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Objective: To explore the characteristics of viral infections in children with diarrhea in Beijing from 2018 to 2022. Methods: Real-time PCR and enzyme-linked immunosorbent assay were used to detect viral nucleic acid of Norovirus (NoV), Sappovirus (SaV), Astrovirus (AstV), Enteric Adenovirus (AdV) or antigen of Rotavirus (RV) in 748 stool samples collected from Beijing Capital Institute of Pediatrics from January 2018 to December 2021. Subsequently, the reverse transcription PCR or PCR method was used to amplify the target gene of the positive samples after the initial screening, followed by sequencing, genotyping and evolution analysis, so as to obtain the characteristics of these viruses. Phylogenetic analysis was performed using Mega 6.0. Results: From 2018 to 2021, the overall detection rate of the above five common viruses was 37.6%(281/748)in children under 5 years old in Beijing. NoV, Enteric AdV and RV were still the top three diarrhea-related viruses, followed by AstV and SaV, accounting for 41.6%, 29.2%, 27.8%, 8.9% and 7.5%, respectively. The detection rate of co-infections with two or three diarrhea-related viruses was 4.7% (35/748). From the perspective of annual distribution, the detection rate of Enteric AdV was the highest in 2021, while NoV was predominant in the other 4 years. From the perspective of genetic characteristics, NoV was predominant by GⅡ.4, and after the first detection of GⅡ.4[P16] in 2020, it occupied the first two gene groups together with GⅡ.4[P31]. Although the predominant RV was G9P[8], the rare epidemic strain G8P[8] was first detected in 2021. The predominant genotypes of Enteric AdV and AstV were Ad41 and HAstV-1. SaV was sporadic spread with a low detection rate. Conclusion: Among the diarrhea-related viruses infected children under 5 years of age in Beijing, the predominant strains of NoV and RV have changed and new sub-genotypes have been detected for the first time, while the predominant strains of AstV and Enteric AdV are relatively stable.
Subject(s)
Child, Preschool , Humans , Infant , Beijing/epidemiology , Diarrhea/epidemiology , Feces , Norovirus/genetics , Phylogeny , Rotavirus/genetics , Virus Diseases/epidemiology , Viruses/geneticsABSTRACT
In this study, a rat morphine drug discrimination model with a fixed ratio (FR) of 10 (FR10) was established using different methods to explore which methods can shorten the modeling time and test the dose-response relationship and median effective dose (ED50) value. Animal welfare and experimental procedures are in accordance with the provision of the Animal Ethics Committee of Shanghai InnoStar Bio-tech Co., Ltd. Forty rats were initially shaped to press lever under a fixed-ratio schedule of food reinforcement. The animals that were successfully trained under a FR10 schedule of food reinforcement were divided into two groups, namely the single-lever + double-lever training group 1 and the double-lever training group 2. In each group, rats were trained to discriminate morphine at 5.6 mg·kg-1 from saline by the intraperitoneal route. After training, different doses of morphine were used to substitute for training dose of morphine, the dose-response curve for morphine were identified in rats, and the ED50 value was calculated. The results showed that, in food training phase: 34 rats successfully entered the discrimination training during food training; in discrimination training phase: 14 animals in group 1 met the discrimination training standard for the first time, which took about (40.71 ± 2.93) days, and there were 13 animals in group 2 that met the discrimination training criteria for the first time, and it took about (51.15 ± 2.55) days. It can be seen that the method of single-lever + double-lever training is better than single-lever training, and the difference is significant compared with group 1 (P ˂ 0.05); in generalization test phase: there are 17 rats completed morphine generalization test, and the percentages of morphine-lever responses produced by the generalization test of different doses of morphine (0, 0.1, 0.5, 1, 3, 5.6, and 10 mg·kg-1) were (9.56 ± 3.13) %, (9.01 ± 5.83) %, (13.82 ± 7.95) %, (29.04 ± 10.13) %, (41.70 ± 10.65) %, (85.36 ± 7.16) %, (94.56 ± 2.76) %, respectively. The results showed that the discriminative stimulative effect induced by morphine dose between 0-10 mg·kg-1 increased in a dose-dependent manner, producing a good dose-response curve, and the ED50 value of morphine was 4.74 mg·kg-1 by linear fitting. The above results showed that, the FR10 morphine drug discrimination model has been successfully established using different methods; the single-lever + double-lever training method is better than the single-lever training, and can relatively shorten the discrimination training cycle.
