Objective:To observe the proteomic changes in vitreous fluid samples from patients with rhegmatogenous retinal detachment combined with choroidal detachment (RRDCD).Methods:A prospective cross-sectional clinical study. Vitreous fluid samples were collected from 35 patients with RRDCD (RRDCD group) and 40 patients with rhegmatogenous retinal detachment (RRD group) who were diagnosed at Wuhan Aier Eye Hospital between November 2021 and December 2023. Prior to vitrectomy, 0.3-0.5 ml of vitreous fluid was collected from the affected eyes. Differentially expressed proteins were analyzed using Data-Independent Acquisition (DIA). Three of these proteins were randomly selected for validation using enzyme-linked immunosorbent assay (ELISA). Bioinformatics analyses, including gene ontology functional enrichment and kyoto encyclopedia of genes and genomes pathway enrichment, were performed to explore the functions of the differentially expressed proteins.Results:Significant differences were observed between the RRDCD and RRD groups in intraocular pressure ( t=-12.795), the number of retinal tears ( t=4.601), the extent of retinal detachment ( χ2=39.642), axial length ( t=0.840), postoperative proliferative vitreoretinopathy incidence ( χ2=4.730), single-surgery reattachment rate ( χ2=7.717), and best-corrected visual acuity ( t=7.033) at 6 months postoperatively ( P<0.05). A total of 237 differentially expressed proteins were identified between the RRDCD and RRD groups, with 63 upregulated and 174 downregulated. These proteins were involved in pathways such as extracellular matrix-receptor interaction, complement activation, coagulation, and lysosomal pathways. ELISA validation results showed that the expression trends of the three selected proteins in the RRDCD and RRD groups were consistent with the DIA proteomic analysis. Compared to the RRD group, proteins such as fibrin, coagulation factors, cathepsins, and trypsin inhibitors were significantly upregulated in the RRDCD group. Conclusions:The protein expression profile in vitreous fluid samples from RRDCD patients show significant alterations compared to the RRD group. These differential changes suggest that RRDCD is closely associated with complement and coagulation cascade activation, lysosomal pathways, and extracellular matrix remodeling.

OBJECTIVE: To study the effect of nanohydroxyapatite (nHA) for promoting surface mineralization of demineralized dentin discs and adsorption of lead ions in simulated sewage water. METHODS: Sixty dentin disks were prepared from freshly extracted teeth with intact crown (including 30 premolars and 30 molars) and treated with 10% citric acid for 2 min to simulate dentinal tubules with dentin hypersensitivity. The etched dentin discs were brushed with distilled water, 0.2 g HA or 0.2 g nHA for 2 min twice a day for 7 consecutive days, after which scanning electron microscopy (SEM) was performed and calcium and phosphorus contents in the dentin discs were detected using EDS. Lead ion adsorption capacities of HA and nHA were tested by mixing 1 mL serial concentrations of HA and nHA suspensions with 50 mL lead ion solutions (1.0 mg/L). After 24 h, the residual lead ion concentration in the supernatant was measured using ICP to calculate lead ion adsorption rate and adsorption capacity of the materials. RESULTS: SEM showed a smooth surface and empty dentin tubules in the acid- etched dentin dics. The dentin dics treated with HA were covered with masses of particles that loosely attached to the surface, and the diameter of the dentin tubules was reduced. In nHA group, the dentin discs showed a fine and homogeneous surface clogging with a tight attachment, and the dentin tubule diameter was obviously reduced. Daily brushing with HA and nHA, especially the latter, significantly increased calcium and phosphorus contents on the surface of the dentin dics ( < 0.000). In lead ion adsorption experiment, the lead ion adsorption rate of HA decreased as its concentration increased with the highest adsorption rate of 83.01%; the adsorption rate of nHA increased with its concentration until the adsorption equilibrium was reached, and its highest adsorption rate was 98.79%. A good liner relationship was found between the adsorption ability and concentration of HA. CONCLUSIONS Compared with HA, nHA has a better capacity for surface mineralization of acid-etched dentin discs and also a better ability of lead ion adsorption.

