1.Construction of a risk prediction model for failure of proximal femoral nail antirotation fixation in intertrochanteric fractures
Zesong TU ; Daxing XU ; Hongbin LUO ; Yusheng WANG ; Xinglun FENG ; Zhonghua PENG ; Shaolong DU
Chinese Journal of Tissue Engineering Research 2025;29(27):5845-5853
BACKGROUND:Intertrochanteric femoral fractures are the main type of fragility fracture in the elderly,and proximal femoral nail antirotation is the preferred surgical option,but the factors associated with postoperative internal fixation failure are controversial.OBJECTIVE:A new"three-column"classification of intertrochanteric femoral fractures was proposed by evaluating patients'imaging data preoperatively and analyzing its interaction with postoperative internal fixation failure.A risk prediction model was developed and validated by using numerical algorithms,which facilitates clinicians to identify and intervene in high-risk patients preoperatively.METHODS:Patients with intertrochanteric femur fractures in Sanshui Branch of Foshan Hospital of Traditional Chinese Medicine between June 2012 and June 2022 were selected.The patients were divided into the internal fixation failure group and the internal fixation maintenance group according to whether they had internal fixation failure after surgery.According to the preoperative radiographs,the proximal femur was divided into three columns:the medial column,the lateral column,and the middle column.Each column had different subgroups.The relationship between the morphological characteristics of the"three columns"and the failure of proximal femoral nail antirotation internal fixation was analyzed,and the independent risk factors for internal fixation failure were screened out by single and then multifactorial logistic regression analyses.A risk prediction model was constructed according to the independent risk factors using R language software.The Bootstrap method was used to resample 1 000 times.The area under the curve,calibration curve,and clinical decision curve were used to evaluate the differentiation,calibration ability,and clinical application value of the model.The Youden index was used to determine the optimal risk cut-off value of the prediction model,according to which the patients were divided into high and low risk groups.The stability and extensibility of the model were evaluated according to the accuracy of its risk prediction ability.RESULTS AND CONCLUSION:(1)The four independent risk factors for postoperative internal fixation failure after surgery were predicted using the"three-column"typing system:medial column(comminuted fracture of the lesser trochanter and femoral talar)[odds ratio=5.385,95%CI(1.961,14.782),P=0.001],medial column(chimney type)[odds ratio=2.893,95%CI(1.167,7.173),P=0.022],lateral column(lateral wall thickness<20.5 mm)[odds ratio=2.804,95%CI(1.078,7.297),P=0.035]and lateral column(lateral wall fracture)[odds ratio=4.278,95%CI(1.670,10.959),P=0.012].(2)The constructed risk prediction model showed good discrimination and accuracy[area under the receiver operating characteristic curve=0.852,95%CI(0.837,0.922)].The calibration curve showed good agreement between the model-predicted risk and the actual risk of occurrence.(3)The clinical decision curve suggested that the model had good clinical applicability when the risk threshold probability was in the range of 0.2-0.82.The risk probability of 28%was the optimal threshold for risk stratification of the model,and the predictive performance of the model was better in patients with different risk groups.(4)The"three-column"typing system constructs a predictive model to calculate the risk probability of postoperative internal fixation failure in patients with intertrochanteric femoral fractures.This method is accurate,simple,and easy to apply clinically,and can be used as a digital tool to guide personalized clinical treatment.
