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Objective:To observe the effect of tetrahydroxy stilbene glycoside (TSG) on the expression of glycogen synthase kinase 3<italic>β </italic>(GSK3<italic>β</italic>), cyclic adenosine monophosphate-dependent protein kinase (PKA) and Serine/threonine phosphatase 2A(PP2A) in the brain of amyloid precursor protein/presenilin-1/Tau (APP/PS1/Tau) triple-transgenic mice dementia model. Method:A total of forty-five 8-month-old APP/PS1/Tau transgenic mice were randomly divided into model group, positive control group (Huperzine-A, 0.15 mg·kg<sup>-1</sup>), low, medium and high dose TSG groups (TSG, 0.033,0.1,0.3 g·kg<sup>-1</sup>), with 9 mice in each group, and another nine C5B7L/6J mice of the same age were selected as normal control group. After 60 days of intragastric administration, the general structure of hippocampal neurons was observed by hematoxylin-eosin (HE) staining, immunohistochemical (IHC) was used to detect the expression of PKA protein in the brain of mice in each group, the mRNA expression levels of GSK3<italic>β</italic>, PKA and PP2A were detected by real time quantitative reverse transcription polymerase chain reaction (Real-time PCR), and protein expression levels of GSK3<italic>β</italic> and PP2A were detected by Western blot. Result:Compared with the normal control group, the apoptosis level of neurons in the model group was significantly increased, the protein and mRNA expression levels of GSK3<italic>β</italic> and PKA were significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01), and the protein and mRNA expression levels of PP2A were significantly decreased (<italic>P</italic><0.05, <italic>P</italic><0.01). Compared with the model group, the apoptosis level of neurons in each treatment group was significantly down-regulated, the protein and mRNA expression levels of GSK3<italic>β</italic> and PKA were significantly down-regulated (<italic>P</italic><0.05, <italic>P</italic><0.01), and the protein and mRNA expression levels of PP2A were significantly increased (<italic>P</italic><0.05, <italic>P</italic><0.01). Conclusion:The mechanism of TSG in the treatment of Alzheimer's disease (AD) may be related to lowering the transcription and expression of GSK3<italic>β</italic> and PKA, increasing the transcription and expression of PP2A.
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The RAS (rat sarcoma) gene is one of the important oncogenes, and its mutation is present in about 30% human tumors. KRAS (kirsten rat sarcoma viral oncogene) is one of the three RAS subtypes, and KRAS mutations are more common than the mutations in other two RAS subtypes. In recent years, with the continuous research, new ideas have been provided for the treatment of cancers via targeting-KRAS. Efforts have been made to develop various KRAS inhibitors. Here, based on the mechanism of action, we classified KRAS inhibitors into two categories: inhibitors that directly target KRAS and inhibitors that indirectly act on KRAS. The representative KRAS inhibitors were summarized and introduced in this paper.
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A method for determination of 9 isoflavones in Puerariae Lobatae Radix was established and the accuracy and feasibility of the method were verified. The relative correction factors of eight isoflavonoids,3'-hydroxy puerarin,puerarinapioside,3'-methoxy puerarin,puerarin 6″-O-xyloside,daidzin,genistin,formononetin and daidzein were determined by HPLC method with puerarin as the internal standard. The contents of 9 isoflavonoids in 11 batches of samples were determined by external standard method and QAMS.The accuracy and feasibility of the methods were evaluated by comparison of the quantitative results between external standard method and QAMS. The reproducibility of the relative correction factors was good under different experimental conditions,and there was no significant difference between the external standard method of the 9 compounds and the content of QAMS method. The results showed that using puerarin as an internal standard to simultaneously determine the 8 isoflavonoids mentioned above is accurate and feasible. Thus,it can be used as quality control of Puerariae Lobatae Radix.
