Deciphering the pathogenicity of COL4A4 heterozygous splicing mutations and the genotype-phenotype correlation in autosomal dominant Alport syndrome / 中华肾脏病杂志

Objective:Through the investigation of the pathogenicity of COL4A4 heterozygous splicing mutations and the genotype-phenotype correlation in autosomal dominant Alport syndrome (ADAS), to better understand the impact of COL4A4 heterozygous splicing mutations on ADAS. Methods:The study was a case series analysis. Patients from 5 ADAS families with COL4A4 heterozygous splicing mutations detected by whole exome sequencing were recruited by three hospitals. In vivo transcriptional analysis and/or in vitro minigene splicing assay were conducted to determine the splicing patterns and assess the pathogenicity of COL4A4 heterozygous splicing mutations. Results:In the five ADAS pedigrees carrying COL4A4 heterozygous splicing mutations, four novel ADAS splicing patterns were described. In pedigree 1-4, most patients presented with continuous hematuria or/and microalbuminuria. Otherwise,the proband in pedigree 4 presented with macroalbuminuria and the proband in pedigree 1 had progressed to chronic kidney disease stage 2 at the age of 70 years old. In pedigree 5, all patients developed end-stage renal disease between 28 and 41 years old. c.735+3A>G detected in pedigree 1 and pedigree 2 and c.694-1G>C detected in pedigree 3 both led to exon 12 skipping in COL4A4, resulting in 42 nucleotides in-frame deletion (c.694_735del). c.2056+3A>G detected in pedigree 4 led to COL4A4 exon 26 skipping, which caused in-frame deletion of 69 nucleotides (c.1988_2056del). c.2716+5G>T detected in pedigree 5 led to a 360 nucleotides large in-frame deletion, including 100 bp sequence at the 3'end of exon 29,the whole sequence of exon 30 and 89 bp sequence at the 5'end of exon 31 (c.2446_2805del). Conclusions:Renal prognosis differs significantly for patients with small in-frame deletions versus large in-frame deletion splicing abnormalities. Determination of the pathogenicity and the splicing patterns of COL4A4 heterozygous splicing mutations using in vivo and in vitro transcriptional analysis may provide renal prognostic information.

Effect of different blood purification modes on short-term complications and prognosis in uremic patients / 中华肾脏病杂志

Objective:To explore the effects of different blood purification modes on short-term complications, quality of life and survival prognosis of patients with uremia.Methods:The patients with uremia who received hemodialysis treatment at two blood purification centers in the First Affiliated Hospital of Dalian Medical University and Renal Care Hospital from January 1, 2017 to December 1, 2019 were enrolled in this study. According to the different blood purification modes, the patients were divided into high-flux hemodialysis (HFHD) group (HFHD group), HFHD+hemodiafiltration (HDF) per month group (HDF once a month group) and HFHD+HDF per week group (HDF once a week group). The three groups were matched with the ratio of 1∶1∶1 on the duration of hemodialysis based on the HDF once a week group. The differences of clinical indicators, medication and rehospitalization between baseline and end points were compared. The patients were followed up to 12 months after enrolled in the study or death. Patients in each group were divided into two subgroups, newly imported group and non-newly imported group based on whether or not they were newly enrolled in HD therapy from January 1, 2017 to December 1, 2019. Kaplan-Meier survival curve and Cox regression model were used to analyze the difference of survival prognosis in non-newly imported patients with different dialysis modes, and the EuroQol-5 Dimensions (EQ-5D-5L) was used to evaluate the difference of quality of life in newly imported patients after different dialysis modes treatment.Results:A total of 139 patients were enrolled, including 43 cases in the HFHD group, 47 cases in the HDF once a month group, and 49 cases in the HDF once a week group. After treatment, the levels of serum creatinine, serum urea nitrogen and serum potassium in the HDF once a week group were significantly lower than those in the other two groups (both P<0.05). Compared with the other two groups, the consumptions of erythropoietin and intravenous iron, the hospitalization times and hospitalization days in the HDF once a week group were significantly decreased, and the level of albumin in the HDF once a week group were significantly increased (all P<0.05). Kaplan-Meier survival curve analysis of non-newly imported hemodialysis patients showed that the survival rate in the HDF once a week group was higher than that in the other two groups (Log-rank χ2=7.020, P=0.030). Multivariate Cox regression analysis showed that HDF was a protective factor for post-dialysis survival in uremia patients ( HR=0.472, 95% CI 0.188-0.836, P=0.023). The total efficacy of EQ-5D-5L of the newly imported patients in the HDF once a week group was significantly higher than that in the other two groups ( F=7.293, P=0.002). Conclusions:The combination of HFHD with HDF per week therapy can significantly improve the short-term quality of life and nutritional status, and reduce the hospitalization frequency, length of hospital stay, the consumption of erythropoietin and intravenous iron and mortality risk in uremia patients.

