OBJECTIVETo investigate mutations of anaplastic lymphoma kinase (ALK) in Chinese children with neuroblastoma (NB).

METHODSGenomic DNA was extracted from 22 cases of paraffin-embedding NB tumor tissues. Gene mutations in the exons 20-26 which were mutational hotspots of ALK were analyzed by PCR-DNA direct sequencing.

RESULTSA novel synonymous mutation C3586T (Leu1196Leu) and a known synonymous mutation C3375A (Gly1125Gly) were found and located at exon 23 and exon 21 of ALK respectively. There were 10 cases (46%) of known synonymous mutation C3375A in 22 cases of NB. The C3375A allelic frequency was 27%. No statistically significant correlation was found between mutation C3375A and clinical parameters of NB such as age, sex, metastasis and tumor differentiation. Mutation was not found in the other 5 exons.

CONCLUSIONSA novel ALK gene synonymous mutation C3586T was identified using PCR-DNA sequencing. A known mutation C3375A in ALK was successfully identified in children, and its incidence is not influenced by the clinical features of childhood NB.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation ; Neuroblastoma ; genetics ; Polymerase Chain Reaction ; Receptor Protein-Tyrosine Kinases ; genetics

OBJECTIVE: To study the types and characteristics of TUBB1 mutation in children with congenital hypothyroidism (CH) and thyroid dysgenesis (TD) in Shandong, China. METHODS: Mutations of the whole coding region of the TUBB1 gene were analyzed for 289 children with CH and TD in Shandong. Whole-genome DNA was extracted from peripheral blood leukocytes. PCR multiplication was performed for the whole coding region of the TUBB1 gene. Sanger sequencing was performed for the PCR products, and a biological information analysis was performed. RESULTS: Among the 289 children with CH and TD, 4 (1.4%) were found to have a c.952C>T(p.R318W) heterozygous mutation in the TUBB1 gene, resulting in the change of tryptophan into arginine at codon 318 of TUBB1 protein. This mutation was evaluated as "potentially pathogenic" based on the classification criteria and guidelines for genetic variation by American College of Medical Genetics and Genomics. CONCLUSIONS A novel mutation is detected in the exon of the TUBB1 gene in children with CH and TD in Shandong, suggesting that the TUBB1 gene may be a candidate pathogenic gene for CH children with TD.
Child ; China ; Congenital Hypothyroidism ; genetics ; DNA Mutational Analysis ; Humans ; Mutation ; Thyroid Dysgenesis ; genetics ; Tubulin ; genetics

OBJECTIVETo investigate the distribution of γ-glutamyl hydrolase gene (GGH) 452C/T genotype and allele frequency in children with acute leukemia (AL) and healthy children.

METHODSBone marrow samples from 92 children with AL and peripheral blood samples from 124 healthy children were obtained to prepare complementary DNAs (cDNAs). The cDNAs were analyzed for a GGH 452C/T polymorphism by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis (RT-PCR-DGGE) and direct sequencing.

RESULTSThe frequencies of the AL patients with TT, CT and CC genotypes were 2.2%, 13.0% and 84.8%, and the frequencies of the control children were 1.6%, 16.9% and 81.5%, respectively. There was no significant difference in GGH genotype or T allele frequency between the two groups (P> 0.05). However, the T allele frequency in Han Chinese children was significantly different from those reported in Japanese, Mexican and African-American populations.

CONCLUSIONThe frequency of 452C/T polymorphism of GGH gene in Han Chinese children has been determined. The results suggested that an ethnic difference may exist.

Acute Disease ; Base Sequence ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genotype ; Humans ; Infant ; Infant, Newborn ; Leukemia ; enzymology ; genetics ; Male ; Polymorphism, Single Nucleotide ; gamma-Glutamyl Hydrolase ; genetics

OBJECTIVETo study the association between methylenetetrahydrofolate reductase (MTHFR) gene polymorphisms and toxicities after high-dose methotrexate (HD-MTX) infusion in children with acute lymphocytic leukemia (ALL).

METHODSMTHFR variants in 52 children with ALL were determined by reverse transcriptase-polymerase chain reaction-denaturing gradient gel electrophoresis and sequencing. Toxicities of children who received HD-MTX chemotherapy were evaluated according to the National Cancer Institute-Common Toxicity Criteria (NCI-CTC).

RESULTSThe children carrying MTHFR 1298AC had a higher risk of developing thrombocytopenia compared with the carriers of the 1298 AA genotype (OR=13.7, 95%CI=1.18-159.36, P=0.036). There was no significant difference in HD-MTX chemotherapy-related adverse effects between the patients with different MTHFR C677T or G1793A genotypes.

