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Objective:To investigate changes in mRNA and protein expression of interleukin-27(IL-27)in glomerular podocyte injury caused by Puromycinonucleoside(PAN), and to explore the mechanism of the protective effect of Tacrolimus(FK506)on glomerular podocyte injury.Methods:Glomerular foot cells from mice were cultured in vitro and divided into 3 groups, which were the control group, PAN group and FK506 group.The morphology of 3 groups of foot cells was observed under a microscope after 8 h, 24 h, and 48 h treatment.The changes in IL-27 concentrations were detected by analyzing the enzyme-linked immunosorbent assay(ELISA)method.The cultured foot cells were then collected.The changes of IL-27 mRNA expression were measured by real-time fluorescence quantitative PCR(qPCR), and the changes of IL-27 protein expression were detected by Western blot. Results:(1)At each time point(8 h, 24 h, 48 h), the cells of the PAN group had smaller volume and different morphology than the cells of the control group, and the cells of the FK506 group was larger and fuller than the cells of the PAN group.(2)The concentrations of IL-27 in the PAN group [(110.00±3.52) ng/L, (302.00±6.23) ng/L, (397.00±8.92) ng/L] were significantly higher than those in the control group [(90.00±5.12) ng/L, (85.00±4.21) ng/L, (88.00±4.20) ng/L] and those in the FK506 group [(96.00±4.17) ng/L, (107.00±4.86) ng/L, (112.00±6.24) ng/L] at 8 h, 24 h and 48 h(all P<0.05). (3)At each time point(8 h, 24 h, 48 h), the IL-27 mRNA expression of the PAN group(1.25±0.11, 1.57±0.08, 1.73±0.13)was significantly higher than that of the control group(1.02±0.02, 1.10±0.04, 0.96±0.02)(all P<0.05). Compared with the control group, the IL-27 mRNA expression of the FK506 group did not significantly increase at 8 h (1.10±0.06), and showed a slight increase at 24 h and 48 h(1.21±0.04, 1.30±0.09). Compared with PAN group, HC506 group were all lower (all P<0.05). (4)At each time point(8 h, 24 h, 48 h), the expression of IL-27 protein in the PAN group(0.94±0.04, 1.56±0.07, 1.63±0.04) was significantly higher than that in the control group(0.83±0.04, 0.85±0.03, 0.83±0.05), there was significant difference(all P<0.05). Compared with the PAN group, the expression of IL-27 protein in the FK506 group(0.84±0.05, 0.89±0.04, 0.91±0.06)was not significantly different at 8 h, but decreased significantly at 24 h and 48 h, there was significant difference ( P<0.05). Conclusions:IL-27 is involved in the pathogenesis of kidney diseases.FK506 can prevent the glomerular podocyte injury by reducing the expression of IL-27.This study provides experimental basis for clinical application of FK506 in the treatment of kidney diseases.
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Objective To investigate the effects of Tacrolimus( FK506 )and Puromycin aminonucleoside ( PAN)on apoptosis and expression of α-actinin-4 mRNA and protein in mouse glomerular podocytes in order to ex-plore the protective effect of FK506 on podocytes. Methods Mouse glomerular podocytes were cultured in υitro,and the control group,PAN group and FK506 group were established. After 8 h,24 h and 48 h of treatment,the cell mor-phology was observed and the apoptosis rate was detected. The cells were collected by real-time PCR and Western blot was used to detect the mRNA and protein expression of α-actinin-4. Results The cell body area of the PAN group was significantly smaller than that of the control group,and the cell area of the FK506 group was larger than that of the PAN group. There was no significant difference in the rate of podocyte apoptosis between those groups at 8 h( all P>0. 05). At 24 h and 48 h,the apoptotic rate of podocytes in PAN group[(8. 21 ± 0. 41)%,(16. 32 ± 0. 17)%]were significantly higher than those in the control group[(4. 28 ± 0. 35)%,(6. 27 ± 0. 28)%],and the differences were significant(all P<0. 05). The apoptosis rate of podocytes in FK506 group[(6. 26 ± 0. 24)%,(13. 32 ± 0. 24)%] were significantly lower than those in PAN group,and the differences were significant(all P<0. 05). At 8 h,there was no significant difference in the expression of α -actinin -4 mRNA and protein( all P >0. 05 ). The expression of mRNA(2. 42 ± 0. 21,3. 78 ± 0. 25)and protein(0. 77 ± 0. 04,1. 22 ± 0. 10)in the PAN group was significantly higher than mRNA(1. 50 ± 0. 22,2. 15 ± 0. 15)and protein(0. 44 ± 0. 03,0. 83 ± 0. 07)in the control group at 24 h and 48 h,and the differences were significant(all P<0. 01). The expression of mRNA(1. 65 ± 0. 24,1. 70 ± 0. 32)and protein(0. 52 ± 0. 05,0. 56 ± 0. 07)in FK506 group was significantly lower than that of PAN group,and the differ-ences were significant(all P<0. 05). Conclusions FK506 can effectively inhibit the damage of PAN on podocytes and stabilize the expression of α-actinin-4,which provides a basis for the clinical application of FK506 in the treat-ment of glomerular diseases.