OBJECTIVE: To explore the genetic basis for a pedigree affected with X-linked recessive mental retardation Claes-Jensen type. METHODS: Genomic DNA was extracted from peripheral blood samples of the patient, his parents (phenotypically normal) and two elder brothers with similar clinical manifestations. Whole exome sequencing was carried out for the proband, and the result was verified by Sanger sequencing. RESULTS: The proband was found to harbor a hemizygous c.1565C>T missense variant in exon 11 of the KDM5C gene. The transition has resulted in replacement of serine by phenylalanine at position 522 (p.Ser522Phe). Sanger sequencing showed that the patient's two elder brothers and mother carried the same variant, which was predicted to be probably damaging by SIFT, PolyPhen2 and Mutation_Taster. The three affected brothers presented with similar clinical phenotypes characterized by mental retardation, speech delay, behavioral problem, self-limited epilepsy responsible to medication, short stature and microcephaly. The mother only had mild cognitive impairment and learning disability. The same variant was not found in their father and was unreported previously. CONCLUSION The c.1565C>T (p.Ser522Phe) of the KDM5C gene probably underlay the X-linked recessive mental retardation Claes-Jensen type in this pedigree.
Aged ; Female ; Histone Demethylases/genetics* ; Humans ; Male ; Mental Retardation, X-Linked/pathology* ; Mutation, Missense/genetics* ; Pedigree ; Phenotype ; Whole Exome Sequencing

Objective:To investigate the clinical and genetic characteristics of developmental and epileptic encephalopathy (DEE) caused by syntaxin-binding protein 1 (STXBP1) gene mutation.Methods:The clinical data, gene variation and treatment outcome of 15 children with STXBP1 encephalopathy admitted to Children′s Hospital Affiliated to Capital Institute of Pediatrics from January 2014 to June 2019 were analyzed retrospectively.Results:Among 15 patients, 11 were male and 4 were female, age ranged from 2 months to 69 months. The clinical manifestations of 14 children were epilepsy and developmental delay (DD) and the remaining one showed developmental delay without seizure. The onset age of epilepsy ranged from two days to 19 months and 11 of them experienced the first attack before 1 year of age. The common seizure types were epileptic spasms and tonic seizures. Seven patients were diagnosed with Ohtahara syndrome or West syndrome. Epileptic form discharges were observed in the interictal electroencephalograms (EEG) of 11 patients, including multifocal discharges, suppression-burst and hypsarrhythmia. The brain magnetic resonance imaging of 7 children were abnormal, including myelin dysplasia, less white matter, lack of corpus callosum or hypoplasia. The follow-up time ranged from 2 months to 57 months, after the last follow-up, 3 cases were seizure free, 6 children showed partial response and the other 5 patients had no response on multitherapy. Six of 8 patients showed good responses to levetiracetam (LEV) monotherapy or in combination with other antiepileptic drugs (AEDs). Vigabatrin (VGB) was applied to 5 patients with epileptic spasms and 4 of them showed response. All patients showed different degrees of developmental delay while four of them showed autistic features. STXBP1 gene mutations were identified in all cases and there were 15 types of gene variations, including 8 missense mutations, 1 nonsense mutation, 5 frame shift mutations and 1 complex mutation. Five novel mutations were unreported before, including c.1193A>G, c.172delG, c.1769C>T, c.1038_1039delCC, c.348_351dupTGAA.Conclusions:Development delay and epilepsy are the major and independent clinical phenotypes in children with STXBP1 encephalopathy. The variation of STXBP1 gene is mainly de novo. Levetiracetam and vigabatrin may be more effective in epilepsy control than other AEDs.

Adult ; Female ; Humans ; Male ; Middle Aged ; Pedigree ; Spastic Paraplegia, Hereditary ; genetics

In order to study the rural elderly chronic patient's seeking healthcare behavior and its influencing factors, the paper uses the 2011 national baseline survey data of the “China Health and Retirement Longitudinal Study ( CHARLS)” and the Multinomial Logit model to conduct an empirical analysis .The study results show that:( 1 ) The rural elderly chronic patients are more likely to choose the county-level and above hospitals and village clin-ics instead of the township health centers .(2) The obvious influencing factors in the healthcare seeking behavior in-clude age , educational background , household per capita annual income , chronic disease severity , self-rated health and region.Therefore, the present study suggests that the treatment of chronic diseases does not transfer to the town -ship health centers .The township health centers should improve their chronic medical service abilities , and actively strengthen their functions in chronic diseases prevention and treatment .

