In order to study the possible role of beta endorphin (? EP) in the pathogenesis of psoriasis, serum ? EP was detected by radioimmunoassay in 51 patients with psoriasis and 32 healthy controls. The results showed that serum ? EP level was markedly increased in patients with psoriasis in comparison with healthy controls. The serum ? EP level was reduced when the skin lesions of the patients were cleared after treatment, however, it was still higher than in healthy controls. Although there was a tendency towards an increase of ? EP in patients with precipitating factors in comparison with those without precipitating factors, their levels were not significantly different. There was a negative correlation between the intensity of pruritus and ? EP levels. It is suggested that ? EP involves in the pathogenisis of psoriasis. Its production and release were controlled by the central nervous system and also affected by the inflammation of skin lesions. There might be a causality between ? EP levels and inflammatory reaction of skin lesions.

Objective To investigate the effects of hyperbaric Oxygen (HBO)exposure on Caspase-3 and Bcl-2 mRNA ex-pressions in both internal carotid artery (ICA)and basal artery (BA)in rabbits.Methods Twenty-four healthy adult New Zealand rabbits were randomly divided into 2 groups:HBO group and the control group,with each group consisted of 12 animals.The rab-bits in the HBO group were exposed to HBO at 2.2 ATA for 60 minutes each day for 3 successive days.The rabbits in the control group were normally fed without any treatment.Real-time PCR was used to detect Caspase-3 and Bcl-2 mRNA expressions in both ICA and BA in 2 groups.Results HBO significantly decreased the Caspase-3 mRNA expression [(0.038 ±0.006 )vs .(1.000 ± 0.225)]and increased the Bcl-2 mRNA expression [(1.877 ±0.1 69)vs .(1.000 ±0.364)].In ICA,HBO similarly decreased the Caspase-3 mRNA expression [(0.41 9±0.091)vs .(1.000 ±0.1 75)]and increased the Bcl-2 mRNA expression [(1.269 ±0.270) vs .(1.000±0.1 1 7)]in BA.All the differences mentioned above were of statistical significance (P <0.01).Conclusion HBO ex-erts an inhibition effect on apoptosis in cerebral vascular endothelial cells.The mechanism may be related to inhibiting the expres-sion of Caspase-3 mRNA and promoting the expression of Bcl-2 mRNA.

OBJECTIVETo summarize the clinical features of those Duchenne and Becker muscular dystrophy (DMD and BMD) patients who are complicated with epilepsy, and try to analyze the genotype- phenotype correlation.

METHODBy a retrospective analysis of 307 patients with DMD and BMD who attended Peking University First Hospital from February 2006 to September 2014,7 patients complicated with epilepsy were identified and their clinical data were collected. The possible mechanism of epilepsy in DMD and BMD patients was proposed after analyzing the genotype-phenotype correlation.

RESULT(1) Among 307 DMD and BMD patients, 7 cases had epilepsy, the prevalence was 2. 28%. (2) The age of onset of epilepsy ranged from 8 months to 11 years. Focal seizure was the most common seizure type (6 cases) , while other seizure types were also involved, such as generalized tonic-clonic seizure. As to epilepsy syndromes, 1 boy was diagnosed as benign childhood epilepsy with centrotemporal spikes (BECT). Six patients were treated with 1 or 2 types of antiepileptic drugs and seizures were controlled well. On follow-up, 6 of the 7 children had normal mental development, while the remaining 1 patient was diagnosed as mild mental retardation. (3) DMD gene mutations of all 7 patients were analyzed. Exons deletions were found in 6 cases while point mutation was found in 1 case.

CONCLUSIONThe prevalence of epilepsy in DMD and BMD patients was higher than the prevalence in normal population. The age of onset of epilepsy varies, and focal seizure may be the most common seizure type. Some patients may also present as some kind of epilepsy syndrome, such as BECT. In most patients, seizures can be controlled well by 1 or 2 types of antiepiletic drugs. No clear correlation was found between genotype and phenotype in DMD and BMD patients who were complicated with epilepsy, probably due to limited number of cases.

