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

Humans ; Infant ; Lead Poisoning, Nervous System, Childhood ; diagnosis ; therapy ; Male

Objective To investigate the effects of Shenfu injection ( SF,a Chinese herbal medicine preparation made of Codonopsis pilosula and Aconitum carmichaeli) on the cell apoptosis of focal cerebral ischemic-reperfusion injured rats and the expression of heme oxygenase-1 (HO-1). Methods Forty-two male Sprague-Dawley rats used for producing unilateral brain ischemia reperfusion model were randomly divided into three groups:sham operation group ( Sham group),ischemia reperfusion group ( IR group),and SF Injection group (SF group).The model of focal cerebral ischemia-reperfusion injury was induced by transient occlusion of middle cerebral artery (ischemia for 2 h,and reperfusion for 3,6 h respectively).In SF group,SF ( 10 mg/kg) was intraperitoneally injected duri(n)g reperfusion.Cell apoptosis rate in brain tissue was detected by the technique of Annexin-V-PI double staining and was counted in flow cytometer.Expression of HO-1 in brain was measured by RT-PCR,while the pathological and ultra structure changes of cerebral tissue were also observed.Results Cell apoptosis rate of brain tissue were significantly higher in IR group than that in Sham group (P ＜0.01 ),while SF group had less significant changes in cell apoptosis rate, HO-1 level of brain tissue than IR group (P ＜ O.01 ).The ultra structure change of brain tissue was less in SF group than that in IR group.Conclusions During early stage of brain IR injury,SF inhibits cellular apoptosis and in turn protects the brain from injury which is attributed to the increase in HO-1 expression induced by SF.

BACKGROUND: The dynamic changes of pneumocyte apoptosis and aspartate-specific cysteine proteases-3 (caspase-3) expression in lung tissue of rats during the process of lung ischemia/reperfusion (I/R) injury and the possible action mechanisms remain unclear.OBJECTIVE: This study was to observe the dynamic changes of pneumocyte apoptosis and caspase-3 expression in the rat lung tissue during the process of lung I/R injury, and to analyze the role of pneumocyte apoptosis and the possible action mechanism.DESIGN: A randomized controlled animal experiment.SETTING: Emergency Center, First Hospital, Nanjing Medical University.MATERIALS: This study was carried out in the Animal Laboratory of the First Hospital of Nanjing Medcial University and Nanjing Center for Radioimmunity between April 2006 and September 2006. Twenty-eight male healthy SD rats of clean grade, with body weight of 250 to 350 g, aged 49 to 76 days, were provided by the Experimental Animal Center of Nanjing Medical University. The involved rats were randomized into experimental group and control group, with 14 rats in each.METHODS: ①Experimental intervention: Rats in the experimental group were created into models of lung I/R injury according to the method of Eppinger et al. They were occluded for 45 minutes at the porta of lung (no systolic and diastolic reactions in lung tissue being considered as successful occlusion), and then they were reperfused (recovery of systolic and diastolic function being considered as successful reperfusion); After that, lung tissues were harvested at 3 and 6 hours after lung I/R injury, 7 rats at each time point. Each rat in the control group was subjected to a thoracotony only, but lung tissues were isolated at the same time point by the same method. ②Experimental evaluation: Apoptotic cells in the lung tissue were detected with a flow cytometer by Annexin-V-PI staining, and apoptosis rate was calculated. Caspase-3 expression in the lung tissue was observed by immunohistochemical method and image analysis. Wet to dry weight ratio(W/D) of lung tissue of rats in the two groups was calculated; the number of injured pulmonary alveoli at I/R 3 hours/that at I/R 6 hours was calculated for quantitative evaluation of injured lung tissue; Patho-morphological changes of lung tissue were observed by haematoxylin & eosin staining under an optical microscope.MAIN OUTCOME MEASURES: ①Pneumocyte apoptosis rate and caspase-3 expression in the lung tissue. ②W/D of lung tissue and quantitative evaluation of injured lung tissue. ③Patho-morphological changes of lung tissue.RESULTS: Twenty-eight rats were involved in the final analysis, without deletion. ①Pneumocyte apoptosis rates in the experimental group at I/R 3 and 6 hours were significantly increased as compared with control group (P＜0.01). In the experimental group, pneumocyte apoptosis rate was decreased a little at I/R 6 hours than at I/R 3 hours (P＜0.05). ②Caspase-3 expression in the lung tissue of rats of experimental group reached its top at I/R 3 hours, and was decreased a little at I/R 6 hours. At each time point, caspase-3 expression in the experimental group was increased as compared with control group (P＜0.01). ③In the experimental group, the number of injured pulmonary alveoli at I/R 3 hours/that at I/R 6 hours and W/D ratios of lung tissues were significantly increased as compared with control group (P＜0.01). In the experimental group, two ratios at I/R 6 hours were higher than those at I/R 3 hours (P＜0.05).④In the experimental group, the structure of pulmonary alveoli was destructed, collapsed and disappeared; lots of inflammatory cell infiltration was found; Patho-morphological changes of injured lung tissue at I/R 6 hours were severer than those at I/R 3 hours. No obvious changes were found in the control group.CONCLUSION: At the early stage of lung I/R injury, the alteration of caspase-3 maybe activate pneumocyte apoptosis and induce the apoptosis of lung tissue, and thereby leads to lung injury.

