paraffin section and enzyme linked immunosorbent assay (ELISA) detecting serum PLA2R-AB and THSD7A-AB concentration to investigate whether there was a correlation between them and clinical indicators.Results Compared with the SMN and MCD groups,the positive rates of PLA2R and PLA2R-AB were significantly higher in IMN groups.Expression PLA2R was detected in 88.4％,47.4％,10％ and 0％ and PLA2R-AB in 82.6％,15％,10％,0％,respectively,of the patients with IMN,HBV-MN,LN-Ⅴ and MCD.Expression THSD7A was detected in 2.3％ of the patients with IMN while not detected in SMN and MCD.THSD7A-AB antibody was negative in all patients.Compared with serum PLA2R-Ab negative individuals,patients with serum PLA2R-Ab positive had lower serum albumin (P ＜ 0.001),higher urine protein excretion (P=0.01).The sensitivity of PLA2R-AB,PLA2R,THSD7A and PLA2R+THSD7A in the diagnosis of IMN were 82.6％,88.4％,2.3％,88.6％,and the specificity was 92％,66.7％,100％,66.7％,respectively.Conclusions PLA2R in renal tissue and serum PLA2R-AB are specific markers for the diagnosis of IMN,which are closely related with the severity of IMN.Expression of THSD7A is only positive in some of IMN patients with negative PLA2R,which can be used as a supplementary examination of IMN patients with negative PLA2R.

Objective To explore the effect of different nasal feedings in enteral nutrition for elderly patients with severe brain hemorrhage. Methods Eighty elderly patients with severe brain hemorrhage were assigned into the observation group and control group according to the odd and even numbers at admission with 40 in each group. The control group was treated with graded injection nasal feeding, and the observation group was treated with continuous infusion nasal feeding. The nutrition indicators and enteral nutrition complications were compared between the two groups. Result The nutrition indicators in the observation group were better than those in the control group and the rate of enteral nutrition complications in the observation group was lower than that in the control group (P<0.05). Conclusion Continuous infusion nasal feeding can improve the nutritional status of elderly patients with severe cerebral hemorrhage and effectively reduce the incidence of complications of enteral nutrition. So it is worthy of clinical application.

OBJECTIVE:To establish a method for the content determination of carbohydrates in Osseous and cucumis extract injection. METHODS:Ion chromatography was conducted on the column of Dionex CarboPac PA20 with mobile phase of water-1 mol/L Sodium acetate solution-250 mmol/L Sodium hydroxide solution(gradient elution)at flow rate of 0.45 ml/min,column tem-perature was 35 ℃,and injection volume was 10 μl,detector was electrochemical analysis detector. RESULTS:The linear ranges of galactose,glucose,mannose,sucrose and fructose were 0.5-10.0μg/ml(r>0.998 0);RSDs of precision,stability and reproduc-ibility tests were lower than 2%;recovery was 94.8%-99.1%(RSD=0.52%-0.89%,n=9). CONCLUSIONS:The method is sim-ple,fast and accurate,and can be used for the determination of carbohydrates in Osseous and cucumis extract injection.

Objective To investigate the renal pathologic changes in both donors and transplant recipients with delayed graft function (DGF).Methods The clinical and laboratory data were retrospectively analyzed in 144 renal recipients with DGF.All the patients received renal biopsy,and donors' biopsy was performed on 131 recipients.The pathological changes were examined under the light microscopy (LM),immunofluorescence (IF) and electron microscopy (EM).Results (1) The incidence of DGF was 10.16%-7.48% during 1994 to 2005,and decreased to 5.35 % during 2006 to 2009.(2) Anuria occurred in 24 cases (16.67 %),oliguria in 24 (16.67%) and hypertension in 68 cases (47.22 %).The enlargement of transplanting kidney and the increased vascular resistance was detected in 79.67 % (98/123 cases) and 45.53 % (56/123 cases) respectively by ultrasound examination.(3) The level of serum creatinine was ranged from 451 to 707 μmol/L.The high level of urinary NAG enzyme was found in 102 cases (70.83 %),proteinuria in 79 recipients (54.86 %) and hematuria in 77 cases (53.47 %).(4) The acute rejection was observed in 66 cases (45.83 %),toxicity of CNI in 22 (15.28 %),IgA nephropathy in 18 (12.50 %),acute tubular necrosis in 8 (5.56 %),and recurrent FSGS in 2 cases (1.39 %).(5) In most recipients (66/109 cases,60.55 %)immunosuppressive regimen altered and renal replacement therapy was given.Conclusion The causes of DGF are complicated.The quality of donors' kidney and early histological changes of recipients are contributed to the development of DGF.It is necessary to perform renal biopsy not only in donors but also in recipients with DGF.And kidney biopsy in transplanted patients was also beneficial to the treatment.