Objective To explore the relationship between X - linked spondyloepiphyseal dysplasia tarda (SEDL) gene escaping X chromosome inactivation( XCI) and SEDL phenotype. Methods RT - PCR was performed on total RNA which was isolated from blood samples of patients, female carriers and controls. Patients and female carriers were selected from the pedigree with SEDL caused by the mutation (IVS2 - 2A→C) of the gene. cDNA was analyzed by polyacrylamide gelelectrophoresis(PAGE). Results PAGE data indicateed that female carriers expressed both normal and mutant SEDL mRNA,meaning the SEDL gene escaping XCI. Family investigation showed carrier females in the SEDL pedigree presented no symptoms. Conclusions The SEDL gene escaping X chromosome in-activation is firstly identified from human body. This may explain that carrier females present no symptoms.

AIM: To prepare and characterize a mouse anti-human CD40 mutant monoclonal mAb. METHODS: Female BALB/c mice of 6-8 weeks old were immunized with CD40 mutant transfectant (L929-CD40mu) as immunogen. The spleen B cells of the mice were fused with Sp2/0 myeloma cells. The hybridoma cells were screened with CD40 mutant transfectant (L929-CD40mu) by FCM. Faststrip analysis was performed to identify Ig subclass of this mAb. The epitope recognized by this mAb was detected by Bio-5C11 competitive assay. Western blot technique was adopted to identify the mAb. The proliferation of tumor cells in vitro was analyzed by MTT assay and apoptosis of tumor cells in vitro was analyzed by PI-annexin V assay. RESULTS: One hybridoma cell line named 10C5 was obtained, which had the property of secreting anti-human CD40 mutant monoclonal antibody continuously and steadily. This mAb specifically recognized human CD40 mutant molecule and induced the apoptosis of tumor cells in vitro. CONCLUSION: One hybridoma cell line which can secret a mouse anti-human CD40 mutant mAb has been prepared successfully. This mAb can inhibit the growth of tumor cells expressing CD40 mutant and induce their apoptosis in vitro.

Objective To investigate the effects of ulinastatin on the expressions of tumor necrosis factor-a (TNF-a) and interleukon-6 (IL-6) in the brain tissue of rats with sepsis. Methods Filly SD rats were randomly divided into control (n=5), sepsis (n=15), ulinastatin pretreatment (n=15) and ulinastatin treatment (n=15) groups. Sepsis was induced in the latter 3 groups by cecal ligation and puncture (CLP), and in the ulinastatin pretreatment and treatment groups, ulinastatin was administered at the dose 25 000 U/kg 2 h before the operation and at 50 000 U/kg 2 h after the operation, respectively. The rats were sacrificed at 3, 6 and 12 h after CLP, and the brain tissues from the left hemisphere was collected for measurement of TNF-a and IL-6 levels by radioimmunity, and those from the right hemisphere was used for pathological examination. Results Compared with control group, the rats in the sepsis group showed obviously increased TNF-a and IL-6 levels in the brain tissues 6 and 12 h after CLP (P<0.05). Ulinastatin treatment before and after the CLP both resulted in significant reduction in TNF-a levels 6 h after CLP in comparison with the levels in the sepsis group (P<0.05), and significant reduction of IL-6 levels occurred till 12 h after CLP (P<0.05). No significant differences in TNF-a and IL-6 levels were noted between ulinastatin pretreatment and treatment groups (P>0.05). Conclusion The inflammatory response caused by elevated TNF-a and IL-6 levels in the brain of septic rats may be an important mechanism of septic encephalopathy. Ulinastatin can reduce TNF-a and IL-6 levels in the brain of septic rats to alleviate sepsis-induced brain injuries, and its therapeutic and prophylactic (at half dose) administration produces similar effects.

AIMTo determine the location and relative quantity of coumarins in the stem of Dendrobium thyrsiflorum Rchb. f. , and to provide a scientific basis for evaluating and utilizing the famous medicinal plant.

