Objective:To investigate the effectiveness and feasibility of whole exome sequencing (WES), as a molecular diagnosis technique, for adult patients with genetic diseases.Methods:The present retrospective analysis included 445 adult patients (ages 18-80 years) with suspected genetic diseases who underwent whole exome sequencing (WES) from August 2021 to December 2022. The pathogenicity classification of each variant was assessed in accordance with the recommendations developed by the American Society of Medical Genetics and Genomics.Results:The overall positive rate of WES among adult patients with suspected genetic diseases was 28.08% (125/445). The highest positive rate was observed in the age group of 41-50 years (34.33%, 23/67). Among the diagnosed genetic diseases, those affecting the cardiovascular system (63.16%, 84/133), nervous system (18.05%, 24/133), and endocrine system (13.53%, 18/133) ranked as the top three. The most common genetic diseases identified through WES in adult patients were hypertrophic cardiomyopathy (18.80%, 25/133), dilated cardiomyopathy (16.54%, 22/133), Marfan syndrome (15.04%, 20/133), epilepsy (9.02%, 12/133), and familial hypercholesterolemia (4.51%, 6/133). The main causative genes identified included FBN1 (14.29%, 19/133), MYBPC3 (9.02%, 12/133), MYH7 (9.02%, 12/133), LDLR (3.76%, 5/133), TTN (3.76%, 5/133), and TNNI3 (3.01%, 4/133).Conclusion:Applying the WES technique in clinical practice can improve the diagnostic rate of adult genetic diseases, especially in adult patients with suspected genetic conditions involving the cardiovascular system, nervous system, and endocrine system.

Objective: To retrospectively analyze the efficacy of aerosol inhalation of N-acetylcysteine(NAC) solution combined with bronchoalveolar lavage in children with Mycoplasma pneumoniae pneumonia(MPP) ． Methods: A retrospective study was conducted on 120 children with MPP． According to different treatment methods ，they were divided into combined treatment group (43 cases) ，atomization group (40 cases) and bronchoalveolar lavage group (37 cases) ．Among them，while receiving conventional treatment，children who were treated with aerosol inhalation of N-acetylcysteine alone were included in the atomization group，and those who were treated with bronchoalveolar lavage alone were included in the bronchoalveolar lavage group，and children treated with the combination of the above two treatments were included in the combined treatment group．The clinical efficacy，imaging recovery and incidence of adverse reactions were compared among all groups. Results: Compared the total effective rate among the three groups，there was no significant difference between the atomization group and the bronchoalveolar lavage group (P＞0. 05) ，which were both lower than that of the combined treatment group (P＜0. 05) ; the difference in the improvement rate of lung imaging among the three groups was statistically significant (P＜0. 05) ， the combined treatment group was the highest，and the BAL group was higher than the atomization group ; There was no significant difference in the duration of fever and pulmonary rales between the combined treatment group and the BAL group (P＞0. 05) ，which were shorter than those in the atomization group (P＜0. 05) ; There was no significant difference in duration of cough，shortness of breath and hospital stay between the atomization group and the BAL group (P＞0. 05) ，which were longer than those in the combined treatment group (P＞0. 05) ．There was no serious adverse reactions in the three groups．There was no significant difference in the three groups(P＞0. 05) . Conclusion Compared with single treatment ，N-acetylcysteine solution combined with bronchoalveolar lavage is more effective and has more advantages in the treatment of Mycoplasma pneumoniae pneumonia in children.

