Objective : To investigate the role of caspase recruitment domain-containing protein 9(Card9) inAspergillus fumigatus(A.fumigatus) keratitis and the effect of its deficiency on macrophage resistance to fungal infection. Methods : (1) C57BL/7 mice aged 6-8 weeks were selected and the mice pretreated Card9 siRNA and Blank siRNA, respectively, and the expression of Card9 in each group was detected by Western blot and RT-PCR. The corneal epithelium of the mice was scraped away 72 hours later, andA.fumigatusspore suspension was injected into the corneal stroma. The corneal scores were recorded at 1 d, 3 d, 5 d and 7 d after infection. The expression of Card9, nuclear factor κB(NF-κB), interleukin 1β(IL-1β), interleukin 6(IL-6) and tumor necrosis factor α(TNF-α) in each group was detected by RT-PCR and immunohistochemical(IHC).(2) Human corneal epithelial cells(HCECs) and human monocytic-leukemia cells(THP-1)in vitro, RT-PCR was used to examine the expression of Card9 gene in the two cells, and a stable cell line of THP-1 cells was constructed using shRNA vectors. The expression of Card9 in the cell line was detected by Western Blot and RT-PCR. The cells were induced into macrophages and stimulation byA.fumigatus, and the expression of Card9, NF-κB, IL-1β, IL-6 and TNF-α was detected by RT-PCR. Results : Card9 expression increased inA.fumigatuskeratitis, mainly distributed in cytoplasm of immune cells. The expression of Card9 in the cornea of mice treated with Card9 siRNA was significantly reduced. After inhibiting the expression of Card9 gene, the expressions of Card9, NF-κB, IL-1β, IL-6 and TNF-α significantly decreased and the changes of IL-1β were most significant. Inin vitrostudies, Card9 exhibited negligible expression in human corneal epithelial cells, contrasting with its pronounced expression in THP-1 cells. After the induction of macrophages, Card9, NF-κB, IL-1β, IL-6, and TNF-α were significantly upregulated under the stimulation ofA.fumigatus. After inhibiting the expression of Card9, the stimulated expression of these factors was significantly reduced, with the most notable change observed in IL-1β. Conclusion Card9 is involved in the inflammatory development and healing process ofA.fumigatuskeratitis. Card9 deficiency can cause functional impairment of macrophages and inhibit the expression of inflammatory factors to a certain extent, in which IL-1β has the greatest effect.

Objective:To analyze the changes of diagnosis and treatment before and after renal biopsy in adult patients with acute kidney disease (AKD), and to explore the value of renal biopsy in the diagnosis and treatment of AKD.Methods:It was a single-center retrospective observational study. The adult patients with AKD who underwent renal biopsy in the Department of Nephrology of the First Affiliated Hospital of Nanjing Medical University from January 1, 2017 to December 31, 2021 were enrolled. Demographic data, general clinical data, laboratory tests, and diagnosis and treatment data before and after renal biopsy were collected to analyze the concordance rate between clinical and pathological diagnoses, changes in treatment after renal biopsy, and bleeding complication.Results:A total of 575 patients diagnosed with AKD by renal biopsy were included in this study, with age of 51 (36, 63) years old and 359 males (62.4%). Among them, there were 293 patients (51.0%) of acute kidney injury, 348 patients (60.5%) of hypertension and 124 patients (21.6%) of diabetes. The peak serum creatinine was 272 (190, 477) μmol/L. The hemoglobin was 106 (86, 126) g/L. The 24-hour urine protein was 2.15 (0.79, 4.82) g. There were 347 patients (60.3%) of acute glomerular diseases, 136 patients (23.7%) of acute interstitial nephritis, 47 patients (8.2%) of thrombotic microangiopathy, and 45 patients (7.8%) of acute tubular necrosis. The most common types of acute glomerular diseases were IgA nephropathy and anti-neutrophil cytoplasmic antibody-associated glomerulonephritis, accounting for 22.3% (128/575) and 12.2% (70/575), respectively. The clinical diagnoses before renal biopsy were consistent with the renal histopathological diagnoses in 454 patients, with an accuracy rate of 79.0%. Following the renal biopsy, the treatment plan involving glucocorticoids or immunosuppressants was adjusted in 394 patients (68.5%). Significant post-biopsy bleeding occurred in 15 patients (2.6%), with 12 patients requiring blood transfusion and 1 patient requiring surgical intervention.Conclusions:Twenty-one clinical diagnoses do not match the pathological diagnoses in adult AKD patients, 68.5% of patients have changes in their treatment plans, and 2.6% of patients have significant hemorrhagic complications after renal biopsy. Clinicians need to carefully consider the benefits and risks and make individualized decisions about renal biopsy.

