Abstract:Objective To observe the expression of calcium-dependent proline-rich tyrosine kinase-2（Pyk2） in myocardium of rheumatic heart disease, the relationship between its role and cardiac fibrosis and clinical significance. Methods The blue myocardium collagen stain were analysed after Masson staining in 30 patients with rheumatic heart disease （RHD group） and 6 normal myocardium specimens （control group）. The contents of hyaluronic acid （HA）, laminin（LN） and type IV collagen（IV-C） were detected by radio-immunity method,and the expressions of Pyk2 protein and messenger ribonucleic acid（mRNA） were explored by immunohistochemistry methods and reverse transcriptase polymerase chain reaction （RT PCR）,then the correlations of these results were statistically analyzed. Results The contents of HA,LN and IV C in RHDgroup increased compared to control group（174.95±76.14μg/L vs. 70.06±15.63μg/L, 153. 86 ± 20. 72μg/L vs. 90.01±14. 11μg/L, 95. 26±7.66μg/L vs. 63. 21±10.62μg/L; P= 0.003, 0. 013, 0. 035）. The Pyk2 absorption and the ratio of Pyk2 mRNA/glyceraldehyde phosphate dehydrogenase （GAPDH） in RHD group were significantly higher than those in control group （0. 325 ± 0. 032 vs. 0.106±0.013, 0.870±0.085 vs. 0.573±0.042; P=0.048, 0.006）.There were positive correlativity between the expression of Pyk2 protein and HA, LN and IV-C （r=0. 611, 0. 743, 0. 829, P〈0. 01）, there were positive correlativity between the expression of Pyk2 mRNA and LN, IV-C （r=0. 794, 0. 766, P〈0.05）. Conclusion Pyk2 may play a key role in the proceeding of cardiac fibrosis in rheumatic heart disease by increasing collagen synthesis in myocardium.
Objective To observe macular vascular density and structural characteristics in children with transfusion-dependent β-thalassemia (TDT). MethodsA retrospective clinical study. From October 2022 to December 2023, 29 TDT children (58 eyes) diagnosed and examined at the Department of Hematology, Affiliated Hospital of Guangdong Medical University were included in the TDT group, along with 29 age- and gender-matched healthy children (58 eyes) as the control group. All participants underwent optical coherence tomography and angiography. Measurements included central macular thickness (CMT), subretinal choroidal thickness (SFCT), choroidal thickness (ChT), choroidal vascularity index, blood flow density in the superficial capillary plexus (SCP), deep capillary plexus (DCP), choriocapillaris layer (CC), and choroidal layer of the macular region, as well as the foveal avascular zone (FAZ) area of the SCP and DCP. A generalized estimating equation was used to compare differences in the above parameters between the two groups. Pearson correlation analysis was employed to examine the relationships between fundus structural parameters, blood flow density, and blood indices. ResultsCompared with the control group, the TDT group showed significantly thinner CMT (χ2=6.044) and ChT at 3.0 mm nasal (χ2=4.451) and temporal (χ2=4.767) to the fovea (P＜0.05). The TDT group also demonstrated reduced blood flow density in the inferior DCP (χ2=5.254), whole CC (χ2=3.996), and superior CC (χ2=5.094), as well as enlarged FAZ area in DCP (χ2=4.286) (P＜0.05). Correlation analysis revealed a negative correlation between SFCT and disease duration (r=−0.357, P=0.006). ConclusionsIn children with TDT, CMT and ChT become thinner and the area of FAZ expands. The blood flow densities of DCP and CC in the macular area decreased.
