Administration, Inhalation ; Adrenal Cortex Hormones ; pharmacology ; Asthma ; drug therapy ; metabolism ; Child ; Child, Preschool ; Drug Interactions ; Drug-Related Side Effects and Adverse Reactions ; Female ; Humans ; Male ; Theophylline ; pharmacokinetics ; pharmacology ; Treatment Outcome

Objective To investigate the prevalence change of drinking water type of endemic fluorosis and the effect of control measures implemented in Huangyuan county of Qinghai province. Methods In 2009, all the endemic fluorosis villages in Huangyuan county were divided into two degrees, light and medium, according to the water fluorosis content before implementing the improving water project, 1 to 2 villages were selected from each degree village, respectively,as monitoring sites, and a total of 3 villages were selected. Source water and tap water samples were collected from each village and water fluoride concentration was determined. Dental fluorosis of all children aged 8 to 12 of monitoring villages was examined, and urine samples were collected by age group of children for determination of urinary fluoride. Clinical skeletal fluorosis of adults over 16 years of age was examined, and 20 copies of adults urine samples were collected to determine urinary fluoride. One village was selected in the 3 villages monitored to conduct X-rays examination of skeletal fluorosis. Water fluoride was tested in accordance with the "Non-metallic Targets Test Methods for Drinking Water" (GB/T 5750.6-2006); urinary fluoride was tested by fluoride ion-selective electrode method (WS/T 89-1996); dental fluorosis was diagnosed using Dean method;adult skeletal fluorosis was diagnosed by "Clinical Diagnostic Criteria for Endemic Skeletal Fluorosis"(WS 192-2008). Results Twelve water samples were assayed, water fluoride was (0.35 ± 0.43) mg/L. The detectable rate of dental fluorosis of 122 children aged 8-12 was 34.43%(42/122) and the geometric mean urinary fluoride was 0.89 mg/L of the 96 children. Of the 834 adults aged 16 and over, clinical detection of skeletal fluorosis was 47.72% (398/836) and geometric mean urinary fluoride was 1.10 mg/L of the 65 cases of adult urine samples assayed, detection rate of X-rays was 31.4% (11/35) in Gangou village of the 35 adults examined.Conclusions In Huangyuan county, water fluoride of the 3 surveyed villages are normal but the endemic fluorosis is still serious. It should strengthen monitoring and analyze the causes and improve prevention measures.

Objective To determine the clinical features,neurophysiological characteristics and cervical magnetic resonance imaging of sensory neumnopathy,and to describe the pathology of skin nerve, sural nerve and spinal dorsal columns.Methods Two patients who died from sensory neuron disease (SND)after infection of digestive tract were discussed including clinical features and ancillary tests which included neurophysiology and pathology of peripheral nerve and spinal dorsal columns.Associated documents are reviewed.Results Early ataxia,widespread sensory symptoms and global loss of deep tendon reflex were the distinctive signs of SND,which was characterized by non-length-dependent abnormalities of sensory nerve action potentials,a hallmark of ganglionopathies.The second patient showed normal cervical magnetic resonance imaging possibly because of short course of disease,while diffuse hyperintensity in the spinal posterior columns of SND was reported.Demyelination of spinal posterior columns and loss of mostly large diameter nerve fibers without regeneration clusters were the main pathological features.Conclusions The distinctive clinical features and neurophysiological characteristics of SND indicate that peripheral sensory nerve fibers are widely damaged.Pathology of spinal posterior columns confirm that central sensory pathway are impaired which allow the localization of the pathologic site to the dorsal root ganglion neurons.Cervical spinal MRI of SND are possibly normal at early phase.