Objective:To evaluate the effect of preoperative plasma D-dimer level on the risk of cardiac surgery-associated acute kidney injury (CSA-AKI). Methods:The clinical data of patients who underwent cardiac surgery with cardiopulmonary bypass in the First Affiliated Hospital of Nanjing Medical University from January 2017 to December 2018 were collected retrospectively. All patients were distributed into two groups (normal D-dimer group with D-dimer level≤0.55 mg/L and elevated D-dimer group with D-dimer level>0.55 mg/L) according to the D-dimer threshold of 0.55 mg/L and the differences of clinical data between the two groups were compared. Kaplan-Meier survival analysis method was used to analyze the difference of the cumulative incidence of CSA-AKI between the two groups. Logistic regression analysis and restricted cubic splines analysis were used to analyze the association between serum D-dimer and the incidence of CSA-AKI. Results:There were 871 patients enrolled in the study with 427 females (49.0%) and age of (56.6±12.3) years, including 215 patients (24.7%) with high D-dimer and 259 patients (29.7%) with CSA-AKI. Compared with the normal D-dimer group, patients with elevated D-dimer had higher baseline serum creatinine, proportion of chronic kidney disease stage 3, international normalized ratio, fibrinogen, proportion of receiving renal replacement therapy and incidence of CSA-AKI (all P<0.05). The prothrombin time, operation time, extracorporeal circulation time, aortic occlusion time and hospital stay in the elevated D-dimer group were longer (all P<0.05), and the preoperative estimated glomerular filtration rate (eGFR) and hemoglobin levels were lower than those in the normal D-dimer group (both P<0.05). There was no statistical difference between the two groups in terms of age, gender, comorbid diseases, cardiac function classification, and hospital mortality (all P>0.05). Kaplan-Meier survival curve results showed that compared with the normal D-dimer group, the risk of CSA-AKI in the elevated D-dimer group was significantly increased (Log-rank χ2=14.227, P<0.001). The multivariate logistic regression showed that after adjusting variables including gender, age, diabetes mellitus, preoperative eGFR, cardiopulmonary bypass time and so on, the higher level of preoperative D-dimer (>0.55 mg/L) was still related to CSA-AKI ( OR=1.476, 95% CI 1.038-2.098, P=0.030). Restricted cubic splines analysis showed that the incidence of CSA-AKI raised when preoperative serum D-dimer concentration increased (non-linear P=0.262). Conclusion:Patients with high preoperative serum D-dimer have an increased risk of CSA-AKI.