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@#【Objective】To screen survival-related differential expression of long non-coding RNA（lncRNA）and its co-expressed genes in breast cancer patients and to verify their expression in breast cancer cells.【Methods】RNA-seq data of 943 cases（837 breast cancer + 106 normal controls）by the TCGA database were screened，and found that long non-coding MAPT-AS1 highly expressed，and breast cancer patients had longer survival. The long non-coding MAPT- AS1 overexpression and interference plasmid was constructed，and the constructed plasmid was transfected into breast cancer cell line T47D，and the stably expressed T47D cell line was screened by puromycin. The expression of long non-coding MAPT-AS1 and its co-expressed genes was verified by the methods of RT-qPCR.【Results】Fluorescence microscopy and RT-qPCR confirmed that the long non-coding MAPT-AS1 overexpression and interference-transfected breast cancer cell line T47D were successfully constructed，and the long non-coding MATS-AS1 interference fragment shRNA3 with the highest interference efficiency was screened. The expression of MAPT ，MAPT- IT1 and NXNL2 in the co-expressed gene was decreased after transfection of the shRNA3 interference fragment ，which was consistent with the expression trend of the long non-coding MAPT-AS1.【Conclusion】The long non-coding MAPT-AS1 overexpression and interference plasmid transfected breast cancer cell line T47D were successfully constructed，and the expression of the co- expressed gene was consistent with the database. The study laid the foundation for further study of the mechanism of action of long non-coding MAPT-AS1 gene in breast cancer.
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OBJECTIVE@#To assess the effects of two surface treatments (sandblasting, SB; microarc-oxidation, MAO) and resin luting on shear bond strength and durability of titanium alloy and composite-resin.@*METHODS@#Eighty cylindrical titanium alloy specimens with a diameter of 10 mm and a height of 8 mm were fabricated by CAD/CAM technique. It was divided into two groups according to the surface treatment methods: sandblasting with Al2O3 particles on the surface of SB specimens; porous ceramic film structure could be formed on the surface of MAO specimens after surface treatment. Each group was classified into SB-resin luting-N group (not used), SB-resin luting-Y group (used), MAO-resin luting-N group (not used), MAO-resin luting-Y group (used) depending on whether or not resin luting was applied. Each specimen was bonded and cured with the Cemerage resin, and the shear bond strength after 0 and 5 000 thermocycling was tested. The results were statistically analyzed. The surface morphology of titanium alloy specimens before and after the shear bond strength test was observed by scanning electron microscopy (SEM).@*RESULTS@#The shear bond strength between titanium alloy and composite-resin was the highest in the SB combined with resin luting group after 0 thermocycling (16.2±1.8) MPa; was the lowest in MAO group after 5 000 thermocycling (8.9±1.5) MPa. The shear bond strength of SB and MAO surface treatment methods combined without resin luting group after 5 000 thermocycling were (10.7±2.2) MPa and (8.9±1.5) MPa, which were statistically lower than those in the thermocycling 0 (P=0.000 and P=0.001). The shear bond strength of SB and MAO surface treatment methods combined with resin luting group after 5 000 thermocycling were (15.5±2.1) MPa and (11.7±1.3) MPa, respectively, which were lower than those in the thermocycling 0 group, but there was no statistical significance (P=0.087 and P=0.234).@*CONCLUSION@#Both the surface treatment methods of SB and MAO combined with resin luting can improve the shear bond strength and durability of titanium alloy and composite-resin. The SB combined with resin luting is more significant. At present, the effect of SB is better than that of MAO due to the limitation of technical parameters of micro-arc oxidation.