Subject(s)
Chromatography, High Pressure Liquid , Drugs, Chinese Herbal , Isoflavones , Plant Roots , Pueraria , Reproducibility of ResultsABSTRACT
The biomass multi-elements self-doped TiO2was synthesized simultaneously by ultrasonic irradiation assisted sol-gel method, and characterized by field emission scanning electron microscopy (FESEM), X-ray diffraction(XRD), X-ray photoelectron spectroscopy (XPS), UV-Vis diffuse reflectance spectroscopy (UV-Vis-DRS), Fourier transform infrared (FT-IR) spectroscopy, and photoluminescence (PL). The characterization results showed that,multiple elements, C, N, P, Cl and K, were doped in the composite TiO2. Compared with pure TiO2,the band gap of the composite catalyst was narrowed by 0.21 eV, and possessed more surface hydroxyl radical and active sites, lower recombination rate of photo-generated carriers, higher crystallinity and higher specific surface area. The photocatalytic ability of the composite catalyst was studied,using methylene blue (MB) as target pollutant. The experimental results showed that, under visible light irradiation,the degradation efficiency of methylene blue was up to 98% after photocatalytic reaction for two hours by the composite catalyst.
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<p><b>OBJECTIVE</b>To investigate the value of serum miR-17-92 cluster in the diagnosis of retinoblastoma (RB).</p><p><b>METHODS</b>Serum samples were collected from 20 children with RB and 20 healthy controls. Quantitative real-time PCR was used to measure the expression of miR-17-92 cluster. The expression of miR-17-92 cluster was compared between children with different stages of RB and the changes in the expression of miR-17-92 cluster after multimodality therapy were analyzed. The receiver operating characteristic (ROC) curve was used to investigate the value of serum miR-17-92 cluster in the diagnosis of RB.</p><p><b>RESULTS</b>Compared with the healthy controls, the children with RB had significantly higher relative expression of miR-17-3P, miR-17-5P, miR-18a, and miR-20a in serum (P<0.05), and miR-18a showed the greatest increase. There were no significant differences in the relative expression of miR-19a, miR-19b-1, and miR-92a-1 between children with RB and healthy controls (P>0.05). There were no significant differences in the expression of miR-17-5P, miR-17-3P, miR-18a, and miR-20a between the children with early-to-moderate stage of RB and those with advanced stage of RB (P>0.05), but there were significant reductions after multimodality therapy (P<0.05). In the diagnosis of RB, the areas under the ROC curve (AUCs) for serum miR-17-3P, miR-17-5P, miR-18a, and miR-20a were 0.770, 0.755, 0.828, and 0.665 respectively, and miR-18a had the largest AUC, with a sensitivity of 90% and a specificity of 65%.</p><p><b>CONCLUSIONS</b>miR-17-3P, miR-17-5P, miR-18a, and miR-20a are highly expressed in the serum of children with RB, and miR-18a may be used as a new marker for the diagnosis of RB.</p>
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Child, Preschool , Female , Humans , Infant , Male , Biomarkers, Tumor , Blood , MicroRNAs , Blood , ROC Curve , Retinoblastoma , Blood , Diagnosis , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To screen out retinoblastoma (RB)-related serum tumor markers by measuring the levels of serum alpha fetoprotein (AFP), carcino-embryonic antigen (CEA), neuron-specific enolase (NSE), carbohydrate antigen 125 (CA125), carbohydrate antigen 153 (CA153), carbohydrate antigen 199 (CA199), and carbohydrate antigen 724 (CA724) in children with RB.</p><p><b>METHODS</b>The levels of seven serum tumor markers (AFP, CEA, NSE, CA125, CA153, CA199, and CA724) were determined in 20 children with RB and 20 healthy children (control) using a chemiluminescent immunoassay.</p><p><b>RESULTS</b>The serum levels and positive rates of NSE, CA153, and CA199 in the RB group were significantly higher than those in the control group (P<0.05). However, there were no significant differences in the levels of AFP, CEA, CA125, and CA724 between the two groups (P>0.05). NSE had the highest sensitivity, but a relatively low specificity for the diagnosis of RB. CA153 and CA199 had a relatively high specificity, but a relatively low sensitivity for the diagnosis of RB.</p><p><b>CONCLUSIONS</b>The serum levels and positive rates of NSE, CA153, and CA199 are high in children with RB. Combined measurement of these three serum tumor markers may have an important diagnostic value for RB.</p>