Objective To investigate the predictive value of nutritional and fluid status measured by bioelectrical impedance methods for the prognosis of acute kidney injury (AKI) patients undergoing continuous renal replacement therapy (CRRT). Methods Patients with severe AKI received CRRT in the First Affiliated Hospital of Nanjing Medical University from September 2016 to September 2018 were enrolled, and divided into death group and survival group according to 28-day survival. Cox regression was used to analyze the association between 28-day survival and lean tissue index (LTI), fat tissue index (FTI), the ratio of extracellular water (ECW) and body cell mass (BCM) (ECW/BCM), and overhydration (OH), respectively. Results A total of 156 patients were included, including 101 males and 55 females. The age was (62.7 ± 15.4) years, with sequential organ failure assessment (SOFA) score of 9.9±3.9. The 28-day mortality rate was 46.2％. The pre-CRRT OH values in the 28-day survival group and death group were 2.95(1.80, 5.50) L and 4.20(2.95, 5.70) L(P=0.016), and ECW/BCM values were 1.00(0.76, 1.18) and 1.07(0.88, 1.25) (P=0.033), respectively. Univariate Cox regression analysis showed that pre-CRRT high OH values (HR=1.08, 95％CI 1.00-1.17, P=0.040) and high ECW/BCM values (HR=3.02, 95％CI 1.46-6.22, P=0.003) were associated with 28-day death. The changes of OH values (HR=0.83, 95％CI 0.72-0.95, P=0.008) and ECW/BCM values (HR=6.79, 95％CI 1.72-26.82, P=0.006) between pre - CRRT and the 7th day after CRRT initiation were significantly associated with 28-day mortality in patients who survived 7 days after CRRT initiation. After adjusting for age, gender, and SOFA scores, multivariate Cox regression analysis showed that the high OH value (HR=1.16, 95％CI 1.06-1.27, P=0.002) and the high ECW/BCM value (HR=2.80, 95％CI 1.30-6.06, P=0.003) before CRRT, the change of OH value (HR=0.82, 95％CI 0.72-0.95, P=0.008) and ECW/BCM value (HR=2.79, 95％CI 1.30-5.98, P=0.009) between the 7th day after CRRT initiation and pre-CRRT, were independently associated with 28-day death, while LTI (HR=0.93, 95％CI 0.86-1.02, P=0.113) and FTI (HR=0.98, 95％ CI 0.92-1.04, P=0.475) before CRRT were uncorrelated with 28-day death. Conclusions In bioelectrical impedance analysis, the high OH value and high ECW/BCM value before CRRT are associated with 28-day mortality in patients with AKI, while the nutritional indicators LTI and FTI before CRRT are not significantly related. The correction of fluid overload by CRRT within 7 days may reduce the risk of 28-day mortality.

Objective To analyze the etiological characteristics of human Brucella strains isolated, and to improve the precision in control and prevention of brucellosis. Methods In 2016, blood samples were collected from patients in Jingyuan County Gansu Province, and tested via the Rose-Bengal Plate Agglutination Test (RBPT) and the tube agglutination test methods,and serological positive blood samples were inoculated to bidirectional blood culture bottle for culturing, and further identified by traditional biological classification method and the Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis species-specific PCR (AMOS-PCR). Multiple-locus variable number tandem repeat sequence analysis (MLVA) -16 was used to detect molecular typing and do cluster analysis. Results The isolated strain was identified by the traditional biological classification method, bacteria could grow in thionine and reddened dye, A and M factors agglutination tests were positive, Bk2phage treatment of bacterial strain cracking, but Tb, Wb phages were not cracked. AMOS-PCR amplification result showed a 731 bp band, which was a strain of Brucella melitensis. The results of MLVA-16 showed that there was a difference in the number of repeats on some Variaable Number of Tandem Repeat(VNTR)sites of the isolated strain. Clustering analysis showed that, the isolated strain was clusted into the same clade with the clustering of Brucella melitensis type 3 from GS-201605 in Gansu. And the clustering was similar compared with that of Zhejiang, Guangdong, Fujian and Yunnan. Conclusion Human brucellosis is a inputting transmission in Gansu Province, there is a genetic variation of genotype 3 sheep Brucella between Gansu Province and other domestic provinces.