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Objective@#To investigate the effects of Tacrolimus(FK506) and Puromycin aminonucleoside(PAN) on apoptosis and expression of α-actinin-4 mRNA and protein in mouse glomerular podocytes in order to explore the protective effect of FK506 on podocytes.@*Methods@#Mouse glomerular podocytes were cultured in vitro, and the control group, PAN group and FK506 group were established.After 8 h, 24 h and 48 h of treatment, the cell morphology was observed and the apoptosis rate was detected.The cells were collected by real-time PCR and Western blot was used to detect the mRNA and protein expression of α-actinin-4.@*Results@#The cell body area of the PAN group was significantly smaller than that of the control group, and the cell area of the FK506 group was larger than that of the PAN group.There was no significant difference in the rate of podocyte apoptosis between those groups at 8 h (all P>0.05). At 24 h and 48 h, the apoptotic rate of podocytes in PAN group[(8.21±0.41)%, (16.32±0.17)%] were significantly higher than those in the control group[(4.28±0.35)%, (6.27±0.28)%], and the differences were significant (all P<0.05). The apoptosis rate of podocytes in FK506 group[(6.26±0.24)%, (13.32±0.24)%] were significantly lower than those in PAN group, and the differences were significant (all P<0.05). At 8 h, there was no significant difference in the expression of α-actinin-4 mRNA and protein(all P>0.05). The expression of mRNA (2.42±0.21, 3.78±0.25) and protein(0.77±0.04, 1.22±0.10) in the PAN group was significantly higher than mRNA(1.50±0.22, 2.15±0.15) and protein(0.44±0.03, 0.83±0.07) in the control group at 24 h and 48 h, and the differences were significant (all P<0.01). The expression of mRNA (1.65±0.24, 1.70±0.32) and protein (0.52±0.05, 0.56±0.07) in FK506 group was significantly lower than that of PAN group, and the differences were significant (all P<0.05).@*Conclusions@#FK506 can effectively inhibit the damage of PAN on podocytes and stabilize the expression of α-actinin-4, which provides a basis for the clinical application of FK506 in the treatment of glomerular diseases.
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Objective To investigate the clinical efficacy and safety of angiotensin-converting enzyme inhibitor (ACEI) and angiotensin receptor blocker (ARB) in treatment of children with Alport syndrome (AS).Methods A total of 22 children with AS in Department of Pediatrics,Guangzhou First People's Hospital and Department of Pediatrics,Shenzhen People's Hospital between January 2013 and December 2017 were selected.But four children were not included in this study since they did not take medication regularly,and the other 18 cases were included in this study.All the 18 children were initially treated with ACEI.The observation time was from 1 to 5 years.If the symptoms were not effectively controlled,treatment plan would be changed to ACEI combined with ARB treatment.The observation time was from 1 to 3 years.The clinical data and laboratory examination results [including 24-hour urine protein (mg/24 h),urine red blood cell count,plasma albumin (Alb),urea nitrogen (BUN),serum creatinine (Scr),total cholesterol (TC)] were collected for retrospective analysis.Results Eighteen patients started their treatment with ACEI inhibitors (Fosinopril).Within 2 years of treatment,the urinary protein and urinary red blood cells in the children decreased to 47.7％ and 41.3％,respectively,and the differences were all statistically significant (all P ＜ 0.05),and the renal function was stable within the normal range.Two years later,7 patients had elevation of urinary protein and urinary erythrocyte elevations and decrease of renal function,and they were treated with ACEI and ARB (Losartan).The other children had no significant change in urine protein and urine red blood cells in the 3 rd,4th,and 5th year,and their renal function was stable.After ACEI treatment alone for 5 years,urinary protein was 47.8％ lower than before treatment,and the difference was statistically significant (P ＜ 0.05);urinary red blood cells decreased to 32.0％ compared with before treatment,and the difference was statistically significant (P ＜ 0.05).Seven patients with ACEI alone had poor efficacy,after the treatment with ACEI combined with ARB,the urinary protein and urine red blood cells were lower in the first year and the renal function improved.There was no significant change in urinary protein and urine red blood cells in the 2nd and 3rd year,and renal function was stable.After ACEI combined with ARB treatment for 3 years,urinary protein decreased to 42.3％ before treatment,and the difference was statistically significant (P ＜ 0.05),and urinary red blood cells decreased to 46.9％ compared with before treatment,and the difference was statistically significant(P ＜ 0.05).Conclusions ACEI treatment of children with AS can reduce urine protein and help delay renal failure.For children with poor efficacy of ACEI treatment,ACEI combined with ARB may have a certain effect.ARB can be used as an adjunctive treatment for patients with AS who have a poor response to ACEI alone.