Anticonvulsants ; therapeutic use ; Child ; Epilepsy ; complications ; drug therapy ; epidemiology ; Exons ; Genotype ; Humans ; Intellectual Disability ; etiology ; Male ; Muscular Dystrophy, Duchenne ; complications ; genetics ; Mutation ; Phenotype ; Prevalence ; Retrospective Studies ; Seizures ; Sequence Deletion

Objective: To explore the injury pattern and features of peripheral nerve in congenital muscular dystrophy patients caused by LAMA2 gene mutation. Method: Seventeen patients genetically or molecular pathologically diagnosed as LAMA2-related congenital muscular dystrophy were recruited in Peking University First Hospital between 2002 and 2015. All the patients received nerve conduction velocity (NCV) and needle electromyography tests. Clinical and laboratory examination data of the patients was retrospectively analyzed. The correlation between the NCV and disease course was determined by Pearson correlation analysis. Additionally, one patient underwent a sural nerve biopsy. Result: Among these 17 identified patients (13 male and 4 female), all of them were diagnosed as congenital muscular dystrophy, and all of them underwent electrophysiological examination at ages between 1 month to 6 years. Electromyogram indicated seventeen patients of myogenic damage, of whom 10 cases were complicated with reduced NCV. Twenty-six of 95 analyzed nerves showed NCV slower than the normal average of contemporary in 17%-47%. Correlation analysis between NCV and the disease course indicated that NCV of median nerves, ulnar nerves, tibial nerves and common peroneal nerves were negatively associated with the disease course (r=-0.737, -0.771, -0.540 and -0.682, respectively; all P<0.05). Sural nerve biopsy revealed peripheral neuropathy changes of myelin. Conclusion There is peripheral nerve injury in LAMA2-related muscular dystrophy patients. It mainly manifests as demyelinating lesions. Moreover, the NCV of peripheral nerve will decrease with the increase of the course of the disease.

For investigating the effect of Jumonji domain-containing protein-3 (JMJD3) on the behavior of lung cancer cell line, A549 proliferation was measured with EDU staining and flow cytometer after JMJD3 expression plasmid and pcDNA3. 1 transfection at 48h. The migration ability of A549 was tested at the same time. The expression of p21 mRNA was measured with RT-PCR. The results showed that JMJD3 transfection increased the EDU positive cells ratio (JMJD3: 40.75% +/- 2.07%, control: 20.97% +/- 1.5%, P < 0.001). G1 phase cell ration also increased after JMJD3 transfection (JMJD3:47. 80% +/- 1.85%, control: 54.60% +/- 0.95%, P = 0.005). The mRNA expression of p21 decreased in JMJD3 group (JMJD3: 35. 89% +/- 3.71%, control: 91.78% +/- 3.74%, P < 0.001). The distances of migration were (0.47 +/- 0.27) cm and (0.96 +/- 0.40) cm after 24h and 48h with JMJD3 tranfection, compared to (0.57 +/- 0.22)cm and (1.08 +/- 0.33)cm in control, respectively (P > 0.05). JMJD3 promoted the proliferation of A549 and decreased the G1 cell numbers, decreased the p21 mRNA, but had no effect on A549 migration.
Adenocarcinoma ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p21 ; genetics ; metabolism ; Humans ; Jumonji Domain-Containing Histone Demethylases ; pharmacology ; Lung Neoplasms ; pathology ; RNA, Messenger ; genetics ; metabolism ; Transfection