Objective To compare different equations for estimated glomerular filtration rate (eGFR) in patients with chronic kidney disease (CKD).Methods Hospitalized patients with CKD from the nephrology department of the First Affiliated Hospital of Nanjing Medical University (Jiangsu Province Hospital) were recruited between December 2014 and May 2015.The calculations of eGFR and 24 h creatinine clearance rate (Ccr) were accomplished in three days after admission.The eGFRs were calculated separately using the 24 h creatinine clearance rate adjusted by the standard body surface area (Ccr_BSA),Cockcroft-Gault equation adjusted by the standard body surface area (eCcr_BSA),CKD-EPI creatinine equation (EPI_Cr),CKD-EPI cystatin C equation (EPI_CysC),CKDEPI creatinine-cystatin C equation (EPI_Cr_CysC),simplified MDRD (MDRD) and China MDRD equations.The EPI_Cr_CysC equation was used as the standard and the precision and accuracy of the other six equations were compared and analyzed.Results A total of 403 CKD participants were enrolled in the study,with 228 male patients and a mean age of (54.9± 18.4) years.The main primary diseases were chronic glomerulonephritis (43.7％) and diabetic nephropathy (13.2％).The median concentration of serum creatinine and cystatin C were 117.5 (69.7,242.4) μmol/L and 1.80 (1.13,3.31) mg/L,respectively.The median values of Ccr_BSA,eCcr_BSA,MDRD,China MDRD,EPI_Cr,EPI_CysC and EPI_Cr_CysC equations were 50.8 (21.1,96.2),51.9 (23.3,93.2),53.6 (23.0,97.4),52.2(22.4,94.1),53.2 (22.1,97.3),35.1 (15.4,67.0) and 49.1 (22.8,82.3) ml · min-1 · (1.73 m2)-1,respectively.There was well agreement among MDRD,China MDRD and EPI_Cr equations,while there were large differences between equations derived from CysC (EPI_Cr_CysC and EPI_CysC) and equations derived only from creatinine (EPI_Cr,MDRD,China MDRD,eCcr_BSA,Ccr_BSA equations).Compared with EPI_Cr_CysC equation (the reference equation),EPI_Cr equation showed the highest accuracy [percentage of other eGFR equation calculations that were ＞ 30％ of the reference equation calculations (1-P30),30.8％] while Ccr_BSA equation showed the lowest (1-P30,42.4％).EPI_CysC equation showed the highest precision [inter-quartile range (IQR) of the difference,11.7 ml·min-1 · (1.73 m2)-1] while Ccr_BSA equation showed the lowest [IQR of the difference,22.8 ml· min-1 ·(1.73 m2)-1].Conclusions The agreement among equations derived only from creatinine is better;while it exhibits some differences between equations with cystatin C and equations derived only from creatinine.The accuracy of EPI_Cr equation is second only to EPI_Cr_CysC equation and it is currently the most suitable eGFR equation for clinical popularization of renal glomerular function assessment.

Objective Acute kidney injury (AKI) is common but usually under-diagnosed in hospitalized patients,of the impact of which on patients is still unclear.The paper was aimed to investigate the impact of delayed recognition of AKI on short-time prognosis of patients through a propensity score matched study.Methods From Oct 2013 to Sep 2014,1401 adult hospitalized patients with AKI in the First Affiliated Hospital of Nanjing Medical University were divided into delayed recognition group and timely-diagnosed group according to propensity score matching (1∶ 1) without replacement method.Primary endpoint was 30-day all-cause mortality,and secondary endpoints included recovery of kidney at discharge,length of hospitalization,length of intensive care unit stay and hospital costs.Results There were significant differences in age,department distribution,complications,stage of AKI,Charlson index,APACHE Ⅱ score,SOFA score between the two groups before matching.After matching,there were no significant difference in demographic data,department distribution,complications,stage of AKI,Charlson index,APACHE Ⅱ score,SOFA score between the two groups except in blood urea nitrogen (P=0.039) and use of diuretics (P=0.018).Delayed recognition of acute kidney injury was not associated with 30-day all-cause mortality in univariate (P=0.711) and multivariate Logistic regression analyses.The secondary endpoints did not differ in two groups.Conclusion Delayed acute kidney injury recognition did not associate with poor short-term outcomes in adult hospitalized patients.

