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Objective Studies on the expression and location of zinc finger protein A20 (A20) and connective tissue growth factor (CTGF) in liver tissues of patients with chronic hepatitis B were conducted, and the relationship between them and liver fibrosis was determined by FibroScan. Methods Studies on A20 and CTGF in liver tissues of 160 patients with chronic hepatitis B were conducted in accordance with the stage of pathological fibrosis and inflammation of the liver, and quantitative immunohistochemistry test was conducted, and statistical analysis was conducted by FibroScan. Results The expressions of A20 and CTGF in liver tissues increased with the aggravation of liver pathological fibrosis and inflammation, and there were significant differences between each stage and the control group (P0.05). There was positive correlation between liver A20 and CTGF, r=0.796 (P<0.05). Conclusions In patients with chronic hepatitis B, A20, CTGF and FibroScan are positively correlated with the degree of liver fibrosis, and A20 and CTGF are also positively correlated with the degree of liver inflammation, which can be used as indicators to evaluate the degree of liver inflammation and fibrosis, and further guide the anti-inflammatory and anti-fibrosis treatment of patients.
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Objective To investigate the plasma levels of methylated DNA in the pregnant women with preeclampsia and its predictive value for the occurrence of preeclampsia.Methods A total of 82 pregnant women with preeclampsia admitted to the hospital from January to December 2022 were included as the obser-vation group,and another 82 healthy pregnant women were included as the control group.Total DNA was ex-tracted,and the relative expression levels of methylated single-intention homolog 2(SIM2),guanine nucleo-tide-binding protein(GNA12),and connective tissue growth factor(CTGF)genes in plasma were detected by real-time fluorescence quantitative PCR(qRT-PCR)after DNA bisulfite modification.The value of methyla-ted DNA in predicting preeclampsia was evaluated by correlation analysis and receiver operating characteristic(ROC)curve.Results The relative expression levels of methylated SIM2,GNA12 and CTGF in plasma in the observation group were significantly higher than those in the control group(P＜0.05),and the relative expres-sion levels of methylated SIM2,GNA12 and CTGF in severe preeclampsia group was higher(P＜0.05).Corre-lation analysis showed that the relative expression levels of methylated SIM2,GNA12 and CTGF in plasma were significantly positively correlated with the occurrence of preeclampsia in pregnant women(P＜0.05).ROC curve analysis results showed that the relative expression levels of plasma methylation SIM2,GNA12,and CTGF,both individually and in combination,had good predictive efficacy in predicting preeclampsia in pregnant women,and the combined detection of the three had the highest predictive efficacy(area under the curve was 0.888,95％CI:0.827-0.949).Conclusion Compared with healthy pregnant women,the relative expression levels of methylated SIM2,GNA12 and CTGF in plasma are higher in pregnant women with pre-eclampsia,which are positively correlated with the occurrence of preeclampsia and the severity of the disease.The relative expression levels of methylated SIM2,GNA12 and CTGF are expected to be important predicting indicators for preeclampsia.
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Objective To investigate the correlation between the expression of purinergic ligand-gated ion channel 7 receptor(P2X7R)and connective tissue growth factor(CTGF)in serum and cognitive function and clinical symptoms in patients with schizophrenia.Methods A total of 160 patients with schizophrenia who were diagnosed and treated in Department of Mental Intensive Care of Wuhan Wudong Hospital from January 2021 to January 2023 were collected as the observation group,and 160 healthy volunteers who underwent physical examinations were collected as the control group for the study.According to the Positive and Negative Symptom Scale(PANSS),patients were evaluated for their clinical and psychiatric symptoms(positive and negative symptoms,general pathological symptoms,and additional symptoms).The patients were grouped into a high score group(PANSS total score≥70 points,n=72)and a low score group(PANSS total score＜70 points,n=88).MATRICS consensus cognitive battery(MCCB)was applied to evaluate the cognitive abilities of patients;enzyme-linked immunosorbent assay(ELISA)was applied to detect serum P2X7R and CTGF levels.Spearman method was applied to analyze the correlation between serum P2X7R,CTGF levels and PANSS scores,and MCCB scores in patients with schizophrenia.Results Compared with the control group,the serum levels of P2X7R(610.71±107.83ng/L vs 384.78±80.62 ng/L)and CTGF(1.85±0.36μg/L vs 1.40±0.21μg/L)in the observation group were increased,with differences of statistical significance(t=21.226,13.658,all P＜0.05).The scores of variety items of MCBB of patients with schizophrenia in the observation group were lower than those in the control group,with differences of statistical significance(t=14.845～24.862,all P＜0.05),the positive symptom score(21.10±3.42score),negative symptom score(23.37±5.03 score),general pathological symptom score(39.48±8.11score),additional symptom score(8.26±1.22 score),and PANSS total score(92.21±12.50score)of schizophrenia patients in the high group were higher than those in the low group(13.65±3.04,15.62±3.91 score,30.14±6.15 score,5.20±0.94score,64.61±5.30score),with differences of statistical significance(t=14.576,10.964,8.280,17.915,18.764,all P＜0.05).The serum levels of P2X7R and CTGF in patients with schizophrenia in the high group were higher than those in the low group,with differences of statistical significance(t=12.233,5.923,all P＜0.05).The levels of serum P2X7R and CTGF in patients with schizophrenia were positively correlated with PANSS score(r=0.464～0.580,all P＜0.05),and negatively correlated with MCCB score[r=-0.603～-0.439,all P＜0.05].Conclusion The serum levels of P2X7R and CTGF in patients with schizophrenia are elevated,they are closely related to the clinical symptoms and cognitive function of patients.