2.Construction of a risk prediction model for failure of proximal femoral nail antirotation fixation in intertrochanteric fractures
Zesong TU ; Daxing XU ; Hongbin LUO ; Yusheng WANG ; Xinglun FENG ; Zhonghua PENG ; Shaolong DU
Chinese Journal of Tissue Engineering Research 2025;29(27):5845-5853
BACKGROUND:Intertrochanteric femoral fractures are the main type of fragility fracture in the elderly,and proximal femoral nail antirotation is the preferred surgical option,but the factors associated with postoperative internal fixation failure are controversial.OBJECTIVE:A new"three-column"classification of intertrochanteric femoral fractures was proposed by evaluating patients'imaging data preoperatively and analyzing its interaction with postoperative internal fixation failure.A risk prediction model was developed and validated by using numerical algorithms,which facilitates clinicians to identify and intervene in high-risk patients preoperatively.METHODS:Patients with intertrochanteric femur fractures in Sanshui Branch of Foshan Hospital of Traditional Chinese Medicine between June 2012 and June 2022 were selected.The patients were divided into the internal fixation failure group and the internal fixation maintenance group according to whether they had internal fixation failure after surgery.According to the preoperative radiographs,the proximal femur was divided into three columns:the medial column,the lateral column,and the middle column.Each column had different subgroups.The relationship between the morphological characteristics of the"three columns"and the failure of proximal femoral nail antirotation internal fixation was analyzed,and the independent risk factors for internal fixation failure were screened out by single and then multifactorial logistic regression analyses.A risk prediction model was constructed according to the independent risk factors using R language software.The Bootstrap method was used to resample 1 000 times.The area under the curve,calibration curve,and clinical decision curve were used to evaluate the differentiation,calibration ability,and clinical application value of the model.The Youden index was used to determine the optimal risk cut-off value of the prediction model,according to which the patients were divided into high and low risk groups.The stability and extensibility of the model were evaluated according to the accuracy of its risk prediction ability.RESULTS AND CONCLUSION:(1)The four independent risk factors for postoperative internal fixation failure after surgery were predicted using the"three-column"typing system:medial column(comminuted fracture of the lesser trochanter and femoral talar)[odds ratio=5.385,95%CI(1.961,14.782),P=0.001],medial column(chimney type)[odds ratio=2.893,95%CI(1.167,7.173),P=0.022],lateral column(lateral wall thickness<20.5 mm)[odds ratio=2.804,95%CI(1.078,7.297),P=0.035]and lateral column(lateral wall fracture)[odds ratio=4.278,95%CI(1.670,10.959),P=0.012].(2)The constructed risk prediction model showed good discrimination and accuracy[area under the receiver operating characteristic curve=0.852,95%CI(0.837,0.922)].The calibration curve showed good agreement between the model-predicted risk and the actual risk of occurrence.(3)The clinical decision curve suggested that the model had good clinical applicability when the risk threshold probability was in the range of 0.2-0.82.The risk probability of 28%was the optimal threshold for risk stratification of the model,and the predictive performance of the model was better in patients with different risk groups.(4)The"three-column"typing system constructs a predictive model to calculate the risk probability of postoperative internal fixation failure in patients with intertrochanteric femoral fractures.This method is accurate,simple,and easy to apply clinically,and can be used as a digital tool to guide personalized clinical treatment.
3.Research on the application of medical dialectics combined with textbook-based problem-oriented teaching in hematology internship teaching
Yuying LI ; Yehui TAN ; Jingnan SUN ; Zhonghua DU ; Sujun GAO
Chinese Journal of Medical Education Research 2024;23(6):804-808
Objective:To explore the application of medical dialectics combined with problem-oriented teaching based on textbooks in hematology internship teaching.Methods:A total of 100 undergraduate students who practiced in the Department of Hematology of the First Hospital of Jilin University from 2022 to 2023 were selected as the research subjects. Students were randomly assigned to a control group and an observation group, with 50 students in each group. The control group received traditional teaching, while the observation group received medical dialectics combined with textbook-based problem-oriented teaching. We assessed the theoretical and operational scores, classroom performance, comprehensive abilities, and teaching satisfaction of two groups of students using t-test and χ 2 test in SPSS 22.0. Results:The theoretical and operational scores of the observation group were (94.26±5.35) points and (92.68±4.72) points, respectively. The theoretical and operational scores of the control group were (86.16±5.42) points and (81.52±5.28) points, respectively. The differences between the two groups were statistically significant ( P<0.001). The recognition rates were significantly higher by students in the observation group than in the control group ( P<0.05) in terms of improving learning efficiency, self-learning ability, understanding and comprehensive analysis of diseases, problem-solving ability, language and organizational expression ability, integration of theory and practice, clinical thinking ability, and independent thinking ability. The satisfaction with teaching was higher in the observation group than in the control group ( P<0.05) in terms of teaching attitudes, teaching methods, teaching arrangements, practicality of teaching content, clear explanation of teaching theories, and outstanding teaching objectives. Conclusions:The medical dialectics combined with textbook-based problem-oriented teaching can improve the assessment scores of medical students, while helping to cultivate their comprehensive abilities and develop good clinical diagnosis and treatment thinking.
