Objective To investigate the effect of pioglitazone on the expression of thrombospondin-1(TSP1)and transfor ming growth factor-β1 (TGF-131)in the kidney of diabetic rats.Methods Forty Sprague-Dawley(SD)rats were randomly divided into 4 groups:normal control(NC),streptozocin-induced diabetic mellitus(DM),diabetic rats treated with pioglitazone(DP)and normal control rats treated with pioglitazone(NP).24-h urine protein quantity,serum creatinine(Scr) and kidney hypertrophy index were measured after treatment for 10 weeks,and the expression of TSP1 and TGF-β1 in the kidney was detected by using RT-PCR and immuohistochemistry.Results As compared with DM group,urine protein level and kidney hypertrophy index were significantly attenuated in DP group(P<0.05),but there was no significant difference in plasma glucose between them(P>0.05).As compared with NC and NP groups,the expression of TSP1 and TGF-β1 was significantly elevated in the kidney of DM and DP groups(P<0.01).Both TSP1 and TGF-β1 expression in DP group was lower than in DM group(P<0.05).Conclusion Pioglitazone can exert a direct protective effect on the kidney in diabetic rats,which is independent of the role of insulin sensitizer,and the mechanism may be associated with the decreased TSP1 expression in the kidney.

Objective To investigate the correlations of ATP-binding cassette transporter A1 (ABCA1) protein expression in renal interstitium with foam cells formation,tubulointerstitial lesion and serum cholesterol in glomerulonephropathy.Methods Five patients with Alport syndrome,28 patients with membranous proliferative glomerular nephritis (MPGN),35 patients with focal segmental glomerulosclerosis (FSGS),36 patients with idiopathic membranous nephropathy (IMN) and 34 patients with IgA nephropathy (IgAN) were enrolled in the study.Expression of ABCA1 protein was detected by immunohistochemical method.The relative area of foam cells in renal interstitium was semi-quantified and the correlations of ABCA1 protein expression in renal interstitium (except for renal tubules) with the relative area of foam cells,integration of tubulointerstitial lesion (TIL) and serum cholesterol were examined using t test and linear correlation analysis.Results In renal interstitium,monocyte-macrophages,foam cells and renal tubular epithelial cells expressed ABCA1 protein.ABCA1 protein expression in renal interstitium with foam cells infiltration was significantly higher as compared to that without foam cells infiltration (P＜ 0.05),but there was no correlation between ABCA1 expression and foam cells area (P＞0.05).In MPGN,FSGS,IMN patients,there was a positive correlation between ABCA1 protein expression in renal interstitium and integration of TIL (P＜0.05).There was a positive correlation between serum cholesterol level and ABCA1 protein expression in renal interstitium (P＜0.05).Conclusions In glomerulonephropathy,cholesterol may promote the ABCA1 expression in renal interstitium.ABCA1 may have double-effects on transportation of cholesterol.ABCA1 may take part in the process of tubulointerstitial lesion.

Objective To explore CT diagnosis of atypical meningioma .Methods 18 cases of atypical meningioma were undergone MR plan scans, among them,17 cases were examined by CT contrast scans. All cases were proved by operation and pathology.Results The tumors appeared as mixed density in 11 cases,cystic in 4 cases,complete enhancement in 3 cases.Conclusion The tumors to be comfirmed at external cerebra is the key in diagnosing atypical meningioma exactly by CT.Atypical manifestations can be seen in a few meningiomas,therefore, it is significant in differential diagnosis of meningioma.

Objective To assess the characteristics of magnetic resonance imaging (MRI) for spinal cavernous angiomas.Methods The examinations of plain scan and contrast enhanced scan of magnetic resonance (MR) were performed in three patients with spinal cavernous angiomas.Results The focus of two cases was located in thorax segment of the spinal cord and one in lower cervical segment.All focuses were single and the shape of spinal cord was normal or slightly thick. MRI characteristic of spinal cavernous angiomas was just like popcorn or mulberry with a jumbled gobbet signal. Low and short T2 signal appeared around the focus. In all cases, there were no obvious contrast enhanced signal in 2 cases and one case with moderate contrast enhanced signal. The diameter of hemorrhage was smaller than that of the spinal cord.Conclusion MRI has higher sensitivity and specificity in the diagnosis of spinal cavernous angioma.

