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Objective:To explore the clinical value of N terminal pro B type natriuretic peptide (NT-proBNP) in diagnosing or predicting heart failure in peridialysis chronic kidney disease (CKD) population.Methods:It was a single-center retrospective study. Patients with peridialysis CKD who visited the Department of Nephrology, First Affiliated Hospital of Zhengzhou University from January 2021 to June 2021 were collected and divided into 4 groups according to the presence or absence of heart failure and the level of left ventricular ejection fraction (LVEF), namely the non-heart failure group, heart failure with reduced ejection fraction (HFrEF) group (LVEF<40%), heart failure with mid-range ejection fraction (HFmrEF) group (40%≤LVEF<50%), and heart failure with preserved ejection fraction (HFpEF) group (LVEF≥50%). The NT-proBNP, echocardiography and other indicators of the 4 groups were compared. The value of plasma NT-proBNP in diagnosing heart failure, HFpEF, HFmrEF and HFrEF was analyzed by drawing receiver operating characteristic curve (ROC curve). Logistic regression analysis was used to analyze the related factors of heart failure in peridialysis CKD patients.Results:A total of 508 patients were included, including 11 cases in the HFrEF group, 29 cases in the HFmrEF group, 152 cases in the HFpEF group, and 316 cases without heart failure. The differences in age, 24-h urine volume, hemodialysis proportion, non-dialysis proportion, serum creatinine, estimated glomerular filtration rate, hemoglobin, serum albumin, C-reactive protein, NT-proBNP, cardiac troponin I, left ventricular internal diameter, LVEF, pulmonary artery systolic pressure, left ventricular end-diastolic volume, E/A value, septal thickness, and left ventricular posterior wall thickness among the four groups were statistically significant ( P < 0.05, respectively). A two-pair comparison (all P values corrected by Bonferroni method) revealed that the 24-h urine volume was higher in the non-heart failure group than in the other three groups (corrected P<0.05, respectively), while the proportion of hemodialysis patients and the levels of NT-proBNP and C-reactive protein were lower in the non-heart failure group than in the other three groups (corrected P<0.001, respectively); the levels of hemoglobin and serum albumin were lower in the HFpEF group than in the non-heart failure group (corrected P<0.001, respectively); troponin I was lower in the non-heart failure group than in the HFpEF group (corrected P<0.001), HFmrEF group (corrected P=0.001) and HFrEF group (corrected P<0.001), and troponin I was lower in the HFpEF group than in the HFrEF group (corrected P=0.008); LVEF was higher in the non-heart failure group than in the other three groups (corrected P<0.001, respectively), and LVEF in the HFpEF group was higher than in the HFmrEF and HFrEF groups (corrected P<0.001, respectively). For patients with peridialysis CKD, the cut-off values of plasma NT-proBNP for diagnosing or predicting heart failure, HFpEF, HFmrEF and HFrEF were 4 943.33 ng/L, 4 976.83 ng/L, 14 964.5 ng/L and 17 847.55 ng/L, respectively. Multivariate logistic regression analysis showed that NT-proBNP (every 500 ng/L increase, OR=1.390, 95% CI 1.287-1.501, P<0.001), LVEF ( OR=0.747, 95% CI 0.656-0.851, P<0.001) and 24-h urine volume (every 100 ml increase, OR=0.842, 95% CI 0.763-0.929, P=0.001) were independently correlated with heart failure. Conclusions:The cut-off value of plasma NT-proBNP for diagnosing or predicting heart failure in peridialysis CKD patients is much higher than that in patients with normal renal function. NT-proBNP, LVEF and 24-h urine volume are independently associated with heart failure in peridialysis CKD patients.