Subject(s)
Child, Preschool , Female , Humans , Infant , Male , Antigens, Tumor-Associated, Carbohydrate , Blood , Biomarkers, Tumor , Blood , CA-125 Antigen , Blood , Phosphopyruvate Hydratase , Blood , Retinal Neoplasms , Blood , Diagnosis , Retinoblastoma , Blood , DiagnosisABSTRACT
Objective To investigate the capability of 99 Tcm?tricine?EDDA/HYNIC?SFSIIHTPILPL ( SP94) as a specific probe for HCC imaging. Methods HYNIC?SP94 peptide was prepared by solid phase synthesis, followed by 99 Tcm labeling with tricine?EDDA as the coligand. After determination of radiochemical purity and stability, cell binding study was carried out by incubating Huh?7 cells with 99 Tcm?tricine?EDDA/HYNIC?SP94 at different specific activities (2.5, 4.0 and 30.0 GBq/μmol). The biodistribution studies and microSPECT/CT imaging were performed in Huh?7 tumor?bearing mice ( study group) and Hela tumor?bear?ing mice ( control group ) . Statistical analysis was by two?sample t test. Results 99 Tcm?tricine?EDDA/HYNIC?SP94 was synthesized with over 95% of labeling yield, which remained stable in saline and FBS up to 12 h. With increasing concentrations of 99 Tcm?tricine?EDDA/HYNIC?SP94, Huh?7 cell binding increased but became gradually saturated. In biodistribution studies, (1.02±0.26) %ID/g of tracer was accumulated in Huh?7 tumors at 0.5 h after injection of 99 Tcm?tricine?EDDA/HYNIC?SP94, higher than that in the HYNIC?SP94 blocking group ((0.34±0.09) %ID/g;t=3.537, P<0.05). Compared to Hela tumors, Huh?7 tumors were clearly visualized by microSPECT/CT, with which better imaging quality could be achieved with higher specific radioactivity. Conclusion 99 Tcm?tricine?EDDA/HYNIC?SP94 could achieve a high labeling effi?ciency and good in vitro stability as a potential diagnostic tracer specifically targeted for HCC.
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The aim of this study was to develop a murine model of brain injury induced by high altitude hypoxic inflammation. In the study, we used a decompression chamber to mimic an acute hypobaric hypoxia, and 8-week-old male C57BL/6 mice were intraperitoneally injected with 5 mg/kg lipopolysaccharide (LPS) to induce inflammatory response. We determined the levels of pro-inflammatory factors (IL-6, TNF-α) and anti-inflammatory factor (IL-10) in mice serum using ELISA assays to confirm the high altitude hypoxic inflammation, and verified the brain injury after the inflammation using hematoxylin-eosin (HE) staining. The results showed that, among four experiment groups (ctrl, acute hypobaric hypoxia, LPS, and acute hypobaric hypoxia plus LPS groups), the acute hypobaric hypoxia plus LPS treatment group displayed the highest levels of IL-6, TNF-α, and IL-10. Meanwhile, the acute hypobaric hypoxia plus LPS treatment group showed the most severe cortex and hippocampus injuries, including cellular swelling, the widened pericellular spaces, angiogenesis, and shrunken neurons with darkly stained pyknotic nuclei, etc. Strikingly, nuclei ventrales posteriors thalami were found to be more sensitive to acute hypobaric hypoxia plus LPS treatment, and their destroy degrees were higher than those neurons in cortex and hippocampus. These results suggested that we established a reliable murine model of brain injury induced by high altitude hypoxic inflammation, and might be useful to the relevant studies.