In order to investigate the effects of connective tissue growth factor (CTGF) antisense oligodeoxynucleotide (ODN) on plasminogen activator inhibitor-1 (PAI-1) expression in renal tubular cells induced by transforming growth factor beta1 (TGF-beta1) and to explore the role of CTGF in the degradation of renal extracellular matrix (ECM), a human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipid-mediated CTGF antisense ODN was transfected into HKC. After HKC were stimulated with TGF-beta1 (5 microg/L), the mRNA level of PAI-1 was detected by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 in the media was determined by Western blot. The results showed that TGF-beta1 could induce tubular CTGF and PAI-1 mRNA expression. The PAI-1 mRNA expression induced by TGF-beta1 was significantly inhibited by CTGF antisense ODN. CTGF antisense ODN also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 protein secreted into the media. It was concluded that CTGF might play a crucial role in the degradation of excessive ECM during tubulointerstitial fibrosis, and blocking the biological effect of CTGF may be a novel way in preventing renal fibrosis.

In order to explore the role of connective tissue growth factor (CTGF) in the pathogenesis of renal tubulointerstitial fibrosis, 48 Wistar rats were randomly divided into sham-operated and unilateral ureteral obstruction (UUO) group. On the postoperative day 1, 3, 7 and 14, the rats were killed and the kidneys were removed. The renal tubulointerstitial injury index was evaluated according to the MASSON staining. The mRNA levels of CTGF, transforming growth factor-β1(TGF-β1), collagen Ⅰ (col Ⅰ ), and plasminogen activator inhibitor-1 (PAI-1) were detected using reverse transcriptional-polymerase chain reaction (RT-PCR). Immunohistochemistry was performed to evaluate the protein expression of the above factors, and the relations among them were analyzed. Quantitative expression of CTGF protein in the kidneys was also assessed using Western blot. The results showed that TGF-β1 mRNA level was increased at first day after UUO, followed by a marked elevation of CTGF mRNA level, which began to increase 3 days after UUO (P＜0.01). With the progression of the disease, the mRNA expression of CTGF, col Ⅰ and PAI-1 was increased progressively. Immunohistochemistry revealed that the CTGF protein expression was significantly increased in fibrotic areas and tubular epithelial cells 3 days after UUO. On the post-UUO day 7, the protein level of CTGF was positively related to the renal tubulointerstitial injury index (r =0.62, P＜0.01), the expression of TGF-β1 (r=0.85, P＜0.01), col Ⅰ (r=0.78, P＜0.01),and PAI-1(r=0.76, P＜0.01). Upon Western blot analysis, CTGF protein expression began to increase 3 days after UUO, and appeared progressively throughout the time course (P＜0. 01, as compared with sham-operated group). It is concluded that CTGF can be induced by TGF-β and mediate various profibrotic actions of this cytokine, such as increasing extracellular matrix (ECM)synthesis and decreasing ECM degradation. The increased expression of CTGF may play a crucial role in the development and progression of tubulointerstitial fibrosis.

OBJECTIVETo analyze the diagnosis and treatment of autoimmune hemolytic anemia (AIHA) in liver transplant recipients.

METHODSA retrospective analysis was conducted of 3 patients who developed AIHA following orthotopic liver transplantation. The results of hemolysis tests and examinations of hemoglobin, white blood cells, platelets, total bilirubin, and alanine aminotransferase before and after treatments were reviewed.

RESULTSThese 3 patients developed AIHA following the transplantation possibly in association with the use of immunosuppressive agents, and the condition was effectively controlled after corresponding treatments.

CONCLUSIONAIHA is a uncommon complication after liver transplantation and can be cured after a definitive diagnosis with corresponding treatments.

