The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

The objective of this study was to develop a rapid and precise detection technique for por-cine Senecavirus A(SVA)employing reverse transcription recombinase polymerase amplification(RT-RAA)in conjunction with CRISPR Cas13a technology.Additionally,the study aimed to opti-mize the assay's reaction conditions to enhance amplification efficiency.Eight RT-RAA primer sets were designed based on the conserved gene sequence of porcine SVA,and a series of reaction condi-tions were evaluated to refine the RT-RAA reaction system.Subsequently,CRISPR-derived RNA(crRNA)sequences were developed and selected to construct the RT-RAA-CRISPR reaction sys-tem.The method's specificity was determined by examining six prevalent porcine pathogenic nucleic acids,while its sensitivity was assessed using SVA cRNA standards quantified by digital PCR.The method's stability and the consistency of clinical sample analysis were also evaluated.The findings revealed that the optimized RT-RAA and CRISPR reaction systems exhibited the highest amplifi-cation efficiency at a reaction temperature of 37 ℃.Among the eight crRNAs,five were identified as exhibiting the strongest detection signals.The formulated RT-RAA-CRISPR Cas13a method demonstrated exceptional specificity,showing no cross-reactivity with other common porcine disea-ses,including ASFV,PRRSV,PEDV,PCV2,CSFV,and PRV.The method achieved high sensitivi-ty,detecting as low as 0.86 copies/μL of SVA,and exhibited stable fluorescence output,robust re-producibility,and the ability to complete clinical sample analysis within 50 minutes.Consistency e-valuation with six positive and 58 negative samples indicated 100％agreement in outcomes.These results substantiate that the study successfully established a rapid and specific RT-RAA-CRISPR Cas13a detection method for the on-site identification of porcine Senecavirus A,demonstrating high specificity and sensitivity,and holds promise for application in SVA monitoring and control initia-tives.

Objective To examine the compliance of and factors affecting target attainment of serum trough concentration of norvancomycin in ICU patients,and the effects of different trough concentrations on clinical efficacy and renal impairment.Methods Adult patients admitted to the Department of Critical Care Medicine of Huangshan People's Hospital and receiving intravenous infusion of norvancomycin from January 2020 to December 2022 were included.The dosing regimens and steady-state trough concentrations of norvancomycin were analyzed.The clinical efficacy and renal impairment were compared between different trough concentration levels.The compliance of trough concentration in critically ill patients with different renal functions was examined.Logistic regression analysis was performed to profile the factors possibly affecting the trough concentration of norvancomycin.Results A total of 97 patients were included.The target serum trough concentration(10-20 mg/L)was reached in only 33.0％(32/97)of the critically ill patients.The serum trough concentration was below the target in 51.5％(50/97)of the patients,above the target in 15.5％of the patients.The clinical cure rate and incidence of renal impairment were significantly different among the three groups of patients with different trough concentrations(P＜0.05).The compliance with target serum trough concentration varied with different renal function tests(P＜0.05).Augmented renal clearance and normal renal function were associated with trough concentrations lower than the target.As renal dysfunction got worse,serum trough concentration was more probably higher than the target(P＜0.05).Univariate analysis showed that daily dose,age,gender,height,weight,body mass index(BMI),APACHE Ⅱ score,sequential organ failure assessment(SOFA)score,blood creatinine,urea nitrogen,procalcitonin,concomitant septic shock,and use of norepinephrine were significantly correlated with trough concentrations of norvancomycin(P＜0.05).Multivariate logistic regression analysis indicated that age,SOFA score,blood urea nitrogen,gender,and norepinephrine use were independent factors affecting the serum trough concentration of norvancomycin(P＜0.05).Conclusions The serum trough concentration of norvancomycin varied with renal function states in ICU patients.It is difficult to achieve the steady-state target trough concentration(10-20 mg/L).The clinical cure rate is lower when the trough concentration is lower than the target.As the trough concentration increases,the incidence of renal impairment increases.Age,SOFA score,urea nitrogen,gender,and norepinephrine use are independent factors affecting the serum trough concentrations of norvancomycin.

Objective: To analysis the expression level and clinical value of serum exosomal miR-744-5p in non-small cell lung cancer (NSCLC). Methods: Retrospective study. A total of 183 subjects, including 92 NSCLC patients and 91 healthy controls, were recruited from Nanfang Hospital Affiliated to Southern Medical University from November 2016 to February 2018. Exosomes were extracted using ExoQuick and the total RNA was extracted from the exosomes.The expression level of serum exosomal miR-744-5p was assessed by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR). The diagnostic efficiency of exosomal miR-744-5p was evaluated by receiver operating characteristic (ROC) curve. Results: The qRT-PCR results revealed that the expression level of serum exosomal miR-744-5p in patients with NSCLC (0.012 2±0.019 7) was significantly lower than that in the healthy control (0.093 6±0.081 9), and the difference was statistically significant (U=553.5, P<0.001). Furthermore, the level of miR-744-5p in patients with early stage (stage Ⅰ and Ⅱ) lung cancer(0.011 7±0.013 9) was significantly different, when compared to healthy controls (U=72.5, P<0.001). The exosomal level of miR-744-5p in samples (0.010 6±0.020 3) was lower than that in squamous cell carcinoma serum (0.017 3±0.017 8) , and the difference was statistically significant (U=394.5, P<0.001). The ROC curve revealed that miR-744-5p expression can be a significant biomarker to discriminate between normal and NSCLC cases, with an area under the ROC curve of 0.933 9 (95%CI=0.899 6-0.968 1, P<0.000 1). MiR-744-5p can effectively diagnose early stage NSCLC, with an area under the ROC curve of 0.943 1 (95%CI=0.883 7-1.000 0, P<0.000 1). Conclusion The serum exosomal miR-744-5p could serve as a non-invasive biomarker for the diagnosis of NSCLC, but further comparative research and expanded verification are needed.