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Objective To evaluate the value of multislice spiral computed tomography angiography (MSCTA) in differential diagnosis between epithelial ovarian carcinoma (EOC) and borderline epithelial ovarian tumor (BOT).Methods The MSCTA images of 39 EOC patients and 23 BOT patients confirmed by surgical pathology were reviewed retrospectively.Main characteristics of tumor vessels were analyzed: the number of feeding arteries, the existence of dilated draining veins, whether the tumor vessels were tortuous, whether the distribution of tumor vessels were disturbed, and whether there were accompanying microaneurysms or arteriovenous malformations (AVMs).Results Two or more feeding arteries of the EOCs and BOTs were 89.7% (35/39) and 8.7% (2/23), respectively.Dilated draining veins were observed in 87.2% (34/39) of the EOCs and 4.3% (1/23) of the BOTs.The tortuosity of tumor vessels was observed in 97.4% (38/39) of the EOCs and 13.0% (3/23) of the BOTs.79.5% (31/39) of the EOCs and 8.7% (2/23) of the BOTs were complicated by microaneurysms, and 74.4% (29/39) of the EOCs and 4.3% (1/23) of the BOTs were complicated by AVMs.The characteristics of tumor vessels were significantly different between the two groups (P<0.01), with relatively high sensitivity and specificity.Conclusion MSCTA can better show the distribution, number and pattern of tumor vessels and is of great value in differential diagnosis between EOC and BOT.
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Objective To evaluate effects of interleukin-36α (IL-36α) on psoriasiform skin lesions and C-C motif chemokine ligand 20 (CCL20) expression in mice.Methods Totally,30 BALB/c female mice were randomly and equally divided into 3 groups:control group treated with topical vaseline cream on the shaved back and intracutaneous injection with phosphate buffer saline (PBS),model group treated with topical imiquimod cream on the shaved back and intracutaneous injection with PBS,experimental group treated with topical imiquimod cream on the shaved back and intracutaneous injection with IL-36α solution.Psoriasis area severity index (PASI) was used to evaluate changes of psoriasiform skin lesions in mice,and light microscopy to observe morphological changes of skin lesions and to measure the thickness of the epidermis.Real-time fluorescence-based quantitative PCR (qRT-PCR) and Western blot analysis were performed to determine the expression of IL-36α in skin lesions in the control group and model group,and qRT-PCR,Western blot analysis and immunohistochemical study to evaluate changes of CCL20 levels in skin lesions.Results The model group showed significantly increased mRNA (△ Ct value:0.0195 ± 0.0059) and protein expression (3.922 ± 0.248) of IL-36α compared with the control group (mRNA:0.0012 ± 0.0004,P ＜ 0.05;protein:0.690 ± 0.025,P ＜ 0.05).The mRNA and protein expression of CCL20 were significantly higher in the experimental group than those in the model group (mRNA:2.152 ± 0.793 vs.0.999 ± 0.178;protein:0.397 ± 0.033 vs.0.145 ± 0.030;both P ＜ 0.05),and higher in the model group than those in the control group (mRNA:0.378 ± 0.075;protein:0.025 ± 0.009;both P ＜ 0.05).Immunohistochemical study showed that the expression intensity of CCL20 in skin lesions significantly increased in the experimental group compared with that in the model group (Z =2.294,P ＜ 0.05).Conclusion IL-36α may aggravate psoriasiform skin inflammation in mice by promoting CCL20 expression.