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BACKGROUND:Recent studies have shown that connective tissue growth factor not only participates in the development of neurons,but also participates in the pathogenesis of neurodegenerative diseases,depression,epilepsy,etc.It can also be used as a therapeutic target to develop related drugs,thereby improving the patients'quality of life. OBJECTIVE:To summarize the biological functions of connective tissue growth factor and the mechanisms involved in neurodegenerative diseases and depression,as well as the progress in intervention with connective tissue growth factor and related emerging treatments. METHODS:The first author searched relevant articles published from January 1996 to December 2022 in PubMed and Web of Science.The key words were"connective tissue growth factor,nervous system,depression,Alzheimer disease,epilepsy,Parkinson disease,epilepsy,amyotrophic lateral sclerosis,FG-3019"in English.After reading,screening and sorting,the articles consistent with the content of the review were collected.Finally,51 articles were selected for review. RESULTS AND CONCLUSION:Connective tissue growth factor participates in multiple biological activities such as fibrosis,cell adhesion,and cell development under different conditions through four different structural domains.Connective tissue growth factor is up-regulated in lesion sites of neurodegenerative diseases,depression and epilepsy.After interfering with the expression of connective tissue growth factor,the symptoms improve or disappear,suggesting that connective tissue growth factor plays an important role in the progression of these diseases.The development of novel therapeutic strategies and intervention targets around connective tissue growth factor is very promising therapeutic research.More research is needed to identify the mechanism of action to transfer from basic medical studies to clinical studies and to achieve safer and more effective treatments.
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BACKGROUND:Numerous clinical and basic studies have shown that electroacupuncture can improve the function of neurogenic bladder after suprasacral spinal cord injury. OBJECTIVE:To observe the effects of electroacupuncture on bladder function and connective tissue growth factor expression in rats with suprasacral spinal cord injury. METHODS:Forty-eight female Sprague-Dawley rats were randomly divided into four groups(n=12 per group):the blank group did not receive any treatment;the sham-operated group only exposed the T8 subvertebral spinal cord;in the model group established,a T8 subvertebral spinal cord transection injury model was established;in the electroacupuncture group,the T8 subvertebral spinal cord transection injury model was established,and electroacupuncture intervention at Ciliao(BL32),Zhongji(RN03)and Sanyinjiao(SP06)was given at 19 days after modeling,20 minutes once a day,for 10 continuous days.After the intervention,the relevant indicators were detected. RESULTS AND CONCLUSION:Urodynamics:Compared with the blank group,the leakage point pressure,maximum bladder capacity and maximum bladder pressure of rats in the model group increased(P<0.05).Compared with the model group,the leakage point pressure,maximum bladder capacity and maximum bladder pressure of rats in the electroacupuncture group decreased(P<0.05).Hematoxylin-eosin staining:Compared with the blank group,the bladder epithelial cells in the model group were arranged in a disordered manner,the lamina propria was destroyed,the detrusor muscle bundles were hypertrophied,the muscle fibers were arranged in a disordered manner,and the tissue edema was obvious.Compared with the model group,the bladder epithelial cells in the electroacupuncture group were arranged in a regular and orderly manner,and the degree of bladder fibrosis and tissue edema was relatively reduced.Masson staining:The degree of bladder detrusor muscle fibrosis was severe in the model group and it was lighter in the electroacupuncture group than in the model group.Transmission electron microscopy:Mitochondria in the bladder tissue in the model group were swollen and vacuolated,the morphology of the detrusor muscle was twisted and distorted,and the muscle gap was widened.Compared with the model group,mitochondria in the electroacupuncture group had a slightly clearer contour and were less vacuolated,and the muscle gap was narrowed.Western blot detection:The protein expression of connective tissue growth factor in the detrusor muscle of the bladder was elevated in the model group compared with the blank group(P<0.05).Compared with the model group,the protein expression of connective tissue growth factor in the bladder detrusor muscle was decreased in the electroacupuncture group(P<0.05).To conclude,electroacupuncture at Ciliao(BL32),Zhongji(RN03)and Sanyinjiao(SP06)acupoints can improve the morphology,structure and function of the bladder in rats with suprasacral spinal cord injury,and the mechanism of action may be related to the down-regulation of connective tissue growth factor protein expression in the detrusor muscle.
