1. Effect of ginsenoside Rh2 on drug resistance sensitivity of human gastric cancer SGC7901/ADR cells
Chinese Traditional and Herbal Drugs 2018;49(17):4113-4117
Objective To observe the effect of ginsenoside Rh2 (G-Rh2) on the proliferation, cell cycle and chemotherapy sensitivity of human gastric cancer cell line SGC7901/ADR. Methods MTT assay was used to detected the effects of G-Rh2 and adriamycin (ADR) on the proliferation of drug-resistant SGC7901/ADR cells and to calculate the reversal fold (RF). Flow cytometry was selected to detect The effects of G-Rh2 on cell cycle. the effects of G-Rh2 on the expression of P-gp and Bcl-2 proteins in drug-resistant SGC7901/ADR cells was detected by Western blotting. Results Compared with the IC50 value (54.52 μmol/L) after the ADR single drug treatment, the IC50 value (30.14 μmol/L) of the cells treated with G-Rh2 and ADR decreased significantly, and RF was 1.81. G-Rh2 combined with ADR arrested the cell cycle in G2/M phase and significantly decreased the protein expression of P-gp and Bcl-2(P < 0.05). Conclusion G-Rh2 combined with ADR could increase the chemosensitivity of SGC7901/ADR cells, which may inhibit the proliferation of gastric cancer cells by blocking cell cycle and increasing apoptosis.
2.Expression pattern of different serotypes of adeno-associated viral vectors in mouse retina.
Journal of Peking University(Health Sciences) 2020;52(5):845-850
OBJECTIVE:
To investigate the expression efficiency of exogenous gene mediated by different serotypes of adeno-associated virus (AAV) vectors in retina, and to compare the expression efficiency of AAV vector and two kinds of promoters commonly used in ophthalmology after transfection into mouse retina, so as to provide the basis for selecting appropriate AAV vector and promoter for gene therapy of retinitis pigmentosa.
METHODS:
AAV2/2, AAV2/5, AAV2/8 and AAV2/9 were prepared. The C57BL/6J mice were injected subretinally with 1 μL purified AAV vectors (1.00×1013 mg/L). Then the mice were killed 2 or 4 weeks after treatment, and the eyes were enucleated for frozen section. The expression of green fluorescent protein (GFP) was observed under the confocal microscope. Two kinds of promoters, CMV and CAG, were selectd, and the expression of AAV2/8-GFP-CMV and AAV2/8-GFP-CAG was observed under confocal microscope.
RESULTS:
No bacterial infection or immune response were seen in the injected mice. 2 weeks after injection, the GFP green fluorescence of AAV2/8 and AAV2/9 in the mouse retina was obvious, which indicated that the GFP green fluorescence of AAV2/8 and AAV2/9 was high after transfection into the mouse retina. In these two serotypes, GFP green fluorescence of AAV2/8 was mainly concentrated in photoreceptor cells while AAV2/8 was expressed in the whole retina, indicating that AAV2/8 was more specific to photoreceptors. Further experiments on AAV2/8 showed that the GFP green fluorescence of the mouse retina was obvious 4 weeks after injection, indicating that the exogenous gene mediated by AAV2/8 could be stably expressed in vivo. For CMV and CAG promoters, CMV promoter was expressed stronger in retinal pigment epithelium (RPE)cells, while CAG promoter was stronger in photorecepters. In photorecepters, CAG promoter was expressed almost the same as CMV promoter, while CMV promoter was stronger in RPE cells.
CONCLUSION
AAV vectors could express transgene robustly in retinal cells; Among several AAV serotypes, AAV2/2 and AAV2/5 showed weaker GFP fluorescence than AAV2/8 and AAV2/9. AAV2/9 showed expression in each layer of the retina including ganglion cells. AAV2/8 was more specific for photoreceptor; CAG promoters had higher specificity for photoreceptors than CMV promoters.
