OBJECTIVE: To investigate characteristics of molecular etiology of children with profound sensorineural hearing loss in Hubei province, and to provide reference for deafness treatment and genetic counseling. METHOD: Three hundred and six children with profound sensorineural hearing loss in Hubei province were enrolled, their genomic DNA were extracted from peripheral blood and a deafness gene test chip was used to screen nine hot spot mutation in the GJB2, GJB3, SLC26A4, and mitochondria 12SrRNA gene. All patients with SLC26A4 gene mutation were given temporal bone CT scan. RESULT: One hundred and thirty-two (43.14%) out of 306 children were found carrying at least one pathogenic gene mutation. The mutation rates of GJB2, SLC26A4 and mitochondria DNA 12SrRNA gene were 29.41% (90/306), 13.72% (42/306) and 0.65% (2/306), respectively. None out of 306 children was detected GJB3 gene mutation. Thirty-six patients carrying SLC26A4 gene mutation were detected enlarged vestibular aqueduct by CT scan. CONCLUSION Mutations of GJB2 and SLC26A4 gene are two major pathogenic gene for genetic hearing loss in children. 235delC mutation is the main mutation type, followed by IVS7-2A> G mutation type. The screening of SLC26A4 gene common mutations contribute to the diagnosis of enlarged vestibular aqueduct syndrome.
Adolescent ; Child ; Child, Preschool ; China ; Connexin 26 ; Connexins ; genetics ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; Genetic Testing ; Humans ; Infant ; Male ; Membrane Transport Proteins ; genetics ; Mutation ; Oligonucleotide Array Sequence Analysis ; Sulfate Transporters

Recombination plasmid pMM085 possessed both immunogens heat-labile enterotoxin(LT) and fimbriae antigen K88 of enterotoxigenic Escherichia coli (ETEC). Althouth vaccine strain MM-3 carrying pMM085 had good effect to protect piglets against diarrhea due to ETEC infections,it was not ideal live vaccine for pMM085 bringing chloramphenicol resistance gene (cat). To solve the problem,the host-plasmid balanced lethal system was introduced which including the replacement of cat gene by asd gene and transformation the new plasmid to the strain X6097 which asd gene was knocked out in its chromosome. Considering pMM085 was a big plasmid (23kb) and traditional genetic manipulations was not easy to carry on,?-Red recombination system was adopt in this work to realize the replacement of cat gene by asd gene. The results indicated that ?-Red recombination system was convenient and efficient to reconstruct big plasmid.

OBJECTIVETo study the effect of qingxin kaiqiao formula and saponin on the learning and memory ability and the expression of the apoptosis signal transducers Abeta and betaAPP in AD rat brain.

METHODThe comparative observation method was adopted for the animal test. Forty male SD rats were randomly divided into five groups, namely the normal group, the model group, the aricept group, the qingxin kaiqiao formula group and the saponin group, with eight rats in each group. Abeta(25-35) (10 g x L(-1)) was injected into their bilateral amygdala to establish the AD rat model. Since the next day, they were intragastrically administered with Aricept (1.67 mg x kg(-1)), Qingxin Kaiqiao decoction (12.67 mL x kg(-1)), saponin (6.30 mg x kg(-1)) and double distilled water filling for 2 weeks to observe their spatial memory ability in a Morris water maze and study the expression of Caspase-3, Abeta and betaAPP in brain tissues by immunohistochemistry.

RESULTEach traditional Chinese medicine groups showed significant improvement in the learning and memory ability of AD rats and notable differences (P < 0.05, P < 0.01) compared with the control group. The qingxin kaiqiao formula group and the saponin group showed a decrease in the expressions of Caspase-3, Abeta and betaAPP in cerebral cortex and hippocampus area, displaying notable differences (P < 0.01, P < 0.05) compared with the control group.

CONCLUSIONqingxin kaiqiao formula and saponin can obviously improve the learning and memory ability of AD rats with by decreasing the expression of Caspase-3, Abeta and betaAPP in cortex and hippocampus.

Alzheimer Disease ; drug therapy ; genetics ; Amyloid beta-Peptides ; genetics ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cerebral Cortex ; drug effects ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Hippocampus ; drug effects ; metabolism ; Learning ; drug effects ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Rats ; Rats, Sprague-Dawley ; Saponins ; administration & dosage ; pharmacology ; Time Factors

OBJECTIVETo analyze and compare molecular mechanisms of active ingredients of honeysuckle (chlorogenic acid, CGA) with bovine lactoferrin (BLF) or bovine serum albumin (BSA).

METHODThe spectral experiment and the computer analog technology were combined to determine the binding parameters, energy transfer parameters and thermodynamic functions between CGA and proteins, study the molecular mechanism, and compare the differences in interactive mechanism between CGA and BLF or BSA.

