Association between Long Interspersed Nuclear Element-1 Methylation and Relative Telomere Length in Wilms Tumor / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>DNA hypomethylation of long interspersed nuclear elements-1 (LINEs-1) occurs during carcinogenesis, whereas information addressing LINE-1 methylation in Wilms tumor (WT) is limited. The main purpose of our study was to quantify LINE-1 methylation levels and evaluate their relationship with relative telomere length (TL) in WT.</p><p><b>METHODS</b>We investigated LINE-1 methylation and relative TL using bisulfite-polymerase chain reaction (PCR) pyrosequencing and quantitative PCR, respectively, in 20 WT tissues, 10 normal kidney tissues and a WT cell line. Significant changes were analyzed by t-tests.</p><p><b>RESULTS</b>LINE-1 methylation levels were significantly lower (P < 0.05) and relative TLs were significantly shorter (P < 0.05) in WT compared with normal kidney. There was a significant positive relationship between LINE-1 methylation and relative TL in WT (r = 0.671, P = 0.001). LINE-1 Methylation levels were significantly associated with global DNA methylation (r = 0.332, P < 0.01). In addition, relative TL was shortened and LINE-1 methylation was decreased in a WT cell line treated with the hypomethylating agent 5-aza-2'-deoxycytidine compared with untreated WT cell line.</p><p><b>CONCLUSION</b>These results suggest that LINE-1 hypomethylation is common and may be linked to telomere shortening in WT.</p>

Identification and analysis of galactose-1-phosphate uridyltransferase gene mutations in children with galactosemia / 中华实用儿科临床杂志

Objective To investigate the mutation of galactose-1-phosphate uridyltransferase gene (GALT gene) of galactosemia children by molecular methods.Methods Two children with galactosemia were investigated.The peripheral blood mononuclear cells were separated and total RNA was extracted.Then,whole cDNAs of GALT were amplified by reverse-transcription polymerase chain reaction;The PCR products were subcloned into T-easy vector and the positive clones were selected and sequenced;meanwhile,the PCR products were also digested by restricted enzymes and identified by restriction fragment length polymorphism.Results Two novel mutations were found in 2 children.In one child,A was changed into G in nucleotide 1006 of GALT gene,which led to amino acid residue M336V mutation.In the other child,A in nucleotide 779 of GALT gene was changed into T and led to amino acid residue H260L mutation.The 2 mutations were both missense mutation and heterozygous mutation.Conclusions Gene diagnosis is an useful method to improve the accuracy of galactosemia diagnosis and will provide valuable references for prenatal diagnosis,hematopoietic stem cell transplantation and gene therapy.

Clinical and genetic characteristics of glucose transporter type 1 deficiency syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>To analyze the clinical and SLC2A1 gene mutation characteristics of glucose transporter type 1 deficiency syndrome.</p><p><b>METHOD</b>The detailed clinical manifestations of six cases were recorded. The laboratory tests including EEG, MRI, blood chemistry, and lumbar puncture were performed. SLC2A1 gene mutations were analyzed by PCR, DNA sequencing and multiplex ligation-dependent probe amplification (MLPA).</p><p><b>RESULT</b>Patient 1, 2 and 3 had classical clinical symptoms including infantile onset seizures, development delay. Patient 4, 5 and 6 had non-classical clinical symptoms including paroxysmal behavior disturbance, weakness, ataxia, lethargy, especially after fasting or exercise, without severe seizures. The plasma glucose levels were normal. The CSF glucose levels decreased in all the six cases, ranged from 1.10 mmol/L to 2.45 mmol/L, the mean level was 1.68 mmol/L. The CSF glucose/plasma glucose ratios decreased, ranged from 0.16 to 0.51, the mean ratio was 0.34. Four patients had normal EEG. Two patients had focal and diffuse epileptiform discharge, and one of them also had paroxysmal occipital or generalized high-amplitude slow waves during awake and sleep time. MRI abnormalities were found in three patients, patient 1 with mild brain atrophy, patient 3 with bilateral ventricle plump, and patient 4 with high signals in T2 in the frontal and occipital white matter, interpreted as hypomyelination. SLC2A1 gene mutations were found in six cases. Patient 1 has large scale deletion in exon 2. In patient 2 to 6, the mutations were c.741 G>A (E247K), 599delA, 761delA, c.1148 C>A (P383H), c.1198 C>T (R400C) respectively. Two patients were treated with ketogenic diet. The seizures disappeared and development became normal. Three patients responded to frequent meals with snacks. One patient refused any treatments, the symptoms continued to exist.</p><p><b>CONCLUSION</b>The clinical manifestations of glucose transporter type 1 deficiency syndrome are varied. The common symptoms included infantile onset seizures and various paroxysmal events. These neurologic symptoms generally fluctuated and were influenced by factors such as fasting or fatigue. This feature could be a very important clue for the diagnosis of GLUT1-DS. Lumbar puncture is recommended in patients with episodic CNS symptoms especially after fasting. GLUT1-DS is a treatable neurometabolic disorder, early diagnosis and treatment may improve the prognosis of the patients.</p>

