InCl3/TfOH-Mediated Convenient Synthesis of 3-Alkylideneoxindoles from 2-Oxindoles with 1,3-Diones, Ketones, or Aldehydes.

Two efficient and convenient methods for the synthesis of 3-alkylideneoxindoles are described in this paper. The InCl3/TfOH-mediated tandem Knoevenagel condensation-deacylation sequence of various 2-oxindoles with 1,3-diones or acetoacetate furnished 3-alkylideneoxindoles in satisfactory to excellent yields (up to >99% yield). Employing the reaction system, the condensation of 2-oxindoles with ketones or aldehydes also proceeded smoothly to produce 3-alkylideneoxindoles. This protocol can be amenable to scale up. The effect of acids on this condensation reaction and intermolecular competition experiments were investigated to understand the aspect of the reaction.

Genomic Elucidation of a COVID-19 Resurgence and Local Transmission of SARS-CoV-2 in Guangzhou, China.

While China experienced a peak and decline in coronavirus disease 2019 (COVID-19) cases at the start of 2020, regional outbreaks continuously emerged in subsequent months. Resurgences of COVID-19 have also been observed in many other countries. In Guangzhou, China, a small outbreak, involving less than 100 residents, emerged in March and April 2020, and comprehensive and near-real-time genomic surveillance of SARS-CoV-2 was conducted. When the numbers of confirmed cases among overseas travelers increased, public health measures were enhanced by shifting from self-quarantine to central quarantine and SARS-CoV-2 testing for all overseas travelers. In an analysis of 109 imported cases, we found diverse viral variants distributed in the global viral phylogeny, which were frequently shared within households but not among passengers on the same flight. In contrast to the viral diversity of imported cases, local transmission was predominately attributed to two specific variants imported from Africa, including local cases that reported no direct or indirect contact with imported cases. The introduction events of the virus were identified or deduced before the enhanced measures were taken. These results show the interventions were effective in containing the spread of SARS-CoV-2, and they rule out the possibility of cryptic transmission of viral variants from the first wave in January and February 2020. Our study provides evidence and emphasizes the importance of controls for overseas travelers in the context of the pandemic and exemplifies how viral genomic data can facilitate COVID-19 surveillance and inform public health mitigation strategies.

Whole-Transcriptome RNA Sequencing Reveals Significant Differentially Expressed mRNAs, miRNAs, and lncRNAs and Related Regulating Biological Pathways in the Peripheral Blood of COVID-19 Patients.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was initially identified in China and currently worldwide dispersed, resulting in the coronavirus disease 2019 (COVID-19) pandemic. Notably, COVID-19 is characterized by systemic inflammation. However, the potential mechanisms of the "cytokine storm" of COVID-19 are still limited. In this study, fourteen peripheral blood samples from COVID-19 patients (n = 10) and healthy donors (n = 4) were collected to perform the whole-transcriptome sequencing. Lung tissues of COVID-19 patients (70%) presenting with ground-glass opacity. Also, the leukocytes and lymphocytes were significantly decreased in COVID-19 compared with the control group (p < 0.05). In total, 25,482 differentially expressed messenger RNAs (DE mRNA), 23 differentially expressed microRNAs (DE miRNA), and 410 differentially expressed long noncoding RNAs (DE lncRNAs) were identified in the COVID-19 samples compared to the healthy controls. Gene Ontology (GO) analysis showed that the upregulated DE mRNAs were mainly involved in antigen processing and presentation of endogenous antigen, positive regulation of T cell mediated cytotoxicity, and positive regulation of gamma-delta T cell activation. The downregulated DE mRNAs were mainly concentrated in the glycogen biosynthetic process. We also established the protein-protein interaction (PPI) networks of up/downregulated DE mRNAs and identified 4 modules. Functional enrichment analyses indicated that these module targets were associated with positive regulation of cytokine production, cytokine-mediated signaling pathway, leukocyte differentiation, and migration. A total of 6 hub genes were selected in the PPI module networks including AKT1, TNFRSF1B, FCGR2A, CXCL8, STAT3, and TLR2. Moreover, a competing endogenous RNA network showed the interactions between lncRNAs, mRNAs, and miRNAs. Our results highlight the potential pathogenesis of excessive cytokine production such as MSTRG.119845.30/hsa-miR-20a-5p/TNFRSF1B, MSTRG.119845.30/hsa-miR-29b-2-5p/FCGR2A, and MSTRG.106112.2/hsa-miR-6501-5p/STAT3 axis, which may also play an important role in the development of ground-glass opacity in COVID-19 patients. This study gives new insights into inflammation regulatory mechanisms of coding and noncoding RNAs in COVID-19, which may provide novel diagnostic biomarkers and therapeutic avenues for COVID-19 patients.