RESULTThe interactive mechanism between CGA and BLF or BSA was a dynamic molecular mechanism, whereas the static quenching mechanism existed between the interaction of CGA and BSA, with differences in the bonding intensity due to difference temperature. The binding distance r between CGA and BLF/BSA was very short, indicating the phenomenon of energy transfer. The results of the molecular modeling showed that the main interaction force between CGA and BLF or BSA was hydrogen bonds, together with Van der Waals' forces and hydrophobic effect.

CONCLUSIONThe computer analog shows consistent results with spectral experiment.

Absorption ; Animals ; Cattle ; Drugs, Chinese Herbal ; chemistry ; metabolism ; Humans ; Hydrophobic and Hydrophilic Interactions ; Lactoferrin ; chemistry ; metabolism ; Lonicera ; chemistry ; Models, Molecular ; Protein Binding ; Protein Conformation ; Serum Albumin, Bovine ; chemistry ; metabolism ; Thermodynamics

OBJECTIVETo show the space structures of Mingmen (GV 4) area and its adjacent tissues.

METHODSA frozen female corpse was cut into about 0.5 mm thick sections with a high-speed steel plane machine and photographed with digital and optical camera. Two 2-D transverse phase picture data sets were obtained and then were sequenced, matched, modified and divided. The 3-D reconstruction software was used to reconstruct the outline form of the lumbar part containing Mingmen (GV 4) to attain transverse phase picture data sets (3 022 sheets, 0.57 mm in thickness). The sagittal, coronal and different oblique plane were cut and the tissue structures in these pictures were analyzed. Thus, the reconstruction data sets were obtained.

CONCLUSIONAfter the structures of Mingmen (GV 4) were reconstructed with computer, the form of Mingmen (GV 4) can be dynamically observed from any angle and the space construction law of gross form of Mingmen (GV 4) can be proved.

Lumbosacral Region ; Software ; Tomography, X-Ray Computed

Objective: To observe the clinical efficacy of Zhen'ai needling method in Nei Jing (Classic of Internal Medicine) for children with allergic rhinitis (AR) accompanied by adenoid hypertrophy (AH). Methods: A total of 74 children who met the screening criteria were divided into a Zhen'ai group and a control group by the random number table method, with 37 cases in each group. The control group was treated with acupuncture at Zusanli (ST 36), Hegu (LI 4), Yingxiang (LI 20), Juliao (ST 3), Yintang (GV 29), Shangxing (GV 23) and Baihui (GV 20). The Zhen'ai group added points of Zhen'ai needling method {Shanglianquan [Extra, located at 1 cun above Lianquan (CV 23)], Tianrong (SI 17) and Lieque (LU 7)} in addition to the points in the control group. The needles were retained for 30 min. The treatment was performed twice a week. The total nasal symptom score (TNSS), sino-nasal outcome test-20 (SNOT-20) and symptom scale for AH (SSAH) were assessed before and after 10 treatments. The clinical efficacy of the two groups was compared after treatment. Results: During the treatment, 5 cases dropped out in the control group and 2 cases in the Zhen'ai group. After treatment, the total effective rate of the Zhen'ai group was 94.3％, versus 93.8％ in the control group. There was no significant difference between the two groups (P>0.05). The markedly effective rate of the Zhen'ai group was 42.9％, versus 12.5％ in the control group, and the difference was statistically significant (P<0.05). After treatment, there were significant intra-group differences in the scores of TNSS, SNOT-20 and SSAH in both groups (all P<0.05); the scores of SNOT-20 and SSAH in the Zhen'ai group were lower than those in the control group, and the differences between the groups were statistically significant (both P<0.05). Conclusion: Both conventional acupuncture and conventional acupuncture plus Zhen'ai needling method can improve clinical symptoms of children with AR accompanied by AH; and conventional acupuncture plus Zhen'ai needling method has a better effect than conventional acupuncture in improving AH symptoms.

AIM: To study the chemical constituents of the roots of Ilex asprella Champ. ex Benth. METHODS: Compounds were isolated by silica gel, ODS, and Sephadex LH-20 column chromatography, and their structures were elucidated on the basis of physicochemical properties and spectroscopic analysis. RESULTS: Four triterpenoid glycosides were isolated and identified as (3β)-19-hydroxy-28-oxours-12-en-3-yl β-D-glucopyranosiduronic acid n-butyl ester (1), ilexasoside A (2), monepaloside F (3), and ilexoside A (4). CONCLUSION Compound 1 is a new triterpenoid glycoside, and compounds 3 and 4 were isolated from this plant for the first time.
Glycosides ; chemistry ; isolation & purification ; Ilex ; chemistry ; Molecular Structure ; Plant Extracts ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Triterpenes ; chemistry ; isolation & purification

OBJECTIVETo investigate the association between genotype and phenotype of microdeletion and microduplication syndromes (MMSs) and the pathogenesis of pathogenic copy number variations (CNVs).

