OBJECTIVE: Investigate common deafness gene mutations in patients with severe and profound non-syndromic hearing loss in Liaoning in order to understand their hereditary etiologies and characteristics at the molecular level. METHOD: Peripheral blood samples were obtained and the DNA templates were extracted from 128 non-syndromic hearing loss patients who are sporadic in clinics. The deafness gene chip was applied to detect hot-spot deafness gene mutations including GJB2, GJB3, SLC26A4 and mitochondrial 12S rRNA. Deafness etiology questionnaires, pure tone audiometry, auditory brainstem response, tympanometry and temporal bone CT were also applied. RESULT: Various types of gene locus mutations were seen in 52 of the 128 patients (40.6%); (1) GJB2 gene mutations (n=22) included c. 235 del C homozygous mutation (n=10), c. 235 del C heterozygous mutation (n=5); c. 176_191 del 16 heterozygous mutation (n=l); c 35 del G heterozygous mutation (n=l); c. 235 del C/c. 299_300 del AT mutation (n=l), c. 235 del C/c. 176_191 del 16 mutation (n=l), c. 35 del G/c. 176_191 del 16 mutation (n=l); c. 299_300 del AT/c. 919-2 A>G mutation (n=l), c. 235 del C/c. 919-2 A>G mutation (n=l). (2) SLC26A4 gene mutations (n=30) included c. 919-2 A>G homozygous mutation (n=6), c. 919-2 A>G heterozygous mutation (n=17), c. 2168 A>G homozygous mutation (n=l), c. 2168 A>G heterozygous mutation (n=2), c. 2168 A>G/c. 919-2 A>G mutation (n=2), c. 919-2 A>G/GJB2 c. 235 del C mutation (n=2); (3) No GJB3 and mitochondrial 12S rRNA mutation. Genetic deafness was confirmed at the gene level in 24 cases (18.8%) and 28 patients (21.9%) were diagnosed as carriers of genetic deafness gene mutations. CONCLUSION Genetic deafness occupies a large population in deaf community in Liaoning. Molecular genetic screening for these mutations and genetic counseling are effective methods to prevent the occurrence of hereditary hearing loss and provide theoretical guidance.
Adolescent ; Child ; Child, Preschool ; China ; Connexins ; DNA Mutational Analysis ; Deafness ; genetics ; Female ; Genetic Testing ; Humans ; Infant ; Male ; Mutation

0.05).Seroquel showed high incidence rate of drowsiness,dizziness and weight gain(26.1%,23.9%,17.3%),chlorpromazine showed high incidence rate of extrapyramidal side effect,pyknocardia and erect collapse(60.9%,39.1%,32.6%).CONCLUSIONS:There was no significant difference of curative effect for the treat-ment of schizophrenia between seroquel-treated group and chlorpromazine-treated group,but seroque showed low side effect and high safety.

To study the effect of Fuzheng Huayu recipe (FZHY) on five types of isozymes of cytochrome P450 (CYP450) of normal and liver fibrosis rats by using the cocktail probe method. Dimethylnitrosamine ( DMN) was injected to induce the liver fibrosis model. After the tail vein injection with Cocktail probe solutions prepared with five CYP450s probe substrates (phenacetin-CYP1A2, omeprazole-CYP2C9, tolbutamide-CYP2C19, dextromethorphan-CYP2D6, midazolam-CYP3A4), the plasma concentrations of the five probe substrates were determined by LC-MS/MS, and the pharmacokinetic parameters were calculated by PK solutions 2. After the oral administration with FZHY, normal rats given phenacetin, omeprazole, tolbutamide and dextromethorphan showed increase in AUC(0-t) and decrease in CL to varying degrees, indicating that FZHY obviously inhibited the activities of CYP1A2, CYP2C9, CYP2C19 and CYP2D6 in normal rats, but with no obvious effect on the activity of CYP3A4. After the oral administration with FZHY, liver fibrosis rats treated with CYP2C9 showed the significant increase in AUC(0-t) and significant decrease in Vd, hut with no obvious changes in the pharmacokinetic parameters of other four types of prove substances, suggesting that FZHY could significantly inhibit the activity of CYP2C9 in rats but had no effect on the activities of CYP1A2, CYP2C19, CYP2D6 and CYP3A4. The changes in the activity of CYP450 isozymes in liver fibrosis rats may be the reason for FZHY's different effects on CYP450 isozymes in normal and liver fibrosis rats.
Animals ; Cytochrome P-450 Enzyme System ; genetics ; metabolism ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Humans ; Isoenzymes ; genetics ; metabolism ; Liver Cirrhosis ; drug therapy ; enzymology ; genetics ; Male ; Mass Spectrometry ; Rats ; Rats, Wistar