Amentoflavone induces apoptosis and suppresses glycolysis in glioma cells by targeting miR-124-3p.

Malignant glioma is the most common type of brain tumor with poor clinical outcome and survival. Therefore, it is imperative to develop novel therapeutic agents for managing glioma. The aim of this study was to investigate the role of amentoflavone (AF), an active flavonoid component in Selaginella tamariscina Spring, in glioma cells and the underlying mechanism of its action. Our results showed that miR-124-3p expression was significantly down-regulated in glioma tissues relative to normal brain tissues. AF decreased cell viability and triggered apoptosis in both glioma cell lines in a dose-dependent manner. AF induced apoptosis and inhibited glycolysis in the glioma cells by upregulating miR-124-3p. Furthermore, AF upregulated miR-124-3p by repressing DNMT1 through Sp1, which in turn was caused by the activation of ROS/AMPK signaling pathway by AF. In conclusion, AF could induce apoptosis and inhibited glycolysis in glioma cells via miR-124-3p. Our findings provide preliminary experimental data that support further investigation on the therapeutic efficacy of AF in glioma.

Autophagy is involved in regulating VEGF during high-glucose-induced podocyte injury.

Podocytes are the major sites of vascular endothelial growth factor (VEGF) production in kidneys. Over-expression of VEGF is involved in the pathogenesis of diabetic nephropathy (DN), and an emerging body of evidence suggests that autophagy plays an important role in DN. In this study, the effect of autophagy on over-expressed VEGF along with its underlying mechanism was investigated in streptozotocin (STZ)-induced diabetic mice and high glucose (HG)-induced podocytes. We found that diabetes caused podocyte foot process effacement and VEGF upregulation significantly. In vitro, high glucose induced VEGF and reduced the podocyte viability. After treatment with rapamycin in podocytes, an autophagy inducer, VEGF activation was significantly abrogated and podocyte injury was ameliorated. In contrast, podocytes treated with 3-methyladenine (3-MA), a potent autophagy inhibitor, had increased VEGF expression. Furthermore, 3-MA significantly increased the production of HG-induced reactive oxygen species (ROS), whereas rapamycin decreased the cellular ROS level. Inhibition of ROS production by N-acetyl-l-cysteine (NAC) effectively reduced the over-expression of VEGF. These studies show the vital role of autophagy in the regulation of VEGF, which presents a protective effect on HG-induced podocyte injury. ROS production may be an important mechanism for mediating this process.