METHODSA total of 50 children with MMSs diagnosed by chromosomal microarray analysis (CMA) from June 2013 to September 2015 were enrolled, and the clinical manifestations and features of pathogenic CNVs were analyzed.

RESULTSThe main clinical manifestations of children with MMSs included mental retardation, developmental delay, short stature, and unusual facies, with the presence of abnormalities in multiple systems. There were 54 pathogenic CNVs in total, consisting of 36 microdeletion segments and 18 microduplication segments, with sizes ranging from 28 kb to 48.5 Mb (mean 13.86 Mb). Pathogenic CNVs often occurred in chromosomes X, 15, and 1.

CONCLUSIONSThe clinical manifestations of MMSs are not specific, and a genotype-first approach can be used for diagnosis. Mode of inheritance, type of recombination (deletion or duplication), size of segment, and functional genes included helps with the interpretation of CNVs of de novo mutations, and in-depth research on rare pathogenesis may become breakthrough points for the identification of new MMSs.

Adolescent ; Child ; Child, Preschool ; Chromosome Deletion ; Chromosome Duplication ; DNA Copy Number Variations ; Developmental Disabilities ; genetics ; Female ; Humans ; Infant ; Intellectual Disability ; genetics ; Male ; Phenotype ; Retrospective Studies ; Syndrome

This study was purposed to explore the clinical characteristics, therapy and prognosis of patients with extranodal NK/T cell lymphoma (ENKL). 47 patients with ENKL from October 1995 to December 2008 in our hospital were analyzed retrospectively. The survival of patients was analyzed by using Kaplan-Meier methods, the prognosis of patients was evaluated by multivariate analysis using COX regression model. The clinical parameters used included CD56, Ann Arbor stage, international prognostic index (IPI) and B symptom. The results showed that the 2-year and 5-year overall survival (OS) rates were 91%, 71% respectively. Multivariant analysis by COX regression showed the CD56 and Ann Arbor stage were independent prognostic factors. Single factor analysis with staging in CHOP chemotherapy group indicated that more than stage IIIE (including IIIE) was a prognostic factor. Single factor analysis with B symptom showed that B symptom also was a prognostic factor. The cumulative survival rate of patients received radiotherapy alone was higher than that of patients got chemotherapy alone, its difference had a statistical significance, but there was no statistical significance between radiotherapy group and chemoradiotherapy group. It is concluded that CD56, Ann Arbor stage and B symptoms are prognostic factors. Radiotherapy alone is better than chemotherapy alone in therapy of ENKL. Chemotherapy combined with radiotherapy can not improve the survival. Improving the status of patients can make prognosis better.
Adolescent ; Adult ; Aged ; Chemoradiotherapy ; Female ; Humans ; Lymphoma, Extranodal NK-T-Cell ; diagnosis ; therapy ; Male ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo review clinical features of four male patients with glutaric academia type I and screen glutaryl-CoA dehydrogenase (GCDH) gene mutations.

METHODSThe 4 patients underwent brain computer tomography (CT) and magnetic resonance imaging (MRI) analyses. Blood acylcarnitine and urine organic acid were analyzed with tandem mass spectrometry and gas chromatographic mass spectrometry. Genomic DNA was extracted from peripheral blood samples. The 11 exons and flanking sequences of GCDH gene were amplified with PCR and subjected to direct DNA sequencing.

RESULTSAll patients have manifested macrocephaly, with head circumference measured 50 cm (14 months), 47 cm (9 months), 46 cm (5 months) and 51 cm (14 months), respectively. Imaging analyses also revealed dilation of Sylvian fissure and lateral ventricles, frontotemporal atrophy, subarachnoid space enlargement and cerebellar vermis abnormalities. All patients had elevated glutarylcarnitine (5.8 umol/L, 7.5 umol/L, 8.3 umol/L and 7.9 umol/L, respectively) and high urinary excretion of glutaric acid. Seven mutations were identified among the patients, among which c.146_149del4, IVS6-4_Ex7+4del8, c.508A>G (p.K170E), c.797T>C (p.M266T) and c.420del10 were first discovered.

CONCLUSIONMacrocephaly and neurological impairment are the most prominent features of glutaric academia type I. Blood tandem mass spectrometry and urine gas chromatographic mass spectrometry analysis can facilitate the diagnosis. The results can be confirmed by analysis of GCDH gene mutations.

Amino Acid Metabolism, Inborn Errors ; diagnosis ; genetics ; metabolism ; Amino Acid Sequence ; Base Sequence ; Brain Diseases, Metabolic ; diagnosis ; genetics ; metabolism ; Glutaryl-CoA Dehydrogenase ; deficiency ; genetics ; metabolism ; Humans ; Infant ; Male ; Molecular Sequence Data ; Mutation ; Sequence Alignment