Objective To compare the mutation sites in human immunodefieiency virus type 1 (HIV-1) vpr gene via of HIV-1 infected individuals from different regions in China with the previous studies, and to provide information for the further study on the relationship between HIV-1 vpr gene mutations and clinical conditions of the patients. Methods Reverse transcription-polymerasc chain reaction (RT-PCR) and nested PCR were used to amplify HIV-1 vpr gene of 398 HIV-1 infected individuals. The amino acid sequences were analyzed to determine polymorphisms, deviation rate and common mutation sites of HIV-1 vpr gene. Meanwhile, the viral load, subsets of lymphocytes and clinical course of patients infected with mutated HIV-1 were analyzed. Results One hundred and fifty three positive samples which were obtained from 398 HIV-1 infected individuals were available for further analysis. The amino acids sequence typing of HIV-1 Vpr were showed that CRF01 AE was 51.63%, subtype C 24.84%, subtype B 17.65%, CRF03_ AB 3.92% and CRF08 BC 1.31%. Eighty four point three percent of 77th amino acid of HIV Vpr sequence was glutamic acid which was significantly different from what overseas researches reported that the R77Q mutation was correlated with long-term non-progression (LTNP) of AIDS. The mutations of the, 63th, 70th, 85th, 86th, 89th and 94th amino acids of HIV Vpr were likely related to the clinical remission of HIV-1 infected individuals. Conclusions M group is the main type of HIV Vpr typing in China, and CRF01 AE is predominant. Some amino acid mutation sites of HIV-1 Vpr are possibly correlated with clinical manifestations of HIV-1 infected individuals.

OBJECTIVE:To screen formulation of Insulin cream and to investigate the effects of insulin cream on wound healing in rabbits.METHODS:The best formulation of insulin cream was selected from 8 kinds of formulations with appearance and stability as index.Rabbit trauma model were induced and divided into matrix group,blank group,Insulin cream groups(high-dose,medium-dose and low-dose).Wound healing rate and healing time were compared among those groups.RESULTS:The fifth formulation possessed good appearance and stability,which included albolene,cetanol,glycerin monostearate,glycerol,etc.Compared with matrix group,the wound healing rates were obviously increased at different time points in Insulin cream groups(P

The rising of evidence-based medicine requires high quality for clinical research and randomized controlled trial is considered as clinical research of the most level of evidence degree and contidence activity.This article illustrates several important and instructive designs of randomized controlled trials,which can give some reference for clinical research on acupuncture.And the developing direction of future clinical research on acupuncture is discussed.

A PubMed search is carried out using the keyword "acupuncture" from in the top four journals: New England Journal of Medicine (NEJM), The Journal of American Medical Association (JAMA), The Lancet, and British Medical Journal (BMJ). The papers with full texts on randomized controlled trials (RCTs) of acupuncture are included up till July 2009, and the corresponding trial design and trial quality in these papers are extracted and analyzed. The results reveal that papers published in these four journals are mostly on randomized controlled trials with good design, high quality and large sample size. Therefore, if Chinese researchers publish papers in these journals, appropriate trial design adhering to the characteristics of acupuncture should be used; the results should be reported according to CONSORT statement and STRICTA recommendations. Moreover, obtaining assistance from foreign research team is also important.
Acupuncture Therapy ; Animals ; Humans ; Periodicals as Topic ; PubMed ; Randomized Controlled Trials as Topic ; Research Design

Animals ; Male ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Respiratory Distress Syndrome, Adult ; chemically induced ; Silicon Dioxide ; chemistry ; toxicity

AIMTo study the enhancing effect of electroporation and iontophoresis on the permeation of insulin through human cadaver skin in vitro.

METHODSUsing side by side two-chamber diffusion cells, the flux of insulin achieved with iontophoresis and electrophoration were compared.

RESULTSThe application of high-voltage pulse combined with iontophoresis resulted in higher flux transdermal permeation of insulin than either one technique alone (P < 0.05). Pulsing at a higher voltage increased the flux of insulin more dramatically than pulsing at a lower voltage (P < 0.01). The transdermal transport of insulin by 90 pulse of 500 V (exponential pulse generater, pulse time: 20-24 ms, pulse frequency: 3 pulse.min-1) followed by iontophoresis led to a quick input and a high steady flux.

CONCLUSIONElectroporation combined with iontophoresis can enhance the permeation of insulin significantly.

Electroporation ; methods ; Humans ; In Vitro Techniques ; Insulin ; administration & dosage ; pharmacokinetics ; Iontophoresis ; methods ; Permeability ; Skin ; metabolism ; Skin Absorption