1.Gene Expression Profiles of Peripheral Blood in a Family with Systemic Lupus Erythematosus
Guangming HAN ; Shunle CHEN ; Nan SHEN ; Jing HUA ; Yuan WANG ; Chunde BAO
Chinese Journal of Dermatology 1995;0(01):-
Objective To investigate the pathogenesis of familial systemic lupus erythematosus (SLE), by analyzing the gene expression profile of peripheral blood in a family with 2 SLE patients and their first-degree relatives. Methods Total RNA was extracted from peripheral blood cells of normal subjects and SLE patients. Then, synthesis double strand cDNA template from total RNA, transcription of cRNA probe with Biotin labeling, hybridization of probe with Microarray, binding of Streptavidin to Biotin, amplification with First Antibody, further amplification with Cy3-Conjugated Second Antibody, detection of Cy3 dye with ScanArray 5000 were performed. With QuantArray microarray analysis software, the scan image information was converted into numeric data. With GeneSpring microarray analysis software, cluster analysis was done to find interested genes. Results Over 3000 target genes were analysed. Fifty-nine genes differentially expressed in familial SLE patients and controls were identified. Among them, 34 genes were up-regulated and 25 genes were down-regulated. These differentially expressed genes identified in two familial SLE patients were almost identical to those found in other sporadic SLE patients. Among 34 expression increasing genes, 22 were up-regulated in SLE sisters and unaffected sisters; among 25 expression decreased genes, 17 genes down-regulated in SLE sisters and unaffected sister. Cluster analysis showed that patients were clearly separated from controls and their unaffected sisters based on their gene expression profile. These results showed that in familial SLE, multiple genes were responsible for susceptibility to SLE, and clinically unaffected relatives shared some lupus susceptibility genes with their clinically affected relatives, in addition environmental factors were probably necessary to trigger disease. Conclusion These results indicate that high-density oligonucleotide microarray has the potential to explore the heredity in SLE families.
2.Recombinant human endostatin improves tumor vasculature and alleviates hypoxia in Lewis lung carcinoma
Fang PENG ; Jin WANG ; Yi ZOU ; Yong BAO ; Wenlin HUANG ; Guangming CHEN ; Xianrong LUO ; Ming CHEN
Chinese Journal of Radiation Oncology 2011;20(1):69-72
Objective To investigate whether recombinant human endostatin can create a time window of vascular normalization prior to vascular pruning to alleviate hypoxia in Lewis lung carcinoma in mice. Methods Kinetic changes in morphology of tumor vasculature in response to recombinant human endostatin were detected under a confocal microscope with immunofluorescent staining in Lewis lung carcinomas in mice. The hypoxic cell fraction of different time was assessed with immunohistochemical staining . Effects on tumor growth were monitored as indicated in the growth curve of tumors . Results Compared with the control group vascularity of the tumors was reduced over time by recombinant human endostatin treatment and significantly regressed for 9 days. During the treatment, pericyte coverage increased at day 3, increased markedly at day 5, and fell again at day 7. The vascular basement membrane was thin and closely associated with endothelial cells after recombinant human endostatin treatment, but appeared thickened, loosely associated with endothelial cells in control tumors. The decrease in hypoxic cell fraction at day 5 after treatment was also found. Tumor growth was not accelerated 5 days after recombinant human endostatin treatment. Conclusions Recombinant human endostatin can normalize tumor vasculature within day 3 to 7, leading to improved tumor oxygenation. The results provide important experimental basis for combining recombinant human endostatin with radiation therapy in human tumors.