Role of serum 25-hydroxyvitamin D in the diagnosis of vitamin D deficiency rickets / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the role of serum 25-hydroxyvitamin D in the early diagnosis of vitamin D deficiency rickets.</p><p><b>METHODS</b>Concentrations of serum 25(OH)D, calcium, phosphorus and alkaline phosphatase were measured in normal control (n=73), suspected rickets (n=45) and confirmed rickets groups (n=65). Receiver operating characteristic (ROC) curve was used to evaluate the diagnostic value of serum 25(OH)D for rickets.</p><p><b>RESULTS</b>Serum 25(OH)D levels in the suspected and confirmed rickets groups were 83±30 and 72±31 nmol/L respectively, which was lower than in the normal control group (112±37 nmol/L) (P<0.01). There was no significant difference between the suspected and confirmed rickets groups (P>0.05). Vitamin D deficiency rates in the suspected and confirmed rickets groups were higher than in the control group (P<0.01). The ROC curve area of serum 25(OH)D for the diagnosis of rickets was 0.760 (95%CI 0.692-0.820, P<0.01), and the optimal operating point was 90.70 nmol/L (sensitivity 68.49%, specificity 72.73%). There was no significant difference in levels of calcium, phosphorus and alkaline phosphatase between the three groups (P>0.05).</p><p><b>CONCLUSIONS</b>Serum 25(OH)D levels in infants with suspected and confirmed rickets are significantly reduced and this may reflect vitamin D deficiency . Therefore, it may be useful to check serum 25(OH)D levels in screening for rickets.</p>

Changes in plasma hydrogen sulfide and nitric oxide levels and their clinical significance in children with Kawasaki disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Cardiac involvement is the most common complication of Kawasaki disease (KD); however, the underlying mechanisms are not understood. The present study was designed to investigate changes in plasma hydrogen sulfide (H(2)S) and nitric oxide (NO) levels in the acute and recovery stages of KD children and to examine their clinical significance.</p><p><b>METHODS</b>Thirty-five KD patients and 32 healthy children were enrolled in the study. KD patients were divided into two subgroups: a non-cardiac involvement group and a cardiac involvement group. Plasma H(2)S levels were measured using the sulfur-sensitive electrode method and plasma NO levels and NO synthase activity were determined using the nitrate reductase method both before and after intravenous immune globulin (IVIG) therapy.</p><p><b>RESULTS</b>Plasma H(2)S levels significantly decreased in KD patients during the acute phase of the disease and NO levels were significantly increased, compared with the control group (P < 0.01). After treatment with IVIG, both plasma H(2)S and NO levels significantly increased (P < 0.01). The plasma levels of H(2)S were significantly lower in the cardiac involvement group compared with the non-cardiac involvement group (P < 0.05).</p><p><b>CONCLUSION</b>H(2)S and NO may play a role in the pathophysiological process of inflammation during the acute phase of KD. Endogenous H(2)S may exert protective effects with respect to cardiac complications in KD.</p>