Subject(s)
Alloys , Composite Resins , Dental Bonding , Materials Testing , Resin Cements , Shear Strength , Surface Properties , TitaniumABSTRACT
Objective:To explores the improvement in survival mechanism of a rat model of enterocoelia heterotopic heart transplant with rat bone marrow mesenchymal stem cells( BMSCs) IDO-overexpressing. Methods:IDO-overexpressing rat BMSCs were produced through transfection of rat BMSCs with IDO gene carried by the lentiviral vector GV308. A rat model of enterocoelia heterotopic heart transplantation was established. This rat model received a cell treatment via its tail veins, as follows: ①Echocardiography was employed to detect the functional changes in the transplanted heart.②The fluorescence intensity of the different parts of the transplanted heart was evaluated using a body imaging system for small living animals.③Receptors rat spleens cells were obtained and used for a flow cytometric detection of the expression levels of CD40,CD86,CD80,MHCⅡ,CD274,CD45RA,CD45RA+CD45RB,and Treg cells. ④A transplanted heart was obtained after injection to evaluate inflammatory cell infiltration through HE staining.⑤Liquid phase chips were used to detect changes in the serum factors IL-1ɑ,IL-4,IL-1β,IL-2,IL-10,IFN-γ,IL-18,TGFβ1, TGFβ2 and TGFβ3 in after injection cells. Results:①After the rat heterotopic heart transplantation model and the corresponding cell treatment were established,after over-expressed IDO-BMSCs treatment 2 days the EF and FS were higher in the transplanted heart than other groups.②The fluorescence intensity of the parts of the transplanted heart was highest in the IDO-BMSC overexpression group as revealed by small animal living body evaluation. ③Two days after the interventions, spleen cells in the over-expressed IDO-BMSCs group showed reduced expression levels of CD40,CD86,CD80,MHCⅡ,CD45RA,CD45RA+CD45RB and increased expression levels of CD274 and Treg cells as revealed by flow cytometry.④Liquid phase chips were used to examine the serum obtained from each group 2 days after the intervention,and the results showed that the expression levels of IL-1α,IL-4,IL-1β,IL-2,IFN-γand IL-18 in the IDO-BMSC overexpression group decrease. By contrast,the expression levels of IL-10,TGFβ1,TGFβ2 and TGFβ3 increase. HE staining results demonstrate that inflammatory cell infiltration was lower in IDO-BMSC overexpression group than in other groups. Conclusion:IDO-overexpressing BMSCs improve the survival of a transplanted heart through effective adjustment of immune DC and T cells,as well as cell factors.
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Objective:To explores the improvement in survival mechanism of a rat model of enterocoelia heterotopic heart transplant with rat bone marrow mesenchymal stem cells( BMSCs) IDO-overexpressing. Methods:IDO-overexpressing rat BMSCs were produced through transfection of rat BMSCs with IDO gene carried by the lentiviral vector GV308. A rat model of enterocoelia heterotopic heart transplantation was established. This rat model received a cell treatment via its tail veins, as follows: ①Echocardiography was employed to detect the functional changes in the transplanted heart.②The fluorescence intensity of the different parts of the transplanted heart was evaluated using a body imaging system for small living animals.③Receptors rat spleens cells were obtained and used for a flow cytometric detection of the expression levels of CD40,CD86,CD80,MHCⅡ,CD274,CD45RA,CD45RA+CD45RB,and Treg cells. ④A transplanted heart was obtained after injection to evaluate inflammatory cell infiltration through HE staining.⑤Liquid phase chips were used to detect changes in the serum factors IL-1ɑ,IL-4,IL-1β,IL-2,IL-10,IFN-γ,IL-18,TGFβ1, TGFβ2 and TGFβ3 in after injection cells. Results:①After the rat heterotopic heart transplantation model and the corresponding cell treatment were established,after over-expressed IDO-BMSCs treatment 2 days the EF and FS were higher in the transplanted heart than other groups.②The fluorescence intensity of the parts of the transplanted heart was highest in the IDO-BMSC overexpression group as revealed by small animal living body evaluation. ③Two days after the interventions, spleen cells in the over-expressed IDO-BMSCs group showed reduced expression levels of CD40,CD86,CD80,MHCⅡ,CD45RA,CD45RA+CD45RB and increased expression levels of CD274 and Treg cells as revealed by flow cytometry.④Liquid phase chips were used to examine the serum obtained from each group 2 days after the intervention,and the results showed that the expression levels of IL-1α,IL-4,IL-1β,IL-2,IFN-γand IL-18 in the IDO-BMSC overexpression group decrease. By contrast,the expression levels of IL-10,TGFβ1,TGFβ2 and TGFβ3 increase. HE staining results demonstrate that inflammatory cell infiltration was lower in IDO-BMSC overexpression group than in other groups. Conclusion:IDO-overexpressing BMSCs improve the survival of a transplanted heart through effective adjustment of immune DC and T cells,as well as cell factors.