3.Efficacy of Shenqi Fuzheng Injection combined with nimodipine in the treatment of convalescent-phase cerebral infarction and its effects on neurocognitive function, hemorheology and T cell subsets
Youkui SHEN ; Yanqiu WANG ; Jun LI ; Qi YE ; Yong DONG ; Guangming BAO
Chinese Journal of Primary Medicine and Pharmacy 2021;28(8):1159-1163
Objective:To investigate Shenqi Fuzheng Injection combined with nimodipine in the treatment of convalescent-phase cerebral infarction and its effects on neurocognitive function, hemorheology and T cell subsets. Methods:A total of 108 patients with cerebral infarction in the convalescent phase who received treatment in Hangzhou Hospital of Traditional Chinese Medicine, China between April 2016 and December 2019 were included in this study. They were randomly assigned to receive either nimodipine treatment (control group, n = 54) or treatment with Shenqi Fuzheng Injection combined with nimodipine (study group, n = 54). Curative effects and changes in neurocognitive function, hemorheology and T cell subsets after treatment relative to before treatment were compared between the control and study groups. Results:Total effective rate in the study group was significantly higher than that in the control group [90.74% (49/54) vs. 75.93% (41/54), χ2 = 4.267, P = 0.039]. After 2 weeks of treatment, whole blood viscosity at a high shear rate, whole blood viscosity at a low shear rate, plasma viscosity in the study group were (4.17 ± 0.24) mPa/s, (9.27 ± 1.98) mPa/s, (1.07 ± 0.19) mPa/s, respectively, which were significantly lower than those in the control group [(4.52 ± 0.31) mPa/s, (13.69 ± 2.13) mPa/s, (1.34 ± 0.23) mPa/s, t = 6.560, 11.169, 6.651, all P < 0.05]. The proportion of CD 3+ cells, CD 4+ and CD 4+/CD 8+ in the study group was (48.59 ± 4.59) %, (44.24 ± 6.17) % and (1.91 ± 0.17) respectively, which were significantly higher than those in the control group [(44.97 ± 5.31) %, (39.55 ± 5.13) %, (1.47 ± 0.22), t = 3.790, 4.295, 11.629, all P < 0.05]. The proportion of CD 8+ cells in the study group was significantly lower than that in the control group [(23.13 ± 5.62) % vs. (26.97 ± 4.26) %, t = 4.001, P < 0.05]. Mini-Mental State Examination score in the study group was significantly higher than that in the control group [(28.87 ± 0.85) points vs. (27.91 ± 1.45) points, t = 4.197, P < 0.05]. National Institute Health of Stroke Scale score in the study group was significantly lower than that in the control group [(9.63 ± 2.19) points vs. (15.27 ± 1.97) points, t = 14.070, P < 0.05]. Conclusion:Shenqi Fuzheng Injection combined with nimodipine can remarkably improve the neurocognitive function, hemorheology and T cell subsets in patients with cerebral infarction in the convalescent phase. The combined method is safe and reliable, and its curative effect is stable.
4.Application of solid-phase extraction column for determination of matrine and oxymatrine in Sophora flavescens.
Xia YANG ; Bao-Lin GUO ; Hong-Yu HU ; Wen-Hua HUANG ; He-Ping QIAO ; Sheng-Ci FAN ; Zha-Gen GUAN
China Journal of Chinese Materia Medica 2013;38(17):2844-2847
A Cleanert Alumina-N-SPE column (0.5 g/6 mL) chromatograpy with 5 mL of chloroform-methanol (7: 3) as eluent, instead of aluminum oxide column (100-200 mesh, 5 g, 1 cm) chromatograpy eluted successively with chloroform and the chloroform-methanol (7:3) (20 mL each), was applied to enrich matrine and oxymatrine in Sophora flavescens. Also, the optimization of the HPLC determination conditions with acetonitrile-ethanol absolute-3% phosphoric acid solution (84: 6: 10) as mobile phase, instead of acetonitrile-ethanol absolute -3% Phosphoric acid solution (80: 10: 10) recorded in Chinese Pharmacopoeia 2010 Edition, was more suitable for determination of matrine and oxymatrine in S. flavescens. This method has advantage of reducing sample handling time and solvent volume and increasing the accuracy and feasibility, which can simplify the procedure for determination of matrine and oxymatrine in S. flavescens.
Alkaloids
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analysis
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isolation & purification
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Chromatography, High Pressure Liquid
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methods
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Drugs, Chinese Herbal
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analysis
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isolation & purification
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Quinolizines
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analysis
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isolation & purification
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Solid Phase Extraction
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methods
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Sophora
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chemistry