Subject(s)
Animals , Male , Mice , Altitude , Brain Injuries , Cerebral Cortex , Disease Models, Animal , Hippocampus , Hypoxia , Inflammation , Mice, Inbred C57BL , NeuronsABSTRACT
<p><b>OBJECTIVE</b>To investigate associations between health-promoting lifestyle and suboptimal health status (SHS) in the population of Guangdong province.</p><p><b>METHODS</b>A cross-sectional survey was conducted in a clustered sample of 24 159 individuals aged 12-80 years from 2012 to 2013. Health-promoting lifestyle was assessed via the Health-Promoting Lifestyle Profile (HPLP-II), and SHS was evaluated using the medical examination report and Sub-health Measurement Scale V1.0 (SHMS V1.0).</p><p><b>RESULTS</b>Of the 24159 participants, subjects with SHS (46.0%) and disease status (35.2%) accounted for a much higher percentage than healthy subjects (18.8%). Regression analyses revealed a significant association between health status and healthy lifestyle (P<0.001). Unhealthy lifestyle was an important risk factor for SHS and disease, especially the former. Compared with the participants with a healthy lifestyle (minimal exposure), after demographic adjustment, subjects with a 'poor' lifestyle (maximal exposure) were at a 43 times higher risk of developing SHS (OR: 42.825, 95% CI: 30.567-59.997), those with a general lifestyle were at a 21 times higher risk of SHS (OR: 21.072, 95%CI: 17.258-25.729), and those with a suboptimal lifestyle had a 4 times higher risk (OR: 4.085, 95%CI: 3.352-4.979). In the general population, the major risk factors for SHS included poor stress management, poor self-actualization, inactive exercise and poor interpersonal relationship.</p><p><b>CONCLUSION</b>s Unhealthy lifestyles are significantly related to an increased risk of SHS. Intervention of unhealthy lifestyles, controlling the risk factors of SHS, and rigorous management of the time window of SHS are necessary to promote the heath status.</p>
Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Humans , Middle Aged , Young Adult , Cross-Sectional Studies , Health Promotion , Health Status , Life Style , Regression Analysis , Risk FactorsABSTRACT
<p><b>OBJECTIVE</b>To determine evaluate the effect of health-promoting lifestyle on the outcomes of suboptimal health status (SHS).</p><p><b>METHODS</b>A prospective population cohort was conducted by consecutively enrolling 5676 college students who took routine health examination from March to May 2013. The participants were assessed for baseline health status and lifestyle and 2972 participants with SHS were followed up for 1.5 years. Exposure was defined as an unhealthy lifestyle. The health-promoting lifestyle was assessed via the Health-promoting Lifestyle Profile (HPLP-II). SHS was evaluated using the medical examination report and Sub-health Measurement Scale V1.0 (SHMS V1.0).</p><p><b>RESULTS</b>Among the 2972 students with SHS, 422 showed recovery of the healthy status at 1.5 year follow-up, 579 showed progression into disease conditions, and 1971 remained in SHS. The participants with recovered health status presented with significant increase of SHMS V1.0 scores by 8.75∓6.95 points compared to the baseline assessment (t=-2.14, P=0.000) in physiological, psychological and social dimensions; they also showed a marked improvement of HPLP-II scores by 14.73 points in 6 dimensions (t=-15.34, P=0.000). Multivariable regression analyses with adjusted demographic variables revealed a significant association between health status and health-promoting lifestyle (P<0.05). Compared with a healthy lifestyle (minimal exposure), a 'poor' lifestyle (the highest level of exposure) was associated with a 30 times higher risk of developing SHS (OR: 30.598, 95% CI: 3.928-238.331), while a 'moderate' lifestyle (a relatively high-level exposure) had a 24 times higher risk of SHS (OR: 23.988, 95%CI: 14.695-39.158), and a suboptimal lifestyle had a nearly 4 times higher risk of SHS (OR: 4.306, 95%CI: 2.767-6.702).</p><p><b>CONCLUSION</b>s SHS may evolve into either a healthy or a disease condition. A unhealthy lifestyle is the important risk factor contributing to the progression of SHS into a disease condition, suggesting the importance of intervention of unhealthy lifestyles in promoting good health.</p>
Subject(s)
Humans , Health Behavior , Health Status , Healthy Lifestyle , Prospective Studies , Regression Analysis , Risk Factors , StudentsABSTRACT