Objective To investigate the role and mechanism of peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) in the activation of aortic valve interstitial cells (AVICs) in aortic stenosis. Methods Isolating primary AVICs and stimulating their activation with transforming growth factor β1 (TGF-β1, 30 ng/mL), the expression of PGC-1α was detected. The activation of AVICs induced by TGF-β1 was observed after overexpression of PGC-1α by adenovirus or inhibition of PGC-1α function by GW9662. The possible downstream molecular mechanism of PGC-1α in AVICs activation was screened. Finally, the phenotype was further verified in primary human AVICs. Results The expression of PGC-1α decreased after the activation of AVICs induced by TGF-β1 (control group: 1.00±0.18; 24 h: 0.31±0.10; 48 h: 0.32±0.06; 72 h: 0.20±0.07; P<0.05). Specific overexpression of PGC-1α by adenovirus inhibited the activation of AVICs induced by TGF-β1 stimulation (periostin: 3.17±0.64 vs. 1.45±0.54, P<0.05; α-smooth muscle actin: 0.77±0.11 vs. 0.28±0.06, P<0.05). On the contrary, inhibition of PGC-1α function by GW9662 promoted the activation of AVICs (periostin: 2.20±0.68 vs. 7.99±2.50, P<0.05). Subsequently, it was found that PGC-1α might inhibit the activation of AVICs through downregulating the expression of calcium/calmodulin-dependent protein kinase (CAMK1δ) (0.97±0.04 vs. 0.74±0.11, P<0.05), and downregulating the expression of CAMK1δ alleviated the activation of AVICs (periostin: 1.76±0.11 vs. 0.99±0.20, P<0.05). The possible mechanism was that the activation of mammalian target of rapamycin (mTOR) signaling pathway was inhibited by reducing the accumulation of reactive oxygen species (ROS) (778.3±139.4 vs. 159.3±43.2, P<0.05). Finally, the protective effect of PGC-1α overexpression was verified in the activated phenotype of human AVICs (periostin: 2.73±0.53 vs. 1.63±0.14, P<0.05; connective tissue growth factor: 1.27±0.04 vs. 0.48±0.09, P<0.05). Conclusions The expression of PGC-1α significantly decreases during the activation of AVICs induced by TGF-β1. The overexpression of PGC-1α significantly inhibites the activation of AVICs, suggesting that PGC-1α plays a protective role in the activation of AVICs. The possible mechanism is that PGC-1α can inhibit the activation of CAMK1δ-ROS-mTOR pathway. In conclusion, interventions based on PGC-1α expression levels are new potential therapeutic targets for aortic stenosis.
ObjectiveTo explore the morbidity rate and risk factors of proliferative diabetic retinopathy (PDR) in type 2 diabetes.MethodsThe clinical data of patients, with PDR in 2739 consecutive cases of type 2 diabetes diagnosed in this hospital from 1994 to 2001 were analyed retospectively. The diagnosis of diabetic retinopathy (DR) was confirmed by ophthalmoscopy and fundus fluorescein angiography (FFA). Blood pressure, fasting and postprandial blood sugar, glycosylated haemoglobin（HbA1c）, total serum cholesterol, triglyceride, creatinine, and albumin excretion rate were measured.ResultsThe morbidity rate of type 2 DR was 27.8%（761/2739）, and the morbidity rate of PDR was 4.2%（114/2 739） occupying 15% of the patients with DR. The duration, fasting blood sugar, glycosylated haemoglobin, blood pressure and albumin excretion rate were much higher than those in the control（P＜0.01, glycosylated haemoglobin P＜0.05）. The independent risk factors of PDR were duration of the disease （r=0.15, P＜0.01） and albumin excretion rate （r=0.08, P＜0.05）. The risk factors of PDR were albumin excretion rate and fasting blood sugar (r=0.13, P＜0.05) in patients with longer duration(≥5 years). The morbidity rate of PDR was 2.3%, 5.9% and 12.4% in patients with duration less than 5 years, 5 to 10 years and over 10 years groups, respectively. The morbidity of PDR of the patients in normal albuminuria, microalbuminuria and overt albuminuria group was 2.1%、5.3% and 18.8% respectively.ConclusionsType 2 diabetes accompanied with PDR is relative to the duration of the diabetes, albumin excretion rate, fasting blood sugar, blood pressure, and glycosylated haemoglobin, in which the duration of the disease, albuminuria and fasting blood sugar are the risk factors of occurance of PDR. (Chin J Ocul Fundus Dis, 2003,19:338-340)
Objective To detect the expression of copine 1 (CPNE1) in the gastric cancer (GC) and investigate its association with prognosis. MethodsThe clinicopathologic data of 121 patients who underwent radical gastrectomy in the Department of General Surgery, The 940th Hospital of Joint Logistics Support Force of Chinese People’s Liberation Army from March 2017 to December 2018 were retrospectively collected. The protein expression of CPNE1 in the GC tissues was detected by immunohistochemical (IHC) staining, and its association with prognosis was analyzed. GC tissues and adjacent tissues samples from 16 patients in the same time were prospectively collected, and the mRNA and protein expressions of CPNE1 were detected by real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) and Western blot (WB), respectively. Multivariate Cox proportional hazards regression model was used to analyze the prognostic factors of GC patients. ResultsIHC staining results showed that the CPNE1 was mainly expressed in the cell membrane and cytoplasm of gastric epithelial cells, and the color showed different degrees of brown. Among the 121 patients, 70 (57.9%) had high CPNE1 protein expression and 51 (42.1%) had low CPNE1 protein expression. The RT-qPCR and WB results of 16 pairs of fresh tissue specimens showed that the expression levels of CPNE1 mRNA and protein in the GC tissues were higher than those in the corresponding adjacent tissues (CPNE1 mRNA mean value: 1.451 vs. 1.100, P=0.048; CPNE1 protein mean value: 0.995 vs. 0.521, P=0.001). The multivariate Cox proportional hazards regression analysis showed that the high protein expression of CPNE1 [HR (95%CI)=1.931 (1.123, 3.321), P=0.017] was the risk factor affecting the prognosis of the patients with GC. ConclusionCPNE1 highly expresses in GC tissues and is associated with a poor prognosis in patients with GC, and it may be a potential tumor biomarker.