Animals
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Dependovirus/genetics*
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Genetic Vectors
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Mice
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Mice, Inbred C57BL
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Retina
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Serogroup
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Transduction, Genetic
3.Comparison Study on Determination of Trace Element Content in Peach Pulp between Wet Digestion Method and Carbonize Acid Dissolution Method
Hongyu HU ; Shuang MEI ; Junliang DU
World Science and Technology-Modernization of Traditional Chinese Medicine 2013;(4):799-803
This study was aimed to determine trace elements in Peach pulp . The wet digestion method and car-bonize acid dissolution method were applied to digest the sample, and flame atomic absorption spectroscopy was used to determine the content of trace elements (Fe, Cu, Mn) in Peach pulp . The results showed that the content is not consistent among different pretreatment methods. However, the metalion content among these three methods are in the order of Fe > Mn > Cu, with the average recovery rate between 92.6% and 119.6%. The RSD is less than 2.86%. It was concluded that this method is with high accuracy and stability as well as reliable accurate re-sults. It also proved that Peach pulp . is rich in Fe, Cu and Mn. It provides some useful information for further pharmacological study of Peach pulp .
4.Severe hypertriglyceridemia caused by autoimmune disease:Case summary and literature review
Shuang ZHENG ; Xingxing REN ; Yaomin HU
Chinese Journal of Endocrinology and Metabolism 2016;32(6):523-526
[Summary]_ Severe hypertriglyceridemia is the third common cause of acute pancreatitis following after alcohol abuse and cholelithiasis. Moreover, it is also an important risk factor of cardiovascular events. However, the cases of severe hypertriglyceridemia caused by autoimmune disease were rare in clinical, which would bring the difficulty for diagnosis and treatment. A better understanding of the clinical characteristics, possible pathogenesis, and corresponding therapy of the disease would be helpful, which would reduce the risk of complications, and finally improve both the survival rate as well as quality of life of these patients.
5.Construction of a full-length cDNA clone of a live attenuated vaccine strain against Japanese encephalitis virus and preliminary study of expressing exogenous gene.
Bing HU ; Shuang YANG ; Zhi-zheng FANG
Chinese Journal of Virology 2014;30(6):652-660
This study aimed to construct full-length cDNA clones of the Japanese encephalitis virus (JEV). SA14-14-2 strain and discuss the feasibility of constructing chimeric viruses for exogenous gene expression based on the JEV genetic skeleton. Long-fragment RT-PCR techniques were applied to amplify JEV cD-NAs, and two amplified fragments with corresponding restriction endonuclease sites at both ends were cloned into the pACYC184 vector sequentially. Using standard molecular techniques, the enhanced green fluorescent protein (EGFP) gene was inserted into the 3' non-coding region of JEV as a reporter gene. After in vitro transcription and transfection procedures, wild-type JEV and chimeric JEV that expressed the EGFP as the reporter gene were successfully rescued. The recovered viruses were characterized by RT-PCR, plaque assays, and direct fluorescence microscopy. After six serial passage generations, the stability of the recovered viruses were studied in terms of virus growth characteristics and structural gene expression. The results showed that cDNA clones of rJEV and rJEV-EGFP were successfully constructed and rescued in BHK-21 cells after in vitro transcription and transfection. Each generation of the recovered viruses was stable and the chimeric virus rJEV-EGFP could stably express EGFP. The findings of this study indicate that both rJEV and rJEV-EGFP could be constructed and rescued in BHK-21 cells, and the JEV SA14-14-2 strain could be obtained as a viral vector to express foreign genes.