Self-assembled GeOX/Ti3C2TX Composites as Promising Anode Materials for Lithium Ion Batteries.

High-capacity germanium-based anode materials are alternative materials for outstanding electrochemical performance lithium-ion batteries (LIBs), but severe volume variation and pulverization problems during charging-discharging processes can seriously affect their electrochemical performance. In addressing this challenge, a simple strategy was used to prepare the self-assembled GeOX/Ti3C2TX composite in which the GeOX nanoparticles can grow directly on Ti3C2TX layers. Nanoscale GeOX uniformly renucleates on the surface and interlayers of Ti3C2TX, forming the stable multiphase structure, which guarantees its excellent electrochemical performance. Electrochemical evaluation has shown that the rate capability and reversibility of GeOX/Ti3C2TX are both greatly improved, which delivers a reversible discharge specific capacity of above 1400 mAh g-1 (at 100 mA g-1) and a reversible specific capacity of 900 mAh g-1 after 50 cycles while it still maintains a stable specific capacity of 725 mAh g-1 at 5000 mA g-1. Furthermore, the composite exhibits an exceptionally superior rate capability, making it a good electrochemical performance anode for LIBs.

Differential microRNA expression in the peripheral blood from human patients with COVID-19.

INTRODUCTION: The coronavirus disease (COVID-19) is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which play important roles in regulating gene expression and are also considered as essential modulators during viral infection. The aim of this study was to elucidate the differential expression of miRNAs in COVID-19. METHODS: The total RNA was extracted and purified from the peripheral blood of ten patients with COVID-19 and four healthy donors. The expression levels of various miRNAs were detected by high-throughput sequencing, and correlation analysis was performed on the target genes that are primed by miRNAs. KEY FINDINGS: Compared with the healthy controls, 35 miRNAs were upregulated and 38 miRNAs were downregulated in the human patients with COVID-19. The top 10 genes were listed below: hsa-miR-16-2-3P,hsa-miR-5695,hsa-miR-10399-3P,hsa-miR-6501-5P,hsa-miR-361-3P,hsa-miR-361-3p, hsa-miR-4659a-3p, hsa-miR-142-5p, hsa-miR-4685-3p, hsa-miR-454-5p, and hsa-miR-30c-5p. The 10 genes with the greatest reduction were listed below: hsa-miR-183-5p, hsa-miR-627-5p, hsa-miR-941, hsa-miR-21-5p, hsa-miR-20a-5p, hsa-miR-146b-5p, hsa-miR-454-3p, hsa-miR-18a-5p, hsa-miR-340-5p, and hsa-miR-17-5p. Remarkably, miR-16-2-3p was the most upregulated miRNA, with a 1.6-fold change compared to that of the controls. Moreover, the expression of miR-6501-5p and miR-618 was 1.5-fold higher in the COVID-19 patients than in the healthy donors. Meanwhile, miR-627-5p was the most downregulated miRNA, with a 2.3-fold change compared to that of the controls. The expression of other miRNAs (miR-183-5p, miR-627-5p, and miR-144-3p) was reduced by more than 1.3-fold compared to that of the healthy donors. Cluster analysis revealed that all of the differentially expressed miRNA target genes were clustered by their regulation of cellular components, molecular functions, and biological processes. Importantly, peptidases, protein kinases, and the ubiquitin system were shown to be the highest enrichment categories by enrichment analysis. CONCLUSIONS: The differential miRNA expression found in COVID-19 patients may regulate the immune responses and viral replication during viral infection.