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Objective:To observe the ocular clinical features of infantile cytomegalovirus (CMV) infection.Methods:A retrospective clinical study. From March 2019 to July 2021, 876 eyes of 438 children with CMV infection who visited Department of Ophthalmology of Henan Provincial Children's Hospital were included in the study. Among them, there were 254 males and 184 females; the age ranged from 3 days to 11 months; the gestational weeks were 28 to 42 weeks; the birth weight was 1 120 to 8 900 g. There were 384 and 54 full-term and premature infants, respectively. Fundus examination was performed in 385 cases (770 eyes) after medical consultation; 53 cases (106 eyes) of premature infants were routinely screened. CMV retinitis (CMVR) was divided into granular type and fulminant type. Patients with CMV-related diseases with moderate to severe symptoms were given intravenous drip and/or oral ganciclovir; patients with severe fundus vasculitis were combined with intravitreal injection of ganciclovir. The follow-up period was from 4 to 28 months, and the characteristics of eye lesions, systemic comorbid diseases and treatment outcomes were observed.Results:There were 516 eyes of 258 cases with normal fundus (58.9%, 258/438); 291 eyes of 180 cases with CMVR (41.1%, 180/438), of which binocular and monocular were 111 (61.7%, 111/180) and 69 (38.3%, 69/180) cases. Among the 291 eyes of CMVR, 281 eyes (96.6%, 281/291) of granular type; yellow-white point-like opacity and/or retinal hemorrhage; 10 eyes (3.4%, 10/291) of fulminant type; fundus Showed a typical "cheese ketchup-like" and vascular white sheath-like changes. Among the 180 children with CMVR, 72 patients (118 eyes) were given systemic intravenous drip and/or oral ganciclovir; 5 patients (10 eyes) were given intravitreal ganciclovir, all of which were fulminant CMVR. At the last follow-up, fundus lesions regressed significantly in 100 eyes of 61 cases; 18 eyes of 11 cases had old lesions or uneven retinal pigment; 108 cases were not treated.Conclusion:The most common fundus manifestation of CMV infection in infants is granular retinitis, and fulminant retinitis is more severe, and the lesions can be significantly regressed after timely antiviral treatment.
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Objective To investigate the correlation between the incidence of thyroid carcinoma and TSH, and to provide evidence for clinical application.MethodsSelect 450 cases of patients with thyroid from August 2014 to August 2016 in our hospital as the object of study in this group, the analysis of gender and age of patients, lymph nodes, tumor, laboratory examination results, to explore the relationship between TSH levels and thyroid cancer.ResultsThyroid patients with TSH was significantly higher than that in benign tumor group (P<0.05);the level of TSH and patients with lesion diameter, tumor stage, lymph node metastasis and capsule invasion was significantly correlated (P<0.05), but the level of TSH and age, gender, tumor number, vascular invasion and no significant correlation;TSH the level of thyroid cancer patients were significantly increased (P<0.05).ConclusionThe level of TSH in serum is closely related to the occurrence and development of thyroid cancer, and is closely related to lymph node metastasis and tumor stage, and the higher TSH level can increase the risk of thyroid cancer.
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Objective To identify the species of sarcosaphagous flies by amplifying cytochrome oxidase subunitI (CO I) gene fragment,combined with morphological characteristics.Methods The DNA of sarcosaphagous flies was extacted by modified Tris balanced phenol-Tris saturated phenol protocol,the amplificationand sequencing of CO Ⅰ fragment were conducted,and the results were then compared with the database for analysis.Results The modified DNA extration method by Tris balanced phenol-Tris saturated phenol could obtain effective DNA of sarcosaphagous flies,and be applied for CO Ⅰ fragment amplification,and thus for identification of the species of sarcosaphagous flies.Conclusion The DNA extracted from sarcosaphagous flies by modified Tris balanced phenol-Tris saturated phenol protocol could be used astemplate for the amplification of CO Ⅰ fragment,and the system could identify the species of sareosaphagous flies after sequencing and alignment with the sequences of database.Compared with traditional identification methods of using morphological characteristics,the current system is more accurate and could be more widely applied.
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Idiopathic membranous nephropathy(IMN) is one of the main pathology of nephrotic syndrome,and has a rising trend year by year.In recent years that IMN is organ specific autoimmune disease.Phospholipase A2 receptor(PLA2R) and thrombospondin type-1 domain containing 7A(THSD7A) are the two main antigens of IMN,which could be used to distinguish the IMN and secondary membranous nephropathy.The anti-PLA2R antibody and anti-THSD7A antibody are associated with diagnosis,severity and activity of the disease.The antibody titer and its change could be used to help deciding the beginning of therapy and the therapitive courses.
