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Objective:To analyze the detection rate of respiratory syncytial virus(RSV)and human rhinovirus(HRV), in different months and age groups, and the clinical characteristics in children in eastern Guangdong from 2019 to 2020.Methods:Pharyngeal swabs were collected from 6 658 children with respiratory tract infections hospitalized in the Second Affiliated Hospital of Shantou University Medical College from January 2019 to December 2020, and respiratory pathogen nucleic acid was detected.The detection rate, month distribution, age group distribution, and clinical characteristics of single RSV as well as single HRV positive cases were analyzed and compared.Results:There were 416 single RSV positive cases(6.25%)and 341 single HRV positive cases(5.12%).The detection rates of RSV was higher than those of HRV, and the difference was statistically significant( χ2=7.880, P<0.05).The detection rates of HRV in March, April, November and December were higher than those of RSV, and the detection rates of RSV in July, August and September were higher than those of HRV, with statistically significant difference( P<0.05).The highest detection rate of RSV was in the age group of ≤6 months with a detection rate of 13.47%(192/1 425), which gradually decreased with age, and the difference was statistically significant( P<0.01).The detection rates of HRV fluctuated between 4.21% and 6.13% in each age group, and the differences among the detection rates of each age group were not statistically significant( P>0.05).All RSV-positive cases showed cough, while 77.13%(263/341)of HRV-positive cases showed cough, with a statistically significant difference( P<0.001).The incidence of wheezing in RSV-positive cases was 37.26%(155/416)compared with 28.45%(97/341)in HRV-positive cases, with a higher incidence of wheezing in RSV than that in HRV, and the difference was statistically significant( P<0.05).In terms of indicators to assess severe pneumonia, RSV-positive cases showed a higher proportion of increased respiratory rate, decreased oxygen saturation or dyspnea than HRV-positive cases, and all differences were statistically significant( P<0.05). Conclusion:The detection rate of single RSV is higher than that of single HRV in children with respiratory infections in eastern Guangdong from 2019 to 2020.The epidemic season of RSV is mainly in autumn, and the epidemic season of HRV is mainly in winter and spring.RSV is more susceptible up to 6 months of age, and the detection rate decreases gradually with age, and there is no significant difference in the detection rate of HRV by age.RSV-positive cases are more likely to have cough and wheeze.RSV-positive cases are more likely to have increased respiratory rate, decreased oxygen saturation, or respiratory distress.
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Snake venom has special pharmacological activities and contains a array of small polypeptides that can antagonize integrins, therefore called disintegrins. Disintegrins can block integrin-dependent platelet aggregation, tumor growth, and tumor metastasis. A disintegrin fraction was isolated and purified from the venom of snake Gloydius brevicaudus (GBV). Its physical and chemical properties were characterized, and its biological activities were investigated. The crude venom of GBV were isolated by Superdex 75 gel filtration chromatography. The anti-platelet aggregation activity of the fractions was screened by the Born method. The fraction that shown anti-platelet activity was further purified with Sephadex G-25 gel filtration, DEAE Sepharose Fast Flow ion exchange chromatography, and Lichrospher C18 reversed-phase chromatography respectively. The purity of the active component was analyzed with SDS-PAGE (Tris-Tricine system) and high-performance liquid-phase chromatography (HPLC), with protein concentration determined by the Bradford method. The molecular weight was evaluated by the gel imaging method and mass spectrometry, and the isoelectric point was measured by disc isoelectric focusing electrophoresis. The protease activity was measured with the Rick method. The phospholipase A activity was determined by the automatic potentiometric titration method. Amino acid sequencing results were subjected to homology comparison using the BLAST program. Seven fractions (Ⅰ-Ⅶ) were isolated from GBV by gel filtration chromatography on Superdex 75 column. The fraction Ⅳ inhibited the platelet aggregation induced by ADP with molecular weight lower than 10 000 