1.Expression of interleukin-8 protein in villi and decidua of patients with recurrent spontaneous abortion
Fang SONG ; Fen HAO ; Shufen YUE ; Shiqi HUANG ; Meixia YANG
Acta Anatomica Sinica 2010;41(2):267-270
ObjectiveTo study the expression of interleukin-8 (IL-8) protein in villi and decidua of patients with recurrent spontaneous abortion(RSA). MethodsThe local and quantitative expressions of IL-8 in villi and decidua of 30 RSA patients were shown and measured by immunohistochemistry and Western blotting. Results IL-8 protein immunohistochemical signals were located in the chorioepithelium cytoplasm and decidual cells plasma in RSA group. While in control group the immunohistochemical staining was negative in decidul cells, but positive in glandular epithelium. The quantitative analysis of IL-8 protein by Western blotting, intensity of the immunostaining was higher in RSA group than that in the control group. Conclusion The higher expression of IL-8 in villi and decidua is releated to RSA, IL-8 might take part in the pathologic process of RSA.
2.Effect of lipoxin A4 on lipopolysaccharide-induced oxidant stress in human renal tubular epithelial cells and possible mechanisms
Fen XU ; Hua HAO ; Hua DAI ; Lixiang LI ; Lei ZENG
The Journal of Practical Medicine 2017;33(1):51-55
Objective The study aimed to investigate the effect of lipoxin A4 (LXA4) on lipopolysaccharide (LPS)?induced oxidant stress in human renal tubular epithelial cells (HK2 cells) and possible underlying mecha?nisms. MethodsHK2 cells were divided into three groups: Control ,LPS and LPS+LXA4 groups. After cells were treated with indicated conditions,morphological changes were observed. The expressions of Nrf2 were detected by immunofluorescence and cells were collected for RT?PCR experiments.Results HK2 cells seemed disrupted and necrotic with the administration of LPS. However ,LXA4 could prevent cells from injury induced by LPS. LPS decreased Nrf2 expression and promoted it to translocate to cytoplasm ,while LXA4 could increase its expression and promote it to translocate to nucleus. Moreover ,LPS could decrease Nrf2 and its downstream molecule mRNA expressions,but LXA4 could reverse this effect. Conclusion Our results demonstrated that LXA4 effectively inhibit?ed HK2 cell oxidant stress via Nrf2 pathway.
3.BML-111, the analogue of lipoxin, inhibits Hela cell proliferation
Hua HAO ; Fen XU ; Liqing WU ; Xinxin ZHANG ; Hua DAI
The Journal of Practical Medicine 2014;(13):2045-2047
Objective To investigate the effect of BML-111 (the analogue of lipoxin) on uterine Hela cell (cervix cancer cell line) proliferation and the underlying mechanism. Methods Hela cells were stimulated by 50, 100, 200 and 400 μg/L BML-111, respectively, and cell viability was determined by MTT assay. Hela cells were divided into three groups:the control group (no treatment), the BML-111(200μg/L) group and the BML-111(200μg/L)plus Boc-2 (10μmol/L)group. Expression and location of P53 protein were detected by immunofluorescence. Expressions of NF-κB p65,P53 and CyclinD1 protein were detected by Western blotting. Results BML-111 (100, 200 and 400 μg/L) could effectively inhibit Hela cell viability compared with the control group (P < 0.05). P53 expression was shown decreased in both the nucleus and the cytoplasm without any change of P53 location , however, Boc-2 could reverse this effect. BML-111 could effectively inhibit P53 and CyclinD1 expression via NF-κB pathway and the effects could also be inhibited by Boc-2. Conclusions BML-111 can effectively inhibit Hela cell proliferation via FPR2 and NF-κB pathway.
4.Optimization of Saccharomyces cerevisiae sp.Strain by1.1b Culture Conditions for Efficient Biosynthesis of D-(-)-mandelate Dehydrogenase
Fen YAN ; Qian WANG ; Zi-Lin LIN ; Yang-Hao GUO ;
Microbiology 1992;0(01):-
The culture conditions of Saccharomyces cerevisiae sp.strain by 1.1 b were optimized for the production of D-(-)-mandelate dehydrogenase which is useful for the asymmetric bioreduction of benzoylformate to form D-(-)-mandelate.The optimum medium(per liter)consistes of 60 g peptone,30 g maltose, 0.5 g MgSO_4,0.01 g ZnSO_4,1.0 g KCl.After optimization of the culture medium,the enzyme production in shake flasks is enhanced from 2.56 to 20.21 U/L.The optimum fermentation conditions were determined as follows:medium volume 100 mL(i.e.,40%for a 250-mL shake flask),pH 6.5,inoculum size 10%,temperature 30℃,and cultivation time 25 h.