METHODSThe stems of one, two and three years old, separately, were collected in February. Location and relative quantity of coumarins in the top, middle and basal parts of each stem sample were determined by using laser scanning confocal microscopy (LSCM). ANOVA and Tukey's test were employed in the statistical analysis.

RESULTSThe coumarins located mainly in vascular bundles, especially on the walls of the outer fiber cells. There was significant difference or great significant difference exist in every part of the stem from different ages. The values of significant difference were as follows: basal part P = 0.004 (< 0.01); middle part P = 0.009 (< 0.01); top part P = 0.036( < 0.05).

CONCLUSIONTwo years old stem could be the best choice when collecting Dendrobium thyrsiflorum in Feburary.

Coumarins ; analysis ; Dendrobium ; chemistry ; Microscopy, Confocal ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Time Factors

OBJECTIVETo study the chemical constituents of Dendobium crystallinum.

METHODCompounds were isolated and purified by silica gel and Sephadex LH-20 column chromatography. Their structures were identified by physicochemical properties and spectral analyses.

RESULTNine compounds were obtained and identified as: 4, 4'-dihydroxy-3, 5-dimethoxybi-benzyl (1), gigantol (2), naringenin (3) , p-hydroxybenzoic acid (4), n-tetracosyl trans-p-cou-marate (5), n-octacosy trans-p-coumarate (6), n-hexacosyl trans-ferulate (7), stigmasterol (8), daucosterol (9).

CONCLUSIONAll these compounds were obtained from this plant for the first time, compounds 1 and 4 were isolated firstly from the genus.

Bibenzyls ; Chromatography ; Dendrobium ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; chemistry ; Guaiacol ; analogs & derivatives ; chemistry ; Magnetic Resonance Spectroscopy ; Parabens ; chemistry ; Sitosterols ; chemistry ; Stigmasterol ; chemistry

OBJECTIVETo explore the methylation status in promoter region of norepinephrine transporter gene (NET, SLC6A2) in heart failure ( HF) patients and its correlation with qi deficiency/blood stasis syndrome (QDS/BSS).

METHODSThirty-six patients with heart failure (NYHA classification III to IV) were recruited in the study (as the heart failure group) and their scores of QDS/BSS were evaluated. Besides, a healthy elderly group (30 cases) and a healthy youth group (30 cases) were also set up. They were recruited from Physical Examination Center of Fujian Provincial Hospital. Pyrosequencing was applied to detect the methylation in promoter region of SLC6A2 gene, and the total methylation index (MTI) of CpG island was calculated. The correlation between the methylation status in promoter region of SLC6A2 and scores of QDS/BSS was assessed using Pearson and Partial analyses. Risk factors were screened and adjusted using Logistic regression.

RESULTSBy one-factor analysis of variance, the total MTI in the HF group (219.72% ± 54.03%) was obviously higher than that in the healthy elderly group (194.47% ± 34.92%) and the healthy youth group (161.60% ± 41.11%) (all P < 0.05). Meanwhile, the total MTI was higher in the healthy elderly group than in the healthy youth group (P < 0.01). By covariance analysis , after controlling age and BMI, the total MTI was higher in the HF group than in the healthy elderly group (P = 0.041), while it was higher in the healthy elderly group than in the healthy youth group (P = 0.016). Age was found to play an essential role in affecting MTI of SLC6A2 gene promoter region among the 3 groups (F = 16.447, P = 0.01). The total MTI was quite lower in the healthy youth group. Results of Partial correlation analysis showed MTI was positively correlated with scores of qi deficiency and blood stasis respectively (r = 0.494 and 0.419 respectively, both P < 0.05). Logistic regression analysis showed after adjusting confounding factors, the relative risk (OR value) of total MTI of SLC6A2 gene in promoter region was 1.038 (95% CI, 1.006 to 1.071, P = 0.020).

CONCLUSIONSAbnormally elevated methylation of the promoter region of SLC6A2 gene is one of risk factors for HF. In addition, the degree of methylation of the promoter region of SLC6A2 gene was positively correlated with the severity of QDS/BSS.