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{L-End}Objective To analyze the changes of seven potential biomarkers in plasma of patients with occupational silicosis (hereinafter referred to as "silicosis"), and explore their clinical value in determining the stage of silicosis. {L-End}Methods A total of 100 male silicosis patients were selected as the silicosis group (63 cases in stage Ⅰ and 37 cases in stage Ⅱ subgroups), and 100 male healthy individuals were selected as the control group using the 1∶1 matched case-control study. Enzyme-linked immunosorbent assay was used to analyze the level of interleukin-17 (IL-17), monocyte chemoattractant protein-1 (MCP-1), matrix metalloproteinase-9 (MMP-9), Krebs von den Lungen-6 (KL-6), connective tissue growth factor (CTGF), platelet-derived growth factor (PDGF), and histone H4 in plasma. Their clinical value for diagnosing silicosis was evaluated using receiver operating characteristic (ROC) curve, discriminant analysis stepwise method, and Fisher discriminant function analysis. {L-End}Results The levels of IL-17, MCP-1, MMP-9, KL-6, CTGF, PDGF, and histone H4 in the plasma of the silicosis group, silicosis stage Ⅰ subgroups, and stage Ⅱ subgroups were higher than those in the control group (all P<0.05). The levels of IL-17, MCP-1, and MMP-9 in the plasma of the stage Ⅱ subgroup decreased (all P<0.05), while the levels of KL-6, CTGF and histone H4 increased (all P<0.05) compared with the stage Ⅰ subgroup. The area under the ROC curve for diagnosing silicosis using these seven potential biomarkers ranged from 0.761 to 1.000 (all P<0.01), with the sensitivity of 0.640-1.000, the specificity of 0.840-0.990, and the Youden index of 0.540-0.990. The Fisher discriminant function was formed by stepwise discriminant analysis, and the results showed that the coincidence rate was 99.5%, and the misdiagnosis rate was 0.5% for diagnosing and staging silicosis with these seven potential biomarkers. The coincidence rate of diagnosing control group, silicosis stageⅠsubgroup and the silicosis stage Ⅱ subgroup was 100.0%, 98.4% and 100.0%, respectively. {L-End}Conclusion IL-17, MCP-1, MMP-9, KL-6, CTGF, PDGF and histone H4 in plasma can be used as biomarkers for the diagnosis of silicosis, and the Fisher discriminant function based on the combination of these seven biomarkers can assist in staging silicosis.
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Objective:To explore the expression of serum connective tissue growth factor (CTGF), glyoxalase Ⅰ (GLO-I) and pyruvate kinase M2 (PKM2) in endometrial cancer and their relationship with clinicopathological characteristics.Methods:A total of 96 endometrial cancer patients in Yuechi County People's Hospital from February 2015 to February 2017 were selected as the research group, 48 patients with endometrial hyperplasia during the same period were selected as the benign control group, and 48 patients with healthy physical examination during the same period were selected as the healthy control group. The serum levels of CTGF, GLO-Ⅰ, and PKM2 in the three groups were analyzed. The correlation between serum levels of CTGF, GLO-Ⅰ and PKM2 in the research group was analyzed, and the relationship between each serum index and clinicopathological characteristics was analyzed.Results:The levels of serum CTGF, GLO-Ⅰ and PKM2 in the research group were higher than those in the benign control group and healthy control group: (184.31 ± 37.14) μg/L vs. (110.45 ± 20.59), (17.28 ± 0.42) μg/L; (95.17 ± 16.56) pmol/L vs. (56.29 ± 10.14), (9.08 ± 0.66) pmol/L; (20.25 ± 6.13) μg/L vs. (13.11 ± 4.58), (9.05 ± 2.74) μg/L; and the levels of serum CTGF, GLO-Ⅰ and PKM2 in the benign control group were higher than those in the healthy control group, there were statistical differences ( P<0.05). The results of Pearson correlation analysis showed that the level of CTGF had positive correlation with GLO-Ⅰ and PKM2 ( r = 0.713, 0.741, P<0.05), and the level of GLO-Ⅰ had positive correlation with PKM2 ( r = 0.823, P<0.05). The results of Spearman correlation analysis showed that the levels of CTGF, GLO-Ⅰ, PKM2 had positive correlation with FIGO stage ( r = 0.609, 0.704, 0.721; P<0.05), myometrial invasion depth ( r = 0.753, 0.695, 0.719; P<0.05), lymph node metastasis ( r = 0.776, 0.744, 0.640; P<0.05); had negative correlation with the degree of differentiation ( r = - 0.711, - 0.720, - 0.668; P<0.05). Conclusions:Serum CTGF, GLO-I, PKM2 expression levels are abnormally elevated in patients with endometrial cancer, which are significantly related to multiple clinicopathological characteristics.