Ultrahigh-Rate Behavior Anode Materials of MoSe2 Nanosheets Anchored on Dual-Heteroatoms Functionalized Graphene for Sodium-Ion Batteries.

MoSe2 is a prospective anode material for Na-ion batteries because of its layered structure and high theoretical capacity, while the unsatisfied electrochemical performance limits its further development. Herein, we report MoSe2 nanosheets anchored on dual-heteroatoms functionalized graphene by a solvothermal method. The heteroatoms and carbon matrix coexist in the form of graphitic-N/pyridinic-N/pyrrolic-N and P-C/P═O bonds, which result in excellent electronic conductivity of the materials and provide abundant active sites for electrochemical process. Results indicated that organic intercalation increased the layer spacing of the materials to facilitate sodium-ion diffusion, and the in situ formed carbon networks improved the conductivity among the layers of the materials and alleviated volume expansion during the continued charge and discharge process. As an anode of Na-ion batteries, the nanosheets materials exhibited ultrahigh rate performance and deliver capacities of approximately 200 mAh g-1 at the current density of 10 A g-1. The ultrahigh-rate performance can be attributed to its unique nanosheets structure, the dual-heteroatoms functionalized graphene, and the considerable pseudocapacitive quality of the material.

[Epidemiological Survey on Intestinal Parasitic Infections in Lhasa City in 2014].

Objective: To investigate the prevalence of parasitic infections in Lhasa, Tibet. Methods: The residents in Chengguan District, Dangxiong County, Dazi County and Qushui County were selected to participate in the study in 2014, using the stratified random sampling method. Infections with the soil-borne nematodes in feces were examined in triplicates using the Kato-Katz method. Further, the Enterobius vermicularis were examined using the cellophane anal swab method in children aged 3-12 years, and the intestinal protozoa were detected by iodine staining. The parasitic infection was analyzed by sex, age, and occupation. Comparisons were made using Chi-square test. Results: A total of 1 015 residents（39.3±18.2 years: age range, 1-89 years） participated in the study, including 391 males（38.6%） and 623 females（61.4%）. Two hundred and forty-nine participants were positive for parasitic infections, with an overall infection rate of 24.5%（249/1 015）. The infection rate was 21.7%（220/1 015） for Taenia sp., 0.2%（2/1 015） for Trichuris trichiura and E. vermicularis respectively, 4.1%（42/1 015） for Entamoeba coli, 0.4%（4/1 015） for Giardia lamblia, 0.2%（2/1 015） for Iodamoeba butschlii. No infection was found for other parasites. There was no significant sexual difference in the infection rate of Taenia (male 21.2%, 83/391; female 22.0%, 137/624), T. trichiura (male 0; female 0.3%, 2/624), E. vermicularis (male 0.3%, 2/391; female 0.2%, 1/624), E. coli (male 5.4%, 21/391; female 3.4%, 21/624), G. lamblia (male 0.8%, 3/391; female 0.2%, 1/624), or I. butschlii (male 0; female 0.3%, 2/624) (P>0.05). The Taenia sp. infection rate in the age groups of 0-17, 18-29, 30-39, 40-49, 50-59 and ≥ 60 years was 10.9%（18/165）, 14.0%（18/129）, 23.0%（43/187）, 25.3%（61/241）, 26.5%（41/155）, and 28.3%（39/138）, respectively, with significant differences between groups of 0-17 and 40-49 years and between groups of 50-59 and ≥ 60 years (P<0.01). The infection rate of E. coli, T. trichiura and I. butschlii was highest in the age group of 50-59 years（6.5%, 10/155; 0.7%, 1/155; 1.3%, 2/155 respectively）. Among various occupations, farmers showed the highest infection rate for Taenia（44.5%, 147/330）, E. coli（6.4%, 21/33）, E. vermicularis （0.6%, 2/330） and I. butschlii（0.3%, 1/330）; housewives had the highest infection rate for T. trichiura（2.5%, 1/40）; and the students had the highest infection rate for G. lamblia (0.7%, 1/134). Conclusion: There are infections with various intestinal parasites in the participants of Lhasa City, among which taenia infection reaches 20.7% while others have a low level of infection.

