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 explore the phenotypic changes of epidermal stem cells (ESCs) differentiating into sweat glands cells (SGCs) in vitro and its mechanisms. MethodsESCs and SGCs were isolated and cultured in vitro, which were identified using immunofluorescence staining. ESCs at passage 2 were divided into 4 groups: ESCs and SGCs co-cultured by Transwell plates in group A, ESCs cultured by simply adding sweat supernatant in group B, ESCs and SGCs co-cultured on Transwell plate adding epidermal growth factor (EGF) (60 ng/mL) in group C, and ESCs and SGCs co-cultured on transwell plate adding PD98059 (10 mmol/L) in group D. The inverted microscope was used for observing the morphology of ESCs, flow cytometry for detecting ESCs positive phenotype, and Western blot for exploring mitogen-activated protein kinase/extracellular signal regulated kinase (MAPK/ERK) pathway. ResultsThe morphology observation and immunofluorescence staining suggested that cultured cells were ESCs and SGCs. The inverted phase contrast microscope observation showed that cells had similar morphological changes, with flat polygonal shape at 9 days in groups A, C, and D; cells had slow morphological change in group B, and had similar change to that of other groups at 12 days. Significant decreasing of β1-integrin expression and increasing of carcino-embryonic antigen (CEA) expression of ESCs were observed in group A when compared with group B, which was inhibited by EGF (group C) and enhanced by PD98059 (group D), and there were significant differences among groups A, C, and D (P<0.05). High level of ERK expression was displayed in 4 groups, but it was significantly lower in group B than the other 3 groups (P<0.05). The expression of phosphorylation ERK was the highest in group A and was the lowest in group C, showing significant difference among 4 groups (P<0.05). ConclusionESCs can be induced to differentiate into SGCs with the phenotypic changes under the condition of co-cultured by Transwell plates. The MAPK/ERK pathway plays a key role in the differentiation of ESCs into SGCs.
Objective To observe the gene mutation and clinical phenotype of patients with retinitis pigmentosa (RP) and cone rod dystrophy (CORD). Methods Thirty-seven patients with RP and 6 patients with CORD and 95 family members were enrolled in the study. The patient’s medical history and family history were collected. All the patients and family members received complete ophthalmic examinations to determine the phenotype, including best corrected visual acuity, slit lamp microscope, indirect ophthalmoscopy, color fundus photography, optical coherence tomography, full-field electroretinogram, and fluorescein fundus angiography. DNA was abstracted from patients and family members. Using target region capture sequencing combined with next-generation sequencing to screen the 232 candidate pathogenic mutations. Polymerase chain reaction and direct sequencing were used to confirm the pathogenic pathogenic mutations and Co-segregation is performed among members in the family to determine pathogenic mutation sites. The relationship between genotype and clinical phenotype of RP and CORD was analyzed. Results Of the 37 patients with RP, 13 were from 6 families, including 4 families with autosomal dominant inheritance, 2 families with autosomal recessive inheritance, and 3 in 6 families were detected pathogenic gene mutations. 24 cases were scattered RP. Six patients with CORD were from four families, all of which were autosomal recessive. Of the 43 patients, 21 patients were detected the pathogenic gene mutation, and the positive rate was 48.8%. Among them, 15 patients with RP were detected 10 pathogenic gene mutations including USH2A, RP1, MYO7A, C8orf37, RPGR, SNRNP200, CRX, PRPF31, C2orf71, IMPDH1, and the clinical phenotype included 10 typical RP, 2 cases of RPSP, 3 cases of Usher syndrome type 2 and 6 cases of CORD patients were all detected pathogenic gene mutations, including 2 cases of ABCA4, 2 mutations of RIMS1 gene, 1 case of CLN3 gene mutation, and 1 case of CRB1 and RPGR double gene mutation. Conclusions RP and CORD are clinically diverse in genotype and clinically phenotypically similar. For patients with early RP and CORD, clinical phenotype combined with genetic analysis is required to determine the diagnosis of RP and CORD.
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.
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.
ObjectiveTo explore the clinical phenotype of patients with acute exacerbation of chronic obstructive pulmonary disease (COPD) by cluster analysis and provide a basis for individualized treatment.MethodsA total of 515 patients with acute exacerbation of COPD admitted to this department from January 2014 to December 2016 were enrolled. The age, duration, smoking index, number of hospitalizations in the past 1 year, hospitalization days, treatment costs and other information were collected for cluster analysis.ResultsThe patients were divided into three categories of phenotype: " mild-glucocorticoid resistance-antibiotic dependent”," mild-glucocorticoid sensitive”, and " serious complication”. The patients with the first two phenotypes had a milder condition and lower hospitalization costs. There were differences in the time and cumulative dose of glucocorticoids in different pathways, antibiotic use time and usage rate. The third phenotype was the most serious, with the highest cost of hospitalization, and may merge or co-exist with other diseases such as cardiovascular disease and digestive tract disease.ConclusionCluster analysis may identify different phenotypes of acute exacerbation of COPD to provide a reference for clinical individualized treatment.
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.
Objective Chronic obstructive pulmonary disease( COPD) is highly heterogeneous. In theory, the patients with same clinical manifestations, treatment response and prognosis can be classified into one phenotype, which may have same biological or physiological mechanisms. In this study the profiles of patients with COPD including body mass index( BMI) , Goddard score, fractional exhaled nitric oxide( FeNO) were analyzed in order to find some special phenotypes.Methods Patients with COPD at stable stage in Ruijin Hospital from May 2011 to February 2012 were evaluated with COPD assessment test ( CAT) in Chinese version, St. George’s Respiratory Questionnaire( SGRQ) , hospital anxiety and depression( HAD) rating scale, pulmonary function test, and 6-minute walking test ( 6MWT) . Baseline data was collected including height, weight, drug use, times of exacerbation, etc. Results A total of 126 patients were recruited. The patients with low BMI had poorer quality of life, lower FEV1 , poorer diffusion function, and higher Goddard score, and was easier to develop anxiety and depression. The patients with high BMI had lower oxygen saturation at rest. We failed to define a certain kind of phenotype according to FeNO. The patients of emphysema phenotype( assessed by Goddard score) had lower BMI, decreased lung diffusion capacity, and poorer quality of life. Conclusion The study can define COPD patients into some special phenotypes( low BMI and emphysema phenotype) , but failed to define a certain kind of phenotype according to FeNO.
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)
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.