Values of the integrated visual and auditory continuous performance test in diagnosing children with attention deficit hyperactivity disorder in clinic and school in China / 中华行为医学与脑科学杂志

Objective To explore the psychometric properties of the Chinese version of integrated visual and auditory continuous performance test (IVA-CPT) ,and to assess the diagnosis value of the Chinese version of IVA-CPT in two samples of children with attention deficit hyperactivity disorder (ADHD) in clinic and school.Methods 112 participants were divided into 4 groups:clinic group, clinic control group, school group and control group from school.The participants were measured by IVA-CPT and the results were compared.Results ①The integrated quotients of IVA-CPT had concordance with the dimensions of the DSM-ⅣV and ASRS.Each integrated quotients of IVA-CPT and each dimension of the DSM-ⅣV and ASRS were negatively correlated(school: r=-0.277, P＜0.05;clinic: r=-0.423, P＜0.05).The score of HI dimension was negatively correlated with integrated attention quotient and integrated inhibition quotient of IVA-CPT (r=-0.480, P＜0.01;r=-0.452, P＜0.01).② The results of the IVA-CPT and the comprehensive assessment had consistency for diagnosis of children with ADHD from the total subjects,the clinic ,and the school.And there were no difference between the sensitivity, the specificity, the coincidence rate, the missed diagnosis rate and the misdiagnosis rate of IVA-CPT when used to diagnose children with ADHD in clinic and school(x2=3.396, P＞0.05).③The IVA-CPT evaluation system had good stability according to the retest of a random sample of 25 subjects(P＜0.01).The pretest and posttest results of each full quotients of IVA-CPT were significantly correlated (P＜0.01).Conclusion The IVA-CPT not only can be used as auxiliary diagnostic tools to diagnose children with ADHD in clinic,it can be used as auxiliary diagnostic tools to diagnose children with ADHD in the community.

Immunogenicity of recombinant Lactobacillus casei expressing VP2 protein of infectious bursal disease virus in chickens / 生物工程学报

In order to determine immunogenicity and protective effect in chickens, we used the IBDV (Infectious bursal disease virus)-Vp2/Lactobacillus casei as antigen transfer system. First, the immunized and control chickens were challenged by IBDV/DQ at lethal dose to determine the protective ratio. Second, chickens were orallyand intranasally vaccinated twice with 10(9) CFU/mL pLA-VP2/L. casei, pLA/L. casei and PBS as negativecontrol and commercial vaccine as positive control. The bursa injury and the lesion score wererecorded post challenge. The level of specific IgG and sIgA in pLA-VP2/L. casei and positive control groups was significantly higher than that in negativecontrol groups. The protection efficacy in pLA-VP2/L. casei oral group was higher than that inintranasal group. The SI. of pLA-VP2/L. casei oral group was significant higher than other groups. The lesion score indicated the pLA-VP2/L. casei was safer than commercial vaccine for bursa. Collectively, the pLA-VP2/L. casei could be a vaccine candidate for IBDV.