[Changes of phosphatase PTEN in neuronal apoptosis in neonate rats with hypoxic-ischemic brain damage].

OBJECTIVE: To examine the changes in expression of phosphatase and tensin homolog deleted on chromosome ten (PTEN) protein, p-PTEN protein and Bim (Bcl-2 interacting mediator of cell death) mRNA in the cortex of neonate rat brains with hypoxic-ischemic brain damage (HIBD) and to explore the mechanisms of neuroprotective effects of PTEN inhibition. METHODS: One hundred and twenty-eight neonate (10 days) SD rats were divided into four groups: hypoxia-ischemia (HI), sham control (Sham), bisperoxovanadium (bpv), and normal saline (NS) group. Rats in the HI group had their right common carotid arteries (CCA) exposed and ligated, and were then exposed to hypoxia in a chamber filled with 8% oxygen (balanced with nitrogen) for 2.5 h. Rats in the sham control group had their right CCA surgically exposed without ligation and exposure to hypoxia. Rats in the bpv treated group received intraperitoneal injections of bpv, 30 min before HI was induced. Instead of bpv, rats in the NS-treated group received intraperitoneal injections of NS. Cerebral cortex samples of the rats were collected 0.5 h and 24 h after hypoxia. Western blot was used to detect the protein expression of PTEN, p-PTEN and Bim. Real-Time PCR was used to detect the level of Bim mRNA. TUNEL staining was used to detect apoptotic cells. RESULTS: No significant changes of PTEN protein were observed in the rats exposed to HI. However, p-PTEN protein decreased in the rats exposed to HI (0.5 h and 24 h) compared with those exposed to sham surgery (P < 0.01). Compared with the sham controls, Bim mRNA and protein increased in the rats exposed to HI (0.5 h, P < 0.01) and then returned to the baseline level 24 h after HI. No significant changes of PTEN protein were observed in the bpv-treated rats. However, p-PTEN protein increased and Bim mRNA and protein decreased in the bpv-treated rats (0.5 h and 24 h, P < 0.01) compared with those in the HI group and NS-treated group. TUNEL positive cells also reduced in the bpv-treated rats (24 h, P < 0.01) compared with those in the HI group and NS-treated group. CONCLUSION: PTEN activities increase in the brains of neonate rats with hypoxic-ischemic brain damage. PTEN activity inhibition can decrease the level of pro-apoptotic protein Bim mRNA, leading to reduction of neuronal apoptosis.

[Effect of integrin ß8 on TGF-ß1 activation in astrocytes with oxygen glucose deprivation].

OBJECTIVE: To study the effect of ß8 expression on transforming growth factor ß1(TGF-ß1) activation in astrocytes with oxygen glucose deprivation (OGD). METHODS: Astrocytes were cultured and then subjected to OGD to generate hypoxia-ischemia (HI) model in vitro. Immunocytochemistry was used to detect the expression and distribution of ß8 in nomoxia cultured cells. ß8 protein expression was quantified by Western blot at 12 hours, 1 day and 2 days after OGD. Astrocytes and luciferase reporter cells (TMLC) were co-cultured. ß8 RNA interference system was established to specifically inhibit ß8 expression in cultured astrocytes. TGF-ß1 activation was then detected in the co-culture system. RESULTS: ß8 was mainly located in the cytoplasm and neurites of astrocytes. OGD resulted in increase of ß8 protein expression at 12 hours after reoxygenation in astrocytes, which was peaked at 1 day after reoxygenation. TGF-ß1 activation was in accordance with ß8 expression in astrocyte-TMLC co-culture system after reoxygenation. After the inhibition of ß8, TGF-ß1 activation was significantly reduced in all time points. CONCLUSIONS: The highly expressed ß8 plays important roles in the regulation of TGF-ß1 activation in neonatal rats with hypoxic-ischemic brain damage.

Destruction of decabromodiphenyl ether (BDE-209) in a ternary carbonate molten salt reactor.

