circ_CSNK1E modulates airway smooth muscle cells proliferation and migration via miR-34a-5p/VAMP2 axis in asthma.

BACKGROUND: Excessive proliferation and migration of airway smooth muscle cells (ASMCs) directly lead to airway remodeling in asthma. However, the role of circular RNAs (circRNAs) in airway remodeling remains unclear. This study aimed to investigate the regulatory role and mechanism of circ_CSNK1E in ASMCs proliferation and migration. METHODS: In this study, RNA-sequencing was used to analyze cicRNAs expression in asthma samples. ASMCs were treated with 25 ng/ml PDGF-BB to establish a model of asthma in vitro. Then, we used RT-qPCR to assess circRNAs, microRNAs (miRNAs) and messenger RNAs (mRNAs) expression. Besides, CCK-8, colony formation, wound healing and transwell chamber assays were carried out to explore cell proliferation and migration. Subcellular localization assay was used to detect the location of circRNA. Next, bioinformatics, luciferase reporter and RIP assays were performed to evaluate the relationship among circ_CSNK1E, miRNA-34a-5p and VAMP2. RESULTS: circ_CSNK1E expression was found to be significantly up-regulated in asthma samples and PDGF-BB-induced ASMCs. Functional experiments revealed that inhibition of circRNA_CSNK1E suppressed proliferation and migration of ASMCs stimulated by PDGF-BB. Next, we found that circRNA_CSNK1E served as a sponge for miR-34a-5p in ASMCs, and miR-34a-5p mimic suppressed proliferation and migration of ASMCs. Moreover, VAMP2 was confirmed as a direct target of miR-34a-5p. At last, inhibition of circRNA_CSNK1E suppressed proliferation and migration of ASMCs stimulated by PDGF-BB through miR-34a-5p/VAMP2 axis. CONCLUSION: Collectively, these findings clarified the importance of circ_CSNK1E/miRNA-34a-5p/VAMP2 axis for the proliferation and migration of ASMCs. These indicated that inhibition of circ_CSNK1E might be a potential target for the treatment of airway remodeling in asthma.

Protective effects of sinomenine against doxorubicin-induced nephrosis in rats.

Sinomenine (SN, 1) is a pure compound extracted from the Sinomenium acutum plant. We investigated the protective effects and mechanism of action of SN in a rat model of doxorubicin (DOX)-induced nephrosis. Nephrosis was induced by a single dose of 5 mg/kg DOX, and DOX-treated rats received a daily i.p. injection of 10 or 30 mg/kg SN, or saline (n = 6). Urine and serum biochemical parameters, serum TNF-α and IL-1ß levels, nephrin, podocin, α-actinin-4, and peroxisome proliferator-activated receptor-α (PPAR-α) protein expression, and renal ultrastructure were examined at day 28. Compound 1 significantly attenuated the effect of DOX on urine and serum biochemical parameters. Electron microscopy demonstrated that 1 suppressed DOX-induced increases in foot process width. Compared with those in control rats, nephrin, podocin, and PPAR-α protein expressions decreased in the glomeruli of DOX-treated rats, and this effect was significantly attenuated by 1. However, no appreciable alterations were observed in the expression level of α-actinin-4. DOX significantly increased serum TNF-α and IL-1ß compared with those in control rats, and 1 significantly reduced the serum levels of TNF-α and IL-1ß. SN ameliorates DOX-induced nephrotic syndrome in rats, resulting in a modulation of renal nephrin, podocin expression, and thereby protecting podocytes from injury.

Overexpression of PGC-1ß improves insulin sensitivity and mitochondrial function in 3T3-L1 adipocytes.

The co-transcription factor peroxisome proliferator-activated receptor γ coactivator-1ß (PGC-1ß) was first identified in 2002. Although the function of PGC-1ß in white adipose tissue (WAT) is largely unknown, it has been studied extensively in the liver, cardiac muscle, and skeletal muscle. Herein, we investigated PGC-1ß overexpression in 3T3-L1 adipocytes. The main findings were as follows: (i) 3T3-L1 adipocytes overexpressing PGC-1ß showed improved insulin sensitivity and elevated insulin-stimulated glucose uptake; (ii) mitochondrial cristae became broader and more ordered, additional smaller mitochondria emerged, mitochondrial DNA increased, and fission 1 protein (Fis1) mRNA expression was greatly elevated; (iii) intracellular ATP levels increased, but no changes were observed in mitochondrial membrane potential, uncoupling protein (UCP) mRNA expression, or reactive oxygen species (ROS) production; and (iv) mitochondrial metabolism factors, namely, acetyl-coenzyme A carboxylase 2 (ACC2) and hexokinase 2 (HK2) were downregulated, while cytochrome c oxidase subunit IV (COX IV) was upregulated. In conclusion, PGC-1ß affects not only insulin sensitivity but also mitochondrial biogenesis and function. We believe that the role of PGC-1ß is distinct from that of PGC-1α in WAT.

[Nephrotic syndrome complicated with intracranial venous thrombosis treated with urokinase: report of 5 cases].