An effective therapeutic regimen for hepatic fibrosis requires a deep understanding of the pathogenesis mechanism. Hepatic fibrosis is characterized by activated hepatic stellate cells (aHSCs) with an excessive production of extracellular matrix. Although promoted activation of HSCs by M2 macrophages has been demonstrated, the molecular mechanism involved remains ambiguous. Herein, we propose that the vitamin D receptor (VDR) involved in macrophage polarization may regulate the communication between macrophages and HSCs by changing the functions of exosomes. We confirm that activating the VDR can inhibit the effect of M2 macrophages on HSC activation. The exosomes derived from M2 macrophages can promote HSC activation, while stimulating VDR alters the protein profiles and reverses their roles in M2 macrophage exosomes. Smooth muscle cell-associated protein 5 (SMAP-5) was found to be the key effector protein in promoting HSC activation by regulating autophagy flux. Building on these results, we show that a combined treatment of a VDR agonist and a macrophage-targeted exosomal secretion inhibitor achieves an excellent anti-hepatic fibrosis effect. In this study, we aim to elucidate the association between VDR and macrophages in HSC activation. The results contribute to our understanding of the pathogenesis mechanism of hepatic fibrosis, and provide potential therapeutic targets for its treatment.
Humans ; Hepatic Stellate Cells/pathology* ; Receptors, Calcitriol ; Liver Cirrhosis/pathology* ; Macrophages/metabolism*

OBJECTIVETo elucidate the usefulness of next generation sequencing for diagnosis of inherited myopathy, and to analyze the relevance between clinical phenotype and genotype in inherited myopathy.

METHODRelated genes were selected for SureSelect target enrichment system kit (Panel Version 1 and Panel Version 2). A total of 134 patients who were diagnosed as inherited myopathy clinically underwent next generation sequencing in Department of Pediatrics, Peking University First Hospital from January 2013 to June 2014. Clinical information and gene detection result of the patients were collected and analyzed.

RESULTSeventy-seven of 134 patients (89 males and 45 females, visiting ages from 6-month-old to 26-year-old, average visiting age was 6 years and 1 month) underwent next generation sequencing by Panel Version 1 in 2013, and 57 patients underwent next generation sequencing by Panel Version 2 in 2014. The gene detection revealed that 74 patients had pathogenic gene mutations, and the positive rate of genetic diagnosis was 55.22%. One patient was diagnosed as metabolic myopathy. Five patients were diagnosed as congenital myopathy; 68 were diagnosed as muscular dystrophy, including 22 with congenital muscular dystrophy 1A (MDC1A), 11 with Ullrich congenital muscular dystrophy (UCMD), 6 with Bethlem myopathy (BM), 12 with Duchenne muscular dystrophy (DMD) caused by point mutations in DMD gene, 5 with LMNA-related congenital muscular dystrophy (L-CMD), 1 with Emery-Dreifuss muscular dystrophy (EDMD), 7 with alpha-dystroglycanopathy (α-DG) patients, and 4 with limb-girdle muscular dystrophy (LGMD) patients.

CONCLUSIONNext generation sequencing plays an important role in diagnosis of inherited myopathy. Clinical and biological information analysis was essential for screening pathogenic gene of inherited myopathy.

Adolescent ; Child ; Child, Preschool ; Contracture ; DNA Mutational Analysis ; Female ; Genetic Diseases, Inborn ; diagnosis ; genetics ; Genetic Testing ; Genotype ; High-Throughput Nucleotide Sequencing ; Humans ; Infant ; Male ; Molecular Diagnostic Techniques ; Muscular Diseases ; diagnosis ; genetics ; Muscular Dystrophies ; congenital ; Muscular Dystrophies, Limb-Girdle ; Muscular Dystrophy, Duchenne ; Muscular Dystrophy, Emery-Dreifuss ; Mutation ; Phenotype ; Sclerosis ; Walker-Warburg Syndrome ; Young Adult

OBJECTIVETo evaluate the characteristics of the serious complications of 17 cases with measles in ICU.

METHODSeventeen cases with measles with serious complications in ICU in Anhui Provincial Children's Hospital were recruited from May 2012 to May 2013. Clinical characteristics, image finding, and prognosis were analyzed retrospectively. IgM antibody was positive in all the 17 cases, which included 9 male cases, 8 female cases, and their age was from 2 months to 10 years.