OBJECTIVE: To observe whether low concentration (1×10-8 mol·L-1) of ouabain (OUA) can increase the contractility in rat cardiocytes and investigate the Na/K pump signal transduction pathways related to positive inotropic action following the low concentration of OUA. METHODS: On enzymatic isolation of rats ventricular myocytes, the Na+/K+ pump current (Ip) was by whole-cell patch-clamp, in order to observe the low concentration of OUA on Ip. The contraction of a single myocyte was assessed by a video-based motion edge-detection system. (1) To detect and compare the potentiations of 1×10-8-1×10-3 mol·L-1 OUA on the contractility in rat cardiocytes. (2) The cardiocytes were pre-treated with PP2 (1 μmol·L-1), NAC (100 μmol·L-1), PD98059(50 μmol·L-1) for 5 min, and the effects of the signals transduction inhibitors on the positive inotropic effect of 1×10-8 mol·L-1 OUA was recorded. RESULTS: The 1×10-8-1×10-3 mol·L-1 OUA increased the contractility of rat cardiocytes (P0.05). CONCLUSION: The 1×10-8-1×10-3 mol·L-1 OUA could increase the contraction amplitude of cardiocytes in rats in concentration-dependent manner. Positive inotropic effect of OUA in low concentration is related to Na/K pump signal transduction. Multiple signal pathways regulate the positive inotropic effect of 1×10-8 mol·L-1 of OUA, including the Src/ROS signal pathway.
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<p><b>OBJECTIVE</b>To investigate the clinical characteristics and changing trends of febrile seizures (FS) in children.</p><p><b>METHODS</b>The clinical data of 1 922 children with FS admitted from 2004 to 2013 were analyzed retrospectively. The clinical characteristics and changing trends of FS in the two five-year periods were analyzed.</p><p><b>RESULTS</b>In 1 922 children with FS, the male/female ratio was 2.27:1. The mean age at onset was 3.0±1.8 years, while the peak age at onset was 1-3 years. There were 1 556 children (80.96%) with simple FS and 366 (19.04%) with complex FS. The number of children with FS in the second five-year period (1 202 cases) increased by 66.9% compared with that in the first five-year period (720 cases). The proportion of children with complex FS was significantly higher in the second five-year period than in the first five-year period (21.13% vs 15.56%; P<0.05). In children with simple FS, there were no significant differences in the age and body temperature at onset, convulsion duration, and the incidence of complications between the two five-year periods (P>0.05). However, children with complex FS had a significantly lower age at onset, a significantly lower body temperature at onset, a significantly longer convulsion duration, and a significantly higher incidence of complications including myocardial injury and hyponatremia in the second five-year period than in the first five-year period (P<0.05).</p><p><b>CONCLUSIONS</b>In the last decade, the number of children with FS and incidence of complex FS increased, and the outcome became worse according to clinical characteristics, suggesting that more attention should be paid to timely diagnosis and treatment of complex FS.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Infant , Male , Age of Onset , China , Epidemiology , Seizures, Febrile , EpidemiologyABSTRACT
Orthostatic hypotension is most common in elderly people, and its prevalence increases with age. Attenuation of the vestibulo-sympathetic reflex (VSR) is commonly associated with orthostatic hypotension. In this study, we investigated the role of glutamate on the vestibulo-solitary projection of the VSR pathway to clarify the pathophysiology of orthostatic hypotension. Blood pressure and expression of both pERK and c-Fos protein were evaluated in the nucleus tractus solitarius (NTS) after microinjection of glutamate into the medial vestibular nucleus (MVN) in conscious rats with sodium nitroprusside (SNP)-induced hypotension that received baroreceptor unloading via sinoaortic denervation (SAD). SNP-induced hypotension increased the expression of both pERK and c-Fos protein in the NTS, which was abolished by pretreatment with glutamate receptor antagonists (MK801 or CNQX) in the MVN. Microinjection of glutamate receptor agonists (NMDA or AMPA) into the MVN increased the expression of both pERK and c-Fos protein in the NTS without causing changes in blood pressure. These results indicate that both NMDA and AMPA receptors play a significant role in the vestibulo-solitary projection of the VSR pathway for maintaining blood pressure, and that glutamatergic transmission in this projection might play a key role in the pathophysiology of orthostatic hypotension.