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Objective To observe the effect of modified Qinghao Biejia Decoction ( QBD) on Th17 cells and renal pathology of MRL/lpr mice with spontaneous systemic lupus erythematosus. Methods Thirty-two female MRL/lpr mice aged 8 to 10 weeks were divided into 4 groups: model group, Chinese medicine group, prednisone group, and combination group, 8 mice in each group. Eight female C57BL/6 mice aged 8 to 10 weeks served as normal control. Mice in Chinese medicine group were given concentrated solution of modified QBD (19.25 g·kg-1·d-1), mice in the prednisone group were given water solution of prednisone acetate (8.75 mg·kg-1·d-1) , mice in the combination group were given the above two kinds of medicine, and mice in the model group and normal control group were given physiological saline. After medication for 7 weeks, spleens and kidneys in all of the groups were taken out for the experiment. Th17 cells in splenic mononuclear cell suspension were detected by flow cytometry, the pathological changes of renal tissue were observed under light microscope, and activity index (AI) of renal tissue in lupus nephritis mice was scored. Results The proportion of Th17 cells in the model group was significantly higher than that of normal control group ( P<0.05) . The proportion of Th17 cells in Chinese medicine group and combination group was lower than that of the model group ( P<0.05) , and prednisone group had higher proportion of Th17 cells than Chinese medicine group ( P<0.05) . Compared with the model group, pathological changes of renal tissue were relieved, and AI scores were decreased in Chinese medicine group, the prednisone group and the combination group ( P<0.05) . Except for the normal control group , AI scores in all groups were positively correlated with the proportion of Th17 cells ( r=0.77, P<0.01) . Conclusion Modified QBD can inhibit the expression of Th17 cells and improve the pathological changes of MRL/lpr mice with lupus nephritis.
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Objective To assess the enhancement feature of intracranial atherosclerotic plaque in the vessel supplying the territory of infarction by using high-resolution MR imaging.To analyze the correlation between the degree of plaque enhancement , time elapsed and the concentration of hypersensitive C-reactive protein ( hs-CRP ).Methods The characteristics of vessel walls and intracranial vascular stenoses were retrospectively analyzed in 81 patients with ischemic strokes.All subjects were imaged with a traditional stroke MR protocol and HR-MRI scanning for plaque on a 3.0 T MRI scanner.According to the elapsed time between infarct and MR examination , all cases were classified into early stage (12 weeks, n=10).The characteristics of vessel walls and degrees of enhancement of atherosclerotic plaques were assessed and the concentrations of hs-CRP in all patients were determined.The Kruskal-Wallis H test was used to compare the degree of enhancement and hs-CRP concentration among the early , middle and late stage.The concentration of hs-CRP was presented as median ( interquartile range ).The Spearman correlation was used to analyze the correlation between elapsed time , hs-CRP concentration and degree of enhancement.Results Fifty-five (55/81) plaques were located at the M1 segments, and the other 26 (26/81) plaques were at the basilar artery.The degree and presence of enhancement from strong to none were 29, 25 and 4 in the early stage;4, 6 and 3 in the middle stage and 0, 4, 6 in the late stage, respectively.The degree and presence of enhancement were significantly different among them (H=16.934,P<0.01).There was a remarkable trend of decreasing degree and presence of enhancement of the atherosclerotic plaque relative to increasing time after the ischemic event(r=-0.792,P<0.01).The serum hs-CRP concentration for early, middle and late stage were 7.0(3.0, 13.0), 2.27(1.0, 3.03) and 1.88(0.50, 4.0)mg/L (H=14.345,P<0.01) , respectively.There was a trend of decreasing hs-CRP concentration relative to the time elapsed ( r =-0.357,P<0.01).The degrees of enhancement of the plaques were parallel to the levels of hs -CRP( r=0.526,P<0.01).Conclusions Enhanced HR-MRI scanning may clearly demonstrate the enhancement characteristics of intracranial atherosclerotic plaques as an indicator of inflammation.It might play an important role to detect risk factors for intracranial plaque rupture and subsequent acute ischemic stroke .
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Purpose To reconstruct perfusion computerized tomography angiography (PCTA) images from the volume data of low-dose brain CT perfusion scan with iterative reconstruction algorithm, to analyze the capability of PCTA on the display of brain arteries, and to explore the methods to reduce the radiation dose for stroke CT examinations. Materials and Methods This was a prospective study, 55 patients (605 arterial segments) with clinical diagnosis of ischemic cerebrovascular disease underwent cranial CT scan, iterative algorithm low-dose brain CT perfusion scan and conventional cranial CTA examinations using a 256-slice spiral CT. 11 segments of the cerebral artery in each case were analyzed using conventional CTA results as the reference standard to assess the display of brain arteries in PCTA. Results Effective dose of CT perfusion scan was 2.12 mSv. Among the 580 vessel segments which CTA showed no stenosis or stenosis0.75 for the consistency test between PCTA and CTA on the display of brain arteries. Conclusion Radiation dose of iterative algorithm cranial CT perfusion scan is significantly lower, and the images reconstructed from the volume data of perfusion CT are highly consistent with the CTA results, thus are able to meet the needs of the clinical diagnosis.