2.Expression of heat shock response protein induced by heat shock in human Tca8113 cells
Lin XIAO ; Xinhua LIANG ; Jing HU ; En LUO ; Zuyi MAO
Journal of Practical Stomatology 1995;0(04):-
Objective: To investigate the expression of heat shock response protein(HSP_ 70 ) in human squamous tongue cancer Tca8113 cells during the recovery periods of heat shock.Methods:Tca8113 cells were subjected to heat shock at 43 ℃ for 30 min, then the cells were cultured for 2,4,6,8,12,24 and 48 h respectively. The expression of HSP_ 70 in the cells was examined with immunohistochemical method, Quantitative analysis was performed by FCM and the cell vitality was detected by MTT method.Results:Heat shock induced HSP_ 70 expression in Tca8113 cells at 43 ℃ for 30 min and the maximum proportion of the positive cells were observed and HSP_ 70 reached the maximum value at 12 h after heat shock(P
3.Separation and Identification of Schizochytrium sp.
Mao-Hong ZHOU ; Lin ZHOU ; Xiao-Wei ZHAO ; Lin-Jing YU ; Na LIN ;
Microbiology 1992;0(04):-
A strain was separeted from the Yueqing bay using pine pollen baiting.The vegetative thallus of the separated strain is oval and unincleate.It possesses a cell wall composed of many compact layers of closely pressed scales, which can be resolved where the cell wall is disrupted.The radiating branched extensions of the thallus, the ectoplasmic net, emerges from the sagenogenetosome.Asexual reproduction is by conversion of the vegetative thallus to many biflagellate zoospores, during which tetrads of cells are formed.It was identified with Schizochytrium sp.based on the features mentioned above.
4.Progress in preparation of small monoclonal antibodies of knock out technique.
Jing LIU ; Xin-min MAO ; Lin-lin LI ; Xin-xia LI ; Ye WANG ; Yi LAN
China Journal of Chinese Materia Medica 2015;40(19):3737-3741
With the application of monoclonal antibody technology more and more widely, its production technology is becoming more and more perfect. Small molecule monoclonal antibody technology is becoming a hot research topic for people. The application of traditional Chinese medicine small molecule monoclonal antibody technology has been more and more widely, the technology for effective Chinese medicine component knockout provide strong technical support. The preparation of monoclonal antibodies and small molecule knockout technology are reviewed in this paper. The preparation of several steps, such as: in the process of preparation of antigen, hapten carrier coupling, coupling ratio determination and identification of artificial antigen and establishment of animal immunization and hybridoma cell lines of monoclonal antibody, the large-scale preparation; small molecule monoclonal antibody on Immune in affinity chromatography column method is discussed in detail. The author believes that this technology will make the traditional Chinese medicine research on a higher level, and improve the level of internationalization of Chinese medicine research.
Animals
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Antibodies, Monoclonal
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chemistry
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genetics
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immunology
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Humans
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Hybridomas
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metabolism
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Immunologic Techniques
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methods
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trends
5.Effect of different processing methods on active ingredient contents and sulfur dioxide residue in Astragali Radix.
Lin JI ; Chun-Qin MAO ; Tu-Lin LU ; Lin LI ; De JI ; Zi-Wan NING ; Jing LIU
China Journal of Chinese Materia Medica 2014;39(15):2819-2822
To compare the differences of the active ingredient contents and the sulfur dioxide residue in Astragali Radix before and after sulfur fumigation and provide a basis for establishing an alternative processing method. Astragali Radix, harvested at the same time in Longxi Gansu, were processed with different methods. high performance liquid chromatography (HPLC) was used to determine the contents of the active ingredients in Astragali Radix and the revised method of the pharmacopoeia of China in 2011 was applied to determine the sulfur dioxide residue. The results show that the three-fold sulfur-fumigation group has the highest level of astragaloside IV and the dried sulfur-fumigation group with 10% water has the lowest level; the content of calycosin-7-O-β-D-glucoside is the highest in naturally dried group and the lowest in the group of sulfur fumigating for 3 times; the sulfur dioxide residue of all sulfur-fumigation groups exceeds certain limit significantly and the group of sulfur fumigating for 3 times reaches the highest level.