Soil contamination by PBDEs has become a significant environmental concern and requires appropriate remediation technologies. In this study, the destruction of decabromodiphenyl ether (BDE-209) in a ternary molten salt (Li, Na, K)2 CO3 reactor was evaluated. The effects of reaction temperature, additive amount of BDE-209 and salt mixture, on off-gas species, were investigated. The salt mixture after reaction was characterized by XRD analysis and a reaction pathway proposed. The results showed that the amounts of C2H6, C2H4, C4H8 and CH4 in the off-gas decreased with increases in temperature, while the CO2 level increased. When the reaction temperature reached 750 °C, incomplete combustion products (PICs) were no longer detected. Increasing BDE-209 loading was not helpful for the reaction, as more PICs were produced. Larger amounts of salt mixture were helpful for the reaction and PICs were not observed with the mole ratio 1: 2000 of BDE-209 to carbonate melt. XRD analysis confirmed the capture of bromine in BDE-209 by the molten salt.

[Clinical application of inhaled nitric oxide in hypoxic respiratory failure of preterm infants].

Inhaled NO (iNO) has been shown to have beneficial effects on decreasing pulmonary inflammation, increasing function of surfactant and improving lung growth in prematurely born animal models. iNO has been gradually applied in the neonatal intensive care unit since its first use for persistent pulmonary hypertension (PPHN) in the early 1990's. Although many research findings have shown the benefits of iNO for hypoxic respiratory failure (HRF) of preterm infants, there is no certain evidence to support the routine use of iNO in premature infants. According to recent literature, the mechanism of iNO therapy, treatment scheme, iNO effectiveness and safety in premature infants were reviewed in this article, so as to provide bases for the clinical use of this treatment.

[Role of the FOXO3a transcription factor in neuronal apoptosis in neonatal rats with hypoxic-ischemic brain damage].

OBJECTIVE: To explore the role and mechanisms of FOXO3a nuclear translocation in neuronal apoptosis after hypoxia-ischemia (HI). METHODS: One hundred and sixty 10-day-old Sprague-Dawly rats were randomly divided into two groups: HI and sham-operated. The right common carotid artery was ligated followed by hypoxia exposure for 2.5 hours in the HI group. The sham-operated group rats were not subjected to carotid artery ligation or hypoxia treatment. Rat cerebral cortex was collected at 0.5, 2, 4, 8 and 24 hours after hypoxia. Western blot was used to detect expression of total FOXO3a protein, pnuclear and cytoplasmic FOXO3a and Bim. TUNEL staining was used to detect apoptotic cells. RESULTS: The nuclear protein of FOXO3a obviously increased from 0.5 to 24 hours after HI in a time-dependent manner compared with the sham-operated group (P<0.01). On the contrary, cytoplasmic protein evidently decreased from 0.5 to 24 hours in the HI group compared with the sham-operated group (P<0.01). Bim protein increased from 0.5 hour, peaked at 2 hours, started to decline at 4 hours (P<0.01), and returned to baseline level at 8 and 24 hours after HI in the HI group compared with the sham-operated group. TUNEL positive cells started to express at 4 hours, and peaked at 24 hours after HI (P<0.01). However, TUNEL positive cells were rarely found in the sham-operated group. CONCLUSIONS: HI induces FOXO3a translocation from cytoplasm to nucleus, and enhances protein expression of its target gene Bim in the neonatal rat brain. The upregulation of Bim expression might be related to neuronal apoptosis.

Abiraterone acetate for treatment of metastatic castration-resistant prostate cancer: final overall survival analysis of the COU-AA-301 randomised, double-blind, placebo-controlled phase 3 study.

