Objective To investigate the phenotypic change and proliferation of fibroblasts in human inflammatory strictured bile duct wall. Methods We observed the density and ultrastructure of fibroblasts, and the histologic structure in human normal bile duct wall and inflammatory strictured bile duct wall by light and electron microscope.Results The results showed that fibroblasts were the main source of extracellular matrix production in bile duct wall. The phenotype of fibroblasts in inflammatory strictured bile duct wall changed obviously, quiescent fibroblasts were activated and transformed to myofibroblasts, with massive proliferation. Conclusion These data suggest that massive proliferation of activated fibroblasts and myofibroblasts is the main source of extracellular matrix overproduction which results in inflammatory bile duct stricture.
ObjectiveTo identify the causative gene and observe the phenotypic characteristics of a family with isolated microphthalmia-anophthalmia-coloboma (MAC). MethodsA retrospective clinical study. One patient (proband) and 3 family members of a family with MAC visited the Henan Eye Hospital from May 2019 to May 2022 were included in the study. The patient's medical history and family history were inquired in detail, and the best corrected visual acuity (BCVA), slit lamp microscope, fundus photography, optical coherence tomography (OCT), ophthalmological B mode ultrasound and axial length (AL) measurement were performed. The peripheral venous blood of the proband, his parents and brother was collected for Trio whole-exome sequencing and pathogenic gene screening. Fluorescence quantitative Polymerase chain reaction was used to verify the suspicious variations. The clinical features of the patient's ocular and systemic also were observed. ResultsThe proband, male, was 3 years old at the first visit. The horizontal pendular nystagmus was detected in both eyes. Vertical elliptical microcornea and keyhole-shaped iris colobomas were detected in both eyes. The objective refraction at first visit (3 years old) was －4.00 DS/－0.50 DC×105° (OD) and －3.50 DS/－1.25 DC×80° (OS). Refraction and BCVA at 6 years old: －6.50 DS/－2.00 DC×110°→0.05 (OD) and －6.00 DS/－1.50 DC×80°→0.2 (OS). The AL at 4 years and 10 months old was 24.62 mm (OD) and 23.92 mm (OS), respectively. The AL at 5 years and 7 months old was 25.24 mm (OD) and 24.36 mm (OS), respectively. Ultrasonography shows tissue defects in both eyes. Fundus photography showed the inferior choroidal coloboma involving optic disc. OCT showed the optic disc in both eyes was abnormal with colobomas around, and the retinal neurosensory layer in colobomas area was disordered and thin; the retinoschisis was visible in the left eye. The proband's parents and siblings have normal phenotypes. Whole exome sequencing reveals a denovo heterozygous deletion of YAP1 gene: YAP1, chr11: 10280247-102100671, NM_ 001130145, loss 1 (EXON: 6-9). The results of bioinformatics analysis were pathogenic variants. Parents and siblings were of the wild type. ConclusionsLoss of heterozygosity in exons 6-9 of YAP1 gene is the pathogenic variation in this family. It can cause abnormal development of anterior segment, chorioretinal colobomas, deepening of axial myopia, even severe macular colobomas and retinoschisis.
OBJECTIVE: To study the gap junction and phenotype of cultured chondrocyte of rabbit, and the gap junction between the chondrocytes in the same cartilage cavities in human femoral head articular cartilage. METHODS: CFDA-AM was added into the medium of the fifth passage of chondrocyte of rabbit in the 96-well plate. The fluorescent in spherical and fibroblast-like chondrocytes was detected separately. The recurrence of the fluorescent in accordant with time in 16 minutes was recorded after blanching the fluorescent with laser. And the fluorescent after blanching of chondrocyte in the cartilage cavities in the proliferative zone of articular cartilage of adult human femoral head was recorded, too. RESULTS: The average fluorescent of the single layer of the fibroblast-like chondrocyte was 83(ranged from 1 to 274), the highest was found in the spherical shaped cell (averaged 2,057, ranged from 340 to 3,538). The recurrence of the fluorescent after the blanching appeared only in the spherical chondrocyte, the gap junctions reappeared only in the spherical chondrocytes, as well as in the cells in the cartilage cavities in the articular cartilage of the human femoral head. CONCLUSION: The appearance of the gap junction is corresponded with the spherical shape, secretion of the cartilage matrix of the chondrocyte. There are gap junctions in the cells in the same cartilage cavities in the articular cartilage of the human femoral head, while no gap junctions in the isolated chondrocytes in the cartilage.