OBJECTIVE: To compare the effect on CYP450 isoenzyme in rats with acute liver injury induced by different chemicals. METHODS: Acute liver injury model of rats induced by tetrachloromethane(CCl4), D-aminogalactose(D-GalN)/lipopolysaccharide(LPS), α-naphthyl isothiocyanate(ANIT) respectively whereas the normal Wistar rats were used as controls. After the tail vein injection with Cocktail probe solutions prepared with five CYP450s probe substrates (phenacetin-CYP1A2, omeprazole-CYP2C9, tolbutamide-CYP2C19, dextromethorphan-CYP2D6, midazolam-CYP3A4), the blood samples were collected from the fundus venous plexus of rat at different time point, the blood drug concentration of the five probe substrates were determined by LC-MS/MS, and the pharmacokinetic parameters were calculated by PK Solutions 2™. Compared with normal rats, the changes of the probe drug pharmacokinetics in different rat models were used as the basis for the evaluation of the metabolic activity of CYP450 isoenzyme. RESULTS: Compared with normal rats, the activities of CYP1A2, CYP2C9, CYP2C19, CYP2D6 of CCl4 group rats were significantly inhibited, and the activities of CYP3A4 was slightly inhibited; the activities of CYP2C9, CYP2D6 and CYP3A4 of D-GalN/LPS group rats were significantly induced, and the activity of CYP2D6 and CYP3A4 was slightly induced, and the activity of CYP1A2 was not significantly affected, but the activity of CYP2C19 was significantly inhibited; the activities of CYP2C9, CYP2C19 and CYP3A4 of ANIT group rats were significantly induced, the activity of CYP3A4 were slightly induced, and the activity of CYP2D6 was not significantly affected, but the activity of CYP1A2 was significantly inhibited. CONCLUSION: There are significant differences in the activities of CYP450 isoenzyme in the rat model of acute liver injury induced by different chemicals.

Objective To explore Toll like receptor 2 (TLR2) expression after partial liver ischemia/ reperfusion (I/R) in BALB/c mice and its relationship with liver function impairment. Methods All the animals were randomly divided into ischemia/reperfusion injury (I/R) and sham operation (S) groups. Total RNA was extracted from the liver samples and the expression of TLR2 mRNA was analyzed quantitatively by real time PCR method. Membrane protein was extracted, and the expression of TLR2 protein was detected by western blot method. Portal vein serum and plasma were taken for further detection of the levels of alanine aminotransferase (ALT), tumor necrosis factor alpha (TNF ?) and endotoxin . Results After 1h of partial liver ischemia and 4 h of reperfusion, the expression of TLR2 mRNA and TLR2 protein were both remarkably upregulated in tissues of ischemic liver lobes in I/R group compared with that in S group (value of TLR2mRNA: 1.06?0.91vs5.08?1.32,P0.05). Conclusions TLR2mRNA and TLR2 protein expression were upregulated in tissues of ischemic liver lobes after partial I/R injury in mice, and were associated with increased levels of portal vein ALT and TNF ?,and impairment of liver function.

Background Leber hereditary optic neuropathy (LHON)is a mitochondrial DNA (mtDNA)hereditary disease,so it is significant to understand the influence of DNA mutation on the occurrence of LHON.Objective This survey was to evaluate the role of mtDNA mutation in the development of LHON.Methods This survey study was approved by the Ethic Committee of Wuhan General Hospital of Guangzhou Military Command and written informed consent was obtained from each subject before the relative medial examination.Seventy-two matrilineal relatives from a family with LHON were collected for a pedigree analysis and mutation screening.Regular eye examination was performed on 11 patients,13 mutant gene carriers and 49 individuals with normal phenotype,and the degree of visual damage was graded as follows: ＞0.3 was normal,0.1-0.3 was mild damage,＜0.05-0.1 was moderate damage,＜0.02-0.05 was severe damage and ＜0.01 was very severe damage.Clinical characteristics of LHON was evaluated.The periphery blood sample of 2-4 ml was collected from individuals to separate the mononuclear cells,and the mtDNA was extracted by modified high salt method.MtDNA was amplified by PCR and the mutation loci was sequenced.Results PCR amplification product sequencing of mutant gene showed that both G11778A and T14502C mutations were detected in 24 of 72 matrilineal relatives,but only 11 of 24 carriers developed LHON.No abnormal clinical findings were seen in the 13 carriers,showing a less 50％ penetrance in this family.There was no G11778A or/and T14502C mutation in the normal phenotype individuals of this family.The onset age for vision impairment in 11 affected matrilineal relatives varied from 8 to 50 years old,with the mean age of 24.36 years old,showing a significantly lower age than that of the 13 carriers (5-72 years old,mean 40.38 years old) (t =2.102,P=0.049).Conclusions This study suggests that the Gl1778A and T14502C mutation in mitochondrial DNA is one of causes in the development of LHON.The primary G11778A mutation together with T14502C mutation in mtDNA is a factor for the occurrence of LHON,hut it is not sufficient to the development of LHON.An effective “second hit” process will play an inducing role for LHON.