4.Protective effect and mechanism of Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea on apoptosis of mice cells with acute liver injury induced by carbon tetrachloride
Dianbo TAN ; Zhonghua LIU ; Qing DU ; Hongliang ZENG ; Yuehui LI
Acta Laboratorium Animalis Scientia Sinica 2024;32(11):1417-1423
Objective Use of silencing information regulatory factor 1(SIRT1)/high mobility group protein B1(HMGB1)/nuclear transcription factor-KB(NF-κB)to investigate the inhibitory effect and mechanism of Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea on hepatocyte inflammatory apoptosis in mice with carbon tetrachloride(CC14)-induced acute liver injury.Methods C57BL/6 mice were randomly divided into a control group;model group;resveratrol group;Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea low-,medium-,and high-dose groups;and a positive drug group.The mice were given continuous intragastric administration of the treatments for 14 days.An acute liver injury model was established by intraperitoneal injection of 0.5%CC14 olive oil solution(5 mL/kg).The levels of alanine transferase(ALT),aspartate transferase(AST),lactate dehydrogenase(LDH)in serum and hydroxyproline(Hyp),malondialdehyde(MDA),and superoxide dismutase(SOD)in the liver were determined by biochemical method.Serum levels of inflammatory tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1 β were determined by enzyme-linked immunosorbent assay.Eosin-hematoxylin and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling(TUNEL)staining were used to examine the pathological morphology and apoptosis in liver tissues.The protein expression of SIRT1,HMGB1,and NF-κB were detected by Western Blot.Results Compared with those of the control group,the model group's serum ALT,AST,and LDH levels and liver tissue Hyp activity were significantly increased;MDA and SOD activities in liver tissue were significantly decreased;the levels of inflammatory factors TNF-α,IL-6,and IL-1 β in liver tissue were significantly increased;and there were obvious signs of pathological injury and hepatocyte apoptosis in the liver tissue.The expression of SIRT1 protein decreased significantly,while the expression of HMGB1 and NF-κB proteins increased,in liver tissue.Compared with those of the model group,the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high group and resveratrol group serum levels of ALT,AST,and LDH and liver tissue Hyp activity were significantly decreased;MDA and SOD activities in liver tissue were significantly increased;the levels of serum inflammatory factors TNF-α,IL-6,and IL-1β were decreased;and pathological injury to liver tissue and the apoptosis of liver cells were significantly improved.The expression of SIRT1 protein in liver tissue was increased,while the expression of HMGB1 and NF-κB protein were decreased in the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high-dose group and resveratrol group(P<0.05 or P<0.01).Conclusions Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea effectively protects against acute liver injury,and its mechanisms may be related to the regulation of the SIRT1/HMGB1/NF-κB signaling pathway and the alleviation hepatocyte inflammatory apoptosis.