Objective To analyze the clinical characteristics and to screen for causative mutations in CRX and GUCY2D genes in children with cone or cone-rod dystrophy. Methods Clinical data and genomic DNA was collected from 18 children with cone or cone-rod dystrophy, aged from 4 months to 8 years. The coding sequence of the cone-rod homeobox (CRX) gene and two exons of the retinal-specific guanylate cyclase GUCY2D gene (exons 2 and 8) were analyzed by using polymerase chain reaction(PCR) and heteroduplex combined with single-strand conformational polymorphism (heteroduplex-SSCP) analysis. Results All of the 18 patients manifested obvious visual impairment. Nystagmus, photophobia and mild ocular fundus changes were found in 13, 8,and 7 cases respectively. Normal fundus was seen in 11 cases. The visual acuity was less than 0.3 in 4 cases and was unable to measure in the other 14 cases because they were too young. Clinical ocular manifestation s between cone and cone-rod dystrophy were overlapped. Mutation in the CRX and G UCY2D genes was not detected in the 18 children with cone and cone-rod dystrophy . Conclusion Visual impairment appeared more early and obvious than fundus changes in children with cone or cone-rod dystrophy. Mutation in the CRX gene may not contribute to this series of patients with cone and cone-rod dystrophy. (Chin J Ocul Fundus Dis, 2001,17:293-295)
Objective To investigate the phenotyping of COPD by cluster analysis and evaluate the value of this method.Methods 168 COPD patients were enrolled from Beijing Tongren Hospital. Demographic and clinical data, such as, sex, age, body mass index ( BMI) , smoking index, course of disease,exacerbation rate, and comorbidities were collected. Pulmonary function test, emphysema scoring by HRCT,dyspnea by MMRC score, COPD assessment test ( CAT) score, six-minute walk test were performed for each patient during the stable stage. Cluster analysis was conducted using SPSS 13. 0. Results According to the GOLD criteria,5, 75, 75, and 13 patients were classified into GOLD stage 1, 2, 3, and 4, respectively. There was no difference among different stages in sex distribution, BMI, smoking index, hypertension, and cerebral infarction incidence( P gt; 0. 05) , but the differences in age, disease course, dyspnea score, six-minute walk distance, BODE score, CAT score, coronary heart disease, exacerbation rate, and HRCT emphysema visual score were significant( P lt;0. 05) . By cluster analysis,168 patients were finally classified into three groups:younger/mild, older/ severe, and older/moderate. The patients with the same GOLD stage appeared indifferent clusters and the patients belonging to different GOLD stages could be in the same cluster. There were significant differences among three groups in age, BMI, exacerbation rate, dyspnea score, CAT score, and comorbidities. The result showed that HRCT emphysema visual score was also an important index todifferentiate clusters, suggesting that emphysema was an important phenotype of COPD. Conclusions Cluster analysis can classify homogeneous subjects into the same cluster, and heterogeneous subjects into different clusters. The results suggest that COPD phenotyping by cluster analysis is clinically useful and significant.
