Objectives:To investigate the genetic etiology of six adult patients with Dilated cardiomyopathy (DCM), and analyze the structure of the identified variants, for providing reference for the diagnosis of DCM.Methods:Six adult patients with DCM (patients 1-6) admitted to the Department of Cardiology of Zhumadian Central Hospital from January 2023 to December 2023 were recruited. Clinical data of the patients were retrospectively collected. And 3 mL of peripheral blood was collected from each patients. Pathogenic variants patients were detected by whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing. The possible functional significance of the identified missense variants was evaluated using software including SIFT, PolyPhen-2 and Mutation Taster. Specific regions of the MYBPC protein encoded by the MYBPC3 gene from different species were aligned using Mutation Taster. The wild-type and mutant MYBPC proteins were constructed using homologous modeling software MODELLER v10.4 and three-dimensional structures were visualized using PyMOL software. The molecular interaction between MYBPC-C5 domain and myosin with or without the mutation was further analyzed using ZDOCK module in Discovery Studio 2019 software. Pathogenicity ratings for the detected variant sites were performed in accordance with the Standards and Guidelines for the Interpretation of Sequence variants by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the ACMG Guidelines). This study was reviewed and approved by the Ethics Committee of Zhumadian Central Hospital (Approval No. 2022092007). Results:The six DCM patients had typical symptoms of heart failure, and echocardiography showed whole-heart dilation and decreased ventricular wall motion. Left ventricular end-diastolic dimension (LVEDD) was 57-74 mm, left ventricular ejection fraction (LVEF) was 35%-43%, and left ventricular fractional shortening (LVFS) was 17%-28%. Variation in DCM related genes, the c. 98473A>T(p.Lys32825*) variation of the TTN gene and the c.1976T>C(p.Ile659Thr) variation of the MYBPC3 gene, were identified in two patients with DCM. Multiple software predicted that both mutations were deleterious. MYBPC3-Ile659Thr mutation affected the highly conserved residue within the C5 domain of MYBPC. Three-dimensional structural analysis of homologous modeling revealed the alterations in amino acid properties and interactions with surrounding amino acids caused by the MYBPC3-Ile659Thr mutation. Further molecular docking analysis showed that the Ile659Thr mutation altered both the hydrogen bond and salt-bridge interactions between the MYBPC-C5 domain and the ligand myosin. Conclusions:Two mutations associated with DCM were identified in this study. The abnormal conformation of the mutant protein further affected its interaction with the ligand myosin, resulting in the phenotype of DCM.

OBJECTIVE: To investigate the genetic etiology of six adult patients with Dilated cardiomyopathy (DCM), and analyze the structure of the identified variants, for providing reference for the diagnosis of DCM. METHODS: Six adult patients with DCM (patients 1-6) admitted to the Department of Cardiology of Zhumadian Central Hospital from January 2023 to December 2023 were recruited. Clinical data of the patients were retrospectively collected. And 5 mL of peripheral blood was collected from each patient. Pathogenic variants of the patients were detected by whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing. The possible functional significance of the identified missense variants was evaluated using software including SIFT, PolyPhen-2 and Mutation Taster. Specific regions of the MYBPC protein encoded by the MYBPC3 gene from different species were aligned using Mutation Taster. The wild-type and mutant MYBPC proteins were constructed using homologous modeling software MODELLER v10.4 and three-dimensional structures were visualized using PyMOL software. The molecular interaction between MYBPC-C5 domain and myosin with or without the mutation was further analyzed using ZDOCK module in Discovery Studio 2019 software. Pathogenicity ratings for the detected variant sites were performed in accordance with the Standards and Guidelines for the Interpretation of Sequence variants by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the ACMG Guidelines). This study was reviewed and approved by the Ethics Committee of Zhumadian Central Hospital (Approval No. 2022092007). RESULTS: The six DCM patients had typical symptoms of heart failure, and echocardiography showed whole-heart dilation and decreased ventricular wall motion, left ventricular end-diastolic dimension (LVEDD) was 59-74 mm, left ventricular ejection fraction (LVEF) was 35%-43%, and left ventricular fractional shortening (LVFS) was 17%-28%. Variations of the DCM related genes, including a c.98473A>T (p.Lys32825*) variation of the TTN gene and a c.1976T>C (p.Ile659Thr) variation of the MYBPC3 gene, were identified in two patients. Multiple software predicted that both mutations were deleterious. MYBPC3-Ile659Thr mutation affected the highly conserved residue within the C5 domain of MYBPC. Three-dimensional structural analysis of homologous modeling revealed the alterations in amino acid properties and interactions with surrounding amino acids caused by the MYBPC3-Ile659Thr mutation. Further molecular docking analysis showed that the Ile659Thr mutation altered both the hydrogen bond and salt-bridge interactions between the MYBPC-C5 domain and the ligand myosin. CONCLUSION Two mutations associated with DCM were identified in this study. The abnormal conformation of the mutant protein further affected its interaction with the ligand myosin, resulting in the phenotype of DCM.