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Objective To investigate the clinical value of blood oxygenation level-dependent functional magnetic resonance imaging(BOLD-fMRI)in the protection of visual function during postoperative radiotherapy for occipital lobe gliomas. Methods Twenty-eight patients with occipital lobe gliomas receiving postoperative radiotherapy from 2014 to 2016 were enrolled as subjects. All patients underwent computed tomography(CT) simulation,conventional MRI,and BOLD-fMRI before radiotherapy. The location and scope of the visual cortex on 3DT1anatomical images were used to guide the labeling of the visual cortex on simulated CT images. A visual cortex protective radiotherapy plan and a conventional radiotherapy plan were made by intensity-modulated radiotherapy. The feasibility of the visual cortex protective radiotherapy plan was evaluated using conformity index(CI),homogeneity index(HI),Dmax,and Dmeanfor planning target volume(PTV)and Dmaxand Dmean to the visual cortex. Results For the two plans, if the effective dose to target volume was guaranteed and the doses to conventional organs at risk were acceptable, there were no significant differences in CI or HI for PTV between them(P=0.874,P=0.602).Compared with the conventional radiotherapy plan,the Dmaxand Dmeanto the ipsilateral visual cortex were reduced by 8.40% and 9.25%, respectively, while the Dmaxand Dmeanto the contralateral visual cortex were reduced by 13.26% and 14.77%, respectively, in the protective radiotherapy plan. Conclusions With a guaranteed prescribed dose to target volume and BOLD-fMRI used as a guide, the visual cortex protective radiotherapy, compared with the conventional plan, can reduce the dose to the visual cortex and protect the corresponding functional areas.
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Objective To analyze the feasibility of incorporation of tracking visual pathway fiber bundles by diffusion tensor imaging ( DTI) in computed tomography ( CT) simulation to develop a protective radiotherapy regimen for cerebral gliomas.Methods A total of 31 patients with cerebral gliomas who were admitted to our hospital from 2013 to 2015 and planed to receive postoperative radiotherapy were enrolled as subjects.All patients underwent CT simulation, conventional or contrast-enhanced magnetic resonance imaging, and DTI.The obtained DTI images of visual pathway fiber bundles were fused with 3DT1 anatomical scans and then imported into the treatment planning system.A protective treatment plan ( setting the entire visual pathway fiber bundles as organs at risk (OARs)) and a conventional treatment plan were made for intensity-modulated radiotherapy ( IMRT) .Comparison of treatment outcomes was made by paired t test.Results There were no significant differences in the conformity index and heterogeneity index of the planning target volume between the two treatment plans ( P=0.875,0.597), both of which had sufficient radiation doses to the target volume and conventional OARs protected.For the patients undergoing the protective treatment plan, the Dmax and Dmean values were reduced to 9.01%and 9.05%, respectively, in the ipsilateral optic tract and to 17.96%and 15.52%, respectively, in the contralateral optic tract;the Dmax and Dmean values were reduced to 5.37%and 5.48%(P=0.000), respectively, in the ipsilateral optic radiation tract and to 12.89%and 11.21%( P=0.000) , respectively, in the contralateral optic radiation tract.Conclusions The protective treatment plan based on CT simulation combined with the display of visual pathway fiber bundles by DTI can reduce the radiation dose to the entire visual pathway fiber bundles, which keeps the risk of visual dysfunction after radiotherapy as low as possible.
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Objective:To evaluate the application value of diffusion tensor imaging (DTI) in guiding the postoperative radiothera-py plan of the gliomas adjacent to the corticospinal tract (CST). Methods:Thirty patients with gliomas adjacent to the CST underwent routine magnetic resonance imaging (MRI) contrast-enhanced scanning and DTI after radiotherapy. Tractography data sets were ac-quired and were fused with the images of corresponding anatomical MRI and computed tomography. The acquired data sets of radio-therapy planning system were imported to assist with the delineation of the target volume, organs at risk, and CST. Two sets of radio-therapy plan, which considered or did not consider the dose protective effect of the CST, were formulated and compared using the treat-ment technique of intensity modulated radiotherapy. Results:The protective radiotherapy and unprotected plans both achieved the thera-peutic dose to the target volume and the protection of the routine organs at risk. In the protective dose (with an optimization program that considered the dose reduction of CST), the maximum and mean radiation doses suffered by the patients' ipsilateral and contra-later-al CSTs were lower compared with the unprotected plan (P<0.05). Conclusion:DTI can identify the location and shape of CSTs, and their relationship with the postoperative radiotherapy target of gliomas. These findings contribute to the formulation of a protective ra-diotherapeutic regimen to keep the CST from the maximum and the mean radiation doses to the largest extent, thereby decreasing the possibility of nerve damage after radiotherapy.