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Objective:To investigate the regulatory effect of miR-26a in radiation-induced heart disease (RIHD) mice.Methods:C57/BL6 mice were used to establish RIHD models. The cardiac function, fibrosis, the expression levels of collagen 1 (COL1) and connective tissue growth factor (CTGF), and miR-26a were detected in RIHD mice. Whether CTGF was the target gene of miR-26a was verified by dual luciferase kit. Moreover, cardiac fibroblasts were transfected with miR-26a up and miR-26a down lentivirus vectors to construct the miR-26a overexpression and underexpression cell models. The expression of CTGF, proliferation, and apoptosis of cardiac fibroblasts were detected.Results:In the RIHD mice, heart function was decreased, myocardial fibrosis was remodeled, the expression levels of COL1 and CTGF were up-regulated, and the expression level of miR-26a was down-regulated. Dual luciferase reporter assay confirmed that CTGF was the target gene regulated by miR-26a. Overexpression of miR-26a could inhibit the expression of CTGF, suppress the proliferation of cardiac fibroblasts, promote cell apoptosis and secrete collagen. Underexpression of miR-26a yielded the opposite results.Conclusion:MiR-26a affects the function of cardiac fibroblasts by targeting CTGF and probably mediates the process of radiation-induced myocardial fibrosis, which may become a new regulatory target of RIHD.
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Objective:To investigate the effects of thrombolytic therapy time on serum inflammatory factors, cathepsin S, connective tissue growth factor (CTGF), left ventricular ejection fraction (LVEF) and left ventricular end diastolic diameter (LVEDD) in patients with acute ST-elevation myocardial infarction.Methods:The clinical data of 119 patients with acute ST-elevation myocardial infarction who received thrombolytic therapy in the People's Hospital of Taierzhuang District of Zaozhuang from January 2019 to May 2020 were retrospectively analyzed. These patients were assigned to three groups according to different time points at which thrombolytic therapy was performed: group A (the time from onset to thrombolytic therapy ≤ 3 hours, n = 27), group B (3 hours < the time from onset to thrombolytic therapy ≤ 6 hours, n = 39), group C (6 hours < the time from onset to thrombolytic therapy ≤ 12 hours, n = 53). Recanalization rate, recanalization time, ST segment resolution rate at 2 and 12 hours, serum levels of inflammatory factors [including interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) and high-sensitivity C-reactive protein (hs-CRP)], cathepsin S, CTGF, LVEF, and LVEDD, and incidence of cardiovascular adverse events. Results:Recanalization time in group C was (148.73 ± 15.37) minutes, which was significantly longer than that in groups A and B [(89.34 ± 8.95) minutes, (98.76 ± 9.33) minutes]. Recanalization rate and ST segment resolution rate at 2 and 12 hours in group C were 45.28%, (40.17 ± 4.77) %, (73.92 ± 8.24) %, respectively, which were significantly lower than those in the groups A and B [96.30%, 79.49%, (47.42 ± 5.12)%; (83.68 ± 9.33)%, (43.56 ± 4.87)%, (78.73 ± 8.44)%] ( t/ χ2 = 248.088, 4.244, 20.204, 11.146, 18.508, 19.861, 6.271, 4.789, 17.995, 10.932, 3.339, 4.111, 4.100, 3.828, 3.100, 2.244, all P < 0.05). At 2 and 12 hours after thrombolytic therapy, IL-6, TNF-α and hs-CRP levels in group C were (23.29 ± 2.12) ng/L, (27.03 ± 2.75) ng/L, (6.49 ± 2.37) mg/L, (22.73 ± 2.05) ng/L, (26.24 ± 2.37) ng/L and (6.01 ± 2.53) mg/L, respectively, which were significantly higher than those in groups A and B ( t = 54.578, 54.578, 10.638, 8.584, 8.735, 5.199, 7.909, 7.171, 3.597, 1.382, 1.584, 1.008, 7.237, 5.190, 4.364, 8.829, 11.114, 2.585, 3.172, 6.815, 2.196, all P < 0.05). At 2 and 12 hours after thrombolytic therapy, cathepsin S and CTGF levels in group C were (29.97 ± 3.98) μg/L, (30.03 ± 4.79) μg/L, (28.05 ± 2.13) μg/L, (28.29 ± 4.31) μg/L, respectively, which were significantly higher than those in groups A and B [(31.74 ± 3.56) μg/L, (29.87 ± 4.91) μg/L; (20.81 ± 2.35) μg/L, (16.94 ± 3.46) μg/L; (30.95 ± 3.79) μg/L, (29.93 ± 4.95) μg/L; (26.37 ± 2.44) μg/L, (21.46 ± 4.79) μg/L, t = 93.870, 68.555, 15.039, 12.562, 6.345, 7.679, 3.096, 1.966, 13.882, 3.514, 11.863, 7.164, 9.239, 4.199, all P < 0.05). At 2 and 12 hours after thrombolytic therapy, LVEF and LVEDD in group C were (42.81 ± 4.77)%, (52.64 ± 4.71) mm, (43.13 ± 5.11)%, (51.57 ± 4.01) mm, respectively, which were significantly lower than those in groups A and B [(42.61 ± 4.58)%, (52.31 ± 4.47) mm, (46.33 ± 4.35)%, (47.75 ± 3.41) mm, (42.73 ± 4.79)%, (52.79 ± 4.76) mm, (44.79 ± 4.44)%, (49.93 ± 3.73) mm, t = 4.285, 9.193, 3.060, 4.214, 1.970, 2.953, 0.333, 1.259, 2.779, 1.626, 4.229, 1.996, 1.404, 2.416, all P < 0.05). The total incidence of cardiovascular adverse events was 7.41%, 12.82% and 33.96% in groups A, B and C, respectively ( χ2 = 4.383, all P < 0.05). Conclusion:The earlier the thrombolytic therapy time after acute ST-elevation myocardial infarction, the higher the recanalization rate and ST segment resolution rate, the milder the inflammatory reaction, atherosclerosis, the better the cardiac remodeling, the better the recovery of cardiac function, and the lower the incidence of cardiovascular adverse events.
