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A 3-year-old male patient was diagnosed with neurofibromatosis type 1(NF1) for two years. The patient has multiple neurofibromas in retroperitoneum, lumbococcygeal paravertebral, lumbosacral spinal canal, and foramina. Due to retroperitoneal mass compression, the child suffered from urological complications such as hydronephrosis, ureterdilation, neurogenic bladder, etc., which seriously affected the urination function and resulted in multiple surgical treatments. Currently, the patient has been treated with mitogen activates extracelluar signal-regulated kinases(MEK) inhibitor selumetinib targeted therapy, and has voluntarily urinated, and his general state is better than before medication. The diagnosis and treatment of this case reflects the importance of multidisciplinary collaboration in the diagnosis and treatment of rare diseases.
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Ferroptosis is a newly discovered regulatory mode of cell death, which is caused by glutathione peroxidase 4 deficiency, abnormal iron metabolism and lipid peroxidation.At present, it is considered that iron metabolism and active oxygen metabolism are the central link of ferroptosis.Ferroptosis involves a variety of physiological and pathological processes, including cancer cell death, neurotoxicity, ischemia-reperfusion injury, and T-cell immunity.Studies have shown that ferroptosis characteristics such as iron overload and lipid peroxidation may occur in different degrees during the development of a variety of nephropathy.Ferroptosis can affect the progression of renal disease by regulating the level of intracellular iron ions and lipid peroxidation.Therefore, effective regulation of ferroptosis is expected to be an important strategy in the treatment of renal diseases.In this paper, the regulation mechanism of ferroptosis and its research progress in kidney disease are reviewed to provide new theories and ideas for the treatment of renal disease.
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Objective@#To investigate the effect of silencing troponin I3 (Tnni3) gene expression on biological property of rat embryonic H9C2 cardiomyocytes.@*Methods@#The rat embryonic H9C2 cardiomyocytes were cultured and divided into 2 groups: control group transfected with negative control small interfering RNA (NC-siRNA group) and experimental group transfected with Tnni3 small interfering RNA (Tnni3-siRNA group). At 48 h, 72 h after transfection, the cells were collected, and real time quantitative polymerase chain reaction (qPCR)was used to detect the mRNA expressions of Tnni3 and Caspase-3, and Western blot was used to detect the protein expressions of Tnni3, Cyclin A1 and Cyclin B1.Annexin V-fluorescein isothiocyanate(FITC) apoptosis detection kit was used to analyze cell apoptosis.Cell proliferation was measured by Cell Counting Kit-8 (CCK-8) solution and cell cycle was detected by flow cytometry.@*Results@#At 48 h post-transfection with Tnni3-siRNA, H9C2 cells exhibited a significant decrease in Tnni3 mRNA (0.27±0.05 vs. 1.00±0.00) and protein (0.18±0.03 vs. 1.00±0.00) compared with those transfected with NC-siRNA, and the differences were statistically significant (t=25.26, 47.40, all P<0.01). Apoptotic cells were observed in the NC-siRNA group and the Tnni3-siRNA group.At 72 h post-transfection, the percentage of apoptotic cells significantly increased in H9C2 cells transfected with Tnni3-siRNA [(11.30±1.85)% vs. (0.33±0.15)%] compared with those transfected with NC-siRNA, an increased expression of Caspase-3 mRNA was also observed in Tnni3-siRNA-transfected H9C2 cells (1.39±0.13 vs. 1.00±0.00), and the differences were statistically significant (t=10.24, 5.19, all P<0.01). Compared with NC-siRNA-transfected H9C2 cells, a time-dependent reduction in cell proliferation was observed in Tnni3-siRNA-transfected H9C2 cells (48 h: 0.32±0.06 vs. 0.46±0.03; 72 h: 0.31±0.01 vs. 0.63±0.04; 96 h: 0.36±0.01 vs 0.75±0.04), and the differences were statistically significant (t=3.62, 13.45, 16.39, all P<0.01). At 72 h post-transfection with Tnni3-siRNA, the percentage of G1 phase, S phase and G2 phase cells was (71.25±3.82)%, (18.28±2.78)% and (9.94±1.09)%, respectively.There was a significant increase in the proportion of G2 phase cells [(9.94±1.09)% vs. (4.54±0.99)%] in H9C2 cells transfected with Tnni3-siRNA compared with those transfected with NC-siRNA, an increased expression of Cyclin A1 protein (1.89±0.09 vs.1.00±0.00) and a decreased expression of Cyclin B1 protein (0.47±0.06 vs.1.00±0.00) were observed in Tnni3-siRNA-transfected H9C2 cells, respectively, and the differences were statistically significant (t=6.35, 17.12, 15.32, all P<0.01).@*Conclusions@#Silencing Tnni3 gene expression in rat embryonic H9C2 cardiomyocytes can induce cell apoptosis, suppress cell proliferation, and led to G2 cell cycle arrest.