Astragalus Plant
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chemistry
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Chromatography, High Pressure Liquid
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Drugs, Chinese Herbal
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chemistry
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Fumigation
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adverse effects
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Sulfur Dioxide
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analysis
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Technology, Pharmaceutical
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methods
6.Selection of Cd~(2+) Binding-peptides and Their Affinities for Heavy Metal Ions
Jing-Shuang HUANG ; Chun-Yan MA ; Xin TONG ; Zhuo-Lin YI ; Liu XU ; Can-Quan MAO ;
Microbiology 1992;0(05):-
Selection and affinities of Cd binding peptides were assayed by phage random dodecapeptide library and affinity chromatography. Two Cd binding peptides were obtained, it was found that the affinities of Cu~ 2+ ,Co~ 2+ ,Zn~ 2+ ,Ni~ 2+ for Cd binding peptides were higher than that of Cd~ 2+ and Cr~ 2+ after detection of the amplified Cd binding peptides displayed phages to different heavy metal-charged resins; the detoxification of E.coli to Ni~ 2+ and Cd~ 2+ was enhanced when infected by Cd binding peptide displayed phages as compared with those of the control. The interaction of Cd binding peptide displayed phages with heavy metals charged resins was also observed under microscope. The work would be of great value and consequences for the study of interaction between heavy metals and proteins(peptides), as well as thedetoxification and bioremediation of heavy metals.
7.Protective Effect of Reduced Glutathione on Kidney against Acute Ischemia-reperfusion Injury
Yan LIN ; Tao HE ; Xiaoyan MAO ; Zhonglin GAN ; Li ZHANG ; Jing HOU
The Journal of Practical Medicine 2016;32(8):1233-1236
Objective To investigate the pretreatment and post-treatment effect of reduced glutathione (GSH) on acute ischemia-reperfusion injury (IRI) of rat kidney. Methods Fifty adult SD rats were divided into 5 groups: control group (Sham group), ischemia reperfusion group (I/R group), GSH pretreatment group (pre-treatment group), and GSH post-processing group (post-treatment group), with ten rates in each group. Animals in pre-treatment group were injected 4% GSH 100 mg/kg intraperitoneally at 24th , 16th , 8th hour and 45th minute before surgery. Animals in post-treatment group were administrated GSH with the same dosage at 45th minute, 6th, 12th and 18th hour after surgery. Creatinine (Cr), urea nitrogen (BUN) level, the total superox-ide dismutase (T-SOD) activity, malondialdehyde (MDA) and nitric oxide (NO) levels in serum were measured at 24th hour after surgery. Histopathological changes were checked by H. E staining. Results Damage on kid-ney structure of animals in pre-treatment group was less than that in I/R group. There was little pathological change on kidney of those in pre-treatment group. Serum Cr, BUN, MDA and NO levels were all decreased but T-SOD activity increased in pre-treatment and post-treatment group when compared with those in I/R group (P <0.05), (P < 0.05). T - SOD activity in post-treatment group was higher than that in pre-treatment group (P <0.05). Conclusion GSH can protect rats against acute renal ischemia-reperfusion injury within 24 hours before and after kidney ischemia-reperfusion , especially after ischemia-reperfusion.
8.Sclerotherapy for faciocervical macrocystic lymphatic malformations in infants
Jing MA ; Yingqin GAO ; Ken LIN ; Fan LOU ; Zhiyong MAO ; Wenjuan ZENG ; Tiesong ZHANG
Chinese Journal of Applied Clinical Pediatrics 2015;30(11):840-843
Objective To investigate the therapeutic effect of Pingyangmycin combined with Triamcinolone Acetonide on the huge faciocervical lymphatic malformations (LMs) in infants.Methods Sixty-seven infants with LMs located in head and neck from January 2009 to June 2014 were retrospectively analyzed in Kunming Children's Hospital.Thirty-five males and 32 females were enrolled,aged from 1 month to 4 years,with a median age of 1.3 years.Computed tomography and ultrasonography were used to evaluate the location,size and extent of LMs before treatment in all the patients.The size of lesion varied from 5.2 cm ×7.5 cm to 9.2 cm × 10.5 cm.All patients were given local injection of Pingyangmycin combined with Triamcinolone Acetonide after puncturing fluid with uhrasonography guiding under general anesthesia.The injection was repeated every 30 d when necessary.Results The number of injections varied from 2 to 5 times,with a median number of 3.9 times.All cases were followed up for 5 to 36 months.Thirty-two cases (62.68%) were cured,improvement in 19 cases (28.36%) and no effect in 6 cases (8.96%).The total effective rate was 91.04%.There was no severe allergic reaction or pulmonary fibrosis.Secondary operation was performed after 6 months in 12 cases.Two post-operative complications were found,1 was minor paralyses of mandibular branch of facial nerve,with mouth askew,the other was trachyphonia,who were both improved after rehabilitation treatment.Conclusions In order to avoid serious complications,the huge LMs may be given local injection of Pingyangmycin combined with Triamcinolone Acetonide after puncturing fluid with ultrasonography guiding.Graded sclerotherapy provides for a less invasive and shorter course of treatment.The complications and risk of secondary resection increase slightly if sclerotherapy has no curative effect.