Current status of anticoagulant treatments and improvements for hemodialysis patients in northern Chinese cities: a five-year comparative study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Anticoagulation treatments are an important aspect of hemodialysis; however, few reports have addressed these treatments. This investigation intends to increase the understanding of the current status and improvements of hemodialysis-related anticoagulation treatments in China.</p><p><b>METHODS</b>In this study, an epidemiological investigation was conducted that examined 842 patients in 2007 and 1 175 patients in 2012 who underwent hemodialysis anticoagulation treatments in seven blood purification centers in northern Chinese cities.</p><p><b>RESULTS</b>Heparin was the most commonly used anticoagulant, although the percentage of use of low-molecular-weight heparin (LMWH) increased from 26.5% in 2007 to 42.1% in 2012. In 2007, there were no significant differences in anticoagulant selection among either patients with various primary diseases or patients with hemorrhage, thrombosis, thrombocytopenia, or a low hemoglobin level. However, compared with patients with other diseases, significantly lower doses of LMWH were administered to patients with hypertension (55.5 U/kg vs. 67.3 U/kg, P < 0.05) or diabetes (58.5 U/kg vs. 67.3 U/kg, P < 0.05), and patients with hemorrhage received lower doses of heparin than the other patients (61.6 U/kg vs. 71.8 U/kg, P < 0.01). In 2012, patients with diabetic nephropathy (51.5% vs. 36.5%, P < 0.01), hemorrhage (43.4% vs. 31.7%, P < 0.01), or a hemoglobin level below 90 g/L (57.2% vs. 37.1%, P < 0.01) experienced significantly higher doses of LMWH administration; patients with hemorrhage received significantly reduced LMWH dosages (50.4 U/kg vs. 57.8 U/kg, P < 0.05), and patients with thrombosis received significantly higher doses of heparin (73.8 U/kg vs. 62.1 U/kg, P < 0.01) or LMWH (57.8 U/kg vs. 52.6 U/kg, P < 0.05). Antiplatelet drugs were administered to 20.4% of the examined patients in 2007 and 20.7% in 2012. In 2012, patients with hypertension (25.9% vs. 18.5%, P < 0.01) and thrombosis (36.6% vs. 16.1%, P < 0.01) had a higher rate of using antiplatelet drugs than patients with other primary diseases and complications. Patients receiving antiplatelet drugs also received higher doses of heparin than patients without using antiplatelet drugs (74.4 U/kg vs. 65.9 U/kg, P < 0.01). However, the use of the drugs was not correlated with thrombocytopenia. The rate at which coagulation indices were determined increased from 45.7% in 2007 to 64% in 2012.</p><p><b>CONCLUSION</b>These findings suggested that hemodialysisrelated anticoagulation treatments in China have gradually become more standardized and individualized.</p>

Fibrinolytic therapy on renal protection in IgA nephropathy patients：a retrospective cohort study / 实用医学杂志

Objective To investigate the protective effect of fibrinolytic therapy on renal function in patients with IgA nephropathy. Methods The clinical data of 84 patients with idiopathic IgA nephropathy were analyzed in a retrospective cohort study. According to different drugs of fibrinolytic therapy, the patients were divided into three groups including control group(n=23), urokinase group (n=32) and fibrinogenase group (n=29). Twenty-four-hour urine protein excretion, serum albumin, estimated glomerular filtration rate (eGFR), prothrombin time and fibrinogen level between the three groups after 6-month and 1-year treatments were compared. Results Compared to the group before treatment, the urine protein excretions were decreased in all 3 groups with 6-month and 1-year treatments (P<0.01). The urine protein excretion in the urokinase group with 6-month and 1-year treatments were lower than that in the control group (P<0.01). The urine protein excretion in the fibrinogenase group with 1-year treatment were lower that in the control group (P<0.05). After 1-year treatment, the eGFR in urokinase group was significantly increased compared with the control group (P<0.05). Conclusion Fibrinolytic therapy can decrease the 24-hour urine protein excretion and improve the renal fuction in those patients with idiopathic IgA nephropathy independent of glucocorticoids and immunosuppressive agents.

Correlation of osteoprotegerin with radial artery calcification in patients with end stage renal diseare / 中国医师进修杂志

ObjectiveTo investigate the correlation of radial artery and serum osteoprotegerin (OPG) expression and vascular calcification in patients with end stage renal disease.MethodsDiscarded radial arteries were gathered from 43 patients with end stage renal disease(end stage renal disease group)during arterial venous fistula plasty,including 24 patients with no calcification,13 patients with mild to moderate calcification and 6 patients with severe calcification,20 patients with dialysis and 23 patients without dialysis.Discarded arteries from 9 patients with upper limb trauma were chosen as control group.The yon Kossa staining,alizarin red staining and OPG immunohistochemical staining were taken and serum OPG level was measured for all the patients.ResultsSerum OPG level was significantly higher in end stage renal disease group than that in control group [(216.47±83.78) ng/L vs.(57.15±23.22) ng/L,P＜ 0.01 ].Serum OPG level in patients with no calcification,mild to moderate calcification and severe calcification[ ( 152.21±32.21 ),(248.31±53.86),(368.44±53.57) ng/L] and OPG staining value ( 1,2,4 scores) were increased gradually as vascular calcification more serious (P＜ 0.01 ).There was a positive relationship between serum OPG and vascular calcification value(alizarin red staining:r =0.517,P ＜ 0.01 ; yon Kossa staining:r =0.677,P＜ 0.01 ).Serum OPG level and OPG staining value in patients with dialysis were higher than those in patients without dialysis [ (283.57±79.02 ) ng/L vs.( 152.46±30.89 ) ng/L,2 scores vs.1 score,P ＜ 0.01 ].ConclusionsThere is relationship between serum OPG level,OPG expression of radial artery and vascular calcification.