Objective:To investigate the effect of sodium arsenite (NaAsO 2) on transcriptional activity of nuclear factor E2-related factor 2 (Nrf2) signaling pathway in mouse lymph node vascular endothelial cell line (SVEC4-10). Methods:In vitro cell culture method was used to treat SVEC4-10 cells for 24 h with different doses of NaAsO 2 [0 (control), 2, 5, 10, 20, 50, 100, 150 μmol/L], and the cell viability was detected by tetrazole compound (MTS) method. The time-response relationship was studied with SVEC4-10 cells treated with 5 μmol/L NaAsO 2 for 0 (control), 2, 6 and 12 h; the dose-response relationship was studied with SVEC4-10 cells treated with 0 (control), 2, 5 and 10 μmol/L NaAsO 2 for 6 h; real-time fluorescence quantitative PCR (RT-qPCR) was used to detect the mRNA expression of Nrf2 and its downstream genes glutamate-cysteine ligase catalytic subunit (Gclc), glutamate-cysteine ligase modifier subunit (Gclm), NAD(P)H dehydrogenase quinone 1 (Nqo1) and metallothionein 1 (Mt1). Establishment of Nrf2 gene stably silenced (Nrf2-KD) cells using SVEC4-10 cells, the interference control (scramble, SCR) cells and Nrf2-KD cells were treated with 0(control), 10 and 20 μmol/L NaAsO 2 for 16 h, and apoptosis was detected by flow cytometry. Results:MTS test results showed that the cell viability of the control, 2, 5, 10, 20, 50, 100, 150 μmol/L NaAsO 2 treatment groups was (100.00 ± 19.53)%, (98.18 ± 9.85)%, (96.09 ± 30.04)%, (90.64 ± 8.74)%, (59.75 ± 12.09)%, (35.43 ± 8.58)%, (26.35 ± 5.89)% and (17.54 ± 4.48)%, respectivily. There was statistically significant difference in cell viability between different dose groups ( F = 18.30, P < 0.05); and the cell viability of the 20, 50, 100, 150 μmol/L NaAsO 2 treatment groups was significantly lower than that of the control group ( P < 0.05). The time-response relationship results showed that there were statistically significant differences in Nrf2, Gclc, Gclm, Nqo1 and Mt1 mRNA level between control, 2, 6 and 12 h treatment groups ( F = 56.69, 85.28, 90.82, 80.46, 758.60, P < 0.05); with extension of arsenic exposure time, the mRNA level of Nrf2, Gclc, Gclm and Mt1 first increased and then decreased, the mRNA level of Nqo1 increased continually; among them, the mRNA level of Nrf2 peaked at 2 h, the mRNA levels of Gclc, Gclm and Mt1 peaked at 6 h, and the mRNA level of Nqo1 peaked at 12 h. The dose-response relationship results showed that there were statistically significant differences in Nrf2, Gclc, Gclm, Nqo1 and Mt1 mRNA levels between control, 2, 5 and 10 μmol/L NaAsO 2 treatment groups ( F = 68.39, 72.26, 30.41, 397.00, 28.88, P < 0.05); with increasing of arsenic exposure dose, the mRNA levels of Nrf2, Gclc, Gclm, Nqo1 and Mt1 increased. The mRNA level of Nrf2 peaked at a dose of 5 μmol/L, and the mRNA levels of Gclc, Gclm, Nqo1 and Mt1 peaked at a dose of 10 μmol/L. Apoptosis test results showed that there were statistically significant differences in the apoptosis rates of SCR and Nrf2-KD cells between control, 10 and 20 μmol/L NaAsO 2 treatment groups ( F = 8.18, 9.66, P < 0.05); compared with the control group, the apoptosis rates of SCR and Nrf2-KD cells in the 20 μmol/L NaAsO 2 treatment group increased ( P < 0.05); and the apoptosis rate of Nrf2-KD cells in the 20 μmol/L NaAsO 2 treatment group was higher than that of SCR cells in the same dose group ( P < 0.05). Conclusions:NaAsO 2 exposure has caused the activation of Nrf2 signaling pathway in mouse lymph node vascular endothelial cell line SVEC4-10 cells, activated the adaptive antioxidant response, and altered transcriptional activity; while silence of Nrf2 has made SVEC4-10 cells more sensitive to NaAsO 2 toxicity.