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Discipline development is the core power of hospital development,which is highly emphasized by Beijing Municipal Administration of Hospitals.To meet the demands of public hospitals reform and requirements of municipal hospitals development,performance evaluation is used as a tool for municipal hospitals in their fine management.This paper introduced the research performance evaluation system and index design of Beijing’s municipal hospitals,with a summary and analysis the operating and practices.
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Based on the worsening relationship between doctors and patients, the paper discusses the causes of the lack of medical ethics and aims to promote culture of traditional Chinese medicine and construct harmonious relationship between doctors and patients by the demands of charity ethics and excellent medical skills in Great Doctors, Great Faith by Shun Simiao.
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Objective To realize the incidence of healthcare-associated infection (HAI),the rate of invasive procedure and related infection rate,use of antimicrobial agents and distribution of pathogens in gereral intensive care units (GICUs) of hospitals in China.Methods HAI cross-sectional investigation data of GICUs reported to China HAI Surveillance Net-work were summarized and analyzed.Results Of 1 313 hospitals,621(47.30%)had GICUs;a total of 5 887 patients were surveyed,1 634 patients developed 1 962 times of infection,HAI prevalence rate and case rate was 27.76% and 33.33%respectively.The main infection sites were lower respiratory tract(70.39%),urinary tract (12.79%)and blood(2.86%). The application rate of urinary catheter,arteriovenous catheter and ventilator was 53.52%(n=3 151),37.05%(n=2 181) and 35.62%(n=2 097)respectively,and related prevalence of urinary tract infection,pneumonia and blood stream infection was 4.67%,20.41%,and 0.60%,respectively,which accounting for 58.57%,30.99%,and 23.21 % of urinary tract infection,pneumonia and blood stream infection respectively,the overall prevalence of above invasive procedure-re-lated infection accounted for 29.97%(588/1 962)of the overall HAI prevalence.A total of 1 795 isolated pathogens causing HAI were detected,the top three were Pseudomonas aeruginosa (20.78%),Acinetobacter baumannii(17.99%)and Klebsiella pneumoniae (11 .64%).The usage rate of antimicrobial agents was 71 .58%(n=4 214), rate of specimens delivered for detection was up to 75.27%(2 553/3 392).Conclusion Prevalence and antimicrobial use in patients in GICUs are all high,all kinds of invasive procedure are frequent,30% of HAI are related to inva-sive procedure;infection caused by non-fermentative bacteria is serious,pathogen detection is well performed.
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Objective To investigate the clinical value of blood oxygen level-dependent functional magnetic resonance imaging (BOLD-FMRI) in guiding the dose reduction and functional protection of the motor cortex during postoperative radiotherapy for brain astrocytoma.Methods This study included 27 brain astrocytoma patients with tumors near the motor cortex,who underwent routine MRI and BOLD-FMRI before postoperative radiotherapy.The location and scope of the motor cortex,which were acquired based on BOLD-FMRI results,were marked on positioning CT images,so that the motor cortex would be dealt with as organs at risk.The dose to the motor cortex was reduced as much as possible while ensuring the therapeutic dose to the target area.Finally,the radiotherapy plan that would reduce the mean received dose of the motor cortex was formulated and compared with the routine plan that could not reduce the received dose.Results With the protective radiotherapy plan,the mean received dose of the motor cortex decreased 0.76%-59.20% (mean 30.78%) on the affected side and 23.33%-68.30% (mean 48.07%) on the unaffected side;the coefficients of variation were 71.41% on the affected side and 36.71% on the unaffected side.Conclusions BOLD-FMRI can help to reduce the received dose of the motor cortex while ensuring the therapeutic dose to the target area when formulating the postoperative radiotherapy plan for brain astrocytoma,thus protecting the motor function and improving patients' quality of life.
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Objective To observe the effects of endoplasmic reticulum stress (ERS) on the activation of monocytes induced by high glucose and explore the underlying mechanism.Methods The monocyte cell line THP-1 was stimulated with high glucose,and then treated with molecular chaperone betaine.The levels of glucose regulation protein 78 (GRP78) and p-JNK,which were associated with ERS were detected by real-time PCR and Western blotting.The proliferation of the cell line was detected by MTT method.Transwell and immunofluorescence were applied to observe the chemotaxis and phenotype of cells respectively.Results The levels of GRP78 and p-JNK of THP-1 cells stimulated by high glucose were significantly increased compared with the normal control group (all P < 0.05).The proliferation and chemotactic were also enhanced (all P < 0.05).The number of cells in M1 phenotype was increased remarkably (P < 0.05).All the indexes above could be rescued by betaine.Conclusion The activation of THP-1 cells can be induced by high glucose through ERS,while molecular chaperone betaine can reverse the activation.