Aged ; Anemia, Hemolytic, Autoimmune ; diagnosis ; etiology ; therapy ; Humans ; Liver Transplantation ; adverse effects ; Male ; Middle Aged ; Retrospective Studies

Objective: To investigate the regulation of JAK2 tyrosine kinase inhibitor ruxolitinib on extracellular matrix metalloproteinase (MMP in JAK2V617F positive myeloproliferative neoplasms (MPN) cells. Methods: ①Forty cases of newly diagnosed JAK2V617F positive MPN patients and 15 healthy volunteers as control in Baoding No.1 Hospital between January 2012 and December 2015 were enrolled in this study. JAK2V617F/JAK2 ratio was detected by real-time-PCR; the expression levels of phosphorylation protein tyrosine kinase 2 (p-JAK2) , MMP-2 and MMP-9 in pathological tissues of bone marrow were detected by immunohistochemistry. The bone marrow cells of JAK2V617F positive MPN patients were treated with ruxolitinib, then the migration ability and MMP-2, MMP-9 gene and protein expression levels were detected. ②The human erythroleukemia cell line HEL cells were treated with different concentrations of ruxolitinib (0, 50, 100, 250, 500, 1 000 nmol/L) . The cell viability was detected by CCK-8 test; cell migration ability was tested by transwell chambers. The mRNA expression levels of JAK2, MMP-2 and MMP-9 were detected by real-time-PCR. The protein expression levels of p-JAK2, MMP-2 and MMP-9 were detected by Western blot. Results: ①The expression levels of p-JAK2, MMP-2 and MMP-9 in the newly diagnosed group were significantly higher than control group respectively [ (78.56±24.55) % vs (41.59±17.29) %, P<0.05; (48.25±18.74) % vs (22.79±13.89) %, P<0.05; (53.29±19.28) % vs (15.56±14.96) %, P<0.05]. Spearman correlation analysis showed the positive correlation of MMP-2 and MMP-9 protein expression levels with JAK2V617F mutation (r=0.526, P=0.001; r=0.543, P=0.001) . ②The proliferation of HEL cells was inhibited by different concentrations of ruxolitinib in time and dose dependent manner. ③Cell migration test showed the number of cells leaked to the low chamber in MPN patients bone marrow cells and HEL cells treated with 5 nmol/L of ruxolitinib group were significantly lower than that without ruxolitinib treatment after 24 h [ (154.7±27.5) vs (320.3±67.3) , t=13.47, P<0.05; (70.7±10.5) vs (135.3±16.7) , t=13.89, P<0.05]. The mRNA and protein expression levels of JAK2, MMP-2 and MMP-9 decreased with the increased concentration of ruxolitinib. Conclusion Ruxolitinib inhibits MPN cell migration and expression of MMP-2 and MMP-9 via JAK2 signal pathway.

In order to investigate the effects of connective tissue growth factor (CTGF) antisense oligodeoxynucleotide (ODN) on plasminogen activator inhibitor-1 (PAI-1) expression in renal tubular cells induced by transforming growth factor β1 (TGF-β1) and to explore the role of CTGF in the degradation of renal extracellular matrix (ECM), a human proximal tubular epithelial cell line (HKC) was cultured in vitro. Cationic lipid-mediated CTGF antisense ODN was transfected into HKC. After HKC were stimulated with TGF-β1 (5 μg/L), the mRNA level of PAI-1 was detected by RT-PCR. Intracellular PAI-1 protein synthesis was assessed by flow cytometry. The secreted PAI-1 in the media was determined by Western blot. The results showed that TGF-β1 could induce tubular CTGF and PAI-1 mRNA expression. The PAI-1 mRNA expression induced by TGF-β1 was significantly inhibited by CTGF antisense ODN. CTGF antisense ODN also inhibited intracellular PAI-1 protein synthesis and lowered the levels of PAI-1 protein secreted into the media. It was concluded that CTGF might play a crucial role in the degradation of excessive ECM during tubulointerstitial fibrosis, and blocking the biological effect of CTGF may be a novel way in preventing renal fibrosis.