Objective To explore the value of Oxford acute severity of illness score in evaluating the severity and prognosis of critical illness patients.Methods All adult patients admitted to the Department of Critical Care Medicine from August 2012 to July 2014 were retrospectively analyzed.The severity in survivors and non-survivors was evaluated by using Oxford acute severity of illness score and APACHE Ⅲ score,and then statistic analysis were performed.Results Of 470 patients,321 (68.297％) were male,the range of age and ((x) ±s) age were 18 to 97 years and (59 ± 18) years respectively,and 123 patients (26.170％) were in non-survivors group and 347 patients in survivors group.The area under the ROC of Oxford acute severity of illness score was 0.760 (95％ CI:0.712-0.808,P ＜ 0.001),and Youden index was biggest when Oxford acute severity of illness score was 30.5.The area under the ROC of APACHE Ⅲ score was 0.844 (95％ CI:0.806-0.882,P ＜ 0.01),and Youden index was biggest when APACHE Ⅲ score was 70.5.Mortality was high (above 70％) as Oxford acute severity of illness score increased (＞ 40),and Spearman r was 0.976 (P ＜ 0.01).Conclusions Oxford Acute Severity of Illness Score was useful to evaluating the severity and prognosis of critical illness patients and it was easy in clinical practice.

Objective To investigate the safety and efficacy of nephron-sparing surgery (NSS)for selective T2 stage renal tumor.Methods The surgical database of 26 patients treated with NSS for clinical T2 stage renal cell carcinomas between March 2010 and May 2013 were collected and analyzed retrospectively.There were 17 males and 9 females,with a mean age of 52 years (39-74 years),mean tumor size of 10.3 cm(7.2-16.5 cm),and mean R.E.N.A.L score of 7.5 (6-10).Patients'demographics,clinical characteristics,oncologic outcomes,renal function were reviewed.Results The renal masses were removed successfully and the surgical margins were negative.There were 21 (80.8％) cases of clear cell carcinoma,4 (15.4％) papillary carcinoma and 1 (3.8％) chromophobe carcinoma.The mean ischemia time was (28.3 ± 12.5) minutes (7 patients were clamp-free).Three patients needed transfusion,one experienced urine fistula and cured by conservative treatment,and one patient's renal function got progressive worsening and required long-term hemodialysis.The average serum creatinine was 121 μ mol/L before and 136 μmol/L after surgery (P =0.06).After a period of 22-47 months' follow-up,no patient had local recurrence or metastasis.Conclusions NSS can be safely performed and provide effective oncologic outcomes for selective patients with clinical T2 stage renal cell carcinomas.R.E.N.A.L nephrometry is an important factor and should be used to evaluate the feasibility of NSS.

Objective To explore the significance of warning score of potential critical disease in predicting changes in patients with traumatic brain injury. Methods The clinic information of 75 patients with traumatic brain injury who were treated in the People's Hospital of Huangshan Affiliated to Wannan Medical College from Jan. to Dec. 2013 were analyzed retrospectively. The warning score of potential critical disease, modified early warning score( MEWS)and glasgow coma score( GCS)of all patients and the rates of changes in patients were calculated. Results Of 75 patients enrolled,60 were survived and 15 were died. Seventy-five patients were performed 448 times of warning score of potential critical disease,MEWS and GCS. The maximum, minimum of warning score of potential critical disease were 24 and 0,and the median score(P25,P75)was 4(2, 7). The maximum,minimum of MEWS were 24 and 0,and the median score( P25,P75 )was 4( 3,7 ). The maximum,minimum of GCS were 8 and 3,and the median score(P25,P75)was 5(4,7). The area under ROC of warning score of potential critical disease was 0. 76(95%CI =0. 66 -0. 86,P ﹤0. 01),Youden index was 0. 42 when score was taken 5. 5. The area under the ROC of MEWS was 0. 71( 95%CI =0. 61 -0. 81,P﹤0. 01),Youden index was 0. 4 when taken 3. 5 score. The area under the ROC of GCS was 0. 51(95%CI=0. 37-0. 63,P=0. 99),Youden index was 0. 27 when score was taken 4. 5. Conclusion The warning score of potential critical disease is effective to predict changes in conditions of patients with multiple injuries and better than MEWS and GCS.