BACKGROUND: Abiraterone acetate improved overall survival in metastatic castration-resistant prostate cancer at a preplanned interim analysis of the COU-AA-301 double-blind, placebo-controlled phase 3 study. Here, we present the final analysis of the study before crossover from placebo to abiraterone acetate (after 775 of the prespecified 797 death events). METHODS: Between May 8, 2008, and July 28, 2009, this study enrolled 1195 patients at 147 sites in 13 countries. Patients were eligible if they had metastatic castration-resistant prostate cancer progressing after docetaxel. Patients were stratified according to baseline Eastern Cooperative Oncology Group (ECOG) performance status, worst pain over the past 24 h on the Brief Pain Inventory-Short Form, number of previous chemotherapy regimens, and type of progression. Patients were randomly assigned (ratio 2:1) to receive either abiraterone acetate (1000 mg, once daily and orally) plus prednisone (5 mg, orally twice daily) or placebo plus prednisone with a permuted block method via an interactive web response system. The primary endpoint was overall survival, analysed in the intention-to-treat population. This study is registered with ClinicalTrials.gov, number NCT00091442. FINDINGS: Of the 1195 eligible patients, 797 were randomly assigned to receive abiraterone acetate plus prednisone (abiraterone group) and 398 to receive placebo plus prednisone (placebo group). At median follow-up of 20·2 months (IQR 18·4-22·1), median overall survival for the abiraterone group was longer than in the placebo group (15·8 months [95% CI 14·8-17·0] vs 11·2 months [10·4-13·1]; hazard ratio [HR] 0·74, 95% CI 0·64-0·86; p<0·0001). Median time to PSA progression (8·5 months, 95% CI 8·3-11·1, in the abiraterone group vs 6·6 months, 5·6-8·3, in the placebo group; HR 0·63, 0·52-0·78; p<0·0001), median radiologic progression-free survival (5·6 months, 5·6-6·5, vs 3·6 months, 2·9-5·5; HR 0·66, 0·58-0·76; p<0·0001), and proportion of patients who had a PSA response (235 [29·5%] of 797 patients vs 22 [5·5%] of 398; p<0·0001) were all improved in the abiraterone group compared with the placebo group. The most common grade 3-4 adverse events were fatigue (72 [9%] of 791 patients in the abiraterone group vs 41 [10%] of 394 in the placebo group), anaemia (62 [8%] vs 32 [8%]), back pain (56 [7%] vs 40 [10%]), and bone pain (51 [6%] vs 31 [8%]). INTERPRETATION: This final analysis confirms that abiraterone acetate significantly prolongs overall survival in patients with metastatic castration-resistant prostate cancer who have progressed after docetaxel treatment. No new safety signals were identified with increased follow-up.

[Research on the Screening Method of Priority Pollutants with Integrated Environmental Socio-economic Indicators: Example of E-waste Dismantling].

While benefiting mankind, electronic information technology has led to the elimination of many electrical and electronic products due to its rapid update and iteration speed. In addition, the normal use in daily life causes the wear and tear of electronic products, resulting in a large amount of electronic waste. With the increase in electronic products, the amount of electronic waste dismantling has also increased yearly. Therefore, it becomes an important to accurately screen the priority control pollutants generated in e-waste process. In view of the current situation of e-waste dismantling pollution and the needs of monitoring and evaluation in China, this study proposed a screening model that combined analysis at levels and systematic clustering methods and performed a comprehensive score of pollutants and cluster analysis on the basis of assigning and scoring the evaluation factors of e-waste dismantling, taking the most potentially dangerous class in the cluster results as the priority control pollutant.

Rock fragmentation indexes reflecting rock mass quality based on real-time data of TBM tunnelling.

Perception of rock condition (RC) is a challenge in tunnel boring machine (TBM) construction due to lack of space and time to observe and detect RC. To overcome this problem, this study aims to extract a new rock fragmentation index (RFI) that can reflect RC from real-time rock fragmentation data of the TBM. First, a comprehensive review of existing rock fragmentation models is conducted, which leads to some candidate RFIs that can reflect RC. Next, these candidate RFIs are investigated using data from 12,237 samples from a well-monitored tunnel boring process of the TBM in a 20,198 m tunnel. Further, a new RFI system is recommended as the parameter involving the optimal models. Finally, a preliminary study of the relationship between these RFIs and RC is carried out, and it is shown that these RFIs can reflect RC to a large extent. In the TBM boring process, these RFIs can be extracted from real-time TBM fragmentation data and used to predict the RC in the field. Therefore, the challenge of RC perception is solved with this new RFI system. The new RFI system offers significant potential for the real-time rock classification, prediction of the surrounding rock collapse potential, and selection of control parameters or support measures during TBM construction. This will be the key to improving TBM construction performance.

