2.Characterization of swine leukocyte antigen class I genes and homology analysis of the similarity to HLA in GGTA1 -/- Wuzhishan minipigs
Yingdi JIANG ; Guomin ZENG ; Ningning SHI ; Xirui LI ; Huili JI ; Dengke PAN ; Shuangbao GUN
Acta Laboratorium Animalis Scientia Sinica 2016;24(4):375-380
Objective This study was aimed to characterize the swine leukocyte antigen( SLA) class I genes of GGTA1 -/ - Wuzhishan minipigs and compare their similarity to human leukocyte antigen( HLA) . It has important implica?tions for understanding the cellular rejection in xenotransplantation. Methods Specimens of ear tissue from six founding GGTA1 -/ - Wuzhishan minipigs were collected, and the SLA class I genes (SLA?1, SLA?3, SLA?2) were amplified by RT?PCR. Purified products were cloned into pEASY?T1 vectors and sequenced, followed by BLAST alignment and using bioin? formatc analysis to characterize the SLA class I genes and compare with the similarity to HLA. Results A total of six al?leles were detected, among them alleles were previously reported (SLA?1?0703,SLA?2?1102, SLA?3?0401, SLA?3?0403), and the other were novel (SLA?1?0401wz01, SLA?2?11wz01). The homology between alleles of SLA class I genes in Wuzhishan minipigs and HLA was from 70?5% to 72?1%. The homology analysis of critical amino acid residues on HLA binding with human CD8 + molecules showed that SLA?1?0401wz01, SLA?1?0703, SLA?2?11wz01, SLA?2?1102 and SLA?3?0401 occurred mutant at amino acid positions 225 and 228 ( T→S,T→M) , whereas the other loci were highly conserved. There was a high homology at amino acid level between SLA?2?11wz01, SLA?2?1102 and HLA class I genes which are NK cell KIRs binding sites. Conclusions The amino acid sequences of SLA class I genes of GGTA1 -/ -Wuzhishan minipigs have a high homology to HLA. From the point of view of cell?mediated xenograft rejection, the amino acid sequences of SLA class I genes of GGTA1 -/ - Wuzhishan minipigs have a high homology to HLA, therefore, Wzhishan minipigs may become a good potential donor for pig?human xenotransplantation.
3.Study on screening potential allergenic proteins from infant milk powders based on human mast cell membrane chromatography and histamine release assays
Ping ZHANG ; Yingdi SHI ; Xiaoshuang HE ; Wei SUN ; Yanni LV ; Xiaofang HOU
Journal of Pharmaceutical Analysis 2019;9(1):55-61
Cow's milk allergy is mainly observed in infants and young children. Most allergic reactions affect the skin, followed by the gastrointestinal and respiratory systems. Conventional diagnosis is based on po-sitive allergy studies and evaluation of parameters including IgE and IgG1 levels, acute allergic skin response and anaphylactic shock reactions. We developed a cell membrane chromatographic (CMC) method based on human mast cells (HMC-1) for screening potential allergens in infant formula milk powders (IFMP). HMC-1 cell membranes were extracted and mixed with silica to prepare cell membrane chromatography columns (10 mm × 2 mm i.d., 5 mm). Under the conditions of 0.2 mL/min flow rate and 214 nm detection wavelength, human breast milk showed no retention. However, IFMP showed clear retention. The retained fractions were collected and analyzed through matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS). Four major milk proteins, i.e., α-casein, β-casein, α-lactalbumin, and β-lactoglobulin A, were identified. Furthermore, these proteins and β-lacto-globulin B showed clear retention on HMC-1/CMC columns. To test the degranulation effects of the five proteins, histamine and β-hexosaminidase release assays were carried out. All five proteins induced HMC-1 cells to release histamine and β-hexosaminidase. Also, we established a reversed phase liquid chromatographic (RPLC) method for the determination of the five proteins in IFMP and the results showed that 90% proteins in IFMP were α-casein and β-casein. We concluded that cow's milk proteins may be potential allergens and caseins cause more β-casein allergic risk than other proteins. This con-clusion was consistent with other studies.