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Objective To investigate the renal expression changes of microRNA-215(miR-215) and its role in diabetic nephmpathy of type 2 diabetic db/db mice. Methods Fourweek-old diabetic db/db mice and norml control group non-diabetic db/m mice were selected.Real-time PCR was used to detect the relative level of miR-215 at the age of 8,12 and 16 weeks.Catenin beta interacting protein 1 (CTNNBIP1) mRNA and protein level were measured by realtime PCR,WesteRN blotting and immunohistochemisty.A lueiferase reporter assay was used to determine whether CTNNBIP1 was a direct target of miR-215. Results (1)With the growth of db/db mice,the major pathological characteristics of kidney included glomerular hypertrophy,segmental mesangial cells proliferation and mesangial matrix expansion.(2)Compared with the db/m mice,the db/db mice of 8,12 and 16 weeks showed obvious increase in body weight(BW),blood glucose (Glu) and 24 hour urinary albumin excretion (UAE) (P<0.05,respectively).(3)Compared with the db/m mice,special miR-215 was highly expressed in the kidney of db/db mice and was up-regulated significantly according to the development of DN (P<0.05).(4)The mRNA and protein expression of CTNNBIPl of kidney were consistently down-regulated in db/db mice than those in controls (P<0.05,respectively). (5)By luciferase reporter,miR-215 could negatively regulate CTNNBIP1 gene by targeting its 3'-UTR sequence (P<0.01). Conclusion High expression level of miR-215 plays a potential role in the initiation and progression of DN by down-regulating the expression of CTNNBIPl.
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Objective To investigate the correlation between plasma proteasome and endothelial dysfunction in patients with uremia. Methods Forty-five uremic patients who did not receive hemodialysis were defined as A group; seventy-five uremic patients who had received hemodialysis for 6 to 12 months were divided into sufficient hemodialysis group (44 cases,B group)and insufficient hemodialysis group (31 cases,C group).The primary disease of these patients was chronic glomerulonephritis.Fifteen healthy people were defined as healthy control group (D group).The diameter of radial artery lumen (DRL),intima-media thickness (IMT),intima-media area (IMA),endothelium-dependent or independent dilation (EDD or EID) of radial artery in right forearm were detected by diasonography.The levels of 20S proteasome,tumor necrosis factor α (TNF-α),C-reaction protein (CRP) and transforming growth factor β 1 (TGF-β1) of plasma and supernatant of cultured human umbilical veins endothelium (HUVEC) were determined by enzyme linked immunosorbent assay (ELISA).20S proteasome activity was analyzed by special substrate.Results Compared with D group,the level and activity of 20S proteasome,as well as TNF-α,CRP and TGF-β1 in A,B and C groups were significantly increased.Compared with A group,these plasma indices levels were significantly decreased in B group but strongly increased in C group.IMT and IMA were elevated,while DRL,EDD and EID were decreased significantly in A,B and C groups when compared with D group.These parameters were worse in C group than those in A and B groups.After co-culture of HUVEC with above mentioned human uremic serum,the level and activity of 20S proteasome and TNF-α were higher in A,B,C groups than that in D group.In A and C groups,there were negative correlations of EDD with the level or activity of 20S proteasome,TNF-α,CRP and TGF-β1,and there were positive correlations of 20S proteasome level or activity with TNF-α,CRP and TGF-β1. Conclusions 20S proteasome level and activity are significantly increased in uremic patients.There is a close correlation between 20S proteasome and endothelial dysfunction of radial artery.
