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This study aimed to obtain the first national estimate of the prevalence of autism spectrum disorder (ASD) in Chinese children. We targeted the population of 6 to 12-year-old children for this prevalence study by multistage convenient cluster sampling. The Modified Chinese Autism Spectrum Rating Scale was used for the screening process. Of the target population of 142,086 children, 88.5% (n = 125,806) participated in the study. A total of 363 children were confirmed as having ASD. The observed ASD prevalence rate was 0.29% (95% CI: 0.26%-0.32%) for the overall population. After adjustment for response rates, the estimated number of ASD cases was 867 in the target population sample, thereby achieving an estimated prevalence of 0.70% (95% CI: 0.64%-0.74%). The prevalence was significantly higher in boys than in girls (0.95%; 95% CI: 0.87%-1.02% versus 0.30%; 95% CI: 0.26%-0.34%; P < 0.001). Of the 363 confirmed ASD cases, 43.3% were newly diagnosed, and most of those (90.4%) were attending regular schools, and 68.8% of the children with ASD had at least one neuropsychiatric comorbidity. Our findings provide reliable data on the estimated ASD prevalence and comorbidities in Chinese children.
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This study aimed to obtain the first national estimate of the prevalence of autism spectrum disorder (ASD) in Chinese children. We targeted the population of 6 to 12-year-old children for this prevalence study by multistage convenient cluster sampling. The Modified Chinese Autism Spectrum Rating Scale was used for the screening process. Of the target population of 142,086 children, 88.5% (n = 125,806) participated in the study. A total of 363 children were confirmed as having ASD. The observed ASD prevalence rate was 0.29% (95% CI: 0.26%-0.32%) for the overall population. After adjustment for response rates, the estimated number of ASD cases was 867 in the target population sample, thereby achieving an estimated prevalence of 0.70% (95% CI: 0.64%-0.74%). The prevalence was significantly higher in boys than in girls (0.95%; 95% CI: 0.87%-1.02% versus 0.30%; 95% CI: 0.26%-0.34%; P < 0.001). Of the 363 confirmed ASD cases, 43.3% were newly diagnosed, and most of those (90.4%) were attending regular schools, and 68.8% of the children with ASD had at least one neuropsychiatric comorbidity. Our findings provide reliable data on the estimated ASD prevalence and comorbidities in Chinese children.
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Objective To explore the characteristics of food preferences in children with autism spectrum disorder,in order to provide some reference for treating the odd diet habits.Methods Comparing food selection styles through a self-made parents questionnaire including 113 kinds of common food in six categories in China in 162 cases of neurotypical children (NC) and 162 cases of children with autism (AC) to investigate the food preference characteristics of autistic children.Results Compared with NC,AC had narrow food repertoire in grain (NC refuse 0.0(1.0),AC refuse 1.0(2.0),P＜0.001),beans(NC refuse 0.0(1.0),AC refuse 1.0(2.0),P＜ 0.05),meat (NC refuse 0.0 (0.0),AC refuse 0.0 (2.0),P＜ 0.05),vegetables (NC refuse 3.0 (5.0),AC refuse 6.0(1.0),P＜0.001) and fruits(NC refuse 0.0(1.0),AC refuse 2.0(5.0),P＜0.001).There was no difference in the food preferences in neurotypical children of different gender.However,AC boys were more selective in the grain (NA girls refuse 0.0 (1.0),AC boys refuse 1.0 (2.0),P＜ 0.05) and vegetables (NA girls refuse 3.5 (5.0),AC boys refuse 7.0(11.0),P＜0.05) than AC girls.Moreover,AC had higher preference to lower acceptance in most of food than NC,but not instant noodles(NC acceptance 71.01％,AC acceptance 81.02％,P＜0.05) and chilli (NC acceptance 20.71％,AC acceptance 28.47％,P＜0.05).Conclusion Compared with NC,AC have narrower food repertoire.On the other hand,AC have significantly higher acceptance of stimulating food like chili and instant noodle.
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Objectlve To investigate the inhibition of Coxsackievirus B3(CVB3)infection in cardiac myocytes cultured by small interfering RNA(siRNA)-mediated RNA interference and to evaluate the feasibility of siRNA as the prophylaxis and therapy for CVB3 infection.Methods Cardiac myocytes were prepared in vitro and infected with CVB3,and transfected with siRNA by lipofectamin and electroporation.The numbers of beating cardiac myocytes were counted under the microscope.Neutral red staining was used to evaluate the mortality of cardiac myocytes.Antiviral activities of these siRNAs were estimated by observing cytopathic effect(CPE),plaque reduction assay,Western blot assay and RT-PCR.Results siRNA-3753,which aimed at sequence in 2B section of CVB3 genome,displayed a stronger inhibition of CVB3 infection through screening in HeLa cells,siRNA-3753,chosen to transfect cultured neonatal mice cardiac myocytes,Wag observed to keep a good states of growing and beating at 24 h after CVB3 infection.Whereas the cytopathic signature of controlled cells became stopping beating,round and finally the cell fell off the culture plate.The results showed that siRNA-3753 could protect cells significantly,98.1%inhibition of CVB3 replication with electroporation transfection and 78.2%inhibition of CVB3 with liposome transfection.Transfection efficiencies were 56.0 3%and 9.0%by electroporation and lipofectamin,respectively.Conclusion siRNA,which aims at sequence in 2B section of CVB3 genome,can inhibit CVB3 infection in cultured cardiac myocytes.
