1. A novel oncolytic adenovirus inhibits hepatocellular carcinoma growth
Journal of Zhejiang University. Science. B 2019;20(12):1003-1013
Objective: To evaluate the inhibitory role of a novel oncolytic adenovirus (OA), GP73-SphK1sR-Ad5, on the growth of hepatocellular carcinoma (HCC). Methods: GP73-SphK1sR-Ad5 was constructed by integrating Golgi protein 73 (GP73) promoter and sphingosine kinase 1 (SphK1)-short hairpin RNA (shRNA) into adenovirus serotype 5 (Ad5), and transfecting into HCC Huh7 cells and normal human liver HL-7702 cells. The expression of SphK1 and adenovirus early region 1 (E1A) was detected by quantitative real-time PCR (qRT-PCR) and western blot, respectively. Cell viability was detected by methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay, and apoptotic rate was determined by flow cytometry. An Huh7 xenograft model was established in mice injected intratumorally with GP73-SphK1sR-Ad5. Twenty days after injection, the tumor volume and weight, and the survival time of the mice were recorded. The histopathological changes in tumor tissues were observed by hematoxylin-eosin (HE) staining and transmission electron microscopy (TEM). Results: Transfection of GP73-SphK1sR-Ad5 significantly upregulated E1A and downregulated SphK1 in Huh7 cells, but not in HL7702 cells. GP73-SphK1sR-Ad5 transfection significantly decreased the viability and increased the apoptotic rate of Huh7 cells, but had no effect on HL7702 cells. Intratumoral injection of GP73-SphK1sR-Ad5 into the Huh7 xenograft mouse model significantly decreased tumor volume and weight, and prolonged survival time. It also significantly decreased the tumor infiltration area and blood vessel density, and increased the percentages of cells with nucleus deformation and cells with condensed chromatin in tumor tissues. Conclusions: GP73-SphK1sR-Ad5 serves as a novel OA and can inhibit HCC progression with high specificity and efficacy.
2.Based on the macromolecules in traditional Chinese medicine exploring the material basis and mechanism of the analgesic effect of Shaoyao Gancao Decoction
Yuan GAO ; Qin GUO ; Huan-huan WANG ; Ying-ying DONG ; Ning ZHAO ; Zheng-shen YAN ; Yu-qin YANG ; Dong BAI
Acta Pharmaceutica Sinica 2023;58(5):1138-1148
In order to study the analgesic effect of Shaoyao Gancao Decoction, this paper discussed material basis and mechanism from the perspective of macromolecules in traditional Chinese medicine. Inspired by the phenomenon of turbidity after boiling Chinese medicine, this experiment took Shaoyao Gancao Decoction as the research object to study the formation process of precipitation during boiling. The results showed that aggregates with a certain shape were formed in the solvent during the boiling process, and the precipitate was obtained by standing and centrifuging. Analysis found that the precipitation was mainly composed of small molecules such as paeoniflorin, albiflorin, liquiritin, glycyrrhizic acid, isoliquiritin and gallic acid, and macromolecules such as protein and polysaccharide. The composition of precipitate was consistent with that of Shaoyao Gancao Decoction, but the analgesic effect of Shaoyao Gancao Decoction after removing the precipitate was significantly reduced. Based on these results, we isolated small molecular compounds, polysaccharides and protein from Shaoyao Gancao Decoction and their contents are 60.4, 700.7 and 207.2 mg·g-1 respectively. We get the ratio, polysaccharide: small molecule = 11.6∶1, protein: small molecule = 3.4∶1, the precipitate is prepared in the state of boiling. The characterization results showed that the particle size of the precipitate will change significantly after co-heating, and the content determination results showed that the content of the six small molecular compounds which was free in solvent was significantly reduced after the formation of the precipitate. The acetic acid writhing experiment proved that the precipitate has a good analgesic effect, and effectively reduced the levels of inflammatory factors prostaglandin E2 and nitric oxide, and increased the level of anti-inflammatory factor interleukin-10. These results proved that the precipitate in Shaoyao Gancao Decoction is an important material basis for analgesic effect, and macromolecules such as protein and polysaccharide are the main components of the precipitate. The study of macromolecules in the precipitate of Shaoyao Gancao Decoction not only provides new ideas and methods for elucidating the pharmacodynamic material basis of Shaoyao Gancao Decoction, but also provides a reference for analyzing the scientificity of traditional decoction.