Adolescent ; Aged ; DNA Methylation ; Heart Failure ; genetics ; physiopathology ; Humans ; Logistic Models ; Medicine, Chinese Traditional ; Norepinephrine Plasma Membrane Transport Proteins ; genetics ; Promoter Regions, Genetic ; Qi

OBJECTIVETo investigate the relative contribution of lifestyle and obesity to the risk of developing type 2 diabetes.

METHODSAll baseline survey data were based on the program Prevention of Multiple Metabolic Disorders and Metabolic Syndrome in Jiangsu Province (PMMJS) which was conducted during April 1999 to May 2004. In the baseline survey, 8685 participants were selected using multi-stage sampling method. Frem March 2006 to November 2007, 4582 participants who had been in the study for at least 5 years were included in the follow-up survey. A total of 3847 participants were followed and of them 3461 non-diabetic subjects were included in this analysis. High fat diet or not, low fiber diet or not, sedentary or not and occupational physical activity classification were defined as lifestyle variables and the incidence of type 2 diabetes at follow-up survey was defined as outcome variable. It was prospectively examined that the separate and joint association of lifestyle and obesity with the development of type 2 diabetes in subjects recruited from PMMJS, using logistic regression model.

RESULTSA total of 162 incident cases of type 2 diabetes during 6.3 years of follow-up in total 3461 participants were documented. The incidence rate was 4.7%. After adjusted for sex, age, family history of diabetes, blood pressure, lipids and fast plasma glucose, risk of type 2 diabetes increased with lighter occupational physical activity (compared with vigorous group, moderate group aRR = 2.15, 95%CI: 1.26 - 3.68; light group aRR = 2.39, 95%CI: 1.12 - 4.87), sedentary lifestyle (aRR = 2.94, 95%CI: 1.90 - 4.54), low fiber diet (aRR = 1.58, 95%CI: 1.01 - 2.53), overweight (aRR = 1.33, 95%CI: 1.01 - 1.90) and obesity (aRR = 1.59, 95%CI: 1.07 - 3.75). In joint analysis of lifestyle and obesity, the impact of sedentary lifestyle (in BMI < 25 group, aRR = 3.42, 95%CI: 1.99 - 5.86; in BMI ≥ 25 group, aRR = 2.41, 95%CI: 1.13 - 5.12) and low fiber diet (in BMI < 25 group, aRR = 1.42, 95%CI: 0.81 - 2.54; in BMI ≥ 25 group, aRR = 2.63, 95%CI: 1.15 - 6.03) on diabetes were independent of overweight and obesity. When stratified by sedentary lifestyle or low fiber diet, there was no association between overweight/obesity and diabetes risk (sedentary aRR = 2.04, 95%CI 0.87 - 4.71, non sedentary aRR = 1.21, 95%CI: 0.82 - 1.78; non low fiber diet aRR = 1.26, 95%CI: 0.87 - 1.84, low fiber diet aRR = 1.88, 95%CI: 0.80 - 4.80).

CONCLUSIONUnhealthy lifestyle, overweight and obesity independently increase the risk of type 2 diabetes. The magnitude of risk contributed by sedentary lifestyle and low fiber diet are much greater than that imparted by overweight and obesity.

Adult ; Aged ; Body Mass Index ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; epidemiology ; Female ; Humans ; Life Style ; Male ; Middle Aged ; Obesity ; epidemiology ; Overweight ; epidemiology ; Risk Factors ; Sedentary Lifestyle