Objective To identify the Brucella strains isolated from sheep blood in Jingyuan County Gansu Province,and to provide scientific basis for Brucellosis control and prevention.Methods In 2016,blood samples from suspected cases of sheep were collected in the home of Brucellosis patient of Jingyuan County,and cultured with vaccination double blood bottles via the enrichment method,and the Brucella strains isolated from sheep were identified by traditional bio-typing,genus specific Brucella surface protein 31 PCR (BSCP31-PCR) and Brucella abortus,Brucella melitensis,Brucella ovis,Brucella suis species-specific PCR (AMOS-PCR),respectively.Results The isolated strain was identified using the traditional biological classification method,bacteria could grow in thionine and fuchsin dyes,A and M factors serum agglutination tests were positive,Bk2 phage lysis test was positive,but Tb and Wb phage lysis tests were negative;and the results of BSCP31-PCR of tested strain was 223 bp band same to Brucella;the results of AMOS-PCR was 731 bp band same to Brucella melitensis.Conclusion Conventional bio-typing test assisted with molecular techniques has confirmed that the isolated Brucella strain from sheep blood is Brucella melitensis type 3 in Jingyuan County Gansu Province.

Objective To investigate the risk factors of fracture patients in CKD5D stage.Methods A total of 124 CKD5D patients according to suffering fracture or not in past or at present were divided into fracture group and non-fracture group.Clinical data of age,sex,height,body mass index(BMI),dialysis duration,dialysis patterns,body mass index(BMI),hypertension,diabetes mellitus (DM) history,drug taking history were recorded.Laboratory examinations included hemoglobin (Hb),hematocrit (Hct),serum creatinine (Scr),blood urea nitrogen (Bun),albumin (A),triglyceride (TG),cholesterol(TC),high-density lipoprotein (HDL),low density lipoprotein (LDL),serum sodium (Na +),serum calcium (Ca2 +),serum phosphorus (P),parathyroid hormone (PTH),alkaline phosphatase (AKP),and 25 (OH) VitD.And bone mass density (BMD) were measured.Results The fracture patients had longer dialysis duration,higher of hemodialysis ratio,higher serum Ca2 +,serum P,serum PTH and AKP,lower T and Z values of bone density than those patients without fracture.Univariate regression analysis of fracture risk suggested that dialysis age,high calcium,high phosphorus,high PTH,high AKP,high 25 (OH) VitD and fracture were positively correlated with peritoneal dialysis,and was negatively correlated with the use of calcium and peritoneal dialysis.The Multivariate regression analysis showed that high PTH,high AKP was positively correlated with fracture occurrence,lumbar vertebra bone density and the use of calcium were negatively correlated with fracture.Conclusion Low bone mineral density is an important risk factor for fracture.The use of calcium can reduce the risk of fracture.High PTH and high AKP should be actively intervened in CKD5D patients.

Objective To investigate the risk factors of fracture patients in CKD5D stage.Methods A total of 124 CKD5D patients according to suffering fracture or not in past or at present were divided into fracture group and non-fracture group.Clinical data of age,sex,height,body mass index(BMI),dialysis duration,dialysis patterns,body mass index(BMI),hypertension,diabetes mellitus (DM) history,drug taking history were recorded.Laboratory examinations included hemoglobin (Hb),hematocrit (Hct),serum creatinine (Scr),blood urea nitrogen (Bun),albumin (A),triglyceride (TG),cholesterol(TC),high-density lipoprotein (HDL),low density lipoprotein (LDL),serum sodium (Na +),serum calcium (Ca2 +),serum phosphorus (P),parathyroid hormone (PTH),alkaline phosphatase (AKP),and 25 (OH) VitD.And bone mass density (BMD) were measured.Results The fracture patients had longer dialysis duration,higher of hemodialysis ratio,higher serum Ca2 +,serum P,serum PTH and AKP,lower T and Z values of bone density than those patients without fracture.Univariate regression analysis of fracture risk suggested that dialysis age,high calcium,high phosphorus,high PTH,high AKP,high 25 (OH) VitD and fracture were positively correlated with peritoneal dialysis,and was negatively correlated with the use of calcium and peritoneal dialysis.The Multivariate regression analysis showed that high PTH,high AKP was positively correlated with fracture occurrence,lumbar vertebra bone density and the use of calcium were negatively correlated with fracture.Conclusion Low bone mineral density is an important risk factor for fracture.The use of calcium can reduce the risk of fracture.High PTH and high AKP should be actively intervened in CKD5D patients.