5.Establishment and validation of a laboratory-based multiparameter model for predicting bone marrow metastasis in malignant tumors
Haocheng LI ; Wei XU ; Zhonghua DU ; Lin SONG ; Dan LIU ; Huihui SHAO ; Chunhe ZHAO ; Weiqi CUI ; Linlin QU
Chinese Journal of Laboratory Medicine 2024;47(11):1248-1255
Objective:To establish and validate the prediction model for bone marrow metastasis (BMM) in malignant tumors by screening out laboratory multiparameters.Methods:This case-control study collected 444 cases of malignant tumor patients who were hospitalized in the First Hospital of Jilin University from March 2018 to March 2024, including 243 cases for model establishment set and 201 cases for model validation set. The model establishment set was divided into BMM positive group (81 cases) and BMM negative group (162 cases), and the model validation set was divided into positive group (67 cases) and a negative group (134 cases). We collected patients′ clinical information such as gender, age, clinical diagnosis, and results of 47 laboratory tests including routine blood analysis, coagulation, liver function, tumor markers, potassium, sodium, chloride, and calcium ion tests, bone marrow morphology, and bone marrow biopsy. BMM was taken as the outcome event, differencial variables were analyzed using inter group comparisons, the correlation among parameters was analyzed using Pearson correlation analysis, the risk factors for BMM were analyzed using multivariate conditional logistic regression analysis, to establish logistic model, followed by efficiency evaluation on BMM predictive model using receiver operating characteristic (ROC) curves.Results:In the model establishment set, Pearson correlation analysis of 28 parameters that differed between the BMM positive and negative groups revealed that the correlation coefficients of 17 parameters, including mean platelet volume (MPV), hematocrit (HCT), hemoglobin (HGB), and prothrombin time (PT), were no more than 0.6 ( P<0.05). Further multivariate conditional logistic regression analysis demonstrated that MPV, HGB, HCT, PT, red cell distribution width (RDW), platelet count (PLT), alkaline phosphatase (ALP), chloride (Cl -), and mean erythrocyte hemoglobin concentration (MCHC) were the risk factors of BMM occurence in malignancy [MPV ( OR=9.929, 95% CI 2.688-71.335), HCT ( OR=8.232, 95% CI 6.223-9.841), HGB ( OR=4.300, 95% CI 1.947-16.577), PT ( OR=3.738, 95% CI 1.359-11.666), RDW ( OR=1.995, 95% CI 1.275-3.807), ALP ( OR=1.025, 95% CI 1.012-1.045), PLT ( OR=1.014, 95% CI 1.002-1.031), MCHC ( OR=0.724, 95% CI 0.523-0.880) and Cl -( OR=0.703, 95% CI 0.472-0.967)]. In the model establishment set, combiation of risk factors provided an AUC of 0.943 (95% CI 0.898-0.987, P<0.001), a sensitivity of 86.3%, and a specificity of 89.2% for BMM prediction. In the model validation set, the AUC was 0.924 (95% CI 0.854-0.960, P<0.001), with a sensitivity and specificity of 86.7% and 83.8%, respectively. Conclusion:This study built and validated a multiple-parameter model for BMM, which may facilitate the timely detection of BMM and provide reference for decision making of bone marrow aspiration.
6.Protective effect and mechanism of Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea on apoptosis of mice cells with acute liver injury induced by carbon tetrachloride
Dianbo TAN ; Zhonghua LIU ; Qing DU ; Hongliang ZENG ; Yuehui LI
Acta Laboratorium Animalis Scientia Sinica 2024;32(11):1417-1423
Objective Use of silencing information regulatory factor 1(SIRT1)/high mobility group protein B1(HMGB1)/nuclear transcription factor-KB(NF-κB)to investigate the inhibitory effect and mechanism of Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea on hepatocyte inflammatory apoptosis in mice with carbon tetrachloride(CC14)-induced acute liver injury.Methods C57BL/6 mice were randomly divided into a control group;model group;resveratrol group;Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea low-,medium-,and high-dose groups;and a positive drug group.The mice were given continuous intragastric administration of the treatments for 14 days.An acute liver injury model was established by intraperitoneal injection of 0.5%CC14 olive oil solution(5 mL/kg).The levels of alanine transferase(ALT),aspartate transferase(AST),lactate dehydrogenase(LDH)in serum and hydroxyproline(Hyp),malondialdehyde(MDA),and superoxide dismutase(SOD)in the liver were determined by biochemical method.Serum levels of inflammatory tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1 β were determined by enzyme-linked immunosorbent assay.Eosin-hematoxylin and terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling(TUNEL)staining were used to examine the pathological morphology and apoptosis in liver tissues.The protein expression of SIRT1,HMGB1,and NF-κB were detected by Western Blot.Results Compared with those of the control group,the model group's serum ALT,AST,and LDH levels and liver tissue Hyp activity were significantly increased;MDA and SOD activities in liver tissue were significantly decreased;the levels of inflammatory factors TNF-α,IL-6,and IL-1 β in liver tissue were significantly increased;and there were obvious signs of pathological injury and hepatocyte apoptosis in the liver tissue.The expression of SIRT1 protein decreased significantly,while the expression of HMGB1 and NF-κB proteins increased,in liver tissue.Compared with those of the model group,the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high group and resveratrol group serum levels of ALT,AST,and LDH and liver tissue Hyp activity were significantly decreased;MDA and SOD activities in liver tissue were significantly increased;the levels of serum inflammatory factors TNF-α,IL-6,and IL-1β were decreased;and pathological injury to liver tissue and the apoptosis of liver cells were significantly improved.The expression of SIRT1 protein in liver tissue was increased,while the expression of HMGB1 and NF-κB protein were decreased in the Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea high-dose group and resveratrol group(P<0.05 or P<0.01).Conclusions Salvia miltiorrhiza Bunge gynostemma pentaphyllum tea effectively protects against acute liver injury,and its mechanisms may be related to the regulation of the SIRT1/HMGB1/NF-κB signaling pathway and the alleviation hepatocyte inflammatory apoptosis.