Change of TGF-β receptor subtype expression in the kidney of rats with unilateral ureteral obstruction / 中华肾脏病杂志

ObjectiveTo investigate the influence of TGF-β receptor subtypes expression and their downstream signaling Smad proteins on rat renal interstitial fibrosis induced by unilateral ureteral obstruction(UUO).MethodsA total of 90 rats were randomly divided into three groups:normal control(CON),sham operation (SOR) and UUO group,and sacrificed 1,3,7,14 and 21 days after operation.Serum creatinine and urea nitrogen were detected to assess renal function.PAS and Masson staining were performed to observe histological damage in the kidneys.Quantitative RT-PCR was used to define expression of mRNA encoding TGF-β receptor subtypes and their downstream signaling Smad proteins in kidney tubular cells.Real-time PCR,Western blotting and immunofluorescence were used to monitor the time-related expression of the TGF-β receptor subtypes and their downstream signaling Smad proteins in kidney.ResultsCompared with the CON group,serum creatinine and urea nitrogen in UUO groups increased at day 3 after operation (P＜0.05) and reached their peak 21 days after operation (P＜0.01).Obvious inflammatory cell infiltration was observed in UUO group 3 days after operation,while renal tubular atrophy and renal interstitial fibrosis were observed in UUO group14 days after operation.The mRNA expressions of ALK-5,ALK-7 and TGF-βR Ⅱ increased significantly in UUO group 3 days after operation (all P＜0.05) and reached their peaks 14 days after operation (all P＜0.01).The mRNA expression of ALK-6 decreased significantly in UUO group 3 days after operation(P＜0.05) and reached its lowest level 14 days after operation (P＜0.01).The changes in the protein level of those receptors were consistent with their mRNA expressions.The protein expressions of Smad2/3 and p-Smad2/3 increased significantly in UUO group at day 3(all P＜0.05) and reached their peak at day 14 after operation(all P＜0.01).ConclusionExpressions of TGF-β receptor subtypes ALK-5,ALK-6,ALK-7,TGF-βR Ⅱ and their downstream signaling Smad2 and Smad3 proteins may influence the progress of renal interstitial fibrosis,tubular atrophy and inflammatory cell infiltration in UUO model rats.

FUT8 siRNA inhibits the activation of transforming growth factor β-Smad2/3 signaling pathway in renal tubular epithelial cells / 中华肾脏病杂志

ObjectiveTo investigate the effect of FUT8-siRNA on transforming growth factor β (TGF-β)-Smad2/3 signalling pathway in renal tubular epithelial cells.MethodsHK-2 cells were divided into six groups:normal group,negative control group,TGF-β1 group,TGF-β1 with FUT8 interference group,TGF-β1 with negative control group,FUT8 interference group.RNAi was performed to silence the expression of FUT8 gene,then immunofluorescent analysis was used to detect the expression of core fucose in the HK-2,immunoprecipitation and lectin blotting were performed to detect the core fucosylation of TGF-βR Ⅱ and ALK-5,and detect the change of Smad2/3 and p-Smad2/3 in HK-2 cells after FUT8 gene was silenced.ResultsCompared with the normal and negative control group,incubation with 5 μg/L TGF-β1 for 48 h could significantly up-regulate the core fucosylation of HK-2 cells,enhance the protein expression of TGF-βR Ⅱ and ALK-5 (P＜0.05),markedly increase the expression level of p-Smad 2/3 (P＜0.05) and cause it to nuclear translocation in HK-2 cells.While FUT8siRNA could inhibit the above up-regulation of TGF-βR Ⅱ and ALK-5(P＜0.05),suppress the increase of p-Smad 2/3(P＜0.05) and its nuclear translocation without disturbing the protein expression of TGF-βR Ⅱ and ALK-5.Conclusion FUT8-catalized core fucosylation of TGF-βR Ⅱ and ALK-5 is needed to fulfill their functions,and blocking core fucosylation of TGF-βR Ⅱ and ALK-5 leads to the inhibition of TGF-β-Smad2/3signalling pathway in HK-2 cells.