Da, suggesting a disintegrin component. A disintegrin named GBV-Ⅳ4 was purified from the fraction by Sephadex G-25 gel filtration, DEAE Sepharose Fast Flow ion-exchange and Lichrospher C18 reverse chromatography. It was homogeneous shown as a single band on SDS-polyacrylmide gel electrophoresis (SDS-PAGE, Tris-Tricine system) with molecular weight 8 746 Da as calculated by Image Master VDS system. The isoelectric point of GBV-Ⅳ4 was 6.3 by disc polyacrylamide gel electrophoresis. GBV-Ⅳ4 exhibited no detectable phospholipase A2 (PLA2) activity with the pH-stat technique or proteinase activity according to the method of Rick. GBV-Ⅳ4 is composed of 70 amino acids with RGD (Arg-Gly-Asp) active region and a molecular weight of 7 442 Dalton as assayed by Mass Spectrography. Characterization of GBV-Ⅳ4 is consistent with meta-chain disintegrin (70 amino acid sequence, six pairs of disulfide bond). Retrieved by Genbank, GBV-Ⅳ4 has high homology with other disintegrins. We concluded that GBV-Ⅳ4 is a novel disintegrin contained RGD. GBV-Ⅳ4 showed dose-dependent inhibition of ADP- or thrombin-induced platelet aggregation with IC50 0.339 or 0.577 μg·mL-1 respectively. In conclusion, a new disintegrin derived from the GBV snake venom and named GBV-Ⅳ4 containing RGD tripeptide sequence could inhibit platelet aggregation.
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A target cell extraction-chemical profiling method based on human alveolar adenocarcinoma cell line (A549 cells) and UHPLC/LTQ Orbitrap MS for screening the anti-lung cancer bioactive compounds from Curcuma longa has been developed in this paper. According to the hypothesis that when cells are incubated together with the extract of Curcuma longa, the potential bioactive compounds in the extract should selectively combine with the cells, then the cell-binding compounds could be separated and analyzed by LC-MS. The bioactive compounds in C. longa are lipophilic components. They intend to be absorbed on the inner wall of cell culture flask when they were incubated with A549 cells, which will produce interference in the blank solution. In this paper, by using cells digestion and multi-step centrifugation and transfer strategy, the interference problem has been solved. Finally, using the developed method, three cell-binding compounds were screened out and were identified as bisdemethoxycurcumin, demethoxycurcumin, and curcumin. These compounds are the main bioactive compounds with anti-lung cancer bioactivity in C. longa. The improved method developed in this paper could avoid the false positive results due to the absorption of lipophilic compounds on the inner wall of cell culture flask, which will to be an effective complementary method for current target cell extraction-chemical profiling technology.
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Objective To explore the character of ruminant thinking individuals in cold executive functions(cool EF) and hot executive functions(hot EF).Methods According to the score of rumination response scale( RRS) ,17 low-level ruminant thinking individuals and 21 high-level ruminant thinking indi-viduals were screened out and finished the classic Stroop test.Results In the cool EF,it was consistent be-tween low-level and high-level ruminant thinking individuals for color naming task response time ((10.61± 23.20)ms vs (10.79±29.32)ms),and there was no significant difference in the classic Stroop test( t=0.21, P>0.05) .In the hot EF,the respone time of the low-level group was longer than that of high-level group on the positive and negative((-5.01±22.20)ms vs (-10.88±20.33)ms;(8.78±29.96)ms vs (-8.68±19.94) ms) ,and the main effect of the emotional Stroop interference scores between positive and negative words was highly significant(F=10.88, P<0.05) .The interactive effect of emotional Stroop interference scores of words × subjects was significant(F=5.70, P<0.05) .The simple effect tests showed that the emotional Stroop interfer-ence scores between high-level and low-level ruminant thinking subjects were significant in the negative group(F=4.69, P<0.05) .And it was also significant between positive and negative words in the low-level group(F=14.63, P<0.05).Conclusion Two types of subjects in the cold EF have no significant difference. High-level ruminant thinking individuals in the cold EF are normal,but impaired in the hot EF that meaning high-level ruminant thinking individuals had bias to negative emotion.These results provide new clues for the intervention of negative emotions caused by ruminants.