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Objective To investigate the effect of silencing troponin Ⅰ3 (Tnni3) gene expression on biological property of rat embryonic H9C2 cardiomyocytes.Methods The rat embryonic H9C2 cardiomyocytes were cultured and divided into 2 groups:control group transfected with negative control small interfering RNA (NC-siRNA group) and experimental group transfected with Tnni3 small interfering RNA (Tnni3-siRNA group).At 48 h,72 h after transfection,the cells were collected,and real time quantitative polymerase chain reaction (qPCR)was used to detect the mRNA expressions of Tnni3 and Caspase-3,and Western blot was used to detect the protein expressions of Tnni3,Cyclin A1 and Cyclin B1.Annexin V-fluorescein isothiocyanate(FITC) apoptosis detection kit was used to analyze cell apoptosis.Cell proliferation was measured by Cell Counting Kit-8 (CCK-8) solution and cell cycle was detected by flow cytometry.Results At 48 h post-transfection with Tnni3-siRNA,H9C2 cells exhibited a significant decrease in Tnni3 mRNA (0.27 ± 0.05 vs.1.00 ± 0.00) and protein (0.18 ± 0.03 vs.1.00 ± 0.00) compared with those transfected with NC-siRNA,and the differences were statistically significant (t =25.26,47.40,all P ＜ 0.01).Apoptotic cells were observed in the NC-siRNA group and the Tnai3-siRNA group.At 72 h post-transfection,the percentage of apoptotic cells significantly increased in H9C2 cells transfected with Tnni3-siRNA [(11.30 ± 1.85) ％ vs.(0.33 ± 0.15) ％] compared with those transfected with NC-siRNA,an increased expression of Caspase-3 mRNA was also observed in Tnni3-siRNA-transfected H9C2 cells (1.39 ±0.13 vs.1.00 ±0.00),and the differences were statistically significant (t =10.24,5.19,all P ＜ 0.01).Compared with NC-siRNA-transfected H9C2 cells,a time-dependent reduction in cell proliferation was observed in Tnni3-siRNA-transfected H9C2 cells (48 h:0.32 ± 0.06 vs.0.46 ± 0.03;72 h:0.31 ± 0.01 vs.0.63 ±0.04;96 h:0.36 ± 0.01 vs 0.75 ± 0.04),and the differences were statistically significant (t =3.62,13.45,16.39,all P ＜ 0.01).At 72 h post-transfection with Tnni3-siRNA,the percentage of G1 phase,S phase and G2 phase cells was (71.25 ± 3.82) ％,(18.28 ± 2.78) ％ and (9.94 ± 1.09) ％,respectively.There was a significant increase in the proportion of G2 phase cells [(9.94 ± 1.09) ％ vs.(4.54 ±0.99) ％] in H9C2 cells transfected with Tnni3-siRNA compared with those transfected with NC-siRNA,an increased expression of Cyclin A1 protein (1.89 ±0.09 vs.1.00 ±0.00) and a decreased expression of Cyclin B1 protein (0.47 ± 0.06 vs.1.00 ± 0.00) were observed in Tnni3-siRNA-transfected H9C2 cells,respectively,and the differences were statistically significant (t =6.35,17.12,15.32,all P＜0.01).Conclusions Silencing Tnni3 gene expression in rat embryonic H9C2 cardiomyocytes can induce cell apoptosis,suppress cell proliferation,and led to G2 cell cycle arrest.
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MicroRNA(miRNA)can reduce the stability of the messenger RNA(mRNA)or inhibit the transla-tion of the messenger RNA by targeting its 3'-untranslated region(UTR),thereby negatively regulate the expression of the target gene.Diverse RNA transcripts including mRNAs;pseudogenes;long non-coding RNAs(LncRNA)and circu-lar RNAs(circRNAs)can competitively combine with the same miRNAs by microRNA response elements(MREs),re-moving or reducing the inhibition of genes targedted by the miRNAs and regulating the expression of the target genes. These RNA transcripts are called competitive endogenous RNA (ceRNA),and this new post-transcription regulation model is called ceRNA hypothesis.CeRNA not only participates in the proliferation,differentiation and aging of normal cells,but also plays an important role in the pathogenesis of tumor and cardiovascular diseases(CVD).This paper re-viewed the studies about ceRNA in cardiovascular diseases in recent years.