OBJECTIVETo investigate relationship between the expression of heparanase (HPSE) and vascular endothelial growth factor-C (VEGF-C) and tumorigenesis, progression in human lung cancer.

METHODSThe expression of HPSE and VEGF-C protein in 65 cases of lung cancer, adjacent tissues of cancer and normal tissues was tested by immunohistochemical SABC method and analysed by clinico-pathological characteristics and prognosis of lung cancer.

RESULTSThe rate of expression of HPSE and VEGF-C protein in tumor tissues (51% and 57%) was significantly higher than that in adjacent tissues of cancer (9% and 12%) and normal tissues (5% and 6%) (chi2 = 34.6, 38.8, 26.7, 28.6; P < 0.01); It was shown that HPSE and VEGF-C protein expression did significantly not correlate with the type (chi2 = 0.39, 0.41, P > 0.05) and grade of the tumor (chi2 = 0.45, 0.04, P > 0.05); but it correlated with the clinical stage (chi2 = 26.6, 20.1; P < 0.01) and survival time of the patients (chi2 = 21.5, 22.2; P < 0.01).

CONCLUSIONSThe results suggest that there be overexpression of HPSE and VEGF-C protein in lung cancer tissues, and which perhaps participate in regulation of tumorigenesis, progression in lung cancer. The expressions of HPSE and VEGF-C protein are used as an useful marker of the biological behavior of lung cancer and as an independent prognosis factor for the patients with lung cancer.

Adult ; Aged ; Female ; Glucuronidase ; metabolism ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Vascular Endothelial Growth Factor C ; metabolism

OBJECTIVETo investigate the effects of hydrogen sulfide preconditioning on myocardial ischemia reperfusion injury in rats.

METHODSSprague-Dawley male rats were divided into 4 groups with 10 in each group: in S group rats received sham operation; in IR group rats were given with NS (1.0 ml/kg iv) 24 h before ischemia; in H group rats were treated with NaHS (0.05 mg/kg iv) 24 h before ischemia; and in D group, NaHS-treated rats received 5-hydroxydecanoate (5-HD) 15 min before ischemia. Rats in IR group,H group and D group were subjected to ischemia by occlusion of coronary artery for 30 min followed by 2 h of reperfusion. At the end of the reperfusion,myocardial infarct size was measured. SAM-s was measured by Western blotting. Plasma SOD activity and MDA were determined at the end of reperfusion.

RESULTSThe infarct size was significantly lesser in H group (25.40 % ± 3.54%) than that in IR group (38.27% ±5.64%,P<0.05). The SAM-s protein expression in myocardium was significantly lower in H group than that in IR group. The plasma MDA content was significantly lower and SOD activity was higher in H group than those in IR group,but there was no difference between IR group and D group.

CONCLUSIONThe hydrogen sulfide preconditioning attenuates myocardial IR injury possibly through down-regulating SAM-s expression,reducing the production of oxygen free radicals and enhancing anti-oxidize effect in rats.

Animals ; Disease Models, Animal ; Hydrogen Sulfide ; pharmacology ; Ischemic Preconditioning, Myocardial ; Male ; Myocardial Reperfusion Injury ; metabolism ; pathology ; prevention & control ; Myocardium ; metabolism ; pathology ; Rats ; Rats, Sprague-Dawley

Objective To investigate the role of toll-like receptor 4 (TLR4) of endothelium or bone marrow derived cells in the acute lung injury (ALI) induced by lipopolyscccharide (LPS) in mice with reciprocal bone marrow transplantation. Method Chimeric mice were produced by reciprocal bone marrow transplantation between TLR4mut/mut and TLR4+/+ mice and divided into 4 groups: WT/WT (recipient/donor),WT/Mutant, Mutant/WT and Mutant/Mutant group. Six to eight weeks following transplantation, LPS was injected inot mice's tail vein in order to produce ALI model,and mice were sacrificed five hours later on.Samples of lung tissues were taken for the following analysis of wet/dry weight (W/D), lung permeabifity index (LPI), myeloperoxidase (MPO),levels of cytokines (TNF-α, IL-1β) and adhesion molecules (ICAM-1). Results Lung injury in the Mutant/Mutant mice was the mildest in the 4 groups. And lung injury in WT/Mutant mice was more serious than that in Mutant/WT mice. levels of MPO and ICAM-1 in WT/Mutant mice were much higher than those of Mutant/WT. In addition,the expression of ICAM-1 in WT/Mutant mice is comarable to that in WT/WT mice. Mutant/WT mice expressed higher levels of TNF-α and IL-1β than WT/Mutant mice. Conclusions Endothelial cell derived TLR4 plays ker-nel role in ALI induced by LPS via lung PMN recruitment,although bone marrow cells derived TLR4 are more im-portant for the release of cytokines.

Microdialysis ; Pharmaceutical Preparations ; metabolism ; Pharmacokinetics