Cloning, Molecular
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DNA, Complementary
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genetics
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metabolism
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Encephalitis Virus, Japanese
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genetics
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metabolism
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Encephalitis, Japanese
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virology
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Gene Expression
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Genetic Vectors
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genetics
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metabolism
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Green Fluorescent Proteins
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genetics
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metabolism
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Humans
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Vaccines, Attenuated
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genetics
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metabolism
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Viral Vaccines
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genetics
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metabolism
6.Effect of Xueshuantong combined with butylphthalide on clinical curative effect, cerebral vascular reserve capacity and serum hs-CRP on acute cerebral infarction
Jianqin HU ; Weilv XIONG ; Shuang SHEN
Chinese Journal of Biochemical Pharmaceutics 2016;36(12):66-68
Objective To observe the efficacy and effect on cerebral vascular reserve and serum hs-CRP of Xueshuantong combined with butylphthalide in patients with acute cerebral infarction.Methods A total of 140 cases with ACI in Huzhou Central Hospital from August 2015 to July 2016 were divided into the observation group and the control group with 70 cases in each group, according to the order of single and double number.The control group was treated with routine western medicine while giving butylphthalide , and the observation group was given Xueshuantong on the basis of the control group.After two weeks of treatment, the efficacy and CVR, BHI and hs-CRP levels were observed.Results After treatment, the total effective rate of the observation group was 92.86%, and the control group was 81.43%, the difference was statistically significant (χ2 =4.080, P<0.05), there was significant differences of CVR, BHI between two groups (Fgroup =5.534, 4.608, P<0.05), the CVR and BHI increased gradually with time (Ftime =11.325, 9.432, P<0.05), the rised amplitude in observation group was higher than the control group (Finteraction =5.742, 5.016, P<0.05), there were significant differences in the levels of hs-CRP between two groups (Fgroup =7.421, P<0.05), and hs-CRP decreased gradually with time (Ftime =10.185, P<0.05), the decrease of observation group was larger than that of control group(Finteraction =4.824, P<0.05).Conclusion Xueshitong combined with butylphthalide in the treatment of ACI is effective , can better improve cerebral vascular reserve capacity and inhibition of inflammatory response, better than the single use of butylphthalide.
7.Quality control of Yixuning Tablet
Shuang HU ; Hong DING ; Yan DU ;
Chinese Traditional Patent Medicine 1992;0(07):-
AIM To develop the quality control methods for Yixuning Tablet (Fructus Lycii,Radix Polygoni Multiflori Preparata,Radix Codonopsis,etc.) METHODS RP-HPLC was performed for the assay of 2,3,5,4′-tetrahydroxy-stilbene-2-O-?-D-glycoside.TLC was used to identify Fructus Lycii,Radix Polygoni Multiflori,and Radix Angelicae Sinensis. RESULTS The good linearity was within the range of 20-180 ?g?mL -1 ( r = 0.999 9 ).The average recovery was 99.0 % ( RSD was 2.8 T《B》 % ).TLC methods were special. CONCLUSION The method is convenient,accurate and sensitive.and suitable for the quality control of the preparation.
8.Investigation and Analysis of the Drug Accessibility Based on the Situation of Drug Use and Storage in Fami-lies in Chengdu
Jianlong YU ; Haiyao HU ; Shuang FENG ; Ming HU ; Naitong ZHOU
China Pharmacy 2016;27(21):2893-2897
OBJECTIVE:To provide reference for further comprehensive evaluation of the improvement of the accessibility of family medicines after the implementation of the new health care reform. METHODS:Sampled as 180 households from 2 urban ar-eas,2 counties and 2 rural areas in Chengdu,a questionnaire survey was adopted based on the WHO’s manual for the Household Survey to Measure Access and Use of Medicines to investigate the situation of drug use,storage and accessibility,and the results were statistically analyzed. RESULTS:Totally 180 questionnaires were sent out,179 were effectively received with effective recov-ery of 99.44%. In the terms of medicines taken for the acute illness,there were more than 60% rural households obtained drugs from the public health care facilities,while more than 60% counties households purchased drugs in public medical and health insti-tutions;in the terms of medicines taken for the chronic illness,the proportion of households reporting a chronic illness who did not take any medicines was low(3.80%);in terms of medicines storage,the proportion of households without reserve drugs was low (13.41%). In terms of drug accessibility,the proportions of respondents who agreed that the location and opening hours of their public healthcare facility were convenient were high;there was no situation for“patients did not take prescribed drugs because med-icines were not available at the healthcare facility and drug store”,while more than 95% respondents who agreed that drugs were available at their public health care facilities and drug stores;the essential medicines in the household use of chronic drugs in Chengdu accounted more than 60%;more than 85% can afford the drug costs,while the proportion of the monthly costs for acute illness and chronic illness in the total month costs was low;the health insurance coverage reached 90% in the survey households, but the proportions of households which could reimburse acute,chronic illness drug cost were lower(14.13% and 12.66%). CON-CLUSIONS:At present,the level of drug accessibility in Chengdu is high;the implementation of National Essential Medicines System has a positive effect on reducing the economic burden of chronic disease family. But there are shortcomings in the Medicare reimbursement levels and antibiotics use,it is suggested to take effective measures to solve it,such as exploring the stable and ef-fective medical insurance fund raising channels,improving the compensation level and the level of insurance reimbursement,formu-late and implement the corresponding antibacterial drugs supervision system and regulate the rational use of antibiotics in residents.