Fronotemporal lobar degeneration: An update / 第二军医大学学报

Frontotemporal lobar degeneration (FTLD) is a non-Alzheimer dementia syndrome characterized by focal atrophy of the frontal and/or temporal lobes. Recently, it has been found that FTLD is related to the degeneration of tau protein, and is closely associated with corticobasal ganglionic degeneration, progressive supranuclear palsy, and motor neuron disease. This article reviews the progress in etiology, genetics, pathology, clinical features, and diagnostic criteria of FTLD.

Unusual patterns of neural tube defects in a high risk region of northern China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the prevalence of different types of neural tube defects (NTDs) in Luliang Prefecture, Shanxi province, where the prevalence of NTDs is unusually high and the correlation between NTDs prevalence and patterns.</p><p><b>METHODS</b>A surveillance population-based birth defects was performed in Luliang Prefecture, Shanxi province.</p><p><b>RESULTS</b>The results of our study showed that the prevalence of NTDs was 2-fold higher in Luliang Prefecture than in other areas of Shanxi province. Unusual patterns of NTDs were found, however, multiple NTDs were relatively common in Luliang Prefecture, accounting for over 13% of all NTDs cases in China.</p><p><b>CONCLUSION</b>The prevalence of NTDs is associated with its patterns.</p>

Recombinant expression of human influenza A virus nucleocapsid protein and its antigenicity analyses / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To prepare anti-recombinant protein antibody from immunized mice with recombinant nucleocapsid protein (NP) of human influenza A3 (IFV-A3) virus expressed in prokaryotic cell, and to explore the feasibility of utilizing anti-recombinant protein antibody to detect influenza A virus.</p><p><b>METHODS</b>NP genes of human influenza A virus were analyzed with computer softwares of ClustalX, Antheprot, et al. to determine the antigenicity in conserved regions. Three different partial NP genes were harvested and cloned into pET-28(c) plasmid, the recombinant plasmids were induced to express partial NP segments in BL21 cells. The recombinant proteins were purified with Ni-agarose by affinity chromatography and immunized BALB/c mice. The polyclonal antisera harvested from mice were analyzed with Western Blot and immunohistochemistry assays to detect the reactions with IFV-A.</p><p><b>RESULTS</b>Three recombinant plasmids were expressed with high yield in BL21 cells, about 15-20 mg/L. Western Blot results indicated that the three prepared antisera (1:2000) positively reacted with NP from IFV-A3-infected cells. And immunohistochemistry assays suggested that anti-NP1, anti-NP2, anti-NP3 antisera positively reacted with IFV-A3 or IFV-A1-infected MDCK cells, with titers of 1:640 to 1:1280.</p><p><b>CONCLUSION</b>The recombinant NP of IFV-A3 would induce polyclonal antibodies with high titers in mice. The polyclonal antibodies would cross-react with IFV-A3 and IFV-A1. It is feasible to predict the antigenicity with systematical bioinformatics analyses and then induce anti-IFV antibodies with high dilutions, and it is possible to be utilized in the early detection and subtyping analyses of IFV-infections.</p>

Meta-analysis of repeated endoscopic sclerotherapy and ligation for prevention of esophageal variceal rehaemorrhagia in cirrhosis / 上海第二医科大学学报