Objective To investigate the genotype and phenotype in patients with leber congenital amaurosis (LCA), and offer accurate genetic counseling and prenatal diagnosis for those families. Methods Three LCA patients and their parents were recruited for this study and received detailed collection of medical history and family history from March to August 2016. The three patients received fundus fluorescein angiography examination and their parents received slit-lamp microscope and indirect ophthalmoscopy examinations. DNA was extracted from the patients and their family members. Whole-exome sequencing method was used for genetic diagnosis and typing of the three LCA patients and their parents. Results The three patients with different clinical features had a definite clinical diagnosis of LCA. Patient 1 showed pale disc, attenuated vessels aroud the optic disc and the salt-and-pepper appearance of the retina, had the homozygous c.744.745insT (p.249, L>Ffs4) mutation inSPATA7. Patient 2 showed optic disc pallor and attenuated retinal vessels, had the heterozygous c.535G>A, p.A179T mutation inWFS1. Patient 3 showed pale disc, atrophic macular and retinal and choroidal degeneration, had the heterozygous mutation in CRB1, RPGRIP1, SPATA7. Conclusion LCA has characteristics of genetic heterogeneity and clinical and phenotypic diversity.
Chronic obstructive pulmonary disease (COPD) is one of the common chronic airway disorders, which accounts for the third to fourth cause of death worldwide. Recently, the focuses of researches are on the multi-factorial risks for development of COPD, mechanisms related to COPD development, early detection and early intervention of COPD, individualized use of long-term maintenance medications as well as phenotypes of acute exacerbation of COPD and their corresponding management. There are huge amount of COPD patients with variety of risk factors or different phenotypes in China, which makes it possible to establish a network for cohort study or real life registration study of COPD. The results will provide new information on the characteristics of COPD in China. Individualized treatment could be recommended according to the phenotypes or endotypes information. All these new findings or progresses could provide impetus for improvement of the ability of research and clinical management of COPD to the worldwide top level.
Abstract: Objective To induce calcification in aortic valvular interstitial cells (VICs) in vitro and observe the shift of cellular phenotype during the process. Methods Porcine aortic VICs were isolated and expanded by collagenase methods. Fluorescent staining was performed to identify the interstitial cells. VICs at 48 passages were used for experiments. The cells were divided into two groups: the experimental group in which cells were cultured in osteogenic media supplemented with βglycerophosphate, vitamin C and dexamethasone, and the control group in which cells were cultured in normal media. After 2 weeks, calcified nodules were quantified. Calcium deposit was stained and measured by Alizarin Red S staining and assay. Real time reverse transcription polymerase chain reaction (RTPCR) was performed to measure expression of alpha smooth muscle actin (α-SMA) and calcification related factors such as osteocalcin, osteopontin and Corebinding factor α1/Runx2 (Cbfα1/Runx2). Results VICs were successfully harvested from porcine aortic valves, identified by positive staining of α-SMA, vimentin and negative staining of Von Willebrand factor (vWF). VICs could calcify after 2 weeks of osteogenic induction with calcified nodules formed. Quantification of calcified nodules and calcification deposit were significantly higher (Plt;0.05) in the experimental group than those in the control group (156.25±17.38 vs. 2.50±1.29, 17.52±2.04 vs. 1.00±0.22). Real Time RT-PCR indicated that expression of α-SMA, as well as calcification related markers like osteocalcin, osteopontin and Cbfα1/Runx2 was much higher in the experimental group than those in the control group (Plt;0.05). Conclusion VICs are activated during the progress of calcification with phenotype shifting to contraction and ossification, which might be the pathological basis of valvular calcification.