ObjectiveThe re-hospitalization and death events of patients heart failure caused by coronary heart disease are characterized by non-independence, heterogeneity, and censored data. A joint frailty model is established to jointly model the events, explore the risk factors affecting the prognosis of patients, and reduce the re-hospitalization rate and mortality of patients. MethodsThe sample included 4 682 patients with heart failure caused by coronary heart disease in two tertiary hospitals from January 2014 and June 2019. The electronic medical record information of patients during hospitalization and their follow-up information were collected. The Cox model, conditional frailty model and joint frailty model were used to analyze patient re-hospitalization and death. ResultsThe joint frailty model identified patients with a higher risk of both relapse and death (θ=0.209, P<0.001). Risk factors for re-hospitalization were advanced age, grade 3 hypertension, mental work, no medical insurance, high cystatin C, low ejection fraction, and low free thyroxine-3 and thyroxine-4. Antiplatelet drugs and statins significantly reduced the risk of re-hospitalization. Risk factors for death were advanced age, New York Heart Association classification Ⅲ to Ⅳ, no medical insurance, mental work, high cystatin C level, high troponin-I level, low free thyroxine-3, and low ejection fraction. Percutaneous coronary intervention, and taking antiplatelet drugs and statins significantly reduced the risk of death. ConclusionThe joint frailty model can simultaneously model recurring and terminal events, and accurately predict them. Our results suggest that thyroid hormone levels and cystatin C levels of patients should be considered more carefully. People with mental jobs should change bad working habits to reduce adverse outcomes.
Objective To observe the influence of the transforming growth factor β1(TGF-β1) on the denervated mouse musclederived stem cells（MDSCs） producing the connective tissue growth factor（CTGF）at different time points in vitro. Methods MDSCs from the primarycultureof the denervated mouse skeletal muscle were isolated and purified by the preplate technique, and they were identified before the culture and after the culturein vitro with TGF-β1 (10 ng/ml) for 24 hours. Then, MDSCs were randomlydivided into 6 groups (Groups A, B, C, D, E and F) according to the different time points, and were cultured in vitro with TGF-β1 (10 ng/ml) for 0, 3, 6, 12, 24 and 48 hours, respectively. The levels of CTGF mRNA in MDSCs were measured by the real time RT-PCR and the expression of CTGF protein was detected by the CTGF Western blot. Results The immunohistochemistry revealed that before the adding of TGF-β1, MDSCs highly expressed Sca-1, with a positivityrate of 96%; however, after the adding of TGF-β1, the positive expression of Sca-1 decreased greatly, with a negativity rate gt;99%. The Western blot test showed that the ratios of CTGF to the average absorbance of βactin in Groups A-F were 0.788±0.123, 1.063±0.143, 2.154±0.153, 2.997±0.136, 3.796±0.153 and 3.802±0.175, respectively. In Groups AD，the absorbance increased gradually, with a significant difference between the abovementioned groups (Plt;0.05). However, in Groups D-F, there was no significant difference between the groups as the promotive tendency became less significant (P＞0.05). The RT-PCR test showed that the △Ct values in GroupsA-F were 1.659±0.215, 1.897±0.134, 2.188±0.259, 2.814±0.263,2.903±0.125 and 3.101±0.186, respectively. In Groups A-D, the increase in the △Ct value was gradual, but the differences were significant between the groups (Plt;0.05). But in Groups E and F, the promotive tendency became less significant（Pgt;0.05）. Conclusion TGF-β1 can promote the production of CTGF inthe mouse MDSCs cultured in vitro and the time-dependent relation exists for 3-12 hours.