Objective To evaluate the efficacy and safety of repeated endoscopic sclerotherapy injection(ESI) and endoscopic variceal ligation(EVL) in eradication of esophageal varices among patients who survive an episode of first variceal hemorrhage with a high risk of rehaemorrhagia and death. Methods The correlated literatures were identified from Medline,Embsae,the Library Cochrance,PubMed and CNKI.RevMan 4.2 Software provided by the Library Cochrance was used for data analysis. Results A total of 4 randomized controlled trials were included.It was demonstrated that there was no significant difference in the esophageal varices eradication rate between repeated ESI and EVL(OR=0.75,95%CI: 0.48-1.15;P=0.19).However,the rehaemorrhagia rate of EVL after emergency hemostasis was significantly lower than that of ESI(OR=2.19,95%CI:1.44-3.31;P=0.000 2).Meanwhile,there was no significant difference in mortality between ESI and EVL(OR=1.34,95%CI:0.82-2.17;P=0.24).Little publication bias was found with funnel plot analysis. Conclusion EVL outperforms ESL in prevention of rehaemorrhagia in treatment of esophageal varices,while does no better than ESL in eradication of esophageal varices and mortality.

Effect of Seasonal Variation on Blood Lead in Health Children / 实用儿科临床杂志

Objective To explore effects of seasonal variation on blood lead in children.Methods Anodic stripping voltammetry(ASV) was used to detect blood lead level in children.The enrolled children were divided into 4 groups as follows:young children(ages 1 month to 3 years);preschool(ages 3 to 6 years);school(ages 6 to 12 years);teenagers(ages 12 to 18 years),and children′s blood lead level and lead poisoning rates were analyzed in the light of seasonality.Results Total 13 233 children were observed,aged from 1 month to 18 years old,8315 males,4918 females.The average blood lead levels were 60 ?g/L,the 5~(th) and 95~(th) percentage was 19,138 ?g/L.The rate of lead poisoning in children was 14.8%,with the majority of low-grade(?~2=116.3125 P

Polymorphisms of exon 17 of insulin-receptor gene in pathogenesis of human disorders with insulin resistance / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the relationship between polymorphisms of insulin-receptor (INSR) gene and insulin resistance in a population-based study in China.</p><p><b>METHODS</b>Polymerase Chain Reaction (PCR) was used to the amplify Exon 17 of INSR gene and all amplified products were analyzed by direct sequencing.</p><p><b>RESULTS</b>Six single-nucleotide polymorphisms (SNPs) were found at the following loci: T to TC at the locus of 10699 (Tyr(984)), G to GC at the locus of 10731 (Glu(994)), Deletion G at the locus of 10798 (Asp(1017)), C to T/TC at the locus of 10923 (His(1058)), C to CA at the locus of 10954 (Leu(1069)), and T to TA at the locus of 10961 (Phe(1071)), which might not change the amino acid sequence. The data were in agreement with the test of Hardy-Weinberg balance (P > 0.05). Among the 345 cases, all clinical indices were higher in males than in females except for HDL cholesterol (P < 0.05). The proportion of insulin resistance in males (64.4%) was higher than that in females (35.6%, OR = 1.83). It implied that the relative risk of developing insulin resistance in males was 1.83 times as high as that in females. The biochemical indices in different loci on Exon 17 showed that the individuals with deletion G on the locus of 10798 had lower TG (P = 0.052) and higher HDL (P = 0.027) than those without deletion G on the same site. Homa-Index was lower in those with deletion G than in those without deletion G (P > 0.05). After sex stratification in analysis, all allele frequencies on the six loci of SNPs of Exon 17 had different distributions between the insulin resistant group and the control group, but P > 0.05.</p><p><b>CONCLUSION</b>SNPs of Exon 17 of INSR gene are unlikely to play a direct role in the pathogenesis of human disorders with insulin resistance.</p>

Study on the relationship between polymorphism of insulin-receptor gene EXON2-2257 and insulin resistance in Chinese people / 中华流行病学杂志