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Objective To investigate the effect and significance of regulating endoplasmic reticulum stress on the expression of histone methyltransferases SET7/9 in the kidneys of db/db mice.Methods Db/db mice were randomly divided into two groups according to random number table method:diabetic nephropathy model group (DN group,n=18) and betaine treatment group (DN+B group,n =18),db/m mice were defined as normal control group (NC group,n =18).At the end of 4,8 and 12 weeks,the expression of GRP78,SET7/9,H3K4me2,and monocyte chemoattractant protein 1 (MCP-1) was determined by real-time fluorescence PCR and Western blotting.24-hour urinary protein excretion rate (UPER) and urine MCP-1 were measured by enzyme linked immunosorbent assay (ELISA).The dynamic changes of blood glucose(BG),serum creatinine (Scr),blood urea nitrogen (BUN) were tested by completely automatic biochemistry analyzer.The morphology of kidney was estimated by special staining of periodic acid-schiff (PAS).Results The levels of BG,BUN,UAER and MCP-1 were significantly higher in DN group than those in NC group (P < 0.05),and were in time-dependent manner.Glomerular basement membrane thickening and mesangial cells proliferation began to emerge in DN group at the end of week 4 and mesangial matrix expansion was more obvious at the end of week 12.The mRNA and protein expression of GRP78 and SET7/9 were elevated significantly in DN group as compared to NC group.The H3K4me2 protein expression level was also increased in time-dependent manner.Compared with the DN group,in DN+B group glomerular lesions attenuated and the GRP78 and SET7/9 expression levels obviously decreased (P < 0.05).Furthermore,the levels of BG,BUN,UPER,MCP-1,H3K4me2 in DN+B group were also reduced (P < 0.05).Conclusion Endoplasmic reticulum stress may be the upstream mechanism of mediating the expression of SET7/9 in the kidneys of DN mice.
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Objective To investigate the effects of MG132 on diabetic nephropathy (DN) rats induced with streptozocin. Methods Seventy-two male SD rats were randomly divided into three groups: normal control group (NC, n=24), DN group (n=24) and DN treated with MG132 group (DN+MG132, n=24). At the end of 4, 8 and 12 weeks, 24 hour urinary protein excretion rate (UPER) was detected. Morphology of kidney was examined by special staining of periodic acid-schiff (PAS). Renal 26S proteasome activity was determined by quantifying the hydrolysis of S-LLVY-AMC in a fluorescence reader. Urinary malondialdehyde (MDA) level and renal SOD and GSH-PX activity were detected by commercial kits. Renal SOD, GSH-PX and p47phox mRNA expressions were determined by real-time fluorescence PCR. Renal p47phox protein expression wasdetermined by Western blotting. Results Compared with NC group, the DN group showed a significant increased of UPER at week 4, 8, 12 (all P<0.05), of mesangium proliferation and mesangial matrix expansion at week 12. In DN+MG132 group, UPER was significantly decreased compared with DN group at the end of 4, 8 and 12 weeks (P<0.05, respectively), and the glomeruler pathological alteration induced by diabetes was attenuated. Increased renal 26S proteasome activity in DN rats was significantly inhibited after MC132 administration (P<0.05). Moreover, renal p47phox mRNA expression in DN group was 155%, 149% and 120% more than those in NC group at 3 time points (all P<0.05), and so was the renal p47phox protein expression, 139%, 152% and 186% more (all P<0.05). Urinary MDA levels in DN group were 1.95-, 2.04-and 2.62-folds more than those in NC group (all P<0.05). In addition, compared with NC group at 3 time points, in DN group, renal SOD activity was decreased by 23.09%, 33.59% and 53.31% (all P<0.05); renal GSH-PX activity was decreased by 28.57%, 33.06% and 48.76% (all P< 0.05); renal SOD mRNA was decreased by 38.09%, 61.44% and 76.53% (all P<0.05); renal GSH-PX mRNA group was decreased by 29.16%, 37.26% and 62.40% (all P<0.05). Compared with DN group, renal p47phox mRNA and protein expression, and urinary MDA levels were significantly lower in DN+MG132 group (all P<0.05); renal SOD and GSH-PX activity as well as mRNA expression were significantly increased in DN+MG132 group (all P<0.05). Conclusions MG132 treatment can provide renoprotection for DN rats effectively maybe through enhancing renal anti-oxidative ability.
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Objective To study the role of endoplasmic reticulum stress in phenotypic change of cultured human glomerular mesangial cells induced by high glucose.Methods Cultured human glomeruar mesangial cells were divided into three groups: control group,high glucose group and high glucose+ 4-phenylbutyric acid (4-PBA) group.Cell number of proliferation was assessed by MTT assay.Cell cycle was measured by flow cytometric analysis.Expression of α-SMA was assessed by immunohistochemistry and was observed by laser scanning confocal microscope.Involved mRNA and protein expression were measured by real-time PCR and Western blotting.Results (1)Cell number of proliferation and S transition proportion in high glucose group significantly increased than that in control group (P < 0.05).High glucose could induce α-SMA expression significantly (P<0.05).4-PBA could significantly inhibit human glomerular mesangial cells proliferation (P<0.05),S transition arrest (P<0.05) and expression of α-SMA (P<0.05) induced by high glucose.(2) Compared with control group,high glucose could significantly increase the expression of glucose-regulated protein78(Grp78 ) mRNA and protein (P< 0.05),which could be inhibited by 4-PBA treatment (P<0.05).(3)High glucose could induce the mRNA and protein expression of TGF-β1 and FN significantly,which could be inhibited by 4-PBA treatment (P<0.05).Conclusion Endoplasmic reticulum stress plays an important role in phenotypic change of cultured human glomerular mesangial cells induced by high glucose.