Humans ; Cardiomyopathy, Dilated/genetics* ; Male ; Adult ; Female ; Carrier Proteins/chemistry* ; Middle Aged ; Mutation ; Exome Sequencing ; Mutation, Missense ; Retrospective Studies ; Myosin Binding Protein C

IL-32 is a multifunctional cytokine with both pro-inflammatory and anti-inflammatory properties.It has been proved that expression of IL-32 increases with progression of gastric mucosal diseases and severity of gastric cancer(GC),thus participating in process of gastric"inflammation-cancer"transformation.However,how IL-32 affects malignant transformation of gastric"inflamma-tion-cancer"and finally leads to adverse outcome of GC invasion and migration is still controversial.In order to better clarify regulatory effect and possible mechanism of abnormal expression of IL-32 on different histopathological stages of gastric"inflammation-cancer"transformation,and to explore new directions and breakthroughs in molecular mechanism of early truncation and treatment of gastric precancerous lesion(GPL),we searched literatures related to IL-32 in six authoritative databases at home and abroad,such as Pubmed,Web of Science and CNKI,in past 30 years.It was found that pathogenicity or protective function of IL-32 in different histo-pathological stages of gastric"inflammation-cancer"transformation depended on its different subtypes,secretory forms,surrounding cytokine environment,disease status and genetic factors.IL-32 may regulate polarization of macrophages through NF-κB,MAPK,COX2,PR3,IDO,NOD,PKCδ,FAK and STAT3,amplify or inhibit chronic inflammatory stimulation of gastric mucosa,and thus participate in process of gastric"inflammation-cancer"transformation.Our new understanding of role of IL-32 in different stages of Cor-rea cascade may contribute to development of cytokine-directed therapy,and therapy aimed at regulating different alternative splicing subtypes of IL-32 and targeting IL-32 signals can be used as a new strategy for medical treatment of GPL and GC in future.

IL-32 is a multifunctional cytokine with both pro-inflammatory and anti-inflammatory properties.It has been proved that expression of IL-32 increases with progression of gastric mucosal diseases and severity of gastric cancer(GC),thus participating in process of gastric"inflammation-cancer"transformation.However,how IL-32 affects malignant transformation of gastric"inflamma-tion-cancer"and finally leads to adverse outcome of GC invasion and migration is still controversial.In order to better clarify regulatory effect and possible mechanism of abnormal expression of IL-32 on different histopathological stages of gastric"inflammation-cancer"transformation,and to explore new directions and breakthroughs in molecular mechanism of early truncation and treatment of gastric precancerous lesion(GPL),we searched literatures related to IL-32 in six authoritative databases at home and abroad,such as Pubmed,Web of Science and CNKI,in past 30 years.It was found that pathogenicity or protective function of IL-32 in different histo-pathological stages of gastric"inflammation-cancer"transformation depended on its different subtypes,secretory forms,surrounding cytokine environment,disease status and genetic factors.IL-32 may regulate polarization of macrophages through NF-κB,MAPK,COX2,PR3,IDO,NOD,PKCδ,FAK and STAT3,amplify or inhibit chronic inflammatory stimulation of gastric mucosa,and thus participate in process of gastric"inflammation-cancer"transformation.Our new understanding of role of IL-32 in different stages of Cor-rea cascade may contribute to development of cytokine-directed therapy,and therapy aimed at regulating different alternative splicing subtypes of IL-32 and targeting IL-32 signals can be used as a new strategy for medical treatment of GPL and GC in future.