3.Clinical analysis of routine testing of vaginal discharge samples in a hospital
Dinghua LIU ; Min BAI ; Jianjiang XUE ; Huan LIU ; Xuemei YU ; Wenming LI
Chongqing Medicine 2015;(19):2654-2656
Objective To analyze the routine testing of vaginal discharge samples and investigate the clinical characteristics of the vaginal micro ecology of a hospital women .Methods The data of 2 533 clinical cases were included .The vaginal infection in different ages and seasons was analyzed .The clinical usefulness of bacterial vaginosis (BV) rapid diagnostic method was estimated . Results The rate of low cleanliness samples (stage Ⅲ + Ⅳ) was significantly higher than that of StageⅠ + Ⅱsamples .Of the 2 533 cases ,the detection rates of fungus and small gram negative or gram variable rods were significantly higher among specific patho‐gens and pathogenic bacteria ,respectively .The rates of low cleanliness samples were highest at age below 20 and in the season of winter (81 .3% ,73 .0% ) ,those of fungus at age from 21 to 30 and in the season of summer (23 .9% ,23 .0% ) ,and those of small gram negative or gram variable rods at age from 31 to 40 and in the season of winter (25 .1% ,37 .3% ) .In comparison with golden method ,the sensitivity and specificity of BV BLUETM method were 80 .0% and 90 .0% ,which reflected the accuracy was 87 .5% . Conclusion Vaginal infection of a hospital women was more common in young group and in the season of winter .Fungus and small gram negative or gram variable rods were the major pathogens .BV BLUETM method could assist in the BV rapid diagnosis ,with potential false positive/negative results being fully validated .
4.Study on the chemical constituents from the herb of Gentianopsis paludosa.
Huan-di WANG ; Cheng-yu TAN ; Yu-guang DU ; Xue-fang BAI ; Hai-feng LU
China Journal of Chinese Materia Medica 2004;29(11):1055-1056
OBJECTIVETo study the chemical constituents from the herb of Gentianopsis paludosa.
METHODColumn chromatogrophy and spectral analysis were used to isolate and identify the constituents.
RESULTSix compounds were isolated and identified as 1,7-dihydroxy-3,8-dimethoxyxanthone (I), 1-hydroxy-3, 7, 8-trimethoxyxanthone (II), 1, 8-dihydroxy-3, 7-dimethoxyxanthone (III), 1-hydroxy-3, 7-dimethoxyxanthone (IV), beta-sitosterol (V), daucosterol (VI).
CONCLUSIONCompounds III-VI were isolated from the plant for the first time.
Gentianaceae ; chemistry ; Plants, Medicinal ; chemistry ; Sitosterols ; chemistry ; isolation & purification ; Xanthones ; chemistry ; isolation & purification
5.Effects of gypsum on the firing of pyrogen-treated thermosensitive neurons in PO/AH of cats.
Yan WANG ; Yu YANG ; Xia BAI ; Li-Hua SUN ; Yu-Huan WU ; Cheng-Liang YANG
Chinese Journal of Applied Physiology 2008;24(3):320-323
AIMTo investigate the possible central mechanism of antipyretic effects of Chinese medicine gypsum.
METHODSGypsum was injected after the fever model was established. The firing rate of thermosensitive neurons in preoptic-anterior hypothalamus(PO/AH) region was recorded by using extracellular microelectrode technique.
RESULTSThe injection of pyrogen evoked decrease in firing rate of the warm-sensitive neurons and increase in the cold-sensitive neurons in the region of PO/AH; the changes of the firing rate of pyrogen- treated warm-sensitive and cold-sensitive neurons could be reversed by the injection of gypsum.
CONCLUSIONThe result may suggest that antipyretic action of gypsum is mediated by its influences on the thermosensitivity neurons in the region of PO/AH.
Action Potentials ; Animals ; Antipyretics ; pharmacology ; Calcium Sulfate ; pharmacology ; Cats ; Fever ; physiopathology ; Hypothalamus, Anterior ; physiopathology ; Male ; Materia Medica ; pharmacology ; Neurons ; physiology ; Preoptic Area ; physiopathology ; Pyrogens
6.A comparative analysis of the methods for titering adenoviruses.
Peng-yu SUN ; Yan-ling ZHANG ; Yu-ming JING ; Xin-ji ZHANG ; Zhe-huan ZHANG ; Cheng-yao LI ; Bai-hong CHEN ; Wan-long TAN ; Hong-wei LI
Journal of Southern Medical University 2011;31(2):234-238
OBJECTIVETo compare different methods commonly used for titering adenovirus and analyze the advantages and limitations of each method.