RESULTAll the 17 patients received mechanical ventilation because of severe respiratory distress within 1 week of onset, of which 14 cases were complicated with acute respiratory distress syndrome (acute ARDS), 6 cases of tension pneumothorax, 3 cases were complicated with bronchitis, laryngeal obstruction III degrees, and totally 7 cases died. The survived 10 patients were followed up for 1 year, 1 patient with localized pneumothorax, bronchopleural fistula, 1 case of mild pulmonary fibrosis, 1 case of acute laryngitis with persistent hoarseness, and mild inspiratory dyspnea were found. The remaining 7 cases fully recovered. Fourteen cases failed to inoculate measles vaccine.

CONCLUSIONA higher risk of death and poor prognosis were found in children with measles who needed to be treated in PICU, especially for ARDS with pneumothorax. Laryngitis needed long time to cure and had weaning difficulties, and the case associated with pleural effusion or bronchopleural fistula must be treated with surgery, and the patients easily develop secondary bacterial infection.

Child ; Dyspnea ; Female ; Humans ; Intensive Care Units, Pediatric ; Lung Diseases ; etiology ; Male ; Measles ; complications ; Pleural Effusion ; Prognosis ; Pulmonary Fibrosis ; Respiration, Artificial ; Retrospective Studies

Objective To investigate the clinical characteristics,diagnosis and treatment of necrotizing fasciitis (NF).Methods The authors reviewed and analyzed clinical manifestations,auxiliary examinations,treatments and prognoses of 14 patients who had been diagnosed with NF and hospitalized in the Children's Hospital of Anhui province between Jan 2007 and Sep 2013.Results Among the patients included in this study,eight cases were male and six cases were female.The average age was (15.86 ± 10.48) month,The time of abnormal temperature was (10.64 ± 5.64) d,hospital day was (29.07 ± 16.30) d,numbers of debridements were (3.07 ± 1.33) times.All patients had septic shock in which 5 cases had multiple organ failure.Diseases were found on hips (5 cases),lower limbs (4 cases),back (2 cases),perineum (2 cases),and neck (1 case).Blood culture showed staphylococcus aureus in six cases (1 case of methicillin-resistant staphylococcus aureus),pseudomonas aeruginosa in four cases and angina group of streptococcus pneumoniae in one case.No obvious bacteria growth was observed in three cases.CT examinations reflected subcutaneous gas formation in 11 cases but skin and subcutaneous tissue edema and fascial thickening in all cases.All cases of NF were further confirmed with soft tissue biopsies.Early symptoms resembled those of cellulitis.As the diseases progressed,other symptoms appeared such as skin ulceration,bullae formation and gas formation in the tissues.All patients were treated with surgical debridements (vacuum sealing drainage continuous drainage in 5 cases),appropriate antibiotic coverage and colloid supporting treatments.Seven patients were healed (50.00％,7/14),four had skin grafts (28.57％,4/14) and 3 died(21.43％,3/14).Six cases were found having limited physical activities in two year follow-up visits.Conclusion NF is a rare but potentially fatal disease.It is commonly found on the perineum,abdominal wall and extremities.NF can easily lead to septic shock and multiple organ failure.Early detection,surgical debridement and proper drainage along with appropriate antibiotic coverage can decrease mortality rates.