4.Progress in the analysis of adulterated additives in traditional Chinese medicines and health care products
Xiaofang HOU ; Yingdi SHI ; Sicen WANG
Journal of China Pharmaceutical University 2019;50(1):33-40
In recent years, cases of illegal addition of chemical substances into the TCMs and health-care products happened regularly. Therefore, it is particularly important to develop fast, sensitive and accurate analysis methods for detection of the adulterated chemical substances. Through literature survey of relevant papers published in 2016-2017, this article summarizes the application of various analytical techniques for adulterated chemical substances to the TCMs and health-care products with useful information for the further development of new methods and technologies in this field.
5.11'-Deoxyverticillin A induces caspase-dependent cell apoptosis in PC3M cells.
Yingdi SHI ; Yingqiu ZHANG ; Yangxiao NI ; Guoli SHI ; Huaiyi YANG
Chinese Journal of Biotechnology 2012;28(1):96-103
Recent years, the incidence and mortality of prostate cancer have increased dramatically in China. At earlier stages, most diagnosed prostate cancers are responsive to androgen depletion treatment, yet, nearly all patients will eventually progress to metastatic androgen-independent prostate cancer (AIPC), which still has no effective therapeutic method or drug to deal with. 11'-Deoxyverticillin A (C42) belongs to the family of epipolythiodioxopiperazines (ETPs), an interesting class of fungal toxins that inhibit farnesyl transferase. Compounds holding such a property have been explored as putative anticancer agents. In this study, using PC3M cells, an AIPC cell line, we investigated the effect of the compound on apoptosis and explored the underlying mechanism. It revealed that C42 markedly enhanced the activity of caspase-3/7 and increased the accumulation of the cleaved PARP, all of which are the markers of apoptosis. It also revealed that C42 either decreased cell viability or inhibited the growth of PC3M cells. Moreover, we observed that the loss of cell viability and cell growth inhibition induced by C42 were both time- and dosage dependent. Taken together, we indicated that C42 can induce caspase-dependent apoptosis in AIPC cells, and the results presented here will broaden our knowledge about the molecular mechanisms by which C42 exerts its anticancer activity, and future work in this direction may provide valuable information in the development of these compounds into effective cancer therapeutic strategies against androgen-independent prostate cancer.
Apoptosis
;
drug effects
;
Caspase 3
;
metabolism
;
Caspase 7
;
metabolism
;
Cell Line, Tumor
;
Disulfides
;
pharmacology
;
Farnesyltranstransferase
;
antagonists & inhibitors
;
Humans
;
Male
;
Mycotoxins
;
pharmacology
;
Piperazines
;
pharmacology
;
Prostatic Neoplasms
;
pathology
6.Morphology and distribution of CD44+/Oct4+colorectal cancer stem cells
Dengcai ZHANG ; Bin LIU ; Lihua ZHANG ; Cailan ZHANG ; Yanli YANG ; Qinjun SU ; Min SHI ; Liang DONG ; Yingdi HA
Chinese Journal of Tissue Engineering Research 2013;(49):8461-8467
BACKGROUND:More and more studies employ CD44 as a specific marker of colorectal cancer stem cells. Oct4 is a transcription factor of embryonic stem cells, and it has been discovered recently that there is a higher expression in primary colorectal carcinoma.
OBJECTIVE:To investigate the quantity, location and distribution of CD44+/Oct4+cells in primary colorectal carcinoma.
METHODS:A total y of 108 cases of human colorectal carcinoma and 18 cases of normal mucosa, 18 cases of adenoma were col ected and made into three tissue microarrays, each containing of 48 dots. The locations of CD44+/Oct4+cells were detected by double-label immunohistochemical staining and hematoxylin-eosin staining. The morphologic features of them were investigated on hematoxylin-eosin staining at the same position.
RESULTS AND CONCLUSION:The results of double-label immunohistochemical staining demonstrated that there were no CD44+/Oct4+cells in normal intestine mucosa and a very smal amount of CD44+/Oct4+cells in adenoma, and double-positive cells could also be seen in colorectal carcinoma. The number of CD44+/Oct4+cells was rare and the cells were scattered or distributed focal y along the basement of gland basal side. The cells with scarce cytoplasm were square, and its nucleus was oval or high cylindrical, deeply stained and homogeneous. The quantity of CD44+/Oct4+cells was negatively correlated with the differentiation of colorectal cancer (r=-0.579, P<0.01), and was associated with the depth of tumor invasion (r=0.236, P<0.05). These findings indicate that CD44+/Oct4+cells may be colorectal cancer stem cells.