Objectives:To investigate the genetic etiology of six adult patients with Dilated cardiomyopathy (DCM), and analyze the structure of the identified variants, for providing reference for the diagnosis of DCM.Methods:Six adult patients with DCM (patients 1-6) admitted to the Department of Cardiology of Zhumadian Central Hospital from January 2023 to December 2023 were recruited. Clinical data of the patients were retrospectively collected. And 3 mL of peripheral blood was collected from each patients. Pathogenic variants patients were detected by whole exome sequencing (WES), and candidate variants were verified by Sanger sequencing. The possible functional significance of the identified missense variants was evaluated using software including SIFT, PolyPhen-2 and Mutation Taster. Specific regions of the MYBPC protein encoded by the MYBPC3 gene from different species were aligned using Mutation Taster. The wild-type and mutant MYBPC proteins were constructed using homologous modeling software MODELLER v10.4 and three-dimensional structures were visualized using PyMOL software. The molecular interaction between MYBPC-C5 domain and myosin with or without the mutation was further analyzed using ZDOCK module in Discovery Studio 2019 software. Pathogenicity ratings for the detected variant sites were performed in accordance with the Standards and Guidelines for the Interpretation of Sequence variants by the American College of Medical Genetics and Genomics (ACMG) (hereafter referred to as the ACMG Guidelines). This study was reviewed and approved by the Ethics Committee of Zhumadian Central Hospital (Approval No. 2022092007). Results:The six DCM patients had typical symptoms of heart failure, and echocardiography showed whole-heart dilation and decreased ventricular wall motion. Left ventricular end-diastolic dimension (LVEDD) was 57-74 mm, left ventricular ejection fraction (LVEF) was 35%-43%, and left ventricular fractional shortening (LVFS) was 17%-28%. Variation in DCM related genes, the c. 98473A>T(p.Lys32825*) variation of the TTN gene and the c.1976T>C(p.Ile659Thr) variation of the MYBPC3 gene, were identified in two patients with DCM. Multiple software predicted that both mutations were deleterious. MYBPC3-Ile659Thr mutation affected the highly conserved residue within the C5 domain of MYBPC. Three-dimensional structural analysis of homologous modeling revealed the alterations in amino acid properties and interactions with surrounding amino acids caused by the MYBPC3-Ile659Thr mutation. Further molecular docking analysis showed that the Ile659Thr mutation altered both the hydrogen bond and salt-bridge interactions between the MYBPC-C5 domain and the ligand myosin. Conclusions:Two mutations associated with DCM were identified in this study. The abnormal conformation of the mutant protein further affected its interaction with the ligand myosin, resulting in the phenotype of DCM.

OBJECTIVE To explore the protective effect of marein against alcoholic fatty liver （AFL） and its potential mechanisms. METHODS AFL mice model was established with strong wine by gavage. The mice were randomly divided into normal control group （n＝9， 0.5% sodium carboxymethyl cellulose solution）， model group （n＝10， 0.5% sodium carboxymethyl cellulose solution） and marein 75 and 150 mg/kg groups （n＝9）. Mice were given relevant medicine intragastrically， once a day， for consecutive 30 days. After the last medication， the levels of triglyceride （TG）， malondialdehyde （MDA）， and superoxide dismutase （SOD） in liver tissue were determined， and hepatic histopathological changes of liver tissue were observed； the protein expression levels of peroxisome proliferator-activated receptor α （PPARα）， carnitine palmitoyltransferase-1 （CPT-1）， and diacylglycerol acyltransferase （DGAT） were determined in liver tissue. BRL hepatocytes injury model was induced by ethanol combined with ferrous sulfate and oleic acid； after treatment with 3， 6 and 12 μmol/L of marein for 24 h， the distribution of lipid droplets， the levels of TG， MDA and SOD and protein expressions of PPARα， CPT-1 and DGAT in hepatocytes were examined. After pretreatment with MK886 （PPARα inhibitor， 10 μmol/L），modeled hepatocytes were treated with 12 μmol/L of marein for 24 h， and the protein expressions of PPARα， CPT-1 and DGAT were determined. RESULTS As the results showed in vivo， compared with the model group， after treatment with 75 and 150 mg/kg of marein， the degree of steatosis was significantly reduced， and the levels of TG and MDA and protein expression of DGAT were significantly decreased（P＜0.05 or P＜0.01）； the levels of SOD， protein expressions of PPARα and CPT-1 were significantly increased（P＜0.05 or P＜0.01）. As the results showed in vitro， after treatment with 3， 6 and 12 μmol/L of marein， the lipid accumulation of hepatocytes was significantly inhibited， and the levels of TG and MDA， protein expression of DGAT were significantly decreased（P＜0.05 or P＜0.01）， while the levels of SOD， protein expressions of PPARα and CPT-1 were significantly increased（P＜0.05 or P＜0.01）. After MK886 pretreatment， the effects of marein on the above protein expressions were abolished. CONCLUSIONS Marein might exert a protective effect against AFL. The mechanisms might be related to inhibiting oxidative stress-mediated injury and improving PPARα-mediated lipid metabolism signaling pathway.