9.Analysis phylogenetic relationship of Gynostemma (Cucurbitaceae).
Shuang-shuang QIN ; Hai-tao LI ; Zhou-yong WANG ; Zhan-hu CUI ; Li-ying YU
China Journal of Chinese Materia Medica 2015;40(9):1681-1687
The sequences of ITS, matK, rbcL and psbA-trnH of 9 Gynostemma species or variety including 38 samples were compared and analyzed by molecular phylogeny method. Hemsleya macrosperma was designated as outgroup. The MP and NJ phylogenetic tree of Gynostemma was built based on ITS sequence, the results of PAUP phylogenetic analysis showed the following results: (1) The eight individuals of G. pentaphyllum var. pentaphyllum were not supported as monophyletic in the strict consensus trees and NJ trees. (2) It is suspected whether G. longipes and G. laxum should be classified as the independent species. (3)The classification of subgenus units of Gynostemma plants is supported.
Gynostemma
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classification
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genetics
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Molecular Sequence Data
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Phylogeny
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Plant Proteins
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genetics
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Sequence Analysis, DNA
10.BRAF V600E mutation and expression of its protein in papillary thyroid carcinoma
Liru DONG ; Hu YANG ; Shuang LI ; Xudong SONG
China Oncology 2017;27(4):251-255
Background and purpose:The BRAF V600E mutation is a highly attractive drug target. Therefore, determining the BRAF gene mutation status of patients is essential in order to assess patients' eligibility for targeted BRAF geneinhibitor therapy. The aim of this study was to validate the utility of immunohistochemistry to rapidly obtain the BRAF gene mutation status. This study aimed to analyze the correlation of the BRAF V600E gene mutation and VE1 protein ex-pression with the clinical pathological characteristics in papillary thyroid carcinoma (PTC).Methods:The mutation status of BRAF V600E was detected by DNA sequencing. Immunohistochemistry was used to detect the expression of BRAF V600E protein in 108 cases of PTC, 54 cases of thyroid adenoma and 54 cases of normal thyroid tissue.Results:The gene mutation rate of BRAF V600E is 67.6%, and VE1 protein expression rate is 64.8% in 108 cases of PTC. The differences were statistically significant compared with thyroid adenoma and goiter (P<0.05), but have no correlation with the clinical pathological characteristics.Conclusion:BRAF V600E gene mutation and VE1 protein expression are useful biomarkers for the pathological diagnosis of PTC. High consistency was observed between the immunohistochemical staining results and the DNA sequencingresults of BRAF V600E gene mutations. Immunohistochemical technique detecting the BRAF V600E protein expression can effectively reflect indirectly BRAF V600E gene mutation status in PTC.BRAF V600E gene mutation has no contribution to the development of papillary thyroid carcinoma.