9.A novel method for culture and identification of primary human retinal microvascular endothelial cells
Yu-xiang, MAO ; Shao-fen, LIN ; Mei-zhen, ZENG ; Jing-yi, TIAN ; Shi-bo, TANG
Chinese Journal of Experimental Ophthalmology 2013;(1):8-12
Background To optimize the culture method of human retinal microvascular endothelial cells is very important for the study of retinal angiogenesis disease.Human retinal microvascular endothelial cells have been successfully cultured in previous studies,but further improvement of the culture method to harvest higher yields and purity cells is still needed.Objective This study was to design a modified method to isolate and purify human retinal microvascular endothelial cells much easily and quickly,and to compare the expression of specific markers of vascular endothelial cells,factor Ⅷ and CD31/CD34 in the cells.Methods The use of human donor eyeballs was approved by the Ethic Commission of Zhongshan Ophthalmic Center of Sun Yat-sen University.The retina tissue from healthy donor was isolated and digested by the two-step digestion method with 2% trypsin and 0.133% collagenase Ⅳ.Human retinal microvascular endothelial cells were collected and plated in 60 mm dishes coated by 0.1% fibronectin and cultured in endothelial cell-specialized medium supplemented with 10% fetal bovine serum,0.3 mg/L β-endothelial cell growth factor (ECGF) and 100 ng/L sodium heparin.During the culturing,the growth situation of the cells was monitored by morphological observation,and immunohistochemical staining was performed to probe vascular endothelial cell-specific membrane protein CD31,CD34 and factor Ⅷ for identification of the cell purity.Results Human retinal microvascular endothelial cells were isolated successfully from the retina by the twostep digestion method.The primary cultured cells adhered to well 72 hours later and achieved confluence with the typical cobblestone appearance 9 to 10 days after cultured.The cells exhibited the blue nuclei and reddish cytoplasm by regular haematoxylin and eosin stain and showed a strong positive response for CD31,CD34 and factor Ⅷ by immunohistochemistry.The positive dye of CD31 and CD34 was lower than Ⅷ factor in both endothelial cells.Conclusions Modified culture method of human retinal microvascular endothelial cells can improve cell culture result and purify target cells.
10.Effects of different processing methods on effective components and sulfur dioxide residue in Gastrodiae Rhizoma.
Zi-Wan NING ; Chun-Qin MAO ; Tu-Lin LU ; De JI ; Jing LIU ; Lin JI ; Huan YANG ; Fa-Qin WANG
China Journal of Chinese Materia Medica 2014;39(15):2814-2818
The contents of adenosine, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxybenzaldehyde, parishin and sulfur dioxide residue were compared in differently-processed Gastrodiae Rhizoma to provide the basis for a reasonable processing method of Gastrodiae Rhizoma. The analysis was performed on a Merck Purospher STAR column (4.6 mm x 250 mm, 5 μm) with a mobile phase consisting of methanol and water (containing 0.1% formic acid) under gradient elution at a flow rate of 1.0 mL x min(-1). The eluates were detected at 270 nm, and the column temperature was 35°C. The content of adenosin, gastrodin, 4-hydroxybenzyl alcohol, 4-hydroxy-benzaldehyde and parishin in processing of boiling or sulfur-fumigated were lower than that of in processing of steaming. Furthermore, the sulfur dioxide residue of sulphur-fumigated groups exceed 400 mg x kg(-1). This stable and reliable method will contribute to the quality control of different processed Gastrodiae Rhizoma.
Drug Contamination
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Drugs, Chinese Herbal
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chemistry
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Gastrodia
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chemistry
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Sulfur Dioxide
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analysis
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Technology, Pharmaceutical
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methods