Efficacy of Children Neuropsychological and Behavioral Scale in Screening for Autism Spectrum Disorders through a Combination of Developmental Surveillance.

OBJECTIVE: This study aimed to explore the clinical value of Children Neuropsychological and Behavioral Scale-Revision 2016 (CNBS-R2016) for Autism Spectrum Disorder (ASD) screening in the presence of developmental surveillance. METHODS: All participants were evaluated by the CNBS-R2016 and Gesell Developmental Schedules (GDS). Spearman's correlation coefficients and Kappa values were obtained. Taking GDS as a reference assessment, the performance of the CNBS-R2016 for detecting the developmental delays of children with ASD was analyzed with receiver operating characteristic (ROC) curves. The efficacy of the CNBS-R2016 to screen for ASD was explored by comparing Communication Warning Behavior with Autism Diagnostic Observation Schedule, Second Edition (ADOS-2). RESULTS: In total, 150 children aged 12-42 months with ASD were enrolled. The developmental quotients of the CNBS-R2016 were correlated with those of the GDS (r=0.62-0.94). The CNBS-R2016 and GDS had good diagnostic agreement for developmental delays (Kappa=0.73-0.89), except for Fine Motor. There was a significant difference between the proportions of Fine Motor, delays detected by the CNBS-R2016 and GDS (86.0% vs. 77.3%). With GDS as a standard, the areas under the ROC curves of the CNBS-R2016 were above 0.95 for all the domains except Fine Motor, which was 0.70. In addition, the positive rate of ASD was 100.0% and 93.5% when the cut-off points of 7 and 12 in the Communication Warning Behavior subscale were used, respectively. CONCLUSION: The CNBS-R2016 performed well in developmental assessment and screening for children with ASD, especially by Communication Warning Behaviors subscale. Therefore, the CNBS-R2016 is worthy of clinical application in children with ASD in China.

[Study on transport of Yangyin Tongnao granules in main effective fractions].

OBJECTIVE: To investigate the absorption characteristics and transportation mechanism Yangyin Tongnao granules in main effective fractions in Caco-2 cell model. METHOD: The safety concentrations of Yangyin Tongnao granules in main effective fractions in Caco-2 cells. A Caco-2 cell model was established to study the transport situations after the compatibility of Yangyin Tongnao granules in main effective fractions, and the content was determined by high performance liquid chromatography (HPLC). RESULT: P(app) of puerarin, ligustrazine and astragaloside were less than 1.0 x 10(-6) cm x s(-1), and their P(app) were hard to be close to atenolol. The oral absorption in descending order is shown as the following: puerarin, ligustrazine, astragaloside. After the compatibility between saponins and flavonoids, P(app) of astragaloside was improved obviously, which promoted the transport from apical (AP) to basolateral (BL); the compatibility of puerarin, ligustrazine and astragaloside showed a significant effect in the efflux of astragaloside and no change in the absorption transport of ligustrazine and puerarin at the same time. There is a great difference in bidirectional transport of representative component of each effective fraction, and P(app)(B --> A) was significantly greater than Papp(A --> B), which suggested that the efflux transport from BL side to AP side had an advantage in the three representative components of the three effective fractions in Caco-2 cell monolayer model. CONCLUSION: Astragaloside, ligustrazine and puerarin may be malabsorptive compounds, and the three compounds may be discharged by the transport protein in small intestine membrane.

Development of a multiplex RT-PCR method for the detection of four porcine enteric coronaviruses.