ObjectiveTo study the phenotype of children with KCNQ2 gene related epilepsy.MethodsForty epilepsy children who were detected with KCNQ2 gene variants were enrolled. Their genotype and phenotype were analyzed.ResultsThirty-six KCNQ2 variants were identified. Twenty variants were novel. Twelve patients had inherited variants, and 28 patients had de novo variants. The age of seizure onset was from one day to 9 months. 80.0% patients had their seizure onset in neonates (32/40). Multiple seizure types were observed. Focal seizure was observed in 38 patients (95.0%). Epileptic spasm was observed in 10 patients (25.0%). Myoclonic seizure was observed in 4 patients. Tonic spasm seizure was observed in 3 patients. In all patients, seizures manifested in clusters. In 28 patients with de novo KCNQ2 variants, 24 had development delay (85.7%), the other 4 patients had normal development. In 12 patients with inherited KCNQ2 variants, one had development delay, the other 11 patients had normal development (91.7%). The most common interictal EEG changes were local epilepsy discharges (31/40). The MRI of brain was abnormal in 14 patients with de novo KCNQ2 variants and developmental delay. The agenesis of corpus callosum was identified in 10 patient (25.0%). Enlargement of subarachnoid spaces in the frontal and temporal region was identified in 11 patients (27.5%). Cortial dysplasia in the bilateral frontal and temporal region was identified in 2 patients. Sulus deepening was identified in 4 patients. Enlargement of bilateral lateral ventricle was identified in 3 patients. In 40 patients with KCNQ2 variants, 3 were diagnosed as benign familial neonatal epilepsy (BFNE), 2 were diagnosed as benign familial neonatal-infantile epilepsy (BFNIE), 3 were benign familial infantile epilepsy (BFIE), 3 were benign infantile epilepsy (BIE), 5 were benign neonatal epilepsy (BNE), 3 wer Ohtahara syndrome (OS), 9 were West syndrome (WS), 12 were unclassified early infantile epileptic encephalopathy (EIEE), one was epilepsy with autism. Sodium channel blockers oxcarbazepine was the most effective among antiepileptic drugs, with a effective rate of 90.9%.ConclusionsMost KCNQ2 variants are missense variants. De novo variants are more common in patients with KCNQ2 variants. The clinical features of patients with KCNQ2 variants including that mainly with seizure onset in neonate, the main seizure type is focal seizures, seizures occur in clusters. Patients with de novo KCNQ2 variants often had developmental delay, and about half of them had frontal and temporal lobe dysplasia and agenesis of corpus callosum. Sodium channel blockers are effective agents for epilepsy patients with KCNQ2 variants.
ObjectiveTo explore the difference in clinical characteristics and airway inflammation in COPD patients with different bronchodilator test results. MethodsA total of 237 COPD patients visited between January 2013 and December 2014 were recruited in the study. The ability to complete daily living questionnaire (ADL),modified Medicine Research Council (mMRC) score,6-minute walk distance,pulmonary function,and cell count in induced sputum were measured in the patients. They were divided into a positive group and a negative group according to the response to bronchodilator test and compared. ResultsThere were 58 cases (24.47%) in the positive group,and 179 cases (75.53%) in the negative group. There were no differences in the cumulative amount of smoking[(44.36±17.51) pack-years vs. (50.15±30.51) pack-years],duration of recurrent cough[(14.1±11.1) years vs. (15.5±11.4) years],history of allergic diseases (22.40% vs. 30.80%),or family history of allergic disease (5.17% vs. 2.23%) between two groups. In the positive group,FEV1%pred[(51.04±13.26)% vs. (44.10±14.66)%] and FVC%pred[(73.81±13.60)% vs. (64.33±15.17)%] were better than those of the negative group (both P<0.05). DLCO%pred[(44.66±13.92)% vs. (40.60±17.31)%] and RV/TLC[(51.80±10.57)% vs. (53.16±11.15)%] had no significant differences between two groups. 43.10% of the patients in the positive group and 61.46% in the negative group felt shortness of breath after walking (P<0.05). The positive group scored 22.6±3.8 points in activities of daily living assessment,1.5±0.9 points in mMRC,436.22±102.83 meters in 6-minute walking test,and 2.7±2.1 points in Borg scale score,which were all better than those in the negative group (all P<0.05). There was no significant difference in cell counting in induced sputum between two groups. ConclusionsA part of COPD patients have positive response to bronchodilator,with better lung function,better ADL score,better mMRC score,and farther 6-minute walking distance. It suggests that a positive bronchodilator response might be a clinical phenotype of COPD.