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 explore effects of zinc on the contents of cycl in D2, cycl in-dependent kinase 4 (CDK4), and their DNA and total cellular protein in human umbil ical cord blood-drived mesenchymal stem cells (hUCBMSCs). Methods hUCBMSCs were isolated and cultured by density gradient centrifugation adherence method in vitro. At the serial subcultivation, the hUCBMSCs were randomly divided into 7 groups. In control group, hUCBMSCs were cultured with DMEM medium (containing 15%FBS). In treatment groups, hUCBMSCs were cultured with DMEM medium (containing 15%FBS plusZnSO4•7H2O). The final concentrations of zinc were 0.5, 1.5, 2.5, 3.5, 4.5, and 5.5 mg/L, respectively. The cellular surface antigens of CD29, CD34, CD44, and CD45 at the 3rd generation of hUCBMSCs were detected by flow cytometry. MTT assay was used to detect cell activity of the 3rd generation of hUCBMSCs. The optimum concentration of zinc was selected by the results of MTT as experimental group. The cell growth curves of experimental group and control group were drown by counting cell. The cell surface antigen, reproductive cycle, and DNA content were detected by flow cytometry motheds. The contents of cycl in D2 and CDK4 were detected by Western blot method. Results The positive expression rates of CD29 and CD44 were more than 70% in hUCBMSCs. The cell activity of 2.5 mg/L treatment group was superior to other treatment groups, as experimental group. At 7, 14, and 28 days, the contents of DNA, total cellular protein, cycl in D2, and CDK4 of hUCBMSCs were significantly higher in experimental group than those in control group (P lt; 0.01). The percentage of hUCBMSCs at S stage and prol iferation index in experimental group were also significantly higher than those in control group (P lt; 0.01). Conclusion Zinc (0.5-4.5 mg/L) has the promoting effect on the hUCBMSCs activity, and 2.5 mg/L is the optimal concentration. Zinc (2.5 mg/L) can accelerate the prol iferation and DNA reproduction of hUCBMSCs and increase the contents of cycl in D2 , CDK4, and cellular total protein.
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 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.
OBJECTIVE: To explore the mechanism of microvascular spasm after limb ischemia-reperfusion. METHODS: The rabbit hindlimb normothermic tourniquet ischemia model was employed. The tendon on the dorsum of the foot was exposed for observation of microvessels. The responses of arterioles on tendon surface to topical application of 10(-6) mol/L noradrenaline (NE) (a vasoconstrictor), 10(-6) mol/l acetylcholine(Ach) (an endothelium-dependent vasodilator) and 10(-4) mol/L nitroglycerin(NTG) (an endothelium-independent vasodilator) were observed at the period of ischemia and following 30 minutes of reperfusion after 2 hours and 5 hours of ischemia by use of intravital microscopy. RESULTS: No significant changes in the responses of arterioles to NE, Ach and NTG were noted following 30 minutes of reperfusion after 2 hours of ischemia compared with pre-ischemia. The constrictor responses of arterioles to NE were still not significantly altered following 30 minutes of reperfusion after 5 hours of ischemia, however, the dilation responses to Ach and NTG were significantly decreased (to Ach P lt; 0.01; to NTG, P lt; 0.05). CONCLUSION: Reperfusion after 5 hours of ischemia significantly impairs both the endothelium-dependent and endothelium-independent vasodilation, meanwhile preserves constrictor responses to NE, these may contribute to the genesis of the vasospasm in ischemia reperfusion.
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.
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.
ObjectiveTo investigate the expression and clinical significance of cyclin dependent kinase 8 (CDK8) and transforming growth factor-β1 (TGF-β1), and their correlation in human colorectal cancer tissues. MethodsThe CDK8 and TGF-β1 mRNA expressions were examined by using real-time quantitative polymerase chain reaction and the protein expresssions of them were detected by immunohistochemistry on a cohort of human colorectal cancer tissues (n=40) and corresponding adjacent tissues (n=40). The correlation between CDK8 and TGF-β1 was also analyzed. ResultsThe expression levels of CDK8 mRNA and TGF-β1 mRNA in colorectal cancer were dramatically increased compared with the corresponding adjacent tissues (P < 0.05). The expression level of CDK8 mRNA was significantly associated with lymphnode metastasis, depth of invasion, and colorectal cancer stage (P < 0.05). However, the expression level of CDK8 mRNA was no correlation with the age, gender of patients, tumor size and differentiation of colorectal cancer in this study. The expression level of TGF-β1 mRNA was significantly associated with lymphnode metastasis, depth of invasion, tumor size and colorectal cancer stage (P < 0.05). However, the expression level of TGF-β1 mRNA was no correlation with the age, gender of patients and differentiation of colorectal cancer. The expressions of CDK8 mRNA and TGF-β1 mRNA in colorectal cancer was positively correlated (r=0.387, P=0.048). ConclusionsThe upregulation of CDK8 and TGF-β1 expression may be related to the occurrence and development of colorectal cancer, and the two may have a synergistic effect, which may be related to the biological behavior and prognosis of colorectal cancer.
ObjectiveTo explore therapeutic mechanisms and clinical application prospects of novel weight-loss medications in patients with obesity complicated by cardiovascular-kidney-metabolic (CKM) syndrome, aiming to provide theoretical support and therapeutic strategies for personalized precision management of CKM syndrome. MethodsRecent domestic and international studies were retrospectively reviewed, focusing on the mechanisms of action, clinical research outcomes, and application progress of novel weight-loss medications, including glucagon-like peptide 1 (GLP-1) receptor agonists, dual glucose-dependent insulinotropic peptide (GIP)/GLP-1 receptor agonists, triple GIP/GLP-1/glucagon receptor agonists, and amylin analogues. Special emphasis was placed on their comprehensive effects on cardiovascular, renal, and metabolic parameters. ResultsNovel weight-loss medications have demonstrated significant weight reduction and multisystem benefits through precise regulation of central appetite pathways, insulin sensitivity, and lipid metabolism. Among these medications, GLP-1 receptor agonists (e.g., semaglutide) and dual receptor agonists (e.g., tirzepatide) have been confirmed in phase Ⅲ clinical trials to effectively reduce cardiovascular event risks, slow renal function deterioration, and markedly improve glycemic control in obese patients with CKM syndrome. Triple receptor agonists (e.g., retatrutide) and combination medication regimen (e.g., CagriSema regimen) have further enhanced weight-loss efficacy, providing novel therapeutic avenues for obesity-related diseases. Additionally, these medications usually require combined application with traditional chronic disease medications, such as sodium-glucose linked transporter 2 inhibitors and renin-angiotensin-aldosterone system blockers, to achieve comprehensive therapeutic outcomes in CKM syndrome patients. However, further studies are needed to address long-term safety in real-world settings, optimization of drug formulations, and application in precision medicine. ConclusionsNovel weight-loss medications offer promising strategies for personalized precision treatment of obesity with CKM syndrome due to their significant weight-loss efficacy and multisystem synergistic effects. Although current clinical trials demonstrate substantial therapeutic potential, the complexity of CKM syndrome and individual patient variability necessitate additional in-depth research to facilitate broader clinical adoption and optimization of these medications.
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.