Objective To explore the application value of pulse induced contour cardiac output (PiCCO) monitoring in diagnosis and treatment of patients with neurogenic pulmonary edema (NPE).Methods With review of literature, the data of 4 patients of severe neurological disease complicated by NPE admitted into Department of Critical Care Medicine of Huangshan People's Hospital Affiliated to Wannan Medical College from 2011 to 2013 were retrospectively analyzed and discussed in their PiCCO hemodynamic characteristics and processes of treatment.Results The PiCCO of 4 patients with NPE showed that the extravascular lung water index (EVLWI) was increased significantly (EVLWI was 12 - 42 mL/kg on admission and 10 - 22 mL/kg after hospitalization for 24 hours), all revealing a high permeability pulmonary edema type. The capacity balance of the first 24 hours in the 4 cases was all of positive balance (+1 130, +1 200, +1 750, +1 120 mL respectively). In the treatment, the supplementary colloid was strengthened, the vasoactive drugs such as, dopamine, dobutamine, milrinone, etc were applied to improve the circulatory oxygenation, then the EVLWI was declined; finally the disease situation in 3 cases was improved and one died.Conclusions The clinical diagnosis and treatment of NPE is complex, and many contradictions appear in the therapeutic course. PiCCO monitoring is valuable in early diagnosis, identification of pulmonary edema type, guidance in fluid supplement and vascular active drug application, and assessment of disease severity and prognosis.

ObjectiveTo explore the risk factors of the occurrence of acute kidney injury (AKI) in critically ill patients, and to investigate the effect of hydroxyethyl starch (HES) on renal function in these patients.Methods A prospective investigation was conducted. Critically ill patients admitted to Department of Critical Care Medicine of People's Hospital of Huangshan, Wannan Medical College from March 2012 to October 2013 were enrolled. For all the patients under observation, the following data were collected: demography, comorbidities, clinical presentation, severity of illness, and the use of blood product and drugs. All patients were divided into AKI group and non-AKI group by means of Acute Kidney Injury Network (AKIN) criteria, then the risk factors of AKI were investigated by means of univariate and multivariate logistic regression analysis. The effect of HES 130/0.4 administration on renal function in critically ill patients was evaluated.Results 314 patients were enrolled for study out of 1 152 patients admitted. Among these patients enrolled, 89 of them were found to suffer from AKI. AKI was classified as stage 1 in 59 patients, stage 2 in 19 patients, and stage 3 in 11 patients. It was shown by the univariate analysis that 12 variables were the risk factors of AKI, including age, hypertension, diabetes mellitus, acute physiology and chronic health evaluationⅡ (APACHEⅡ) score, sequential organ failure assessment (SOFA) score, coagulation SOFA score, neurological SOFA score, cardiovascular SOFA score, blood pH on intensive care unit (ICU) admission, blood glucose on ICU admission, accumulating dose of HES, and presence of shock (P< 0.05 orP< 0.01). However, HES administration and daily maximum dose of HES were not the risk factors of AKI in critically ill patients (bothP> 0.05). Using the multivariate logistic regression analysis, it was shown that total SOFA score [odds ratio (OR) = 1.20, 95% confidence interval (95%CI) = 1.09-1.32,P< 0.001], hypertension (OR = 2.44, 95%CI = 1.22-4.89,P= 0.012), blood glucose level on ICU admission (OR= 1.85, 95%CI = 1.32-2.59,P< 0.001), and presence of shock (OR = 3.81, 95%CI = 1.93-7.53,P< 0.001) were independent predictors of AKI in critically ill patients, however, the cumulative dose of HES was not independent risk factor for AKI (OR = 0.77, 95%CI = 0.68-0.87,P< 0.001).Conclusions Total SOFA score, hypertension, blood glucose level on ICU admission, and presence of shock were independent risk factors for AKI in critically ill patients. HES administration may not be a causative factor of an increased risk of AKI in the ICU.

Objective To explore the value of waring score of potential critical disease in predicting changes in condition of patients with multiple injuries. Methods From January 1, 2013 to July 31, 2013, all patients with multiple injuries were included prospectively. Patients were observed as soon as ICU admission. The waring score of potential critical disease and MEWS of all patients and the rates of changes in condition of patients were calculated then statistic analysis was performed. Results Of 50 patients enrolled, 44 were survived and 6 were died and 295 changes were found. The maximum , minimum median (P25, P75) of waring score of potential critical disease were 22, 0, 5 (3, 7). The maximum, minimum median (P25, P75) of MEWS were 12, 0, 4 (2, 6). The area under the ROC of waring score of potential critical disease was 0.880 (95% CI, 0.813-0.947, P < 0.001). Youden index was the biggest when waring score of potential critical disease was 6.5. The area under the ROC of MEWS was 0.767 (95% CI, 0.661-0.873, P < 0.001). Youden index was the biggest when MEWS was 5.5. Conclusion The waring score of potential critical disease was effective to predict changes in conditions of patients with multiple injuries and better than MEWS.