METHODSFour recombined adenoviruses (Ad-G-AT2R-EGFP, Ad-CMV-EGFP, Ad-mif-shRNA-EGFP and Ad-CBA-GFP) were amplified and purified, and each was titered by optical absorbance, real-time PCR, green fluorescent protein (GFP)-labeled method, immunoassay, and cytopathic effect (CPE). The results were then comparatively analyzed.
RESULTSNo significant difference was found in the titer amounts derived from GFP-labeled method, immunoassay, and cytopathic effect method (P>0.1). A positive correlation was noted in the titer amounts determined by real-time PCR and immunoassay (r=0.965), even though the value (vg/ml) obtained by real-time PCR was 10 times higher than that by immunoassay (ifu/ml).
CONCLUSIONGFP-labeled method and immunoassay allow rapid determination of the adenoviral titer. Real-time PCR can not directly determine the real infectious titer of the adenovirus, but the result is well correlated to that of immunoassay and reflects, though indirectly, the actual infectious titer of adenovirus. Considering the procedural convenience and shorter time consumption, real-time PCR is still a practical method for adenoviral titration.
Adenoviridae ; isolation & purification ; metabolism ; physiology ; Capsid Proteins ; metabolism ; DNA, Viral ; isolation & purification ; Green Fluorescent Proteins ; metabolism ; Real-Time Polymerase Chain Reaction ; methods ; Viral Plaque Assay ; methods ; Virus Replication
7.Isolation and identification of a canine coronavirus strain from giant pandas (Ailuropoda melanoleuca).
Feng Shan GAO ; Gui Xue HU ; Xian zhu XIA ; Yu Wei GAO ; Ya Duo BAI ; Xiao Huan ZOU
Journal of Veterinary Science 2009;10(3):261-263
Two giant pandas (Ailuropoda melanoleuca) died of unknown causes in a Chinese zoo. The clinical disease profile suggested that the pandas may have suffered a viral infection. Therefore, a series of detection including virus isolation, electron microscopy, cytobiological assay, serum neutralization and RT-PCR were used to identify the virus. It was determined that the isolated virus was a canine coronavirus (CCV), on the basis of coronavirus, neutralization by canine anti-CCV serum, and 84.3% to 100% amino acid sequence similarity with CCV. The results suggest that the affected pandas had been infected with CCV.
Amino Acid Sequence
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Animal Diseases/*virology
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Animals
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Animals, Zoo/*virology
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Coronaviridae Infections/*veterinary/virology
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Coronavirus, Canine/genetics/*isolation & purification
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Fatal Outcome
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Female
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Male
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Molecular Sequence Data
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Sequence Alignment
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Sequence Homology, Amino Acid
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Ursidae/*virology
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Viral Proteins/chemistry
8.Anatomical observation on oral part of the facial artery and facial vein and its clinical application
Dongqin YANG ; Lei YU ; Huan BIAN ; Feng TANG ; Yang TAN ; Xueqin BAI ; Yating FU ; Yuexuan HU ; Lan HUANG ; Jing CHEN ; Longhai WU ; Jingping ZHANG ; Yan ZENG ; Xiaobo WANG ; Maocheng RAN
Journal of Regional Anatomy and Operative Surgery 2015;(3):267-269
Objective To observe the oral part of the facial artery and facial vein and to provide anatomical data for clinical applica-tion. Methods The origin, branches, course, diameter, position of oral part of facial artery and facial vein were observed on 32 fixed cada-ves (64 sides). Results The position relation between the facial artery and facial vein is non-constant. Measure the distance from inferior border of mandible to corner of the mouth, angulus mandibulae, mental protuberance midpoint. It is (5. 49 ± 0. 63) cm, (2. 50 ± 0. 89) cm and (6. 20 ± 1. 68) cm in the left side respectively, and (5. 69 ± 0. 72) cm, (2. 56 ± 1. 08) cm and (6. 85 ± 1. 86) cm in the right side re-spectively. The diameter of facial artery in inferior border of mandible is (0. 33 ± 0. 08) cm in the left side and (0. 38 ± 0. 07) cm in the right side;while the diameter of facial vein is (0. 40 ± 0. 12) cm in the left side and (0. 42 ± 0. 18) cm in the right side. The facial artery and facial vein are not concomitant and they are not asymmetry also. The position of superior labial artery arteries is constant, but the position of inferior labial artery arteries have more variations. Conclusion The branches, course, diameter and position of oral part of facial artery and facial vein have a number of variations. The superior labial artery arteries could be positioned more easily than inferior labial artery arter-ies. Being familiar with their distribution is of great importance for clinical application.