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<p><b>OBJECTIVE</b>To observe the inhibitory effect of gefitineb on the proliferation and its inducing effect on the apoptosis of mouse I-10 Leydig testicular cancer cells in vitro.</p><p><b>METHODS</b>We treated I-10 Leydig testicular cancer cells of mice with gefitineb at 0, 1.25, 2.5, 5, 10, 20, and 40 µmol/L. Then we determined the inhibitory effect of gefitineb on the growth of the cells by MTT, detected their early and late apoptosis by Annexin V-FITC/propidium iodide double staining and Hoechst 33258 nuclear staining, respectively, and observed the expressions of apoptosis-related proteins Bcl-2, Bax and caspase 3/9 by Western blot.</p><p><b>RESULTS</b>Compared with the blank control group, gefitineb significantly inhibited the proliferation of the I-10 cells at 10 and 20 µmol/L (P < 0.05). The survival rate of the cells was (32.4 ± 2.8)% (P < 0.01) and their early and late apoptosis rates were (26.7 ± 4.2)% and (59.33 ± 10.2)% in the 40 µmol/L group, significantly different from those in the control (P < 0.05 and P <0.01). In comparison with the blank control group, gefitineb at 10, 20, and 40 µmol/L increased the expression of pro-apoptotic protein Bax by (41.9 ± 7.1), (60.1 ± 9.8), and (69.0 ± 11.3)% (all P < 0.05), decreased that of apoptosis-inhibitory protein Bcl-2 by (50.3 ± 8.9), (63.9 ± 6.9), and (88.7 ± 13.9)% (all P < 0.05), and elevated that of the cleft proteins caspase-3 by (69.0 ± 6.9)% (P < 0.05), (71.5 ± 8.1)% (P < 0.05), and (110.9 ± 14.2)% (P < 0.01) and caspase-9 by (51.8 ± 4.9), (54.7 ± 6.7), and (43.8 ± 11.8)% (all P < 0.05).</p><p><b>CONCLUSION</b>Gefitineb can increase the cytotoxicity of I-10 Leydig testicular cancer cells of mice and induce their apoptosis via the mitochondria-mediated apoptosis signaling pathway.</p>
Subject(s)
Animals , Male , Mice , Antineoplastic Agents , Pharmacology , Apoptosis , Apoptosis Regulatory Proteins , Metabolism , Caspase 3 , Metabolism , Caspase 9 , Metabolism , Cell Proliferation , Cell Survival , Leydig Cell Tumor , Drug Therapy , Metabolism , Pathology , Neoplasm Proteins , Metabolism , Neoplasms, Germ Cell and Embryonal , Drug Therapy , Metabolism , Pathology , Quinazolines , Pharmacology , Testicular Neoplasms , Drug Therapy , Metabolism , Pathology , bcl-2-Associated X Protein , MetabolismABSTRACT
In order to establish a rapid and accurate method for the detection of Ebola virus (EBOV), the primers used in SYBR Green I real-time RT-PCR were designed based on the EBOV NP gene sequences published in GenBank. The SYBR Green I real-time RT-PCR was established and optimized for the detection of EBOV. The EBOV RNA that was transcribed in vitro was used as a template. The sensitivity of this method was found to reach 1.0 x 10(2) copies/microL and the detection range was 10(2) - 10(10). No cross reaction with RNA samples from Marburg virus, Dengue virus, Xinjiang hemorrhagic fever virus, Japanese encephalitis virus, Influenza virus (H1N1 and H3N2) and Porcine reproductive and respiratory syndrome virus E genomic RNA was found. The method would be useful for the detection and monitoring of EBOV in China.