Porcine enteric coronaviruses are pathogens that cause viral diarrhea in pigs and are widely prevalent worldwide. Moreover, studies have shown that some porcine enteric coronaviruses can infect humans and poultry. In order to effectively monitor these viruses, it is necessary to establish a multiple detection method to understand their prevalence and conduct in-depth research. Common porcine enteric coronaviruses include Porcine epidemic diarrhea virus (PEDV), Porcine transmissible gastroenteritis virus (TGEV), Porcine delta coronavirus (PDCoV), and Swine acute diarrhea syndrome coronavirus (SADS-CoV). Pigs infected with these viruses have the common clinical symptoms that are difficult to distinguish. A quadruplex RT-PCR (reverse transcription-polymerase chain reaction) method for the simultaneous detection of PEDV, PDCoV, TGEV and SADS-CoV was developed. Four pairs of specific primers were designed for the PEDV M gene, PDCoV N gene, TGEV S gene and SADS-CoV RdRp gene. Multiplex RT-PCR results showed that the target fragments of PDCoV, SADS-CoV, PEDV and TGEV could be amplified by this method. and the specific fragments with sizes of 250 bp, 368 bp, 616 bp and 801 bp were amplified, respectively. This method cannot amplify any fragment of nucleic acids of Seneca Valley virus (SVV), Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Atypical Porcine Pestivirus (APPV), and has good specificity. The lowest detection limits of PDCoV, PEDV, TGEV and SADS-CoV were 5.66 × 105 copies/µL, 6.48 × 105 copies/µL, 8.54 × 105 copies/µL and 7.79 × 106 copies/µL, respectively. A total of 94 samples were collected from pig farms were analyzed using this method. There were 15 positive samples for PEDV, 3 positive samples for mixed infection of PEDV and PDCoV, 2 positive samples for mixed infection of PEDV and TGEV, and 1 positive sample for mixed infection of PEDV, TGEV, and PDCoV. Multiplex RT-PCR method could detect four intestinal coronaviruses (PEDV, PDCoV, TGEV, and SADS-CoV) in pigs efficiently, cheaply and accurately, which can be used for clinical large-scale epidemiological investigation and diagnosis.

Chuanxiongzine-astragaloside IV decreases IL-1ß and Caspase-3 gene expressions in rat brain damaged by cerebral ischemia/reperfusion: a study of real-time quantitative PCR assay.

The purpose of this study was to establish an absolute quantitative method to detect IL-1ß and Caspase-3 gene expressions in rat brain after cerebral ischemia-reperfusion (I/R) using real-time PCR. Rats were randomized into the following groups: sham operation group, model group (cerebral I/R group), astragaloside IV (AST IV) group, chuanxiongzine-AST IV group and nimodipine group (n = 10 in each group). The rats in all the groups except sham operation group were subjected to cerebral I/R treatment. Sham operation and model groups were treated by normal saline (5 mL/kg). AST IV, chuanxiongzine-AST IV and nimodipine groups received 20 mg/kg AST IV, 10 mg/kg chuanxiongzine plus 20 mg/kg AST IV, and 10 mg/kg nimodipine treatments, respectively. The administrations of drugs were performed with intraperitoneal injections at 0 and 12 h, 1 d, 2 d, 3 d, till to 7 d after I/R. A real-time quantitative PCR assay was developed for absolute quantification of the expressions of IL-1ß and Caspase-3 genes. The absolute quantification approach relies on the construction of an accurate standard curve. Thus, two plasmids which contained rat IL-1ß and Caspase-3 genes respectively were constructed. The cloned circular plasmids were then quantified using a spectrophotometer and used as standards. Standard curves were generated, and the copy numbers of IL-1ß and Caspase-3 mRNA isolated from I/R-damaged brain tissue were also calculated by SYBR Green I dye method using specific primers. The results showed that melting curves exhibited sharp peaks, and PCR product also generated prominent band with expected size in agarose gel electrophoresis, which validated the optimization of the selected primer sets of IL-1ß and Caspase-3 genes. The optimal annealing temperatures of IL-1ß and Caspase-3 genes were 59 °C and 61.2 °C, respectively. Real-time PCR results showed that the expression of IL-1ß and Caspase-3 genes in the model group was significantly elevated compared to that in the sham operation group. However, compared to those in the model group, IL-1ß and Caspase-3 gene expressions were obviously decreased in AST IV, chuanxiongzine-AST IV and nimodipine groups. Especially in chuanxiongzine-AST IV group, those two genes showed the most significant expression down-regulation. These results suggest the absolute quantitative method established in the present study is capable of detecting the changes of IL-1ß and Caspase-3 gene expressions in rat brain damaged by I/R.