Subject(s)
Aged , Animals , Humans , Rats , Blood Pressure , Denervation , Excitatory Amino Acid Antagonists , Glutamic Acid , Hypotension , Hypotension, Orthostatic , Microinjections , N-Methylaspartate , Nitroprusside , Pressoreceptors , Prevalence , Receptors, AMPA , Receptors, Glutamate , Reflex , Sodium , Solitary Nucleus , Vestibular NucleiABSTRACT
Input signals originating from baroreceptors and vestibular receptors are integrated in the rostral ventrolateral medulla (RVLM) to maintain blood pressure during postural movement. The contribution of baroreceptors and vestibular receptors in the maintenance of blood pressure following hypotension were quantitatively analyzed by measuring phosphorylated extracellular regulated protein kinase (pERK) expression and glutamate release in the RVLM. The expression of pERK and glutamate release in the RVLM were measured in conscious rats that had undergone bilateral labyrinthectomy (BL) and/or sinoaortic denervation (SAD) following hypotension induced by a sodium nitroprusside (SNP) infusion. The expression of pERK was significantly increased in the RVLM in the control group following SNP infusion, and expression peaked 10 min after SNP infusion. The number of pERK positive neurons increased following SNP infusion in BL, SAD, and BL+SAD groups, although the increase was smaller than seen in the control group. The SAD group showed a relatively higher reduction in pERK expression when compared with the BL group. The level of glutamate release was significantly increased in the RVLM in control, BL, SAD groups following SNP infusion, and this peaked 10 min after SNP infusion. The SAD group showed a relatively higher reduction in glutamate release when compared with the BL group. These results suggest that the baroreceptors are more powerful in pERK expression and glutamate release in the RVLM following hypotension than the vestibular receptors, but the vestibular receptors still have an important role in the RVLM.
Subject(s)
Animals , Rats , Blood Pressure , Denervation , Glutamic Acid , Hypotension , Neurons , Nitroprusside , Pressoreceptors , Protein KinasesABSTRACT
<p><b>OBJECTIVE</b>To investigate the preventive effect of behavioral therapy plus flunarizine in children with migraine.</p><p><b>METHODS</b>Ninety pediatric patients with migraine between January 2011and January 2014 were randomly divided into treatment group (45 cases) and control group (45 cases). The treatment group received behavioral therapy in addition to oral flunarizine, while the control group received oral flunarizine alone. All patients were followed up for 3 months to evaluate the therapeutic effect by the Pediatric Migraine Disability Assessment Score (PedMIDAS) and improved Bussone headache index.</p><p><b>RESULTS</b>There were no significant differences in PedMIDAS (P>0.05) and improved Bussone headache index (P>0.05) between the control and treatment groups before treatment. Significant differences were observed in PedMIDAS (16±8 vs 20±10; P<0.05) and improved Bussone headache index (25±18 vs 37±21; P<0.05) between the two groups after 3 months of treatment.</p><p><b>CONCLUSIONS</b>Preventive treatment of behavioral therapy plus oral flunarizine shows a better clinical efficacy than oral flunarizine alone in children with migraine and holds promise for clinical application.</p>
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Adolescent , Child , Female , Humans , Male , Behavior Therapy , Combined Modality Therapy , Flunarizine , Therapeutic Uses , Migraine Disorders , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To study the clinical features and risk factors of co-morbid tic disorder (TD) in children with attention deficit hyperactivity disorder (ADHD).</p><p><b>METHODS</b>A total of 312 children with ADHD were involved in this study. Subtypes of co-morbid TD, incidences of TD in different subtypes of ADHD (ADHD-I, ADHD-HI and ADHD-C) were observed. Thirteen potential factors influencing the comorbidity rate of TD in ADHD were evaluated by univariate analysis and multiple logistic regression analysis.</p><p><b>RESULTS</b>Forty-two of 312 children with ADHD suffered from co-morbid TD (13.5%). Comorbidity rate of TD in children with ADHD-C (24.1%) was significantly higher than in those with ADHD-HI (10.9%) and ADHD-I (8.8%) (P<0.05). There were 21 cases (50.0%) of transient TD, 12 cases (28.6%) of chronic TD, and 9 cases (21.4%) of Tourette syndrome. The univariate analysis revealed 6 factors associated with comorbidity: addiction to mobile phone or computer games, poor eating habits, infection, improper family education, poor relationship between parents and poor relationship with schoolmates. Multiple logistic analysis revealed two independent risk factors for comorbidity: improper family education (OR=7.000, P<0.05) and infection (OR=2.564, P<0.05).</p><p><b>CONCLUSIONS</b>The incidence of co-morbid TD in children with ADHD is influenced by many factors, and early interventions should be performed based on the main risk factors.</p>