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Objective: To investigate the effect of electroacupuncture (EA) on rat with diet induced obesity (DIO) and to explore the possible neurochemical mechanisms using the technique of immumohistochemisty. Methods:To establish DIO rat model by feeding the animals with high fat diet for 14 weeks. DIO rats were randomly divided into 4 groups: (1) 2Hz EA group, (2) 100Hz EA group, (3) restrain control group,(4) diet resistance (DR) group,(5) DIO group and (6) normal control group. EA treatment: (1) The acupoints used were Zusanli and Sanyinjiao on both legs. (2) The intensities of stimulation were 0.5, 1.0 and 1.5mA for 10 mins each. EA treatment was administered 3 times per week. Food intake and body weight were measured daily for 4 weeks. (3) The changes of the expression of ? melanocyte stimulating hormone (? MSH) in the hypothalamic arcuate nucleus (ARC) were measured with immunohistochemical semiquantitative analysis. Results: (1) The food intake and body weight of 2 Hz EA group and 100 Hz EA group were decreased significantly compared with the restrain control group and DIO group. (2) The number of ? MSH positive cells in hypothalamic ARC in 2 Hz EA and 100 Hz EA group was significantly higher than that in restrain control group and DIO group. The number of ? MSH positive cells in hypothalamic ARC in DIO group is significantly lower than those in DR group or normal control group. Conclusion: A decrease of ? MSH level in hypothalamus may be associated with diet induced obesity. The therapeutic effect on obesity produced by EA may be accounted for by the stimulation of pro opio melanocortin neurons in hypothalamic ARC to release ? MSH, which inhibits food intake , resulting in a decrease of body weight.
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Objective: To observe the effect of 2 Hz electroacupuncture (EA) on long term depression (LTD) of synaptic transmission in the spinal dorsal horn in rats with neuropathic pain, so as to explore the central mechanisms of the antinociceptive effects of 2 Hz electroacupuncture on neuropathic pain. Methods: The neuropathic pain models were produced by tight ligation of the L5/L6 spinal nerves in Sprague Dawley rats. The C fiber evoked field potentials in the spinal dorsal horn were recorded with extracellular recording techniques. The parameters of the electroacupuncture were as follows: frequency of 2 Hz, wavelength of 0.6 ms, intensity of 1,2,3 mA lasting 10 min for each intensity, stimulation time of 30 min. The positive stimulating electrode was placed in acupoint “sanyinjiao” and the negative electrode in “zusanli”. Results: (1) 2 Hz electroacupuncture significantly decreased the amplitudes of C fiber evoked field potentials in the spinal dorsal horn in rats with neuropathic pain to (49.4?0.6)% of the control, compared with that (100.1?1.2) % of the control before EA (unpaired t test, P
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Single domain antibodies against telomerase protein hTERT were prepared by technique of displayed on the surface of recombinant bacterio phages. Total RNA of spleen lymphocytes were extracted from mice immunized recombinant hTERT and transcripted to cDNA. First-strand cDNA was used as a template, heavy chain variable region genes against hTERT were amplified with VHfor and VHback primers by PCR technique. Amplification reaction yielded a fragment about 350 base pairs in length. Amplified cDNA were cloned into the vehide of bacteriophage PCANTAB 5E, the phagemid containing VH gene transformed into competent E. coli TG1, in the presence of helper phage M13K07, VH-g3 fusion proteins were display on the surface of recombinant phages. The phage carrying VH genes that encode binding activities could be detected directly with DOT BLOT. Single domain antibodies were generated successfully and had binding activities with hTERT. Results suggest that phage display technique be a new way of making antibodies. VH genes were cloned successfully, which could provide possibility for futher preparing single-chain antibodies(ScFv) anti-hTERT.