CONCLUSIONSMTHFR A1298C polymorohism may associate with the toxicity of HD-MTX chemotherapy in children with ALL.

Antimetabolites, Antineoplastic ; adverse effects ; Child ; Child, Preschool ; Female ; Genotype ; Humans ; Male ; Methotrexate ; adverse effects ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Polymorphism, Genetic ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo explore the prophylactic effect of acupuncture Neiguan (PC 6) on nausea and vomiting after laparoscopic operation.

METHODSOne hundred patients with laparoscopic gastrointestinal operation were randomly divided into an acupuncture group and a control group, 50 patients in each group. The operation was carried out with the combined infusion and inhalation anesthesia. The patients in the acupuncture group were being punctured at bilateral Neiguan (PC 6) before anesthesia and during the operation. The needles were extracted after operation, and the acupoints were covered with opaque tape. In contrast, the patients in the control group only accepted tape covering without acupuncture. After operation, all patients were given the self-controlled intravenous analgesia, and followed up at 6 h, 12 h, 24 h, 48 h for recording the incidence rate of the nausea, retching and vomiting, then scoring with VAS.

RESULTSAt 6 h, 12 h, 24 h, 48 h after operation, in the acupuncture group, the incidence rates of the nausea were 12.0%, 6.0%, 6.0% and 2.0%, and the incidence rates of the retching were 0, 0, 2.0% and 2.0%, respectively; in the control group, the incidence rates of the nausea were 28.0%, 20.0%, 12.0% and 2.0%, and the incidence rates of the retching were 2.0%, 6.0%, 2.0% and 0, respectively. At 6 h, 12 h after operation, the incidence rates of the nausea and retching in the acupuncture group were lower than those of the control group (P < 0.05, P < 0.001). The vomiting was not happened in both groups. There was no difference between the two groups according to the scoring with VAS.

CONCLUSIONAcupuncturing at Neiguan (PC 6) can reduce the incidence rates of the patients' nausea and retching after laparoscopic operation, especially in 24 h.

Acupuncture Therapy ; Adult ; Aged ; Analgesics ; adverse effects ; Female ; Humans ; Laparoscopy ; Male ; Middle Aged ; Nausea ; prevention & control ; therapy ; Postoperative Complications ; therapy ; Vomiting ; prevention & control ; therapy

BACKGROUNDInflammation plays a pivotal role in cardiac remodeling, especially in myocardial fibrosis. Abnormal growth of cardiac fibroblasts is critically involved in the pathophysiology of cardiac hypertrophy/remodeling. Previous study has demonstrated that many inflammation stimulating factors trigger transforming growth factor-β (TGF-β) induction and reactive myocardial fibrosis. Activin A (ACT A) is a member of TGF-β superfamily, and follistatin (FS) is an activin-binding protein, i.e. an antagonist of ACT A. Our previous studies have shown that ACT A-FS imbalance occurs in rats with heart failure (HF), and overexpression of ACT A can lead to ventricular remodeling, and resultant HF. Low expression of FS after myocardial infarction further exacerbated HF. The pathogenic change resulting from overexpression of ACT A is consistent with that of overexpression of angiotensin II (AngII). Ventricular remodeling includes cardiocyte remodeling and myocardial interstitial collagen deposition and fibrosis. Therefore, the present study was designed to investigate the effects of inflammatory factors on the ACT A-FS and the secretions of cardiac fibroblasts in order to explore in depth the mechanism of myocardial fibrosis.

METHODSA rat model with HF was established, and the results showed that there was a greater degree of cardiac fibrosis in HF rats. In addition, we found that there was an imbalance of the ACT A/FS system in HF rats, which was characterized by increased levels of ACT A. Further, primary rat cardiac fibroblasts were cultured and the MTT assay was performed to determine the effect of the inflammatory factor-bacterial endotoxin lipopolysaccharide (LPS) on cardiac fibroblast proliferation.

RESULTSThe results showed that LPS can stimulate the cardiac fibroblasts to proliferate in a dose-dependent manner. Cellular immunohistochemical staining showed that the rat cardiac fibroblasts themselves could express ACT A and FS proteins, and stimulation by LPS could apparently promote the cultured primary rat cardiac fibroblasts to secrete ACT A, but inhibit the secretion of FS. The results also showed that ACT A promoted, in a dose-dependent manner, the proliferation of the cultured primary rat cardiac fibroblasts, and the expression of collagen types I and III. Moreover, ACT A promoted, in a dose dependent manner, the cardiac fibroblasts to secrete nitric oxide (NO), and unregulated the expression of inducible nitric oxide synthase (iNOS) mRNA.