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Objective To explore the effect of reduced alpha - cardiac actin 1(ACTC1)gene expression on the rat embryonic cardiomyocytes H9C2 cell apoptosis and its mechanism. Methods The rat embryonic cardiomyocytes H9C2 cell was cultivated;the rat embryonic cardiomyocytes H9C2 cell was transfected with ACTC1 - small interfering RNA(siRNA),and at 24 h,48 h,72 h after transfection,the cells were collected for extraction and purification of RNA, the real - time quantitative PCR(qPCR)method was used to detect the expression level of ACTC1 gene;and the termi-nal deoxynucleotidyl transferase - mediated dUTP - biotin nick end labeling assay(TUNEL)method was used to ex-plore the effect of reduced ACTC1 gene expression on the rat embryonic cardiomyocytes H9C2 cell apoptosis. Western blot was used to detect the expression of Cyto C,cysteine - containing aspartate - specific proteases(Caspase)- 3, Caspase - 8,Caspase - 9,Bcl - 2 and Bax. Results The expression of ACTC1 mRNA detected by qPCR decreased compared with that of the scramble siRNA group in 24 h,48 h,72 h(0. 80 vs. 1. 00,0. 20 vs. 1. 00,0. 25 vs. 1. 00),and in the ACTC1 - siRNA group decreased significantly at 48 h,72 h,and the difference was statistically significant(t =4. 245,P < 0. 05);TUNEL positive cells rate significantly increased in the ACTC1 - siRNA group(80％)compared with that in the scramble siRNA group(20％),and the difference was statistically significant(P < 0. 05);Western blot also confirmed that the expression of Caspase - 3,Caspase - 9,Cyto C and Bax/ Bcl - 2 were accordingly increased (0. 91 ± 0. 12 vs. 0. 59 ± 0. 01,0. 48 ± 0. 09 vs. 0. 24 ± 0. 03,0. 92 ± 0. 03 vs. 0. 45 ± 0. 01,2. 25 ± 0. 26 vs. 1. 16 ± 0. 12),and the differences were statistically significant(t = 2. 821,7. 336,2. 420,0. 798,all P < 0. 05);but the expre-ssion of Caspase - 8 had no obvious change,and the difference was not statistically significant (P > 0. 05 ). Conclusions Reduced ACTC1 gene expression can induce the rat embryonic cardiomyocytes H9C2 cell apoptosis perhaps mainly through endogenous mitochondrial signal transduction pathways.
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Objective To analyze the expression of TBX1 gene in kidney tissues in patients with clear cell renal cell carcinoma(ccRCC)and in?vestigate its molecular genetics mechanism during the tumor development. Methods Real?time quantitative polymerase chain reaction(qRT?PCR)was used to detect the expression of TBX1 mRNA in 12 cases of clear cell renal cell carcinoma tissues and the corresponding normal kidney tissues adjacent to carcinoma .The protein expression of TBX1 was assayed by Western blot in both groups. Results Both TBX1 mRNA level and the protein level were significantly up?regulated in ccRCC tissues compare to those in normal kidney tissues adjacent to carcinoma(all P<0.05). Conclusion Over?expression of TBX1 gene might be a potentially pathogenic mechanism of ccRCC.
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In recently years,the incidence rate of obesity - related glomerulopathy(ORG)is increasing. Adi-pose tissue as a new endocrine organ releases inflammatory factors which can lead to glomerular artery endothelial damage,mesangial expansion and extracellular matrix accumulation and podocyte lesion,etc. Studies have confirmed that the chronic inflammation may play a key role in the pathogenesis of ORG. Now,the molecular mechanisms of chro-nic inflammation and its effect in the development of ORG were summarized.