BACKGROUND:To construct tissue engineering cartilage would open up a novel way for the repair of cartilage damage in avoidance of the disadvantages of traditional therapeutic method.OBJECTIVE: To probe the techniques for the isolation of mesenchymal stem cells (MSCs) from bone marrow, as well as the in vitro differentiation into chondrocytic phenotype in a specific culture fluid.DESIGN:A complete randomized experimentSETTING:The Department of Traumatic Orthopedics and Hand Surgery of the First Affiliated Hospital of Guangxi Medical University, and Teaching and Research Faculty of Histology and Embryology of Guangxi Medical University.METHODS: The experiment was carried out at Guangxi Medical University between August 2002 and April 2003. Twenty SD neonatal weaning rats were selected. Bone marrow was aspirated from the bones of rat limbs and was isolated by gradient centrifugation in Percoll, and MSCs could be obtained in combination with adherent screening method, which were then cultured in DMEM-LG with 15% fatal bovine serum (FBS) in the incubator of 37℃ with 5% CO2 for 10-14 days. The passage cells were induced in DMEM-HG with 15% FBS (containing TGF-β1 10 μg/L, 10-7 mol/L dexamethasone, 50 mg/L VitC).MAIN OUTCOME MEASURES :The morphology, growth, as well as proliferation and specific expression of chondrogenic matrix of in vitro cultured MSCs due to specific induction.RESULTS: Totally 20 SD rats were observed and analyzed with no loss SCs grew in visible symmetric colonies, displaying a long-spindle shape,and the morphological characteristics of marrow-derived MSCs had no obvious changes during passage-culture, but its proliferation time was found from a shuttle fibroblastic appearance to polygonal shape, displaying posiHC staining of type Ⅱ collagen of cartilage specific matrix.bronectin adherent screening technique is a convenient, effective and practical method to separate and collect MSCs from rat bone marrows in chondrogenic phenotype when induced by a specific medium and can secrete cartilage specific matrix, and they can be the optimal seed cells for cartilage tissue engineering.

Objective To explore the expression of leptin and its receptors in liver tissue of nonalcoholic steatohepatitis(NASH)patient and its significance in pathogenesis.Methods Samples of 59 liver biopsies of NASH patients were divided into three groups G,S and F according to their degree of inflammation and steatosis and stage of fibrosis.The expression of leptin and its receptors including Ob-R,the short form of Ob-R(Ob-Ra)and the long form of Ob-R(Ob-Rb)were semi-quantitatively determined at the protein and/or mRNA expression levels by using immunohistochemistry,in situ hybridization and image analysis.Results The expressions of leptin in NASH liver tissues were found in KC,HSC and MNC,which were mainly situated in zone 3 near steatotic hepatocytes and perisinusoidal or portal fibrosis areas.Leptin mRNA(Ob)positive cells were distributed in hepatocytes in zone 3.The expression of leptin at protein and mRNA levels in the NASH liver tissues was significantly increased in NASH compared with normal liver tissues(P

Objective To investigate the expressions of pivotal cytokines such as TNF-? and TGF-?_1, and leptin in human nonalcoholic lipoid hepatitis (NALH), and to explore their potential roles in the pathogenesis of NALH. Methods Liver tissue obtained from 59 patients pathologically diagnosed as nonalcoholic lipoid hepatitis and 10 healthy adult liver tissues were examined in the present study. The respective expression of TNF-?, TGF-?_1 and Leptin was qualitatively and semi-quantitatively determined at the protein expression levels by using immunohistochemical method and image analysis. Results In NALH liver tissue the expression of TNF-? appeared mainly in inflammatory cells and lining cells of liver sinusoids in zone 3, lipogranulomas and portal tract, where obvious inflammation was found. The expression was parallel to different degrees of inflammation in NALH, significant correlation was found between degree of inflammation and fibrosis and extent of expression (P

Objective To evaluate the changes of hepatic blood perfusion of liver cirrhosis patients and the role of pericardial devascularization (PCDV)on them.Methods Hepatic artery and portal vein perfusions of 22 liver cirrhotic patients (LC) pre and post P CDV compared with control were evaluated with rheohepatogram(RHG) and ultrasonogram(USG).Results Compared with control,both hepatic and portal effective perfusion of LC were decreased on RHG, PCDV could increase portal perfusion of LC patients without significant effect on hepatic artery.USG showed that portal vein diameter and blood quantity of LC increased and PCDV didn't influence liver perfusion;No correlation between RHG and USG parameters were seen by STATA analysis. Conclusions RHG is an effective way to evaluate comprehensive blood perfusion of cirrhotic livers,the effect of operation on blood perfusion and prognosis.