OBJECTIVE: To explore the effect of urokinase and low molecular weight heparin in children with nephrotic syndrome complicated with intracranial venous thrombosis. METHODS: Urokinase and low molecular weight heparin were administered to the 5 patients intravenously. The initial dose of urokinase was 2000 - 4000 U/(kg.d), the initial pulse dose was 20 000 - 40 000 U given within 15 - 30 minutes, and the left was infused by using a pump, from the second day 2000 U/(kg.d) urokinase was infused daily for 3 to 7 days. During the treatment thrombin time (TT), activated partial thromboplastin time (APTT) were tested 3 times every week, with particular attention to bleeding. Low molecular weight heparin 100 - 120 AXaIU/kg, 1 or 2 times per day was hypodermally injected for a course of two weeks. Anti-platelet drugs: long-term oral administration of dipyridamole 3 - 5 mg/(kg.d) was applied 2 - 3 times every day for 3 months. RESULTS: The clinical symptoms disappeared after one month of the combined therapy of urokinase, low molecular weight heparin and dipyridamole in 5 cases of nephrotic syndrome complicated with intracranial venous thrombosis in children, the plasma viscosity returned to normal in 1 month, activated partial thromboplastin time, prothrombin time, fibrinogen degradation products returned to normal in 1 to 2 months, venous thrombosis disappeared after 1 to 3 months in head CT or MRI examination, showing the cerebral venous sinus thrombosis complete recanalization without relapse cases in follow-up. CONCLUSION: The early application of urokinase and low molecular heparin and anti-platelet coagulation drugs was effective. The early diagnosis, treatment and prevention of intracranial vein thrombosis in patients with nephrotic syndrome is important.

[Clinical and pathological study of 47 cases with Alport syndrome].

OBJECTIVE: To analyze the clinical and pathological features of children with Alport syndrome (AS). METHODS: A series of 47 patients with AS from unrelated families hospitalized from Jan. 1990 to Jan. 2007 were involved in this study. The clinical and histopathological data were collected and analyzed. RESULTS: Of the 47 cases, 32 were male and 15 female, M/F: 2.1:1. The patient's age ranged from 15 months to 13 years, mean 9 years. Thirty-nine of the 47 cases had positive family history, X-linked dominant inheritance AS was diagnosed in 37 cases, autosomal recessive inheritance AS in 2 cases. Gross hematuria or microscopic hematuria were found in 59.3% of the cases as the first manifestations, while 29.8% showed edema or proteinuria. The major clinical manifestations were isolated hematuria (23.4%), hematuria and proteinuria (36.2%), nephrotic syndrome (29.8%), and renal failure (10.6%). Hematuria and proteinuria existed in all the cases, while only 7 to 13 years children had nephrotic syndrome and renal failure. Of the 47 patients, 33 (70.2%) showed mesangial proliferative glomerulonephritis (MsPGN) under the light microscope, 13 (27.6%) focal segmental glomerulosclerosis (FSGS), 1 (2.1%) membrane proliferative glomerulonephritis (MPGN). For immunofluorescence, there was IgM (40.4%) as the dominant deposition in 19 patients, IgA in 9 (19.1%), IgG in 9 (19.1%), and 10 (21.4%) were negative. Thirty-nine cases showed typical glomerular basement membrane (GBM) pathological changes under electron microscope, while thin basement membrane in 8 cases; 46 showed abnormal skin and/or renal alpha-chain distribution. CONCLUSION: For Alport syndrome, number of male patients was higher than that of female patients. There was a significant difference among different age groups. Hematuria might be present throughout the course, while urine protein increases gradually. MsPGN was the dominant pathological change. The GBM pathological changes in younger children is not typical, so the immunofluorescence test of alpha-chain in collagen IV should be used as an important diagnostic method.

[Cyclosporin A treatment of 83 children with nephrotic syndrome of different pathological types].

OBJECTIVE: To evaluate the efficacy of cyclosporin A (CyA) therapy in 83 children with nephrotic syndrome of different pathological types. METHODS: Eighty-three children enrolled in this study were all hospitalized children with idiopathic nephrotic syndrome, aged 3 to 14 yrs (average 8.3 yrs) and included 52 males and 31 females. There were 35 cases with steroid-dependent, 17 with steroid resistant and 26 with frequent relapses. CyA was given to each patient with dosage of 5 mg/(kg.d) during the corticosteroid was diminished. The renwal biopsy was performed in all patients before the administration of CyA. The duration of CyA therapy lasted for about 3 to 6 months. The plasma concentration of CyA was monitored. RESULTS: Eighty-three children with nephrotic syndrome of different pathological types were treated with CyA, including 42 cases of minimal change nephrotic syndrome (MCNS), 31 cases of mesangioproliferative glomerulonephritis (MsPGN), 5 cases of membranoproliferative glomerulonephritis (MPGN) and 4 cases of focal segmental glomerular sclerosis (FSGS). All the 83 patients tolerated well to the CyA treatment. Forty-five cases got complete remission, 23 partial remission, 15 cases no change after one month treatment with CyA in the hospital. The overall response rate was 82%. Patients with different renal pathological types showed different responses. Among them, MCNS and MsPGN exhibited the best response rates of 86% and 84%, respectively; MPGN cases showed a lower response rate and FSGS cases showed the lowest rate. The response time was 7 to 45 days. The blood concentration of CyA was monitored for 1 week and 2 weeks after the drug was given. The effective drug concentration was maintained at 100 to 200 microg/L, and the course lasted for 3 to 6 months. During the follow-up of 83 cases, in 17 of 68 cases the disease relapsed when therapy was tapered or discontinued. The relapse rate was 25%. The results indicated that CyA would be effective to the relapsed cases. The serum creatinine increased temporarily after administration of CyA in 5 cases, N-acetyl-beta-D-glucosaminidase (NAG) in 8 cases and eventually reached the normal range after the adjustment of dosage. The side effects included anorexia, nausea, vomiting and so on. CONCLUSION: CyA is one of the effective substitutes for the treatment of nephrotic syndrome, especially for the cases with MCNS and MsPGN. And CyA could control refractory nephrotic syndrome effectively and rapidly. The clinical effect was related to the blood concentration of CyA and pathological types.