Development of a lung cancer image database and visualization toolkit / 生物医学工程学杂志

Lung cancer is the most common tumor and one of the malignant tumors with the lowest livability after diagnosis, as is known so far. Large-scale image database is the foundation of developing computer-aided diagnosis methods, education and training in lung cancer diagnosis to improve medical diagnostic efficiency and to reduce the doctors' burden. In this study, aiming at improving the low data storage efficiency and solving the lacking of tool for data visualization and data retrieval existing in the use of traditional Lung Image Database Consortium (LIDC) from the lung cancer database, we developed a new lung cancer image database platform including an improved data model, a data integration tool, an image and annotation visualization tool and a data retrieving component. Firstly, the data format in LIDC was analyzed and an improved information model was provided to manage and manipulate large amount data stored in it. Next, some tools such as data integration component, DICOM, image and annotation visualization tool, and data query were designed and implemented. The study demonstrated that the lung cancer image database platform had the capacity of data collection, visualization, and query, and could promote diagnose lung cancer research.

Calcitriol pulse therapy combined with resin adsorption in treatment of severe secondary hyperparathyroidism / 中国医师进修杂志

Objective To investigate the effects of oral calcitriol pulse therapy combined with resin adsorption in treatment of severe secondary hyperparathyroidism.Methods Thirty-two hemodialysis patinents were randomly divided into three groups:oral calcitriol pulse therapy(CP group,12 cases),oral calcitriol pulse therapy combined with resin adsorption group(CP+RA group,10 cases),and resin adsorption group(RA group,10 cases).Serum immunoreactive parathyroid hormone(iPTH),albumin,urea,creatinine,hemoglobin levels were detected before treatment and 1,2,3 months after treatment respectively.Serum calcium and phosphorus levels were also detected every half a month.Results The clearance rate of resin adsorption Was decreased significantly every time(P<0.05).Serum iPTH in CP+RA group decreased significantly 2 months after treatment.Serum iPTH3 months after treatment in CP+RA group[(598.50 ±317.45)ng/L]was less than that in CP group[(1008.67±436.85)ng/L]and RA group[(1464.60±411.27)ns/L](P<0.05).Conclusion Oral calcitriol pulse therapy combined with resin adsorption can clear iPTH safely and effectively.

Quantitative Analysis of Feature Parameter in Brain Tumor CT Perfusion Image / 中国医学物理学杂志

Objective:The abstraction of characteristics in CT perfusion image is crucial in the clinical diagnosis of brain infarction and cerebral tumor.Methods:After describing the relevant parameters of CT cerebral perfusion imaging,such as cerebral blood flow (CBF),cerebral blood volume (CBV),the mean transition time (MTT) and the permeability surface (PS).With using Visual C ++ programming and CT scan images sequence obtained,the time-density curves of the brain tumor region were visualized.The more accurate hemodynamic parameters were calculated through an improved algorithm basing on the deconvolution and measuring method of the length,angle,area and other parameters.Conclusion:The quantitative analysis of feature parameter is of great benefit to clinical doctor in making timely diagnosis and treatment of brain tumor.