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Objective:To investigate the efficacy of atenolol combined with perindopril in the treatment of chronic heart failure in older adult patients and its effects on ventricular function, serum connective tissue growth factor (CTGF), and nexin.Methods:120 older adult patients with chronic heart failure who received treatment in the Department of Cardiology, the First People's Hospital of Fuyang District from January 2016 to January 2018 were included in this study. They were randomly assigned to receive either basic treatment + atenolol treatment (control group, n = 60) or atenolol combined with perindopril treatment (observation group, n = 60). Clinical efficacy and clinical symptom, serum CTGF, nexin level and ventricular function pre- and post-treatment, as well as adverse reactions were compared between the control and observation groups. Results:After treatment, effective rate in the observation group was significantly higher than that in the control group (91.6% vs. 78.3%, χ2 = 4.183, P = 0.041). After treatment, CTGF and nexin levels in the control and observation groups were decreased compared with before treatment (both P < 0.05). After treatment, CTGF and nexin levels in the observation group were (4.42 ± 0.46) μg/L and (0.82 ± 0.03) μg/L, respectively, which were significantly lower than those in the control group [(4.82 ± 0.51) μg/L, (0.98 ± 0.04) μg/L, t = 18.153, 4.511, 19.335, 24.787, all P < 0.05]. After treatment, left ventricular end diastolic diameter and left ventricular end systolic diameter in the control and observation groups were deceased compared with before treatment (both P < 0.05). After treatment, left ventricular end diastolic diameter and left ventricular end systolic diameter in the observation group were (48.73 ± 4.41) mm and (41.13 ± 4.15) mm, respectively, which were significantly lower than those in the control group [(56.01 ± 4.67) mm, (47.45 ± 4.17) mm, t = 5.700, 8.799, 8.317, 8.351, all P < 0.05]. After treatment, left ventricular ejection fraction in the control and observation groups was significantly increased compared with before treatment. After treatment, left ventricular ejection fraction in the observation group was significantly higher than that in the control group [(44.86 ± 4.59) % vs. (39.05 ± 4.69) %, P < 0.05]. Conclusion:Atenolol combined with perindopril in the treatment of older adult patients with chronic heart failure can reduce clinical symptoms, improve ventricular function, decrease serum CTGF and nexin levels and is highly safe. Therefore, this method is worthy of clinical application.
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Objective:To investigate the effect and mechanism of bone marrow mesenchymal stem cell (BMSC)-derived exosomal microRNA-1297 (miR-1297) on hippocampal neuron damage in depressed rats.Methods:BMSCs and BMSCs-derived exosomes were prepared and identified. Rats were first injected with corticosterone to establish the model of depression, and then injected with BMSCs-derived exosomes. Superoxide dismutase (SOD), malondialdehyde (MDA), lactate dehydrogenase (LDH), TNF-α and IL-1β in rat serum samples, hippocampal tissues and neurons were detected. Expression of miR-1297 in hippocampal tissues and neurons was detected by RT-qPCR. A rat hippocampal neuron injury model was established to investigate the role of BMSC-derived exosomes and miR-1297 in neuronal apoptosis and proliferation. The targeting relationship between miR-1297 and connective tissue growth factor (CTGF) was analyzed using dual luciferase reporter genes.Results:In the hippocampus of depressed rats, the expression of miR-1297 was low, while the expression of CTGF was elevated. Exosomes derived from BMSCs can inhibit the expression of CTGF by up-regulating the level of miR-1297, thereby inhibiting neuronal cell apoptosis in the hippocampus of depressed rats, while increasing the level of SOD, and reducing inflammatory damage, and ultimately improving the behavioral function of depressed rats.Conclusions:Depressed rats showed decreased expression of miR-1297 and increased expression of CTGF. BMSC-derived exosomes inhibited CTGF expression through up-regulating miR-1297, thereby improving hippocampal neuron damage in rats with depression.