Objective To observe the state of endemic flurosis in Qinghai province in 2008 and to provide scientific basis for endemic flurosis control and prevention. Methods All the endemic fluorosis villages in Qinghai province was divided into slight,medium and heavy degree according to the water fluoride content before improving water,and 2,4,4 villages were collected from each degree,respectively. Water samples were collected from each direction of east,west,south,north and centre,and fluoride concentration were determined from each surveyed villages with unimproved-water. At the same time,3 copies of the peripheral water samples and a sample of source water were collected to determine fluoride concentration. In all the village children aged 8 to 12 were tested for dental fluorosis by Dean method. Six copies of the urinary fluoride were randomly sampled in different age groups. The fluorine content in urine was determined with F-ion selective electrode. The situation of clinical skeletal fluorosis of adults over 16 years of age was examined,and 20 adults in the villages of medium and heavy levels were filmed for skeletal fluorosis. Results The detectable rate of dental fluorosis of children aged 8-12 years old was 43.59% (245/562),that of skeletal flurosis of adults aged above 16 years was 50.32% (1582/3144) and detectable rate of X-ray was 29.45% (48/163). Fluoride content in urine was 1.52 mg/L. Conclusions Endemic fluorosis is still very serious in Qiaghai province in a rising trend,so the control measures need to be strengthen.

BACKGROUNDSince an effective method for generating induced pluripotent stem cells (iPSCs) from human neural stem cells (hNSCs) can offer us a promising tool for studying brain diseases, here we reported direct reprogramming of adult hNSCs into iPSCs by retroviral transduction of four defined factors.

METHODSNSCs were successfully isolated and cultured from the hippocampus tissue of epilepsy patients. When combined with four factors (OCT3/4, SOX2, KLF4, and c-MYC), iPSCs colonies were successfully obtained.

RESULTSMorphological characterization and specific genetic expression confirmed that these hNSCs-derived iPSCs showed embryonic stem cells-like properties, which include the ability to differentiate into all three germ layers both in vitro and in vivo.

CONCLUSIONOur method would be useful for generating human iPSCs from NSCs and provide an important tool for studying neurological diseases.

Cell Differentiation ; genetics ; physiology ; Cells, Cultured ; Cellular Reprogramming ; genetics ; physiology ; Humans ; Immunohistochemistry ; Induced Pluripotent Stem Cells ; cytology ; metabolism ; Kruppel-Like Transcription Factors ; metabolism ; Neural Stem Cells ; cytology ; metabolism ; Octamer Transcription Factor-3 ; metabolism ; Proto-Oncogene Proteins c-myc ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; SOXB1 Transcription Factors ; metabolism

OBJECTIVETo explore the relationship between gene polymorphism of GABAA receptors and childhood autism by detecting rs140682, rs2081648 and rs140679 site of single nucleotide polymorphism (SNP) in GABAA receptors gene.

METHODSA total of 94 children with autism and 124 normal children were enrolled in a hospital from November 2010 to May 2011. Childhood autism rating scale (CARS) and autism behavior checklist (ABC) were used to evaluate or investigate the case group. After collecting venous blood and extracting the genome DNA, the allele and genotype of SNP rs140682, rs2081648 and rs140679 site in GABAA receptors gene were detected by PCR-RFLP. The allele and genotype of case group and control group were analyzed by χ(2) test, while the score of scales was analyzed by Spearman rank correlation analysis.