Augmentation index's change and its influencing factor in patients on continuous ambulatory peritoneal dialysis / 中国医师进修杂志

Objective To analyze the factors influencing augmentation index (AI) by examining the AI changes in patients who received continuous ambulatory peritoneal dialysis (CAPD) therapy, and evaluate the artery elastic function situation. Methods The height, weight, blood pressure, blood indicators were detected in 46 CAPD patients (CAPD group) and 42 controls (control group). AI was managed by OMRON HEM-90OOAI, and the influencing factor was analyzed. Results Systolic blood pressure(SBP),diastolic blood pressure(DBP), pulse pressure, central aortic pressure and AIp75 (AI value, when pulse was 75 beats/min) in CAPD group were significantly higher than those in control group. Within CAPD patients,the AIp75, in female was significantly higher than that in nude [(96.67±8.95 )% vs (89.89±6.46)% ,P =0.022]. Simple correlation showed positive relationship in gender, age, SBP, DBP, pulse pressure and central aortic pressure with AI; and negative relationship in height, weight and weight index with AI; while there was no relationship between AI and all kinds of chemical examination indicators or dialysis years.Multiple linear regression analysis evidenced that gender,age, height, DBP and central aortic pressure of CAPD patients had linear regression relation with AI. Conclusions It indicates a decreased artery elasticity and an increased stiffness occurs in CAPD patients. AI has tight correlation with age, height, DBP and central aortic pressure, and is an non-invasive factor thai partially reflects the arterial elasticity. Central aortic pressure is relevant with aortic elasticity and rigidity closely. Measuring central aortic pressure is very imporrant for preventing cardiovascular accident.

Expression of tissue inhibitor of metalloproteinases 1 in chronic renal failure rats with arterial calcification / 中华肾脏病杂志

Objective To examine whether tissue inhibitor of metalloproteinases-1 (TIMP-1) is involved in arterial calcification of chronic renal failure (CRF) rats. Methods CRF model was induced in male Wistar rats by garage daily with 2% adenine 250 mg/kg. The calcification of aorta, femoral artery, renal artery and coronary artery was evaluated histomorphometrically by van Kossa-stained sections at 2-, 4-, 6- and 8-week respectively. RT-PCR and Western blot were used to observe the expressive levels of TIMP-1 mRNA and protein. Expressions of TIMP-1, osteopentin (OPN) and core binding factor α1 (Cbfα-1) protein were analyzed by immunhistochemistry. Results Serum urea nitrogen, creatinine, inorganic phosphate, calcium-phosphorus product and intact parathyroid hormone (iPTH) increased significantly in the model animals compared with control group after 2 weeks (P<0.01). Medial calcification was found in above four arteries of model groups after 6 weeks. RT-PCR and Western blot showed that TIMP-1 expression of model group was significantly higher than that of control group (P< 0.05), and obviously elevated in a time-dependent manner. The expression of TIMP-1 and OPN in calcified aortic smooth muscle cells increased obviously (P<0.05), and positive immunostaining of Cbfα-1 was found. The expression of TIMP-1 was positively correlated with OPN and Cbfα-1 (r=0.317, P=0.000; r=0.485, P=0.000). Conclusions The pathology of arterial calcification in CRF rats induced by adenine is similar to CRF patients, which may serve as a useful model of CRF with arterial calcification. The up-regulation of TIMP-1 seems to participate in the formation and development of vascular calcification in CRF.