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@#Drug transportation is impeded by various barriers in the hypoxic solid tumor, resulting in compromised anticancer efficacy. Herein, a solid lipid monostearin (MS)-coated CaO2/MnO2 nanocarrier was designed to optimize doxorubicin (DOX) transportation comprehensively for chemotherapy enhancement. The MS shell of nanoparticles could be destroyed selectively by highly-expressed lipase within cancer cells, exposing water-sensitive cores to release DOX and produce O2. After the cancer cell death, the core-exposed nanoparticles could be further liberated and continue to react with water in the tumor extracellular matrix (ECM) and thoroughly release O2 and DOX, which exhibited cytotoxicity to neighboring cells. Small DOX molecules could readily diffuse through ECM, in which the collagen deposition was decreased by O2-mediated hypoxia-inducible factor-1 inhibition, leading to synergistically improved drug penetration. Concurrently, DOX-efflux-associated P-glycoprotein was also inhibited by O2, prolonging drug retention in cancer cells. Overall, the DOX transporting processes from nanoparticles to deep tumor cells including drug release, penetration, and retention were optimized comprehensively, which significantly boosted antitumor benefits.
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Objectives: Assessment ofplasma level of connective tissue growth factor in congestive heart failure children,assessment of its diagnostic and prognostic role and correlate its level with clinical and echocardiographic assessment of congestive heart failure.Methods: Connective tissue growth factorlevel in the plasma was measured in 40 children; 20 of them have congestive heart failure, and 20 are healthy then,correlated with clinical parameters. Results: The diagnostic and prognostic value of itwas evaluated. Wecompareditslevels in both patientsand healthy children. We found that connective tissue growth factor level was significantly increased in diseased children. Fractional shortening and ejection fraction correlated negatively with the plasma levelof connective tissue growth factor. Heart rate, respiratory rate and calibrated integrated backscatter correlated positively with connective tissue growth factor. Connective tissue growth factorwas significantly correlated with the class of heart failure according to Ross classification.Conclusions: Plasma connective tissue growth factor has a promising diagnostic and prognostic value as a biomarker for congestive heart failure in children with high sensitivity and specificity.
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BACKGROUND: Chronic nerve compression leads to muscle atrophy and fibrosis. Previous studies mainly focus on its pathogenesis. However, little is known about the dorsal root ganglia(DRG) responses to chronic nerve compression injury. OBJECTIVE: To investigate the effect of chronic sciatic nerve compression on fibrosis of the DRG. METHODS: Animal models of chronic sciatic nerve compression were made in rats according to the method described by Mackinnon. L4-6 ipsilateral and contralateral DRG were harvested 3 weeks post injury. Real-time RT-PCR, immunofluorescence and western blot were performed to determine the expression levels of transforming growth factor-β, connective tissue growth factor, and collagen type I in ipsilateral and contralateral DRG. RESULTS AND CONCLUSION: Three weeks after injury, the m RNA and protein expression of transforming growth factor-β, connective tissue growth factor and collagen type I were increased significantly in the ipsilateral DRG as compared with the contralateral DRG(P < 0.05). Transforming growth factor-β and connective tissue growth factor mainly expressed in DRG neurons and axons, while collagen type I formed a net structure that surrounded DRG neurons and axons. These findings indicate that chronic sciatic nerve compression can induce fibrotic changes in the DRG that appears to be associated with an increase in transforming growth factor-β and connective tissue growth factor expression in DRG neurons.
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Objective To explore the potential effects of neutrophil extracellular traps (NETs) on rheumatoid arthritis synovial fibroblasts (RA-FLSs).Methods The synovial tissues of RA patients were isolated and cultured in vitro.Peripheral blood neutrophils were extracted from healthy volunteers and used to stimulate NETs' formation,following with NETs' extraction.MTS proliferation assay was used to evaluate the effect of NETs on the proliferation of RA-FLSs.QRT-polymerase chain reaction (q-PCR) was used to determine the expression of connective tissue growth factor (CTGF) mRNA in cells treated with NETs-stimulated RA-FLSs for 60 h.The results were processed using paired sample t-test and one-way analysis of variance (ANOVA).Results The isolated and purified neutrophils could form NETs by in vitro stimulation.The concentration of extracted NETs-DNA was 58.5 ng/μl (1×106 cells).Compared with the control group (0 μl NETs),NETs could promote the proliferation of RA-FLSs.With the increase of NETs' concentration,the proliferation of RA-FLSs was also enhanced (F=99.519,P＜0.05).Compared with the control group (0 μl NETs),10 μl NETs could significandy promote the proliferation of RA-FLSs (t=-12.226,P＜0.01).Pretreatment of NETs with DNase Ⅰ inhibited its effect on promoting the proliferation of RA-FLSs (t=-2.376,P=0.049),NETs stimulated the upre-gulation of CTGF mRNA expression in RA-FLSs [(30.7±0.5),t=12.13,P＜0.01].Conclusion NETs can promote the proliferation of RA-FLSs and stimulate the up-regulation of CTGF mRNA in RA-FLSs in vitro.