RESULTSThe age of the case group was 5.12 ± 0.32, and it was 5.25 ± 0.27 in the control group (P < 0.05). In case group, the frequency of genotype CC, CT and TT of rs140682 site was 44, 41 and 9, while it was 48, 65, and 11 in control group (P > 0.05), respectively. The frequency of genotype AA, AG and GG of rs2081648 site was 8, 58 and 28 in case group, while it was 12, 49 and 63 in control group (P < 0.05), respectively. In case group, the frequency of genotype CC, CT and TT of rs140679 site was 15, 36 and 43, while it was 18, 59 and 47 in control group (P > 0.05), respectively. It was revealed by Spearman rank correlation analysis that of rs2081648 site, there was a positive correlation between genotype AG and sensation factor (S), social intercourse factor (R), and language factor (L) of autism behavior checklist (ABC) (r values were 0.149, 0.165 and 0.155, all P values < 0.05). A negative correlation between genotype GG and S, R, L and self-help factor (V) was proved (r values were -0.140, -0.173, -0.158 and -0.135, all P values < 0.05). There was a positive correlation between allele A and R and L factors (r values were 0.153 and 0.137, all P values < 0.05), while a negative correlation between allele G and R and L factors (r values were -0.153 and -0.137, all P values < 0.05). In case group, 42 children were diagnosed with mild-to-moderate autism, while 52 children were severe autism. There was no statistically significant correlation between allele or genotype of SNP rs140682 and rs140679 site and the degree of autism (P > 0.05). There was a positive correlation between allele A and genotype AG and the degree of autism (r values were 0.147 and 0.616, all P values < 0.05), while a negative correlation between allele G and genotype GG and the degree of autism (r values were -0.159 and -0.616, all P values < 0.05).

CONCLUSIONThe SNP rs2081648 site which located in GABAA receptors gene may be related to autism. No evidence for significant association between rs140682 and rs140679 site and autism was found.

Alleles ; Autistic Disorder ; genetics ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Male ; Polymorphism, Single Nucleotide ; Receptors, GABA-A ; genetics

OBJECTIVES: To explore the relationship between the change rules of volatile organic compounds （VOCs） in rat muscle and postmortem interval （PMI）. METHODS: A total of 120 healthy rats were divided randomly into 12 groups （10 for each group）. After the rats were sacrificed by cervical dislocation, the bodies were kept at （25±1） ℃. Rat muscle samples were separately obtained at 12 PMI points, including 0, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10 d. The VOCs in rat muscles were collected, detected and analyzed by headspace solid-phase microextraction （HS-SPME） coupled to gas chromatography-mass spectrometer （GC-MS）. RESULTS: In total, 15 species of VOCs were identified, including 9 aromatic compounds, 3 sulfur compounds, 2 aliphatic acids and 1 heterocyclic compound. The species of VOCs increased with PMI： no species were detected within 1 day, 3 species were detected on day 2, 9 on day 3, 11 on day 4, 14 from day 5 to 7, and 15 from day 8 to 10. Total peak area of 15 species of VOCs was significantly correlated to PMI （adjusted R²=0.15-0.96）： the regression function was y=-17.05 x²+ 164.36 x-246.36 （adjusted R²=0.96） from day 2 to 5, and y=2.24 x+101.13 （adjusted R²=0.97） from day 6 to 10. CONCLUSIONS The change rules of VOCs in rat muscle are helpful for PMI estimation.
Animals ; Autopsy ; Gas Chromatography-Mass Spectrometry/methods* ; Muscles/pathology* ; Rats ; Solid Phase Microextraction ; Volatile Organic Compounds/chemistry*

Objective To analyze the status of quality indicators(QI) on specimen acceptability and establish preliminary qual ity specification.Methods Web based External Quality Assessment system was used to collect data of laboratories partici pated in "Medical quality control indicators in clinical laboratory" from 2015 to 2017,including once in 2015 and 2017 and twice in 2016.Rate and sigma scales were used to evaluate incorrect sample type,incorrect sample container,incorrect fill level and anticoagulant sample clotted.The 25th percentile (P25) and 75th percentile (P75) of the distribution of each QI were employed to establish the high,medium and low specification.Results 5 346,7 593,5 950 and 6 874 laboratories sub mitted the survey results respectively.The P50 of biochemistry (except incorrect fill level),immunology and microbiology reach to 6σ.The P50 of clinical laboratory is 4 to 6σ except for incorrect sample container.There is no significant change of the continuous survey results.Based on results in 2017 to establish the quality specification,the P25 and P75 of the four QIs is 0 and 0.084 4 ％,0 and 0.047 6 ％,0 and 0.114 2 ％,0 and 0.078 4 ％,respectively.Conclusion According to the results of the survey,most laboratories had a faire performance in biochemistry,immunology and microbiology,and clinical laboratory needs to be strengthened.Laboratories should strengthen the laboratory information system construction to ensure the actual and reliable data collection,and make a long time monitoring to achieve a better quality.