<p><b>OBJECTIVE</b>To understand the role of insulin-receptor gene in the development of insulin resistance on a population-based study in China.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) was used to amplify the EXON2 of the insulin-receptor gene and all amplified products were analyzed by direct sequencing.</p><p><b>RESULTS</b>Three genotypes of single nucleotide at the site of 2257 in EXON2 of the insulin-receptor gene were identified. In 237 of the 345 cases (68.7%), homozygote genotype of CC phenotype was found with a gene frequency of 0.825; 13 cases (3.77%) showed homozygote genotype of TT phenotype with a gene frequency of 0.175 and the rest 95 cases (27.54%) showed heterozygote genotype of CT phenotype. Data were in agreement with the test of Hardy-Weinberg balance (chi(2) = 0.2898, upsilon = 3 - 2 = 1, 0.5 < P < 0.75). The serum level of triglyceride and the HOMA-IR index, the status of insulin resistance assessed by homeostasis model assessment (HOMA), were lower in the TT or CT genotype group than that in the CC genotype group (P < 0.05). Comparing with CC genotype, the proportion of CT genotype (18.4%) in the insulin resistance group (large than 75th percentile of HOMA-IR) was significantly lower than that in the control group (30.6%) (P = 0.022, OR = 0.493). The proportion of TT genotype (2.3%) in insulin resistance group was lower than that in the control group (4.3%, P = 0.297). logistic analysis revealed that after adjusting possible confounding factors such as taking anti-hypertensive and anti-diabetic medications, serum triglyceride and high density lipoprotein cholesterol (HDL-C), hypertension, BMI, smoking history, the OR value of people in the insulin resistance group with CT genotype was 0.448 (95% CI: 0.214 to 0.940) compared to the group with CC genotype.</p><p><b>CONCLUSION</b>The hybridization CT genotype at the site of 2257 in EXON2 of insulin-receptor gene might have a candidate gene to serve as a protective factor for insulin resistance.</p>

A novel KCNQ1 mutation in Chinese with congenital long QT syndrome / 中华儿科杂志

<p><b>OBJECTIVE</b>Congenital long QT syndrome (LQTS) is an inherited disorder of cardiac repolarization characterized by prolongation of QT interval and polymorphic ventricular tachycardia torsade de pointes (TdP) in the electrocardiogram (ECG). Clinical symptoms include recurrent syncope, seizure or even sudden death. It is caused by mutations of at least seven genes, six of them encoding ion channels that determine the duration of ventricular action potentials. One of these genes, KCNQ1, encodes an alpha-subunit of cardiac slowly activated delayed rectifier potassium channel. Patients carrying mutations of KCNQ1 are named as LQT1, which accounts for 42% of patients with LQTS. This study sought to analyze the clinical data of Chinese with LQTS and to screen for the mutations of KCNQ1.</p><p><b>METHODS</b>The universally accepted phenotypic criteria of LQTS was used for identification of probands. There were six families with LQTS. They were enrolled in this study. Clinical and ECG data of each family member were recorded. Genomic DNA was prepared from peripheral blood lymphocytes. Polymerase chain reaction-single strand conformation polymorphism analysis was used to screen for mutations throughout the whole coding region of KCNQ1 and DNA sequencing was performed to determine the exact mutation site.</p><p><b>RESULTS</b>There were totally 13 patients in the six LQTS families. Five were male and eight female. One suffered from sudden death, 10 had syncope and 2 were asymptomatic. Eleven of the 13 patients had ECG data. Their QT and QTc (mean +/- SD) were (0.460 +/- 0.058) s and (0.516 +/- 0.058) s, respectively. TdP was observed in 3 patients (27%) during the syncope attack. By PCR-SSCP analysis, two novel KCNQ1 deletion mutations 356-357 Delta QQ and 626-631 Delta GSGGPP were identified in 7 patients of 2 families. None of 50 normal individuals carried these mutations, indicating these two mutations were likely to cause the disease. In addition, P448R was found in one affected and some unaffected members in other two families and in 7 of 50 (14%) normal individuals, indicating that this might be a polymorphism. All the three mutations located in C-terminal domain of KCNQ1 protein.</p><p><b>CONCLUSIONS</b>Two novel deletion mutations and one novel polymorphism of KCNQ1 gene were identified among 6 Chinese families with LQTS.</p>