7.CAG stimulating regimen in treatment of adult early T cell precursor acute lymphoblastic leukemia complicated with fusarium infection: report of 1 case and review of literature
Cong WANG ; Gaoling ZHANG ; Zhonghua DU ; Wei HAN ; Xiaoxia ZHAO ; Sujun GAO ; Qiuju LIU
Journal of Leukemia & Lymphoma 2023;32(3):166-170
Objective:To investigate the clinical effect of CAG stimulating regimen for refractory adult early T cell precursor acute lymphoblastic leukemia (ETP-ALL) complicated with fusarium infection and the clinical features as well as antifungal strategy of cutaneous fusarium infection.Methods:The diagnosis and treatment of 1 adult patient diagnosed as ETP-ALL complicated with cutaneous fusarium infection in the First Hospital of Jilin University in September 2020 were retrospectively analyzed, and related literatures were reviewed.Results:VICP chemotherapy regimen showed no effectiveness in this patient who was presented with persistent agranulocytosis complicated with cutaneous fusariosis infection. After amphotericin B therapy for infection, he achieved the stable disease and successfully underwent CAG stimulating regimen salvage treatment. The minimal residual disease turned into negative after consolidation chemotherapy based on the myeloid regimen. Finally this patient survived from haploid allogeneic hematopoietic stem cell transplantation after consolidation chemotherapy and fusarium was under the control by using posaconazole as secondary prevention therapy.Conclusions:CAG stimulating regimen can be recommended as reinduction therapy for relapsed/refractory ETP-ALL. Sequential therapy of amphotericin B followed by posaconazole can be a useful antifungal strategy for fusarium infection.
8.Study on the risk signal mining related to brigatinib based on the US FDA Adverse Event Reporting System
Zhonghua FU ; Zihan GUO ; Mengmeng WANG ; Qiong DU ; Qing ZHAI
Adverse Drug Reactions Journal 2023;25(1):34-39
Objective:To explore the risk signals of brigatinib-related adverse events (AEs) and provide reference for the safe use in clinical practice.Methods:The US FDA Adverse Event Reporting System database was searched and AE reports on brigatinib as the primary suspect drug from April 1, 2017 to March 31, 2022 were collected. AEs were standardized and classified according to the preferred terms (PT) and system organ class (SOC) of Medical Dictionary for Regulatory Activities 24.0. Reported odds ratio ( ROR) and proportional reporting odds ratio ( PRR) methods were used to mine the AE risk signals of brigatinib. An AE with reports ≥3, ROR≥2, 95% confidence interval ( CI) lower limit of ROR>1, or reports ≥3, PRR≥2, and χ2>4 was defined as a positive signal. Positive PT signals were analyzed using descriptive method. Results:A total of 1 564 AE reports were included in the analysis, involving 672 PTs. After analysis using ROR and PRR methods, 52 PTs with positive risk signals were obtained, involving 16 SOCs. The top 10 PTs in report amount were fatigue, diarrhea, nausea, cough, abnormal serum creatine phosphokinase, dyspnea, headache, rash, vomiting, and hypertension, all of which were common AEs in the instructions. The top 10 PTs in signal intensity were pituitary infarction, radiation necrosis, elevated amylase, esophageal varices, early saturation, elevated lipase, abnormal serum creatine phosphokinase, pulmonary toxicity, prolonged activated partial thromboplastin time, and photosensitivity. Among them, the PTs ranked 1st, 2nd, 4th, 5th, 8th, and 10th were not recorded in the label. Pneumonia and interstitial lung disease (ILD) were serious AEs, with 31 and 8 reports, respectively. In the 52 PTs, 28 were not included in the drug label, involving 12 SOCs. Conclusions:The main adverse reactions of brigatinib were diarrhea, nausea, cough, and abnormal serum creatine phosphokinase and serious adverse reactions such as pneumonia and ILD were both reported, which were consistent with the common AE recorded in the drug label. In addition, brigatinib might cause pituitary infarction, radiation necrosis, pulmonary toxicity, photosensitivity, etc., which should be vigilant in clinical practice.