ObjectiveTo analyze the expression and clinical significance of cyclin-dependent kinase 1 (CDK1) in lung adenocarcinoma by bioinformatics.MethodsBased on the gene expression data of lung adenocarcinoma patients in The Cancer Genome Atlas (TCGA), the differential expression of CDK1 in lung adenocarcinoma tissues and normal lung tissues was analyzed. The expression of CDK1 gene in lung adenocarcinoma was analyzed by UALCAN at different angles. Survival analysis of different levels of CDK1 gene expression in lung adenocarcinoma was performed using Kaplan-Meier Plotter. Correlation Cox analysis of CDK1 expression and overall survival was based on clinical data of lung adenocarcinoma in TCGA. Gene set enrichment analysis was performed on gene sequences related to CDK1 expression in clinical cases. The protein interaction network of CDK1 from Homo sapiens was obtained by STRING. CDK1-related gene proteins were obtained and analyzed by the web server Gene Expression Profiling Interactive Analysis (GEPIA).ResultsBased on the analysis of TCGA gene expression data, CDK1 expression in lung adenocarcinoma was higher than that in normal lung tissues. UALCAN analysis showed that high CDK1 expression may be associated with smoking. Survival analysis indicated that when CDK1 gene was highly expressed, patients with lung adenocarcinoma had a poor prognosis. Univariate and multivariate Cox regression analysis of CDK1 expression and overall survival showed that high CDK1 expression was an independent risk factor for survival of patients with lung adenocarcinoma. Gene set enrichment analysis revealed that high CDK1 expression was closely related to DNA replication, cell cycle, cancer pathway and p53 signaling pathway.ConclusionCDK1 may be a potential molecular marker for prognosis of lung adenocarcinoma. In addition, CDK1 regulation may play an important role in DNA replication, cell cycle, cancer pathway and p53 signaling pathway in lung adenocarcinoma.
ObjectiveTo improve the knowledge of a rare disease named pyridoxine-dependent epilepsy.MethodsHigh-throughput sequencing and Sanger sequencing were used to validate the genes of epilepsy. Mutation gene validation was performed on two probands and their parents. Analyze clinical manifestations, electroencephalogram (EEG), imaging and prognostic features of the two probands.ResultsProbands 1, seizure onset at 4 months, progress as drug-refractory epilepsy, manifested as seizures types origin of multi-focal lesions. Head MRI and fluorodeoxyglucose-positron-based tomography (FDG-PET) were both normal. Gene detection showed that Aldehydedehydrogenase (ALDH7A1) gene has a complex heterozygous mutation contain c.1442G> and c.1046C> T.Proband 2, seizure onset at 5 months, manifested as a tonic-clonic seizure. Intermittent EEG and head MRI were both normal. Genotyping revealed ALDH7A1 gene contain a complex heterozygous mutation c.1547A> G and c.965C> T. Two cases were both seizure free by vitamin B6 therapy and gradually reduce the antiepileptic drugs.ConclusionsPyridoxine-dependent epilepsy may be late onset, some patient can be atypical and early experimental treatment can help to identify and the diagnosis should be confirmed by gene test.
ObjectiveTo investigate the clinical electrophysiological characteristics of Cyclin-dependent kinase-like 5 gene induced developmental epileptic encephalopathy (CDKL5-DEE). MethodsThe clinical data and series of video EEGs of children with CDKL5-associated developmental epileptic encephalopathy (CDKL5-DEE) who were admitted to the Children’s Medical Center of Peking University First Hospital from June 2016 to May 2024 were retrospectively analyzed. Results A total of 16 patients with CDKL5-DEE were enrolled, including 13 females and 3 males. All patients had de novo variants of CDKL5 gene, including 6 cases of missense variants, 5 cases of frameshift variants, 4 cases of nonsense variants, and 1 case of large fragment deletion. The age of onset was 8 days (d) after birth ~1 year (y) and 10 months (m), and the median age was (85.94±95.76) days. Types of seizures at onset: 4 cases of tonic seizures [age of onset 10~52 days, median age (25.5±15.84) days]; There were 5 cases of focal seizures [age of onset 8 d~8 m, median age (77.76±85.97) d]. There were 4 cases of epileptic spasmodic seizures [age of onset 3 m~1 y 10 m, median age (6.25±3.49) m]; There were 2 cases of bilateral tonic-clonic seizures [age of onset 30~40 days, median age (35.00±5.00) days]; focal concurrent epileptic spasm seizures 1 case (age of onset 2 m). A total of 59 VEEG sessions were performed in the pediatric EEG room of Peking University First Hospital for 4 hours. All the results were abnormal, including 26 normal background, 25 slow rhythm difference with background, and 8 no background. The interictal was 16 posterior or focal discharges, 19 multifocal discharges, 17 generalized or accompanied by focal/multifocal discharges, and 7 hypsarrhythmia; The ictal was 33 epileptic seizures, 6 myoclonic seizures, 5 focal seizures, 2 tonic-clonic seizures, 2 atypical absence seizures, 2 tonic seizures, 1 myoclonic sequential focal seizure, 1 focal sequential epileptic spasm, and 1 hypermotor-tonic-spasms. The background of patients within 6 months of age was normal, and the background abnormality increased significantly with age. generalized discharges are evident after 2 years of age between seizures. Conclusion CDKL5-DEE seizures have an early onset and are refractory to medications. Epileptic spasms are the most common type of seizure in every patient and long-lasting, with generalized seizures increasing markedly with age. EEG is characterized by a normal background within 6 months. With the increase of age, the background and interictal discharges have a tendency to deteriorate.