Objective:To analyze the clinical characteristics of patients with different types of coronavirus disease 2019 (COVID-19).Methods:A total of 272 eligible COVID-19 patients who were admitted to Guangzhou Eighth People′s Hospital, Guangzhou Medical University from January 22 to February 15, 2020 were retrospectively enrolled. General characteristics, the first laboratory examination and imaging data of these patients were collected. According to the clinical classification, there were 236 cases in non-severe group (mild+ common type) and 36 cases in severe group (severe+ critical type). Comparisons between groups were performed by t test, chi-square test or rank-sum test when appropriate. Results:There were 23 males and 13 females in the severe group, 103 males and 133 females in the non-severe group, and the difference was statistically significant ( χ2=5.149, P=0.023). The age of severe group was (60.5±11.2) years, which was higher than that of non-severe group (46.8±15.7) years. The difference was statistically significant ( t=6.43, P<0.01). The lymphocyte (LYM) count, platelet (PLT) count and arterial partial pressure of oxygen (PaO 2) in the severe group were 0.90(0.55, 1.10)×10 9/L, 170.00(143.50, 198.00)×10 9/L and 73.50(69.70, 83.00) mmHg(1 mmHg=0.133 kPa), respectively, which were all lower than those in the non-severe group (1.42(1.09, 1.95)×10 9/L, 187.00(148.00, 230.00)×10 9/L and 96.00(83.20, 108.00) mmHg, respectively). The differences were all statistically significant ( Z=5.59, 2.00 and 5.00, respectively, all P<0.05). The levels of creatine kinase (CK), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), C reaction protein (CRP) and procalcitonin (PCT) in the severe group were 123.00(79.00, 212.00) U/L, 32.10(27.00, 47.40) U/L, 305.50(216.00, 396.00) U/L, 37.02(23.92, 63.66) mg/L and 0.09(0.05, 0.19) μg/L, respectively, which were all higher than those in the non-severe group (68.00(48.00, 103.00) U/L, 20.10(16.70, 26.20) U/L, 179.00(150.00, 222.00) U/L, 26.55(18.11, 36.96) mg/L and 0.04(0.03, 0.06) μg/L respectively), and the differences were all statistically significant ( Z=3.89, 5.60, 5.12, 2.85 and 5.43, respectively, all P<0.01). No significant differences were observed in white blood cell count, creatine kinase isoenzyme and blood lactate between the two groups ( Z=1.53, 0.41 and 1.00, respectively, all P>0.05). Conclusion:Gender, age, LYM count, PLT count, PaO 2, CK, AST, LDH, CRP and PCT could be used to provide reference for clinical classification of COVID-19 patients.