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Objective: To investigate the effect of X-ray irradiation on epithelial-mesenchymal transition (MET) in human colorectal cancer SW480 cells and its involved potential signaling pathway. Methods: SW480 cells were irradiated with different doses of X-ray (0, 2, 4, 6 and 8 Gy). The migration and invasion abilities of SW480 cells after irradiation were detected by Transwell method. The expression levels of E-cadherin (E-cad), vimentin, K-ras and Smad3 mRNAs and proteins in SW480 cells were detected by real-time fluorogenic quantitative-PCR and Western blotting, respectively. Results: The migration and invasion abilities of SW480 cells after irradiation with X-ray were increased as compared with that of the control cells (no irradiation) (P < 0.05). The mRNA and protein expression levels of E-cad and K-ras were significantly decreased while the expression levels of vimentin mRNA and protein were significantly increased after irradiation as compared with those of the control cells (all P < 0.05). The expression level of Smad3 mRNA and protein in transforming growth factor-p1 (TGF-p1)/Smad3 signaling pathway were also increased (both P < 0.05) after irradiation. Conclusion: The results of this study support that X-ray irradiation can induce EMT of human colorectal cancer SW480 cells, and this effect may be related to the activation of TGF-β1/Smad3 signaling pathway.
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Objective: To develop an analysis method based on N-propylethylenediamine (PSA)-HPLC-QQQ-MS/MS for the determination of 12 kinds pesticide residues in Fritiliariae Thunbergii Bulbus and to quantify them in 20 batches of Fritiliariae Thunbergii Bulbus produced in various places in Zhejiang province. Methods: Fritiliariae Thunbergii Bulbus samples were extracted with acetonitrile and purified by the small column of PSA. The prepared samples were analyzed by HPLC-QQQ-MS/MS in multiple reaction monitoring (MRM) mode, and the pesticides were qualitified by the internal standard method. Results: All the 12 pesticides showed good linearities in their reasonable range (r = 0.9986-0.9998), and the average recoveries of all the pesticides were in the range of 68.1%-108.0% at three spiked levels of 90, 300, and 900 ng/mL. The RSD values were in the range of 1.1%-6.1%, and the LODs of each pesticide were all in the range of 0.08-1.0 μg/kg. Conclusion: The method is suitable for the multiresidues analysis of pesticides in Fritiliariae Thunbergii Bulbus simultaneously and the quality of Fritiliariae Thunbergii Bulbus is basically good for the safty although it contains a trace of several pesticide residues.
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<p><b>OBJECTIVE</b>To investigate the change of expression level of metastasis suppressor gene Kiss-1 in the colorectal cancer cell line SW480 after radiation, and to determine its association with the proliferation and apoptosis of SW480 cells.</p><p><b>METHODS</b>SW480 cells were divided into control group (0 Gy) and study groups (2, 4, 6, 8 Gy). Cells in the study groups were irradiated by 6-MV X-ray radiation for 48 hours. Immunohistochemistry and real-time PCR methods were used to investigate the influence of radiation on Kiss-1 gene expression of SW480. Colony formation assay was used to detect the proliferation of SW480. Flow cytometry-Annexin- V/PI assay was used to observe the change of the apoptosis rate.</p><p><b>RESULTS</b>Compared with the control group, Kiss-1 protein expression increased after radiation of 6, 8 Gy (P<0.05), but no significant changes were observed after radiation of 2, 4 Gy(P>0.05). Kiss-1 gene mRNA level increased after radiation of 2, 4, 6 Gy, while no obvious change was observed for 8 Gy radiation. The apoptosis rates increased for 4, 6, 8 Gy radiation(P<0.05), however, there was no significant difference for 2 Gy radiation (P<0.05).</p><p><b>CONCLUSION</b>Radiation may increase Kiss-1 gene expression in SW480 cells, which results in decreases proliferation and increases apoptosis in residual surviving cells.</p>