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Objective To explore the relationship between transforming growth factor-β1 (TGF-β1) and obesity-related glomerulopathy (ORG),and to analyze the possible mechanism for ORG and the new approach to its treatment.Methods Based on their body weight,30 SD rats were randomly divided into 2 groups : the normal control group (15 rats) fed with common food and the ORG model group (15 rats) fed with fat-enriched diets.The rats were sacrificed at the end of the 8th week,and their kidneys were taken out.Immunohistochemistry was used to detect TGF-β1 protein expression.Real time (RT)-polymerase chain reaction (PCR) was used to extract and detect the expression of TGF-β1 mRNA,and Western blot was applied to examine the expression of TGF-β1 protein.The findings were analyzed by using SPSS 13.0 software.Results Compared with the control group, qualitative TGF-β1 expression in ORG model group were significantly increased detected by immunohistochemistry mainly in renal tubules and interstitium.The average absorbance value of the control group and the model ORG group was 0.040-0.013,0.171 ± 0.084, respectively.The difference was statistically significant(P ＜ 0.05).The expression of TGF-β1 mRNA detected by RT-PCR was also increased compared with that of the control group(4.4 vs 0.6).The difference was statistically significant(P ＜ 0.05).The protein expression of TGF-β 1 examined by Western blot showed that it was more than that in the control group(4.3 vs 0.4).The difference between the control group and ORG model group was statistically significant(P =0.002).Conclusions The expression of TGF-β 1 in kidneys of ORG model rats increased, which not only indicates it can participate in ORG's occurrence and development, but also provide the basis to find out the mechanism and the approach to treatment.
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Objective To explore the possible mechanisms of the protective effect of leflunomide on kidneys by observing the effects of leflunomide on rat kidney tissue of focal segmental glomerulosclerosis (FSGS) expression level of transforming growth factor (TGF)-β1.Methods Twenty-four SD rats were randomly divided into 3 groups:normal control group (n =8),FSGS model group (n =8) and leflunomide treatment group (n =8).Unilateral nephrectomy 1 week after repeated injection of doxorubicin established FSGS model.Since 2 weeks after surgery,the treatment group had been given the leflunomide suspension 5 mg/(kg · d) orally,while normal control group and model group had been given the same amount of solvent orally.In the 8th week of the experiment,the rats were sacrificed and the specimens were collected,so serum creatinine,blood urea nitrogen,total cholesterol,albumin and 24 hours urinary protein were recorded; renal tissue was taken for pathological examination and calculation of glomerular sclerosis index (GSI) was made;immunohistochemical detection of TGF-β1 expression in the kidney was performed;The expression of TGF-β1 was examined by Western blot.Results Compared with normal control group,FSGS model group and leflunomide treatment group rats' 24 hours urinary protein excretion,serum creatinine,blood urea nitrogen,and cholesterol significantly increased while serum albumin significantly reduced severe renal pathological changes and TGF-β1 protein expression was significantly increased,and the differences were statistically significant (all P ＜ 0.05) ; Compared with the FSGS model group,in the 8th weekend of the experiments,the treated rats' 24 hours urinary protein excretion,relevant serum biochemical indicators of renal pathological changes had different degrees of improvement in renal tissue and TGF-β1 protein expression was decreased,so the differences above were statistically significant(all P ＜ 0.05).Conclusions Leflunomide may reduce the FSGS kidney tissue fibrosis by inhibiting the expression of TGF-β1,and thus protect the kidneys.
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The morbidity of obesity-related glomerulopathy(ORG) is on the increase in recent years.Studies have demonstrated that the chronic inflammation may play a key role in the pathogenesis of obesity related metabolic dysfunction.LipoxinA4 (LXA4) is an important anti-inflammatory lipid mediator,which is well known as the stop signal of the inflammatory reaction that can promote the resolution of inflammation.This review will provide a survey of recent advances on ORG and LXA4.
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ObjectiveTo investigate the association between R405Q polymorphism of GLI1 gene and tetralogy of fallot(TOF).Methods In the case-control study,the R405Q polymorphism of GLI1 gene in 112 children with TOF and 200 healthy controls were detected with polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).The distribution of genotype and allele frequency at R405Q polymorphism site were analyzed and to investigate its relationship with the risk of TOF.ResultsThe distribution of genotype frequency at R405Q polymorphism site was not different between TOF group and the healthy control group(x2 =5.317 ,P = 0.07) .However, the distribution of allele frequency at R405Q polymorphism site was significantly different between TOF group and the healthy control group (x2 = 6.790, P = 0.009) , and the relative risk for TOF in A allele carriers was higher than that in G allele carriers (OR = 1.561,95％ CI 1.116 ～ 2.185) Conclusion The R405Q polymorphism of GLI1 gene is associated with TOF and people with A allele have higher risk with TOF.GLI1 gene might be the genetic susceptibility gene of TOF.