9.Contrast Researches of Liver Biopsy and Ultrasound in the Diagnosis of Fatty Liver
Lin ZHUANG ; Ming-Fen HU ; Xing-Mei PANG ; Bao-Li BAI ; Lin WANG ; Hong-Jie YANG ; Yun-Li LI ; Zhi-Jian DONG ; Wen-Qing HUAN ; Qing-Qing WANG ; Wei-Qin YU ; Xiao-Lin KUANG
Journal of Kunming Medical University 2018;39(1):85-91
Objective To evaluate the significance of liver biopsy and B ultrasonograpgy in the diagnosis of fatty liver. Methods The results of 62 patients with liver steatosis diagnosed by liver puncture biopsy but not by B-ultrasonograpgy were contrastively analyzed and combined with liver function, blood lipids, blood glucose, and body mass index. Results The 62 cases which were not diagnosed as fatty liver by B-ultrasonograpgy were proved to be 5%-33%liver steatosis after liver puncture biopsy. Among the 62 cases, 23 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo did not see abnormalities, 18 cases showed the liver parenchyma echo slightly was enlarged, 17 cases showed the liver parenchyma echo density was a bit enhanced and 4 cases were diffuse liver damage,which respectively were 37.01%、29.03%、27.42%and 6.45%. Pathologically it indicated that 45 cases were 5%≤liver steatosis≤19%. Among the 45 cases, 18 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo was not seen abnormalities, 8 cases showed the liver parenchyma echo slightly was enlarged, 17 cases showed the liver parenchyma echo density was a bit enhanced, and 2 cases were diffuse liver damage, and the change of ultrasound was mainly showed by the liver parenchyma echo not seen abnormalities and the enhanced liver parenchyma echo density. Pathologically it indicated that 17 cases were 20%≤liver steatosis≤33%, 6 cases were indicated by the B-ultrasonograpgy that the liver parenchyma echo did not see abnormalities, 5 cases showed the liver parenchyma echo slightly was enlarged, 5 cases showed the liver parenchyma echo density was a bit enhanced, and 1 case was diffuse liver damage, and the change of ultrasound was mainly showed by the liver parenchyma echo not seen abnormalities, the slightly enlarged liver parenchyma echo and the enhanced liver parenchyma echo density. By analyzing the influence to the ultrasound changes by patients' liver function, body mass index, blood fat and blood sugar, and with logistic regression analysis through a disorderly classification, it was found that the larger value of the glutamine transferase, alkaline phosphatase, body mass index, triglyceride and low density lipoprotein cholesterol, the higher possibility of diffuse liver damage, and the higher level of fatty degeneration, the higher possibility of diffuse liver damage. Conclusion In the diagnosis of fatty liver, when the fatty degeneration is below 1/3, B-ultrasonic examination can't show characteristic changes of fatty liver. It should be closely observed or take liver puncture biopsy to make a definite diagnosis of fatty liver.
10.Monitoring and analysis of activity concentrations of 7Be and 210Pb in atmospheric aerosol in Beijing from 2017 to 2020
Huan WANG ; Qinghua MENG ; Yongzhong MA ; Yun LOU ; Bin BAI ; Weijie ZHU ; Yuxia KONG ; Hongfang WANG ; Hong ZHANG ; Zechen FENG ; Shuguang ZHAI ; Jun YU
Chinese Journal of Radiological Medicine and Protection 2021;41(9):690-694
Objective:To monitor and analyze the activity concentrations of 7Be and 210Pb in atmospheric aerosol in Beijing from 2017 to 2020. Methods:Totally 1 074 atmospheric aerosol samples were collected by high flow air sampler (SnowWhite) including 275 in spring, 266 in summer, 262 in autumn and 271 in winter and subjected to analysis by low background high-purity germanium gamma-ray spectrometery (ORTEC) to determine the activity concentrations of 7Be and 210Pb. Results:The activity concentrations of 7Be in atmospheric aerosol were from 0.56 to 14.84 mBq/m 3, with an avearage of 6.84 mBq/m 3, and from 0.01 to 9.37 mBq/m 3 for 210Pb, with an avearage 3.19 mBq/m 3. The differences in activity concentrations of 7Be and 210Pb in atmospheric aerosol among differernt seasons were statistically significant ( F=32.66, 93.93, P<0.05). Activity concentration of 7Be was highest in spring, followed by autumn, and lowest in summer and winter. Activity concentration of 210Pb ranged from winter, autumn, spring to summer in descending order. Conclusions:The activity concentrations of 7Be and 210Pb in atmospheric aerosol in Beijing from 2017 to 2020 fluctated within the range of normal level.