CONCLUSIONSThese results suggest that the inflammatory mediator LPS can promote ACT A-FS imbalance in cardiac fibroblasts, mainly overexpression of ACT A. Overexpression of ACT A promotes the proliferation and the secretion of collagens in cardiac fibroblasts through autocrine/paracrine stimulation of NO, and is involved in the pathological process of myocardial fibrosis.

Activins ; genetics ; metabolism ; Animals ; Cell Proliferation ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Female ; Fibroblasts ; cytology ; drug effects ; Follistatin ; genetics ; metabolism ; Immunohistochemistry ; Lipopolysaccharides ; pharmacology ; Myocardium ; cytology ; Nitric Oxide ; metabolism ; Rats ; Rats, Wistar ; Real-Time Polymerase Chain Reaction ; Ventricular Remodeling ; drug effects

Alkaloids ; chemistry ; isolation & purification ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Antiviral Agents ; pharmacology ; Carbolines ; chemistry ; isolation & purification ; pharmacology ; Humans ; Molecular Structure ; Urochordata ; chemistry

OBJECTIVETo investigate the changes of EEG approximate entropy (ApEn) in rats during focal cerebral infarction.

METHODSTwenty-four Sprague-Dawley (SD) rats were randomly divided into infarction group (n=12) with middle cerebral artery occlusion and sham-operated group (n=12). The EEG data (ApEn) was recorded in the bilateral areas (C3, C4) of the rats with focal cerebral infarction before the infarction and immediately and at 5, 15, 30, and 60 min after the infarction. The same measurement was carried out in the sham-operated group.

RESULTSIn the sham-operated group, ApEn in C3 and C4 showed no obvious differences at the time points (P>0.05), but in the infarction group, ApEn in C3 and C4 increased significantly after the infarction. ApEn in the ischemic area (C4) was significantly lower than that in the non-ischemic area (C3) (P<0.05). The bilateral ApEn decreased with the passage of time. ApEn in the ischemic area (C4) was significantly lowered at 30 min after the infarction in comparison with that before infarction (P<0.05). In the sham-operated group, ApEn showed no significant difference between C3 and C4. ApEn was comparable between the two groups before the operation.

CONCLUSIONApEn can help monitor the occurrence of focal cerebral infarction of rats, and may be used to assess the extent of cerebral ischemia after infarction.

Animals ; Cerebral Infarction ; physiopathology ; Electroencephalography ; Entropy ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the methylation status of 5' CpG island of fragile histidine triad (FHIT) gene in plasma and the expression of FHIT protein in cancer tissue of cervical cancer patients.

METHODSMethylation-specific PCR (MS-PCR) was employed to examine methylation of FHIT gene in 151 plasma samples before treatment. The immunohistochemistry was used to the expression of FHIT protein in cancer tissues.

RESULTSCpG island methylation of FHIT was detected in 31.13% of plasma samples. The expression of FHIT protein was decreased or discarded in 59.60% of cervical cancer tissues. Among them 47.78% was included in methylation positive samples.

CONCLUSIONCpG island methylation of FHIT gene in plasma plays an important role on cervical cancer, which results in decreased expression of FHIT protein. It can be used to diagnose and evaluate the effect of treatment to cervical cancers.

Acid Anhydride Hydrolases ; blood ; genetics ; metabolism ; Adult ; Aged ; DNA Methylation ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasm Proteins ; blood ; genetics ; metabolism ; Polymerase Chain Reaction ; Uterine Cervical Neoplasms ; blood ; genetics ; metabolism

Objective To explore the spatiotemporal distribution pattern, and identify risk cluster of esophageal cancer in Huai’an City so as to provide evidence for control and prevention of esophageal cancer. Methods Data of esophageal cancer incidence at township level in Huai’an City from 2011 to 2015 was collected. Spatial autocorrelation and local indications of spatial autocorrelation (LISA) were implemented to evaluate the spatial pattern of esophageal cancer incidence. Spatial scan statistics was used to examine spatio-temporal clustering of risk areas. Results The average incidence of esophageal cancer in Huai’an from 2011 to 2015 was 67.12/10 million, the incidence of male was significantly higher than that of female. The results of Moran’s I values implyed the spatial autocorrelation at township level. The results of LISA indicated that there were local hot spots and cold spots. The significant high-risk clusters included townships in Huai’an County, Huaiyin County and Jinhu County. The low-risk clusters were located in the main urban area and Xuyi County. Conclusions There are significant spatio-temporal aggregation for the distribution of incidence of esophageal cancer in Huai’an City and same spatiotemporal high-risk clusters between male and female. Our findings have a foundation to explore the multi-factorial etiology of esophageal cancer and have vital practical value for health services and policies implementation.