Subject(s)
Animals , Female , Mice , Antibodies, Monoclonal , Genetics , Bacteriophages , Genetics , Cloning, Molecular , DNA-Binding Proteins , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin Variable Region , Genetics , Mice, Inbred BALB C , Telomerase , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To develop monoclonal antibodies against the catalytic subunit of human telomerase hTERT for its expression detection of human tumors.</p><p><b>METHODS</b>A dominant epitope in hTERT (peptide hTERT(9))was automatically synthesized based on Fmoc method, and was used to immunize BALB/c mice. Hybridomas were generated and screened by ELISA for specific monoclonal antibodies, and the characterization of which were performed by Western blotting and immunohistochemical staining.</p><p><b>RESULTS</b>Antigenic peptide hTERT(9) was synthesized and confirmed by MALDI-TOF-MS and HPLC analysis. Three hybridoma cell lines secreting anti-hTERT(9) antibodies designated as H4, G8 and A11 were established after primary screening and consequent three rounds of limited dilution. Both of H4 and G8 were IgM, while A11 was IgG1 in isotyping. The competitive assay showed that the antibodies were hTERT(9) specific, and the affinity of G8 was stronger than that of H4 and A11 assayed by affinity ranking. However, in Western blotting, both of H4 and G8 stained an about 123 000 protein band with HeLa and 293 cell extracts but not with normal 2BS cells. Besides, positive staining presented in the nucleus of HeLa, while 2BS was non-reactive immunohistochemically. The sections from paraffin-embedded blocks of 127 cases of human cancer, 40 of precancerous and 19 of benign tumors were in situ stained by G8 antibody, the results showed that the human cancer tissues were 80.31% (102/127) positive in specific nuclear reaction, on the contrary, only a minority of precancerous lesions present weak positive (17.5%, 7/40), and negative in benign tumors (0/19).</p><p><b>CONCLUSIONS</b>The monoclonal antibodies developed against synthetic peptide were hTERT-specific and could recognize both the native and the denatured form. Thus their use in immunoblotting or immunohistochemistry for detecting the telomerase hTERT expression of cancer cell and tissues was promising.</p>
Subject(s)
Animals , Female , Humans , Mice , Antibodies, Monoclonal , Allergy and Immunology , Binding, Competitive , Blotting, Western , Methods , Catalytic Domain , DNA-Binding Proteins , HeLa Cells , Immunohistochemistry , Methods , Mice, Inbred BALB C , Neoplasms , Pathology , Telomerase , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To develop a recombinant single domain antibody against hTERT, human telomerase catalytic subunit.</p><p><b>METHODS</b>A previously prepared His-tagged hTERT fusion protein was used as the antigen, and the variable regions in heavy chain (VH) of immunized mice were RT-PCR amplified and cloned into the pCANTAB 5E, a phagemid vector. By transfection, the display library of mouse VH was developed. The candidate clones were selected by affinity panning, and soluble VH were obtained after expression in E. coli, HB2151. The resultant single VH antibodies were characterized on their binding potentials by western blotting.</p><p><b>RESULTS</b>An about 350 bp VH fragment was amplified from spleen cells of mice immunized by His-tagged hTERT and expressed by phage displayed as VH library. The size of the library was 8 x 10(4). After three rounds of affinity panning, 4 independent clones were chosen and consequently expressed as soluble single domain antibodies (Mr = 16 000). In Western blot analysis, the single domain antibody from 2 of 4 clones proved to react with the His-tagged hTERT fusion protein (Mr = 167 000) without dependence of His-tags and also detect the native hTERT (Mr = 127 000) extracted from the human HeLa cancer cell line. DNA sequencing showed both of the single domain antibodies were encoded by the heavy chain variable region of the mouse.</p><p><b>CONCLUSIONS</b>The single domain antibodies developed were hTERT recognizable and hTERT specific, thus providing a basis for application of recombinant single domain antibody in inhibition of telomerase activity and anticancer therapy.</p>
Subject(s)
Animals , Humans , Mice , Amino Acid Sequence , Antibodies, Monoclonal , Genetics , Allergy and Immunology , Base Sequence , Cloning, Molecular , Complementarity Determining Regions , Genetics , DNA-Binding Proteins , HeLa Cells , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin Variable Region , Genetics , Molecular Sequence Data , Sequence Analysis, DNA , Telomerase , Allergy and ImmunologyABSTRACT
@#Surface transcutaneous electric nerve stimulation(TENS) was applied to treat the spasiticity resulting from spinal cord injury(SCI) in the acupoints of the subjects with SCI in order to oberve thetranscient and long-term effect of the treatment Fand its mechanism will be discussed in this paper. The results show that: 1 )more significant effect in relieving the spasiticiy was got by stimulating in high frequencyfor 30minutes,while less effect in low frequency(2Hz) ; 2)the effect in high frequency was partially ostracized by naloxone; 3)the effect in high frequency lasted for a long time i 4)compared with lumbar and sacralstimulation,the acupoint stimulation lasted for the longest time. This study also gets a hypothesis that highfrequency TENS in acupoints improves dynorphin in CSF to release and significantly relieve the spasiticityby kappa opioid receptor which decreases the excitment of anterior motor neuron.
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The effects of formaldehyde fixation on the binding capacity of opiate receptors were studied with radioreceptorassay and in vitro receptor autoradiography. Incubation with 1% paraformaldehyde for 30 minutes or 12 hours has no significant influence on the binding capacity of opiate receptors of rat brain P_2 membranes, and incubation with 2% or 4% paraformaldehyde for 30 minutes also did not alter the binding capacity of opiate receptors significantly, but 12 hour incubation with 2% or 4% paraformaldehyde would change the binding capacity significantly. The saturation curve of [~3H]-etorphine binding with opiate receptors in formaldehyde fixed brain tissue sections coincided with that of unfixed brain tissue sections. The opiate receptors were successfully demonstrated with in vitro receptor autoradiography in 1% paraformaldehyde fixed spinal cord sections. These results indicate that formaldehyde fixed tissue are applicable to in vitro receptor autoradiography.