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Adolescent , Child , Female , Humans , Male , Attention Deficit Disorder with Hyperactivity , Comorbidity , Logistic Models , Risk Factors , Tic Disorders , EpidemiologyABSTRACT
Establishment of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) with co-expression E2 Epitope of Classical Swine Fever virus (CSFV) is a crucial step to develop a genetic engineered vaccine against PRRSV and CSFV. Reverse genetic manipulation could be adopted as a com monly used technique. In this study, we focus on using nonessential regions of NSP2 (aa480-532 and aa508-532) as viral vector to express E2 Epitope of CSFV. A neutralizing epitope of classical swine fever virus (CSFV) E2 protein was inserted into the two nonessential region of nsp2 by the method of mutant PCR, basing on the infectious clone of HuN4-F112 vaccine strain. The co-expressed full-length cDNA clones (psk-HuN4-F112-delta508-532 + E2 and psk-HuN4-F112-delta480-532 + E2) were assembled by cloning and splice of the gene fragments. The completely assembled full-length cDNA clones were confirmed by sequence and Swa I enzyme digestion. Capped RNAs were transcribed in vitro from a full-length cDNA clone of the viral genome and transfected into BHK-21 cells by liposome to acquire the rescued virus. The rescued recombinant viruses were passaged on MARC-145 cells. The successfully rescued viruses were tested by RT-PCR, digestion, and genome sequence. The results showed that these rescued viruses could be distinguished from the parental virus (HuN4-F112) with the mutant genetic marker (Mlu I enzyme site of virual genome at 14 667nt was created by synonymous mutation) and the inserted nsp2 gene region. The results of IFA showed that the inserted E2 epitope could be expressed by the recombinant viruses and the E2 epitope gene was stable during the viral serial passage. The results of plaque assay and viral growth curve showed that the recovery viruses possessed similar characterses of viral growth to those of the parental virus. In summary, the full-length infectious cDNA clones containing the marker gene were constructed and the marker recombinant viruses were rescued. The results suggested that these stable infectious clones could be used as an important tool for development of novel vaccine against PRRSV.
Subject(s)
Base Sequence , Cysteine Endopeptidases , Genetics , Epitopes , Genetics , Molecular Sequence Data , Porcine respiratory and reproductive syndrome virus , Genetics , Viral Envelope Proteins , Genetics , Allergy and Immunology , Viral Vaccines , Allergy and ImmunologyABSTRACT
Contribution of the vestibular end organ to regulation of arterial pressure was quantitatively compared with the role of baroreceptors in terms of baroreflex sensitivity and c-Fos protein expression in the rostral ventrolateral medulla (RVLM). Baroreflex sensitivity and c-Fos protein expression in the RVLM were measured in conscious rats that had undergone bilateral labyrinthectomy (BL) and/or baroreceptor unloading. BL attenuated baroreflex sensitivity during intravenous infusion of sodium nitroprusside (SNP), but did not significantly affect the sensitivity following infusion of phenylephrine (PE). Baroreflex sensitivity became positive following sinoaortic denervation (SAD) during infusion of PE and attenuated sensitivity during infusion of SNP. Baroreflex sensitivity also became positive following double ablation (BL+SAD) during infusion of PE, and attenuated sensitivity during infusion of SNP. c-Fos protein expression increased significantly in the RVLM in the sham group after SNP administration. However, the BL, SAD, and SAD+BL groups showed significant decreases in c-Fos protein expression compared with that in the sham group. The SAD group showed more reduced c-Fos protein expression than that in the BL group, and the SAD+BL group showed less expression than that in the SAD group. These results suggest that the vestibular system cooperates with baroreceptors to maintain arterial pressure during hypotension but that baroreceptors regulate arterial pressure during both hypotension and hypertension. Additionally, afferent signals for maintaining blood pressure from the vestibular end organs and the baroreceptors may be integrated in the RVLM.