Objective To investigate the application of sandwich teaching method based on CBL principle in clinical teaching of pediatric surgery combined with network database. Methods A total of 147 undergraduate students in clinical medicine were selected and randomly divided into experimental group (84 persons) and control group (63 persons). The experiment group used sandwich teaching based on CBL principle + bedside demonstration + network resource library, while the control group adopted tradi-tional teaching + bedside teaching. The teaching effect was evaluated through theoretical and practical ex-aminations and quantitative feedback questionnaires. The t-test of independent samples was performed in two groups, and the scores of the questionnaires were matched by paired t-test. Results The scores of net-work test, case discussion, summary and written test in the experimental group were better than those in the control group [(37.56 ±1.32) vs. (35.10 ±1.25); (27.61 ±0.94) vs. (24.71 ±1.51); (27.02 ±1.11) vs. (26.29±1.30); (88.28±1.97) vs. (83.60±4.58)], the difference was statistically significant. The question-naire survey showed that the students in the experimental group were better than the control group in the learning efficiency, team cooperation, independent thinking, theory connection, and so on. Conclusions The sandwich teaching method based on CBL principle, combined with the network database, is better than the traditional method in the clinical teaching of pediatric surgery, and can promote the theoretical and overall teaching effect, which is worth popularizing.

The purpose of this study is to analyze the family's clinical data of 22 children who were given an intended clinical diagnosis of Duchenne muscular dystrophy (DMD), and to explore the clinical value of next-generation sequencing (NGS) in the molecular diagnosis of DMD. The probands were simultaneously tested by NGS for a gene panel associated with hereditary neuromuscular disease and multiplex ligation-dependent probe amplification (MLPA) for the Dystrophin gene. The exon deletion/repetition mutations of the Dystrophin gene determined by both methods were compared and the point mutations of the Dystrophin gene were verified by Sanger sequencing. Dystrophin gene mutations were found in all the 22 probands, including 14 exon deletion/repetition mutations and 8 point mutations/minor variations. The results of MLPA detection were consistent with those of NGS. The results of Sanger sequencing showed that the point mutations and minor variations determined by NGS were correct. One missense mutation (c.6290G>T), 1 nonsense mutation (c.3487C>T) and 4 minor deletion-induced frameshift mutations (c.1208delG, c.7497_7506delGGTGGGTGAC, c.9421_9422delAA and c.8910_8913delTCTC) had not been reported in the Human Gene Mutation Database, and thus were considered as novel mutations of the Dystrophin gene. The results of this study showed that NGS can detect variations in the Dystrophin gene, including exon deletion/repetition, point mutation, minor deletion and intron mutation. Therefore, NGS is of certain clinical value in the molecular diagnosis of DMD and is worthy of recommendation.
Dystrophin ; Exons ; High-Throughput Nucleotide Sequencing ; Humans ; Muscular Dystrophy, Duchenne ; Mutation

Objective To study the impact on dynamic change of waist circumference(WC) through follow-up data on the incidence of hypertension in several cohort groups.Methods In this prospective study,2778 free-hypertension subjects were recruited from a program “Prevention of Multiple Metabolic disorders and metabolic syndrome (MS) in Jiangsu province” (PMMJS) to evaluate the risk of hypertension in relation to WC dynamic change on normal WC or abnorminal obesity group.Dynamic change of WC was measured by WC D-value,which was expressed by data on the difference of WC between baseline and the first follow up.Study outcome was defined as incident hypertension during the first to the second follow up period in this study.The association between dynamic change of WC and incident hypertension was analyzed by using Cox proportional hazards regression model.Results There were 2778 participants,660 subjects developed hypertension during the follow-up,regardless of the normality of the baseline WC,the risk of hypertension increased across the tertiles of WC,while the incidence of hypertension was higher in non-control group than that in control group.In populations with abdominal obesity and normal WC at baseline,RRs (95％CI) of hypertension were 1.95 (1.19-3.19) and 2.38 (1.89-2.99) in subjects with abdominal obesity seen at the first follow up period,compared to subjects with normal WC in the same period.After adjustment for gender,age and other hypertension related risk factors,in populations with abdominal obesity and normal WC at baseline survey,RRs (95％ CI) of hypertension were 4.36 (1.69-9.74) and 1.44( 1.03-2.35 ) respectively,for the non-control group.Conclusion WC dynamic change was associated with hypertension,WC control while WC reduction was important for early prevention on hypertension.