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Objective: To investigate the effect of transforming growth factor β 1 (TGF-β 1) induced proliferation of ligamentum flavum cells and ligamentum flavum hypertrophy and its effect on connective tissue growth factor (CTGF) expression. Methods: The ligamentum flavum tissue in lumbar intervertebral disc herniation was extracted and the ligamentum flavum cells were isolated and cultured by collagenase pre-digestion method. Morphological observation, immunofluorescence staining observation, and MTT assay were used for cell identification. The 3rd generation ligamentum flavum cells were divided into 5 groups. The cells of groups A, B, C, and D were respectively sealed with 3 ng/mL TGF-β 1, 50 ng/mL CTGF, 3 ng/mL TGF-β 1+CTGF neutralizing antibody, and 50 ng/mL CTGF+CTGF neutralizing antibody. Serum free DMEM was added to group E as the control. MTT assay was used to detect the effects of TGF-β 1 and CTGF on the proliferation of ligamentum flavum cells. Western blot was used to detect the expression of CTGF protein. Real-time fluorescence quantitative PCR (qRT-PCR) was used to detect the expression of collagen type Ⅰ, collagen type Ⅲ, and CTGF genes. Results: The morphological diversity of cultured ligamentum flavum cells showed typical phenotype of ligamentum flavum fibroblasts; all cells expressed collagen type Ⅰ and vimentin, and some cells expressed collagen type Ⅲ; MTT identification showed that with the prolongation of culture time, the absorbance ( A) value of each generation of cells increased gradually, and the A value of the same generation of cells at each time point was significantly different ( P0.05). After cultured for 24 hours, MTT assay showed that the A value of cells in groups A and B was significantly higher than that of group E ( P0.05); after neutralizing antibody was added, the mRNA relative expression of collagen type Ⅰ and collagen type Ⅲ in group D was inhibited and was significantly lower than that in group B, but still significantly higher than that in group E ( P0.05). Conclusion: TGF-β 1 can promote CTGF, collagen typeⅠ, collagen type Ⅲ gene level and protein expression in ligamentum flavum cells, and TGF-β 1 can synergistically promote proliferation of ligamentum flavum cells through CTGF.
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@#AIM: To investigate the effect of connective tissue growth factor(CTGF)on the proliferation and migration and transformation in Tenon's capsule fibroblasts(Tfb)of primary open angle glaucoma(POAG)patients.<p>METHODS: Tfb were cultured from Tenon's tissues in POAG patients <i>in vitro</i>. The free-serum DMEM-F12 containing 1.0, 10.0, 100.0ng/mL of CTGF was added into medium for 24h and 48h in different experimental group respectively, and only equal volume of free-serum DMEM-F12 was added in the negative control group. After 24h, the cell proliferation was analyzed through MTT assay, and migration was evaluated by crutch method. After 48h, Semi-quantitative RT-PCR was used to observed the mRNA expression of α-smooth muscl actin(α-SMA), and expression of α-SMA protein was examined by immunochemistry.<p>RESULTS: The proliferation values <i>A</i> of the cells in 1.0, 10.0, 100.0ng/mL of CTGF group respectively were 0.436±0.009, 0.643±0.009, 0.679±0.006, and 0.423 ±0.156 in the negative control group. The migrated cell number was 34.600±3.507, 70.400±2.074, 80.000±2.915 in different concentrations of CTGF group respectively, and 31.000±3.536 in the negative control group. And also in different experimental groups respectively, the absorbance ratio of α-SMA/β-actin was 0.873±0.161, 1.213±0.312, 1.352±0.376, and 0.851±0.158 in the negative control group, the expressing levels <i>A</i> of α-SMA protein in Tfb were 0.110±0.026, 0.141±0.017, 0.175±0.027, and 0.108±0.020 in the negative control group. The statistics of the above experimental data showed that, comparing with the negative control group, the 10.0 and 100.0ng/mL CTGF groups was statistically significant different(<i>P</i><0.05), but there was no statistical different between the 1.0ng/mL CTGF group and the negative control group(<i>P</i>>0.05). <p>CONCLUSION: The proliferation, migration, and phenotypic transformation of Tfb can be promoted in CTGF group in POAG patients. These findings suggest that CTGF may play a role in the development of filtering bleb scarring.