Subject(s)
Animals , Rats , Arterial Pressure , Baroreflex , Blood Pressure , Denervation , Hypertension , Hypotension , Infusions, Intravenous , Nitroprusside , Phenylephrine , Pressoreceptors , SalicylamidesABSTRACT
<p><b>OBJECTIVE</b>To investigate the clinical results of distal tibia fractures accompanying with anterior soft tissue injury by posteromedial approach.</p><p><b>METHODS</b>From August 2008 to August 2010,31 patients (21 males and 10 females, ranged in age from 24 to 68 years with an average of 46 years) with distal tibia fractures accompanying with anterior soft tissue injury were treated with open reduction and internal fixation through posteromedial approach. Of the 31 cases, 14 cases were open fractures (including 5 cases Gustilo-Anderson type I, 5 type II, 2 type III A, and 2 type III B) and 17 cases were closed fractures (including 3 cases Tcherne-Oestern type I, 12 type II, 2 type III). According to typing of AO, type 43A were in 26 cases and type 43C1 were in 5 cases. Operation time, intraoperative blood loss, fracture healing time, AOFAS ankle score, and complications were recorded to evaluate clinical effects.</p><p><b>RESULTS</b>All the patients were followed up from 12 to 36 months with an average of 21 months. The mean operation time was 70 min (ranged, 40 to 110 min) and the mean blood loss was 100 ml (ranged, 50 to 200 ml). Average healing time for closed fractures was 13 weeks (ranged, 10 to 18 weeks), while open fractures was 19 weeks (ranged, 15 to 29 weeks). Cross-foot of 40 occurred in 1 case and pes valgus of 30 in 1 case at final follow-up; and AOFAS ankle scores were from 69 to 100 with an average of 88.4 +/- 9.7. Of them, 10 cases obtained excellent results, 17 good, 4 fair.</p><p><b>CONCLUSION</b>Postermedial approach is an effective method for distal tibia fractures especially accompanying with local soft tissue injury or bad skin condition.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Follow-Up Studies , Fracture Fixation, Internal , Methods , Soft Tissue Injuries , General Surgery , Tibial Fractures , General SurgeryABSTRACT
<p><b>OBJECTIVE</b>To investigate therapeutic effects of vacuum sealing drainage (VSD) in the treatment of soft tissue defect combined with tendon and bone exposure.</p><p><b>METHODS</b>From October 2007 to February 2011, 397 patients (412 feet) with open ankle fracture and dislocation combined with soft tissue defected were treated by VSD. There were 301 males and 96 females with an average age of 36 years (ranging age from 20 to 73 years). According to AO classification, 74 feet were type I, 211 feet were type II, 108 feet were type III and 19 feet were type IV. The mean time from injury to operation was 5.6 h ( 2 to 12 h). The mean treatment time of was 10 months (4 to 19 months).</p><p><b>RESULTS</b>One hundred and forty-one patients were primarily healed, 97 patients were sutured at stage II. Split-thick skin grafting was performed at stage II was performed in 103 patients; free flap transplantation was performed in 25 patients. Three of the 34 patients with infection were removed steel plate; Eviscerate flap coverage wound was performed in 14 patients caused by the first metatarsal bone exposure; Toe amputation were performed in 22 cases caused by toes necrosis. Tarsometatarasl joints perforators' surgery was performed in 10 patients with forefeet necrosis. Thirty hundred and six patients were followed up from 3 to 20 months (averaged 10 months). The wounds healed well.</p><p><b>CONCLUSION</b>VSD for soft tissue defects caused by ankle injury is a simple and effective method, but can not replace debridement and transfer flap.</p>