Objective:To establish a hypergravity loading model with a high-acceleration centrifugal loading device and to investigate the effects of different hypergravity loading and icariin on osteoblast adhesion and cytoskeleton.Methods:MC3T3-E1 cells were seeded in the dishes of cell culture at a density of 2×10 5/cm 2. And the experiment was divided into 6 groups: control group (without icariin and loading); simple administration group (only icariin); 10 G loading group (only loading); 10 G administration group (with icariin and loading); 40 G loading group (only loading); 40 G administration group (with icariin and loading). The experimental loading group was loaded with MC3T3-E1 cells using a high-acceleration centrifugal loader. And continuous loading for 3 d, 30 min per d. The control group and the simple administration group were exposed to normal gravity, and the remaining conditions were not different from the experimental group. Icariin was used at a concentration of 10 -7 mol/L in all administration groups, and the experiments were carried out according to the method of preventive administration. At the same time, the related molecular biological techniques such as alizarin red staining, alkaline phosphatase (ALP) activity measurement, CCK-8 cell proliferation experiment, cytoskeleton phalloidin staining, qPCR and Western Blot were used to detect the effects of icariin on osteoblasts adhesion protein integrin α5 and integrin β1 and cytoskeleton protein F-actin under hypergravity extreme mechanical environment. Results:All models were successfully prepared. The alizarin red staining: The icariin could significantly promote the formation of osteoblastic calcified nodules. And the 10 G loading could also promote the mineralization of osteoblasts and increase the number of mineralized nodules, while the mineralization and the number of mineralized nodules of osteoblasts are significantly reduced in 40 G loading. ALP activity test: The OD values of simple administration group, 10 G loading group and 40 G loading group were 0.246, 0.331 and 0.163, respectively. Compared with 0.207 in the control group, the differences were statistically significant ( P<0.05). The 10 G administration group and the 40 G administration group were 0.373 and 0.180, and the differences were statistically significant ( P<0.05). The results of CCK-8 proliferation experiments: The OD value of simple administration group were 0.650, which was statistically significant compared with 0.551 of control group ( P=0.031). The 10 G loading group and 40 G loading group were 1.193 and 0.245, and their differences with the control group were both statistically significant ( P<0.05). The OD value of 10G administration group and the 40 G administration group were 1.300 and 0.310, which were significantly different from the respective loading groups ( P<0.05). Phalloidin staining: 10 G loading significantly increased the number of cells, but the changes in cells morphology and skeleton were not obvious. 40 G loading significantly inhibited the increase of the number of cells, meanwhile, made the pseudopods of cells more shorter and even disappeared. 40 G loading made the seriously damage of the cytoskeleton and even cause the cells to death. Icariin had no effect on the cells morphology, but it did has a certain repair effect after the cells loading. The results of qPCR and Western Blot experiments all confirmed that the expressions of integrin α5, integrin β1 and F-actin were up-regulated after icariin treatment. 10 G loading could promote the expression of integrin α5, integrin β1 and F-actin, and 40 G loading significantly inhibited the expression of the mRNA and proteins. Conclusion:Both 10 G condition and icariin can promote the development, cell adhesion and the cytoskeleton's stability of osteoblasts, while 40 G has a significant inhibitory effect.

Objective:To study the correlation between hypercoagulant status of patients with hip fracture and the level of microparticles (MPs) in their peripheral blood, and to explore the feasibility of removing MPs to correct hypercoagulant status in patients with hip fracture.Methods:Sixty-five patients from December 2018 to September 2019 with hip fracture were included. There were 24 males and 41 females with the average age of 75.6±9.8 years old (range 58-95 years). Among them, 27patients (43.1%) were femoral neck fracture and38 patients (56.9%) were intertrochanteric fracture.All patients were diagnosed with X-ray and CT. Meanwhile, about 20 healthy people in the physical examination center included as controls in our study. There were 8 males and 12 females with the average age of 72.3±6.5 years old (range 56-81years). 