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Objective • To investigate the effects of silencing connective tissue growth factor (Ctgf) gene on the growth, cell cycle and the expression of TGF-β1, Smad3 and Smad7 of rat hepatic stellate cell line HSCT6. Methods • The recombinant lentivirus vector pCDH/Ctgf-shRNA of Ctgf gene was constructed by RNA interference. The recombinant vector was packaged to obtain highly infectious pCDH/Ctgf-shRNA lentivirus particles for HSCT6 infection. The expression of green fluorescent protein (GFP) in the transfected HSCT6 cells was observed under fluorescence microscope. The effects of Ctgf-shRNA lentivirus on the growth of HSCT6 cells were tested by CCK-8. The effects of Ctgf-shRNA lentivirus on the cell cycle of HSCT6 cells were analyzed by flow cytometry (FCM). The effects of Ctgf-shRNA lentivirus on the expression of mRNA of Ctgf, Tgf-β1, Smad3 and Smad7, and their proteins in HSCT6 cells were detected by real-time PCR and Western blotting, respectively. Results • The lentiviral vector pCDH/Ctgf-shRNA has been constructed successfully. The HSCT6 cells transfected by Ctgf-shRNA lentivirus significantly expressed GFP under fluorescence microscope. The results of CCK-8 confirmed that the growth of HSCT6 cells transfected by Ctgf-shRNA lentivirus was slower than that of controls and the differences were statistically significant after being cultured for 72 h (P<0.05). The results of FCM revealed that the growth of HSCT6 cells transfected by Ctgf-shRNA lentivirus was blocked in the S phase of cell cycle. The results of real-time PCR and Western blotting showed that the Ctgf-shRNA lentivirus effectively silenced Ctgf gene, down-regulated the expression of genes and encoding proteins of TGF-β1, and Smad3 of HSCT6 and up-regulated the expression of genes and encoding proteins of Smad7 of HSCT6 cells. The differences between transfected cells and controls were statistically significant (P<0.05). Conclusion • Silencing Ctgf gene can effectively inhibit the growth of HSCT6 cells, down-regulate the expression of TGF-β1 and Smad3 and up-regulate the expression of Smad7. The inhibition of the growth of HSCT6 cells may be closely related to interference of the TGF-β1/Smads (Smad3 and Smad7) signaling pathway.
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OBJECTIVE: To observe the effect of herbal cake-partitioned moxibustion on renal function and expression of connective tissue growth factor (CTGF), integrin-linked kinase (ILK) and bone morphogenetic protein-7 (BMP-7) in chronic renal failure (CRF) rabbits, so as to reveal its mechanisms underlying improvement of CRF. METHODS: Twenty-four male New Zealand rabbits were randomly divided into control, model, medication and herbal cake-partitioned moxibustion (moxibustion) groups (n=6 rabbits in each group). The CRF model was established by gavage of suspension of Adenine (150 mg·kg-1·d-1) for 21 days. Herbal cake-partitioned moxibustion was applied to "Mingmen"(GV4) and bilateral "Shenshu"(BL23), "Pishu"(BL20) and for 5 moxa-cones every time. Rabbits of the medication group was treated by gavage of Losartan Potassium (2.33 mg·kg-1·d-1). All the treatments were conducted once daily,12 times a course for consecutive 3 courses with a two-day rest after each course of treatment. Serum creatinine (Scr), blood urea nitrogen (BUN) and 24 h urine protein contents were detected by using an automatic biochemical analyzer. The expression levels of CTGF, ILK and BMP-7 proteins and mRNA in the kidney tissue were determined by Western blot and quantitative real time-PCR, separately. RESULTS: Following modeling, Scr and BUN and 24 h urine protein contents were significantly increased in the model group in comparison with the control group (P0.05). CONCLUSION: Herbal cake-partitioned moxibustion can improve renal function in CRF rabbits, which may be related to its effects in suppressing the expression of ILK and CTGF, and in up-regulating the expression of BMP-7 in the kidney tissue.
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Objective: To investigate the mechanism of p38 mitogen activated protein kinase (MAPK) signaling pathway in regulating the hyperplasia and hypertrophy of human lumbar ligamentum flavum via transforming growth factor β 1 (TGF-β 1)/connective tissue growth factor (CTGF). Methods: The lumbar ligamentum flavum tissue taken from patient with lumbar intervertebral disc herniation was isolated by collagenase-predigested explant cultures. The ligamentum flavum cells were treated with the extracellular regulated protein kinase pathway blocker PD98059, c-Jun N-terminal kinase pathway blocker SP600125, and p38 pathway blocker SB203580, and then the mRNA expressions of CTGF, collagen type Ⅰ, and collagen type Ⅲ were detected by real-time fluorescence quantitative PCR (qRT-PCR). The ligamentum flavum cells were divided into 4 groups, and transfected with small interfering RNA (siRNA), p38 siRNA, siRNA+3 ng/mL TGF-β 1, and p38 siRNA+3 ng/mL TGF-β 1 in groups A, B, C, and D, respectively. After 24 hours of transfection, immunofluorescence staining was performed to observe the expressions of p38 and phosphorylation p38 (p-p38); the relative mRNA expressions of CTGF, collagen type Ⅰ, and collagen type Ⅲ in each group were detected by qRT-PCR; the protein expression of CTGF in each group was detected by Western blot. Results: p38 pathway blocker SB203580 could significantly reduce the relative mRNA expressions of CTGF, collagen type Ⅰ, and collagen type Ⅲ ( P<0.05). After 24 hours of transfection, immunofluorescence staining showed positive staining with p38 and p-p38 expressions in groups A, C, and D and negative staining in group B. Compared with group A, the relative mRNA expressions of CTGF, collagen type Ⅰ, and collagen type Ⅲ and relative protein expression of CTGF in group B decreased significantly ( P<0.05), while those in groups C and D increased significantly ( P<0.05); and those indicators significantly increased in group C than in group D ( P<0.05). Conclusion: TGF-β 1/CTGF based on the p38 MAPK signaling pathway play an important role in the occurance and development of hypertrophy of human lumbar ligamentum flavum.