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

Objective:To establish the ultra-high performance liquid chromatography (UPLC) fingerprint and high performance liquid chromatography (HPLC) content determination method of Curcumae Radix standard decoction; To predict the quality markers of Curcumae Radix standard decoction combined with network pharmacology.Methods:UPLC method was used to establish the fingerprint of Curcumae Radix standard decoction, and the common peaks were determined. Combined with chemical pattern recognition techniques such as similarity analysis and clustering analysis, Curcumae Radix standard decoction from different producing areas was studied, and curcumol was used as an index to determine the content of 24 batches of Curcumae Radix standard decoction. At the same time, network pharmacology was used to predict potential of curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one.Results:A total of 24 batches of Curcumae Radix standard decoction from different habitats were compared and analyzed, and 10 common peaks were calibrated. The similarity of 24 batches of samples ranged from 0.982 to 0.999. Clustering analysis and principal component analysis divided them into three categories. Heat map analysis showed that peak 8 (curcumol) and peak 9 ((1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one) were the main components. The content of curcumol in 24 batches of Curcumae Radix standard decoction was 0.69-1.87 mg/g; curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β- (3-oxobutyl) bicyclo [4.1.0] heptan-3-one may regulate the neuroactive ligand-receptor interaction signaling pathway, calcium signaling, and excitation by regulating neuroactive ligand-receptor interaction signaling pathway, calcium signaling, and excitation. It was preliminarily predicted that curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one were potential quality markers of Curcumae Radix.Conclusion:Curcumol and (1S, 6β)-1β-Methyl-4-(1-methylethylidene)-7β-(3-oxobutyl) bicyclo [4.1.0] heptan-3-one are potential quality markers of Curcumae Radix standard decoction, and the established fingerprint can be used for the quality control of Curcumae Radix standard decoction.

Objective To explore the injury pattern and features of peripheral nerve in congenital muscular dystrophy patients caused by LAMA 2 gene mutation.Method Seventeen patients genetically or molecular pathologically diagnosed as LAMA 2-related congenital muscular dystrophy were recruited in Peking University First Hospital between 2002 and 2015.All the patients received nerve conduction velocity (NCV) and needle electromyography tests . Clinical and laboratory examination data of the patients was retrospectively analyzed .The correlation between the NCV and disease course was determined by Pearson correlation analysis.Additionally, one patient underwent a sural nerve biopsy .Result Among these 17 identified patients (13 male and 4 female), all of them were diagnosed as congenital muscular dystrophy , and all of them underwent electrophysiological examination at ages between 1 month to 6 years. Electromyogram indicated seventeen patients of myogenic damage , of whom 10 cases were complicated with reduced NCV.Twenty-six of 95 analyzed nerves showed NCV slower than the normal average of contemporary in 17%-47%.Correlation analysis between NCV and the disease course indicated that NCV of median nerves, ulnar nerves, tibial nerves and common peroneal nerves were negatively associated with the disease course (r=-0.737, -0.771, -0.540 and -0.682, respectively; all P<0.05).Sural nerve biopsy revealed peripheral neuropathy changes of myelin .Conclusion There is peripheral nerve injury in LAMA 2-related muscular dystrophy patients .It mainly manifests as demyelinating lesions .Moreover, the NCV of peripheral nerve will decrease with the increase of the course of the disease .

Objective To evaluate the predicting value of circulating miRNAs for sepsis secondary to pneumonia in elderly patients.Methods From April 2016 to January 2017,44 cases with sepsis secondary to pneumonia,52 elderly patients with pneumonia and 21 healthy older adults as control were involved in this study.The expression levels of MiRNA-150 5p,miRNA-25-3p,miRNA-122 5p and miRNA-223-3p in plasma were evaluated by fluorescence quantitative PCR.The demographic characteristics,sequential organ failure assessment (SOFA)scores,prognosis and days stayed in ICU were recorded.The area under the receiver operating charaeteristic(ROC)curve was used to calculated the specificity and sensitivity of miRNA in identifying sepsis-associated pneumonia.Results There were significantly differences among levels of circulating miRNA-223-3p in pneumonia,sepsis and healthy control groups(F =36.441,P =0.000),△CT values were 2.39 ± 1.36,1.44± 1.43,and 4.58 ± 0.91,respectively.The relative expression levels of miRNA-223-3p in the three groups were significantly different (P =0.000),which were 0.189 (0.107,0.367),0.361 (0.221,0.735),and 0.044 (0.022,0.061),respectively.The AUC of miRNA-223-3p for predicting sepsis from pneumonia was 0.964(95 ％CI =0.925 1.000).At a cutoff value of 2.759,miRNA-223-3p yielded a sensitivity of 82.9％ and a specificity of 100.0％.Conclusions MiRNA-223-3p expression is up-regulated in patients with sepsis secondary to pneumonia compared to that of patients with pneumonia,and it could be used to predict sepsis associated pneumonia.