AIM:To investigate the effects of human induced pluripotent stem cell-derived exosomes (hiPSC-exo) on cell viability, capillary-like structure formation , and senescence in endothelial cells exposed to high glucose .METHODS: Exosomes were isolated from the conditional medium of hiPSCs and confirmed by transmission electron microscopy , nanoparticle tracking analysis , and Western blot analysis using Alix and CD63 as markers.hiPSC-exo were labeled with PKH26 for tracking.Cultured HUVECs were treated with high glucose (33 mmol/L) with or without hiPSC-exo (20 mg/L) for 48 h, and cell viability, capillary tube formation, and senescence were assessed .RESULTS:hiPSC-exo showed a typical cup shape and could be taken up by human umbilical vascular endothelial cells (HUVECs) in a concentration-dependent manner.When exposed to high glucose, viability and tube formation in HUVECs was signifi-cantly reduced, whereas the proportion of senescent cells was higher compared to that in control HUVECs (P<0.01).Furthermore, hiPSC-exo restored cell viability and capillary-like structure formation , and reduced senescence in HUVECs exposed to high glucose (P<0.01).However, hiPSC-exo had minimal effects on normal HUVECs.Therefore, stem cell-derived exosomes can promote cell proliferation, enhance capillary-like structure formation , and reduce senescence in endothelial cells exposed to high glucose . CONCLUSION:Our study highlights the role of exosomes derived from hiPSC and may provide a new strategy for maintaining vascular health, preventing vascular aging , and avoiding pathological vascular remodeling that occurs in many diseases .

OBJECTIVE To investigate the pharmacodynamics of chronopharmaceutical drug delivery of erlotinib in lung cancer model nude mice and its potential mechanism. METHODS A nude mouse model of human lung adenocarcinoma HCC827 cell subcutaneously implanted tumor was established and subsequently the nude mice were randomly divided into 6 erlotinib groups and a model group, with 10 nude mouse per group. Erlotinib groups were respectively gavaged with 5 mg.kg-1 erlotinib at 08:00, 12:00, 16:00, 20:00, 24:00 and morrow 04:00, while the model group was given the same volume fraction of captisol. The tumor volume and tumor mass were measured and the tumor growth inhibitory rate was calculated. The mRNA expression of epidermal growth factor receptor (EGFR), mitogen-acti-vated protein kinase (MAPK), cyclin-dependent kinase inhibitor 1A(P21Waf1) and the related protein level were detected by real-time quantitative PCR and Western blotting. RESULTS Compared with model group, the tumor volume and tumor mass of mice at the dosing time of 08:00 and morrow 04:00 were significantly decreased(P<0.05). Compared with dosing time at 20:00 group〔(0.70±0.36)g〕, the tumor mass of 08:00 group〔(0.30±0.17)g〕 and morrow 04:00〔(0.39±0.29)g〕 group was significantly decreased(P<0.05). The mRNA expression of EGFR and MAPK in the tumor group of 08:00 was lower than in group of 20:00(P<0.05), while the mRNA expression of P21Waf1 was significantly higher than that of model group( P <0.05). Compared with model group, the protein expression of p-EGFR and p-MAPK in tumor 08: 00 and morrow 04: 00 group was negative-regulated significantly (P<0.05). CONCLUSION The antitumor effect of erlotinib on the human lung adenocarcinoma implanted tumor nude mice model presents rhythmicity. The dosing time at 08:00 is the most effective. lts mechanism is likely related to EGFR/ MAPK/ P21Waf1 signal transduction pathway.

The mesaconitine and its major metabolites in the rat urine were identified by liquid chromatography and electrospray ionization tandem mass spectrometry. The rat urine was collected for consecutive 24 hours from the rat following intragastric infusion of mesaconitine, subsequently which were enriched and purified using solid phase extraction. The metabolites of mesaconitine in the rat urine were analyzed by the liquid chromatography and electrospray ionization tandem mass spectrometry. It is shown that the parent drug mesaconitine and its metabolites were found in the rat urine, such as hypo-mesaconitine glucuronic acid conjugate, 10-hydroxy-mesaconitine, 1-O-demethyl mesaconitine, deoxy-mesaconitine and hypo-mesaconitine. Among the five of metabolites, the hypo-mesaconitine glucuronic acid conjugate (m/z 766) was first discovered as the aconitine in rats phase II metabolites, which revealed a new way of mesaconitine metabolism in rats.