OBJECTIVETo diagnose muscular dystrophy using Western blot (WB) by improving the method of the protein extraction.

METHODFirstly,we compared the effect of different sample buffer solutions and processing Methods on the extraction of muscle protein in rats,then selected the appropriate extracting method and the process of the muscular protein.

RESULTSWe put the selected sample buffer into the micro-sample,then mixed. The concentration of the extracting protein was much more,and the loss during the process was much less. We extracted enough protein in 62 cases. The protein bands were showed clearly by WB,and the abnormal protein bands were shown in some patients. Compared with the Results of immunohistochemical staining detected the severe abnormal expressions of Dys-R,Dys-C,and Dys-N in the specimens,we did not detect the corresponding target band in WB. We detected the target protein band of the specimens were abnormal position,light or normal staining in WB,while Dys were mildly expressed in immunohistochemical staining.

CONCLUSIONSThe improved protein extraction method can save the muscle tissue,and the protein bands can be used for diagnosing the muscular dystrophy. For clinically suspected patients with dystrophinopathy,if normal or mild deficiency is shown by immunohistochemistry,WB should be applied to detect the dystrophin protein band.

Animals ; Blotting, Western ; Dystrophin ; Humans ; Immunohistochemistry ; Muscular Dystrophies ; Protein Transport ; Rats ; Staining and Labeling

ObjectiveTo preliminarily predict the active components， action targets， and signaling pathways of Arnebia euchroma in the treatment of melanoma based on network pharmacology and molecular docking， and to verify its possible mechanism of action in in vitro experiments. MethodThe active components and related targets of A. euchroma were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform （TCMSP）SwissTargetPrediction and literature， and the targets related to melanoma from the GeneCards， Online Mendelian Inheritance in Man （OMIM）， and Comparative Toxicogenomics Database （CTD）. Following the construction of the protein-protein interaction （PPI） network of active components and related targets of A. euchroma and melanoma-related targets using STRING， Cytoscape 3.8.2 was used for screening and analyzing the nodes in the network of A. euchroma against melanoma. The intersections were subjected to gene ontology （GO） and Kyoto encyclopedia of genes and genomes （KEGG） enrichment analysis using DAVID 6.8. Acetyl alkannin， the active component in A. euchroma， was docked to the target by AutoDock Vina 1.1.2. The in vitro experiments were then carried out to verify the anti-melanoma effect of A. euchroma. ResultA total of 271 common targets of A. euchroma and melanoma were harvested， among which 23 were key targets， including matrix metalloproteinase-9 （MMP-9） and Janus kinase 2 （JAK2）. As revealed by KEGG enrichment analysis， A. euchroma mainly acted on Janus kinase/signal transduction and activator of transcription （JAK/STAT）， tyrosine kinase receptor （ErbB）， and vascular endothelial growth factor （VEGF） signaling pathways to resist melanoma. According to molecular docking， acetyl alkannin exhibited a good docking activity with JAK2， STAT3， VEGF， MMP-9， and E-cadherin receptors. The results of Western blot and Real-time quantitative polymerase chain reaction （Real-time PCR） showed that acetyl alkannin at different doses inhibited the protein and gene expression of JAK2， STAT3， VEGF， MMP-9， and E-cadherin in A375 cells （P<0.05）. ConclusionA. euchroma alleviates melanoma via multiple targets and multiple pathways， and it may exert the therapeutic effects by affecting the expression of such key target proteins as JAK2， STAT3， VEGF， MMP-9， and E-cadherin and inhibiting the invasion and metastasis of melanoma cells. This study has provided an experimental basis for the treatment of tumor with A. euchroma.