ERK1/2 pathway play a important role in the apoptosis of glomerular mesangial cells induced by TIMP-1 / 中国实用内科杂志

Objective To investigate the role of ERK1/2 signal pathway in the apoptosis of rat mesangial cells inhibited by TIMP-1.Methods The human TIMP-1 sense and antisense plasmid were transferred into rat mesangial cells through liposome,while mesangial cells transfected with pcDNA3 and non-transfected mesangial cells were used as control.After being stimulated with high glucose(HG 25mmol/L D-glucose)and PD98059(50 ?mol/L)for 24 hours,the apoptosis rate was measured by flow cytometry.The expression of ERK1mRNA was assayed by RT-PCR.The protein expression of p-ERK1/2 in the cell was analyed by ELISA.Results(1)The apoptosis rates of the non-transfected group,transfect pcDNA3 group,sense group and antisense group in the culture medium of high glucose withact PD98059,were(12.10?2.21)%,(11.90?3.34)%,(5.50?0.50)%and(20.70?3.41)%,respectively.The apoptosis rate of the sense group was significantly lower than that without transfection(P

Relationship between serum or urine TIMP-1 levels and the stages of CKD / 中国实用内科杂志

Objective To investigate the relationship between serum or urine tissue inhibitor of metalloproteinases-1 (TIMP-1)levels and the stages of chronic kidney disease (CKD).Methods Serum and urine level of TIMP-1 were measured by enzyme-linked immunoadsorbent assay (ELISA) in 122 CKD patients and 40 healthy controls.Serum cystatin-C (CystC),parathyroid hormone (PTH),low-density lipoprotein (LDL)and other markers were determined simultaneously.Correlationanalysis between TIMP-1 level and degree of renal function were performed.Results Serum and urine TIMP-1 of the patients in CKD group was significantly higher than that in control group (P

Influence of continuous venovenous hemofiltration on patients with multi pleorgan dysfunction syndrome / 中国实用内科杂志

Objective To study the influence of continous ven ovenuous hemofiltration on patients with multiple organ dysfunction syndrome.Methods 64 patients with MODS were divided into CVVH group(34) and IHD group(30)between 1999-12~2003-12.The serum BUN,Cr,iron,blood pressure,ar rhythmia were monitored during treatment and KT/V was also calculated after each therapy.APACHE-Ⅱ score was assessed before and after the treatment and the pr ognosis of each patient was recorded as well.Results CVVH group had more stable hemodynamics than IHD group; there was no significant difference in the removal of BUN and Cr in the two grou ps.The survival time of CVVH group was longer than IHD,and APACHE-Ⅱ score of C VVH group was lower than that in IHD group,17.23?6.86 verse 20.91?5.77 ( P

Effect of TIMP-1 on expression of FN and type Ⅳ collagen in rat glomerular mesangial cells / 中华肾脏病杂志

ObjectⅣe To investigate the effect of sense and antisense TIMP-1 on the expression of FN and type Ⅳ collagen of rat glomerular mesangial cells in vitro. Methods Two recombinant retroviral vectors, pLXIN-TIMP-1 (PLT)encoding sense TIMP-1 and pLXIN-ATIMP-1 (PLA)encoding antisense TIMP-1 were constructed by using DNA recombining techniques. These two vectors were then introduced into the PA317 packaging cell line with lipofectin DOTAP. The high-titer retroviral supematants containing sense or antisense TIMP-1 were used to infect rat glomerular mesangial cells. PCR and Northern blot were used to detect the integration and expression of human TIMP-1. Northern blot and ELISA were employed to investigate the expression of endogenous TIMP-1, FN, and Ⅳ collagen. Results Both sense and antisense TIMP-1 were successfully integrated into rat mesangial cells and expressed the sense and antisense ITMP-1 RNA. Overexpression of TIMP-1 induced by sense TIMP-1 caused upregulation of FN and Ⅳ type collagen in protein level, in contrast, supprssion of TIMP-1 expression by antisense TIMP-1 caused downregulation of FN and Ⅳ type collagen protein; but neither sense nor antisense TIMP-1 infection had effects on the RNA level of FN and Ⅳ type collagen. Conclusion TIMP-1 suppresses the degradation of ECM in rat glomerular mesangial cells and antisense TIMP-1 may be available for renal fibrosis in the future.