2 ml of anticoagulant whole blood was taken on an empty body in the morning, and purified microparticlesby whole-blood density gradient centrifugation in whole blood were identified by electron microscope and nanoparticle tracking analyzer. After cell free plasma (CFP) was obtained by whole-blood density gradient centrifugation, the number of whole annexin V (AV) positived MPs and these MPs which from platelet (PMPs) was determined by flow cytometry. The activated clotting time (ACT) was determined by coagulation and platelet function analyzer to evaluate the degree of hypercoagulability. Then, Logistic analysis was performed on risk factors associated with hypercoagulability to determine whether the level of MPs was an independent risk factor for hypercoagulability, and the correlation between ACT value and MPs level was analyzed. Finally, the four coagulation items of each sample CFP before and after MPs removal were determined by automatic coagulation analyzer.Results:Under electron microscopy, MPs presented vesicular appearance,with a complete double-layer membrane structure, the size was in the range of 100-1 000 nm, and the morphology was not uniform. there were irregular vesicular and circular vesicular general shapes. The average size of MPs in peripheral blood of patients with hip fractures was 239.7±4.0 nm. The mean size of MPs distribution in the control group was 247.7±3.3 nm, and there was no statistically significant difference in MPs diameter between the two groups. The average level of circulating AV +MPs in patients with hip fracture was 564±171/μl, and the average level of PMPs was 326±104/μl. In the control group, the average level of AV +MPs was 252±82/μl, the average level of PMPs was 192±41/μl, the difference between AV +MPs and PMPs was statistically significant ( P<0.05). The average ACT level of patients with hip fracture was 324±94 s, while the average ACT level of the normal population was 535±76 s, and the difference between the two was statistically significant ( P<0.05). Single factor logistic regression analysis showed that the levels of APTT, PMPs and AV +MPs may be risk factors for hypercoagulability, and multivariate logistic regression analysis showed that AV +MPs is an independent risk factor for hypercoagulability.It has a highly negative correlation with ACT ( r=-0.822, P<0.05). The results of four coagulation items determined by CFP were PT 10.8±0.46 s, APTT 30.6±1.56 s, Fib 3.08±0.36 g/L, INR 0.98±0.04 and TT 19.3±0.62 s. After the removal of MPs, the coagulation function was PT 10.8±0.52 s, APTT 32.4±3.0 s, Fib 2.90±0.33 g/L, INR 0.99±0.05 s, and TT 19.9±0.63 s. There was no statistically significant difference before and after coagulation function. Conclusion:There is a hypercoagulable state in patients with hip fracture, moreover, the level of AV +MPs is an independent risk factor for hypercoagulability, which is highly correlated with ACT, and MPs has no significant effect on the classic four factors of coagulation.

Objective To investigate the role and mechanism of nico-tinamide adenine dinucleotide phosphate oxidase 4 (NAPHD oxidase 4,Nox4)-mediated reactive oxygen species (ROS) generation on high-dose dexamethasone (DEX) induced apoptosis in osteoblasts.Methods According to culture conditions,3rd passage of murine osteoblastic MC3T3-E 1 cells were divided into control group,Dexamethasone group,Dexamethasone+NAC (N-acetyl-L-cysteine) group,NAC group,Dexamethasone+DPI (Diphenyleneiodonium) group and DPI group.24 hours after culture,the morphology of osteoblasts was observed by inverted phase contrast microscope.Cell viability was determined by MTT assay.The generation of ROS in osteoblasts was measured using a fluorescent probe DCFH-DA.The apoptosis of each group was observed through Hoechst staining.The mRNA level and protein expression of Nox4 were detected by real-time quantitative PCR and Western Blot.In addition,after silence of Nox4 with small interfering RNA (siRNA),the ROS generation was further detected by a fluorescent probe DCFH-DA.Results After treatment with 1000 nmol DEX for 24 hours,compared to control group,the results of inverted phase contrast microscope and MTT showed that osteoblasts in DEX group exhibited more obvious signs of shrinkage and deformation with decreased cell viability.After intervene with NAC and DPI,morphology of osteoblasts was good with increased viability of osteoblasts.Compared to control group (5.86％± 0.28％),the production of ROS in DEX group (45.14％±1.49％) was significantly increased (P=0.000).The apoptotic rates in DEX group (29.60％± 1.52％) was significantly increased compared with control group (4.12％±0.67％) (P=0.000).Compared to DEX group,the production of ROS generation in DEX+NAC group (28.06％±1.61％) and DEX+DPI group (23.70％±1.28％) was significantly decreased (P=0.000).It presented that NAC or DPI significantly decreased the formation of ROS.Compared to DEX group,the apoptotic rate in DEX+NAC group (8.94％± 1.47％) and DEX+DPI group (12.96％±2.03％) was significantly decreased (P=0.000).It presented that NAC or DPI significantly decreased osteoblast apoptosis.In addition,the Nox4 mRNA level in DEX group was 2.67-fold compared with control group (t=-10.301,P=0.009).The difference had statistically significance.The protein expression of Nox4 in DEX group was 2.37-fold compared with control group (t=-15.542,P=0.004).The difference has statistically significance.After silence of Nox4 by siRNA,the generation of ROS in DEX+Nox4 siRNA group (14.53％± 1.00％) was decreased by 16.92％ compared with DEX group 31.45％±0.72％ (P=0.000).The difference had statistically significance.Conclusion Nox4-mediated ROS generation plays an important role in osteoblasts apoptosis induced by high-dose dexamethasone.It provided us the new target in the management of Nox4 to provide possible therapy for steroid-induced avascular necrosis of the femoral head (SANFH).