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Objective To identify the rale of NKX2-5 gene in cardiomyocyte differentiation and its mechanism.Methods P19 cells were divided into transfected and non-transfected groups.In the transfected group,P19 cells were with stable expression of NKX2-5 gene.The P19 cells were cultured in suspension for 4 days,and the formed aggregates were transferred to Petri dish for adherent culture.On days 4,8,12,and 16 of the adherent culture,the expressions of ct-saicomeric actin(α-SA)and cardiac troponin T(cTnT)were detected with double-labeling immunofluorescence and Western blot.The ultrastruetural changes were observed on day 16.Results In the transfected group,no expression of α-SA and cTnT was found on day 4,and the expression of these 2 proteins or co-expression existed on days 8,12,and 16.There were early cell junction and myofilament-like structure in the cytoplasm of some cells in the transfected group.In the non-transfected group,these 2 proteins were negative,and no differentiated cell was found.Conclusion Stable expression of NKX2-5 gene can induce cardiomyocyte differentiation from P19 cells,but the P19 cells with stable expression of JVKX2-5 gene is not suitable to be an in vitro model of cardiac development.
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Objective To investigate the effect of 4-phenylbutyric acid(4-PBA)on the renal pathogenesis of rats with streptozotocin-induced diabetes and its mechanism. Methods Fifty-four male SD rats were randomly divided into three groups:normal control group(NC group,n=18),diabetic nephropathy group(DN group,n=18),diabetic nephropathy plus 4-PBA treatment group(4-PBA group,n=18).At the end of 4,8 and 12 weeks,index of kidney weight/body weight ratio(KI)were measured and calculated.Serum creatinine (Scr),blood urea nitrogen(BUN),urinary MDA levels,urinary SOD activity,and 24 hour urinary protein excretion ram(UAER)were detected by HITACHI automatically.Morphology of kidney wag examined by special staining of periodic acid-schitt (PAS).The p47phox and nitrotyrosine (NT) expression in kidney were determined by real-time fluorescence PCR and Western blotting. Results Compared with the NC group, the DN group rats showed a significant increase of KI(P<0.05), UAER(mg/24 h) (4.92±0.70 vs 0.26±0.07, 5.29±0.83 vs 0.28±0.08, 5.54±0.81 vs 0.29±0.04,respectively, P<0.05]for indicated time, mesangial cells proliferation and mesangial matrix expansion at 12 week. However,4-PBA treatment could significantly inhibit the increase of KI (P<0.05), decrease UAER (mg/24 h) (3.71±0.37, 3.47±0.36, 3.28±0.40, respectively, P<0.05]for indicated time, and prevent the glomeruler pathological alteration induced by diabetes. Moreover, the mRNA expression of p47phox in the kidney of DN group was 154.72%, 148.60% and 91.95% more than that of NC group (all P<0.05) for indicated time. The protein expression of p47phox was 118.00%, 140.10% and 177.82% more than that of NC group (all P<0.05), and the protein expression of NT was 45.29%,59.13% and 89.28% more than that of NC group (all P<0.05). In addition, urinary MDA levels in DN group were 2.05-, 2.26- and 2.43- folds of NC group, and urinary SOD activities were decreased by 64.78%, 71.29% and 79.32% of NC group. Compared with the DN group, the mRNA and protein expression of p47phox, and protein expression of NT in 4-PBA group were decreased markedly (all P<0.05) at the end of 8 and 12 weeks. The urinary MDA level was decreased, and the urinary SOD activity was increased significantly in rats with diabetes after 4-PBA treatment for indicated time (all P<0.05). Conclusion 4-PBA treatment can significantly inhibit the renal pathogenesis of rats with diabetes through inhibition of oxidative stress.