9.Study on the risk signal mining related to brigatinib based on the US FDA Adverse Event Reporting System
Zhonghua FU ; Zihan GUO ; Mengmeng WANG ; Qiong DU ; Qing ZHAI
Adverse Drug Reactions Journal 2023;25(1):34-39
Objective:To explore the risk signals of brigatinib-related adverse events (AEs) and provide reference for the safe use in clinical practice.Methods:The US FDA Adverse Event Reporting System database was searched and AE reports on brigatinib as the primary suspect drug from April 1, 2017 to March 31, 2022 were collected. AEs were standardized and classified according to the preferred terms (PT) and system organ class (SOC) of Medical Dictionary for Regulatory Activities 24.0. Reported odds ratio ( ROR) and proportional reporting odds ratio ( PRR) methods were used to mine the AE risk signals of brigatinib. An AE with reports ≥3, ROR≥2, 95% confidence interval ( CI) lower limit of ROR>1, or reports ≥3, PRR≥2, and χ2>4 was defined as a positive signal. Positive PT signals were analyzed using descriptive method. Results:A total of 1 564 AE reports were included in the analysis, involving 672 PTs. After analysis using ROR and PRR methods, 52 PTs with positive risk signals were obtained, involving 16 SOCs. The top 10 PTs in report amount were fatigue, diarrhea, nausea, cough, abnormal serum creatine phosphokinase, dyspnea, headache, rash, vomiting, and hypertension, all of which were common AEs in the instructions. The top 10 PTs in signal intensity were pituitary infarction, radiation necrosis, elevated amylase, esophageal varices, early saturation, elevated lipase, abnormal serum creatine phosphokinase, pulmonary toxicity, prolonged activated partial thromboplastin time, and photosensitivity. Among them, the PTs ranked 1st, 2nd, 4th, 5th, 8th, and 10th were not recorded in the label. Pneumonia and interstitial lung disease (ILD) were serious AEs, with 31 and 8 reports, respectively. In the 52 PTs, 28 were not included in the drug label, involving 12 SOCs. Conclusions:The main adverse reactions of brigatinib were diarrhea, nausea, cough, and abnormal serum creatine phosphokinase and serious adverse reactions such as pneumonia and ILD were both reported, which were consistent with the common AE recorded in the drug label. In addition, brigatinib might cause pituitary infarction, radiation necrosis, pulmonary toxicity, photosensitivity, etc., which should be vigilant in clinical practice.
10.Genistein attenuates LPS-induced inflammatory injury of rat dorsal root ganglion neuron via down-regulating HDAC6.
Songlin ZHOU ; Junqing HUANG ; Ke LI ; Shuaigang DU ; Bin YANG ; Zhonghua GUO
Journal of Central South University(Medical Sciences) 2022;47(6):707-716
OBJECTIVES:
Neuropathic pain (NP) is a chronic pain caused by somatosensory neuropathy or disease, and genistein (Gen) might be a potential drug for the treatment of NP. Therefore, this study aims to investigate the effect of Gen on lipopolysaccharide (LPS)-induced inflammatory injury of dorsal root ganglion neuron (DRGn) in rats and the possible molecular mechanism.
