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 explore the mechanism of metformin on osteogenic differentiation of rat jaw bone marrow mesenchymal cells(BMSCs)under high sugar and high fat environment.Methods:BMSCs of male SD rats were isolated and cultured in vitro and divided into 4 groups:Control group(C group),high sugar and high fat group(H group),control cells and H group cells were respectively stimulated with metformin(CM group and HM group).EDU staining was used to detect the cell proliferation;after osteogenesis and adi-pogenesis induction of BMSCs,alkaline phosphatase staining,alizarin red(O)staining and oil red O staining were respectively used to detect the osteogenic and adipogenic differentiation of the cells.The protein expression of Runx2 and OCN was detected by Western blot and immunofluorescence staining respectively.Results:Compared with the H group,in the HM group the cell proliferation ability was enhanced,the expression levels of Runx2 and OCN were increased(P＜0.05),the osteogenic differentiation ability was significantly in-creased,and the adipogenic differentiation ability was significantly decreased.Conclusion:In vitro,metformin promotes the prolifera-tion and osteogenic differentiation of rat jaw BMSCs cultured with a high sugar and high fat environment by increase of the expression of Runx2 and OCN.

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 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 To investigate the changes of clinical and laboratory manifestations of rheumatic fever (RF) in recent ten years by reviewing the medical data of 315 patients with RF. Methods Three hundred and fifteen in-patients and out-patients with RF during 1985-1995 (group A) and 1997-2007(group B) were selected. Their manifestations were compared. Results Male/female ratio was about 1:2. Carditis and polyarthritis were common manifestations. Compared with group A, the rate of low-grade fever and carditis increased and the rate of heart failure, positive rate of C reaction protein and antistreptolysin O decreased in group B. In group B, 61.4% patients fulfilled the updated Jones diagnostic criteria. 76.2% fulfilled the 2002-2003 WHO criteria. The sensitivity and specificity of peripheral blood lymphocyte procoagulant activity (PCA) for the diagnosis of rheumatic carditis was 79.1% and 71.4% respectively. That of the anti-streptococcal group A polysaccharide (ASP) antibodies was 70.3% and 70% respectively. Five to ten years follow-up clinical data were available for 35 cases since Dec. 1997. The recurrent rate of RF was 62.8%. Only 1/3 cases received regular secondary prevention. Recurrence rate of patients with regular secondary prevention was significantly lower than that of patients without regular secondary prevention. Conclusion Mild earditis has been increasing during last ten years. PCA and ASP are valuable tests for diagnosing rheumatic carditis. More emphasis should be paid to atypical cases, early diagnosis and regular secondary prevention in order to improve prognosis.