METHODS:
The DRGn of 1-day-old juvenile rats were taken for isolation and culture. The DRGn in logarithmic growth phase were divided into a control group, a LPS group, a tubastatin hydrochloride (TSA)+LPS group, a Gen1+LPS group, a Gen2+LPS group, a Gen2+LPS+TSA group, a Gen2+pcDNA-histone deacetylase 6 (HDAC6)+LPS group, and a Gen2+pcDNA3.1+LPS group. The LPS group was treated with 1 μg/mL LPS for 24 h; the TSA+LPS group, the Gen1+LPS group, the Gen2+LPS group were treated with 5 μmol/L TSA, 5 μmol/L Gen, 10 μmol/L Gen respectively for 0.5 h, and then added 1 μg/mL LPS for 24 h; the Gen2+TSA+LPS group was treated with 10 μmol/L Gen and 5 μmol/L TSA for 0.5 h and then added 1 μg/mL LPS for 24 h; the Gen2+pcDNA-HDAC6+LPS group and the Gen2+pcDNA3.1+LPS group received 100 nmol/L pcDNA-HDAC6 and pcDNA3.1 plasmids respectively, and 24 h after transfection, 10 μmol/L Gen was pretreated for 0.5 h, and then added 1 μg/mL LPS for 24 h. Real-time RT-PCR was used to detect the HDAC6 mRNA expression in DRGn; CCK-8 method was used to detect cell viability of DRGn; flow cytometry was used to detect cell apoptosis of DRGn; ELISA was used to detect the levels of IL-1β, IL-6, and TNF-α in DRGn culture supernatant; Western blotting was used to detect the protein expression of HDAC6, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and NF-κB p65 in DRGn.
RESULTS:
Compared with the control group, the expression levels of HDAC6 mRNA and protein, the expression levels of TLR4 and MyD88 protein in DRGn of LPS group rats were significantly up-regulated, the ratio of p-NF-κB p65/NF-κB p65 was significantly increased, and the activity of DRGn was significantly decreased, the apoptosis rate was significantly increased, and the levels of IL-1β, IL-6 and TNF-α in the DRGn culture supernatant were significantly increased (all P<0.05). Compared with the LPS group, the expression levels of HDAC6 mRNA and protein, TLR4 and MyD88 protein expression levels in DRGn of the TSA+LPS group, the Gen1+LPS group, the Gen2+LPS group and the Gen2+TSA+LPS group were significantly down-regulated, the ratio of p-NF-κB p65/NF-κB p65 was significantly decreased, the activity of DRGn was significantly increased, the apoptosis rate was significantly decreased, and the levels of IL-1β, IL-6 and TNF-α in the DRGn culture supernatant were significantly decreased (all P<0.05), and the above changes were most obvious in the Gen2+TSA+LPS group. Compared with the Gen2+LPS group, the expression levels of HDAC6 mRNA and protein, TLR4 and MyD88 protein expression levels in DRGn of the Gen2+pcDNA-HDAC6+LPS group were significantly up-regulated, the ratio of p-NF-κB p65/NF-κB p65 was significantly increased, the activity of DRGn was significantly decreased, and the apoptosis rate was significantly increased, and the levels of IL-1β, IL-6 and TNF-α in the DRGn culture supernatant were significantly increased (all P<0.05).
CONCLUSIONS
Gen can alleviate LPS-induced DRGn inflammatory injury in rats, which might be related to down-regulating the expression of HDAC6 and further inhibiting the activation of TLR4/MyD88/NF-κB signaling pathway.
Animals
;
Ganglia, Spinal
;
Genistein/pharmacology*
;
Histone Deacetylase 6/metabolism*
;
Interleukin-6/metabolism*
;
Lipopolysaccharides
;
Myeloid Differentiation Factor 88
;
NF-kappa B/metabolism*
;
Neurons/metabolism*
;
RNA, Messenger
;
Rats
;
Toll-Like Receptor 4/metabolism*
;
Tumor Necrosis Factor-alpha/metabolism*

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