5.STUDIES OF HEPG2 CELLS INFECTED WITH HGV RNA GENOME
Hao REN ; Fen-Lu ZHU ; Zhong-Tian QI ;
Microbiology 1992;0(01):-
In order to observe the replication and expression of HGV RNA genome in HepG2 cells and establish a cell model of HGV infection, HGV RNA genome was prepared in vitro and transfected HepG2 cells with lipofec-tamin. HGV RNA-positive supernatants were used to infect fresh HepG2 cells. RT-PCR, immunohistochemistry and Western blot assays were carried out to detect the replication and expression of HGV in HepG2 cells. Both positive and negative strands of HGV RNA could be detectable in cell culture supernatants and cells at 24h post-transfection. During the culture periods of 90 days, the cells were maintained by changing the medium every 3 or 5 days, and cultured for more than 20 passages. Both strands of HGV could be detectable in culture supernatants and cells. Immunohistochemistry and Western blot results also confirmed that HGV E2 protein could be expressed in the infected HepG2 cells. HGV RNA could also be detectable in the frozen-thawed HepG2 cells infected with HGV RNA genome. Therefore, HGV RNA genome can replicate and express in HepG2 cells, this HGV RNA genome transfected cells model could be used as a cell model in the studies of replication and infection of HGV.
6.Inhibition of MEK sensitizes human breast carcinoma cells to endoplasmic reticulum pathway's apoptosis
Fen YANG ; Hao LIU ; Xudong ZHANG ; Zhiwen JIANG
Chinese Pharmacological Bulletin 1986;0(06):-
Aim To investigate the inhibition of MEK/ERK pathway affecting the sensitivity of human breast carcinoma cells SK-BR-3 to endoplasmic reticulum(ER) stress-induced apoptosis and wish to find new targets for human breast carcinoma chemotherapy.Methods Different concentrations(0,1.5,3,6,9 and 12 ?mol?L-1) tunicamycin(TM) treated human breast carcinoma cells SK-BR-3 for 48 h,then propidium iodide(PI) staining measured apoptotic cells in Flow Cytometry(FCM).Different times(0,6,12,24 and 36 h) of TM(3 ?mol?L-1) treated SK-BR-3 cells,Western blot measured proteins GRP78,ERK1/2 and pERK expression.MEK inhibitor U0126(20 ?mol?L-1) pretreated cells for 1 h before treatment with TM(3 ?mol?L-1) in different concentrations and times,measured above identical indexes and compared with their diversities of treatment with U0126 or not.Results TM induced apoptotic cells
7.Expressions of OPN, COX-2 and CyclinD1 in breast infiltrating carcinoma and their relationship with clinicopathological features
Hua HAO ; Fen XU ; Liqing WU ; Lixiang LI ; Lei ZENG ; Xinxin ZHANG ; Dan GONG ; Yuqing YANG
The Journal of Practical Medicine 2014;(17):2734-2736
Objective To observe the expressions of OPN, COX-2 and CyclinD1 in breast infiltrating carcinoma and evaluate their relationships with clinic pathological features. Methods Expression of the above three indexes were detected from 70 breast cancinoma patients by immunohistochemistry. The relationships among them and clinicopathological features were analyzed. Results The positive expression rates of OPN were 78.8% in cases (≤45 years old) and 73.0% in cases (> 45 years old); the positive expression rates were 79.3%(tumor diameter ≤ 3 cm) and 73.2% (tumor diameter > 3 cm); the positive expression rates were 77.8%, 73.8% and 78.9% in cases ofⅠgrade, Ⅱgrade and Ⅲ respectively, the positive rates had no statistical significances(P > 0.05). The expression rates of OPN in cases of breast infiltrating carcinoma without and with axillary node metastasis were 62.5% and 93.3%, in cases at stage Ⅰ~Ⅱ and Ⅲ ~Ⅳ were, 68.0% and 95.0% respectively, the positive rates had statistical significances(P < 0.05). The expression of OPN was negatively correlated with ER and PR while positively correlated with CerbB2, COX-2 and CyclinD1. Conclusions OPN plays an important role in the invasion and metastasis of breast carcinoma coordinated with COX-2 and CyclinD1.
10.Development of Multiplex Real-time PCR for Detection of Toxigenic Vibrio cholerae and Virbio parahaemolyticus
Wei ZHANG ; Jin-Cao PAN ; Dong-Mei MENG ; Xin-Fen YU ; Hao-Qiu WANG ; Wei ZHENG ;
Microbiology 1992;0(05):-
A multiplex real-time PCR was developed to detect ctxA of Vibrio cholerae, gyrB and tdh of Vibrio parahaemolyticus simultaneously. The multiplex real-time PCR were evalidated by detection for the three genes in 47 toxigenic V. cholerae O1 and O139 strains (ctxA+; O1=3, O139=44), 25 non-toxigenic V. cholerae strains (ctxA-; O1=12, O139=6, non-O1 and non-O139=7), 116 V. parahaemolyticus strains with or without tdh (73 or 43) and 9 other bacteria strains. The specificity and sensitivity of the multiplex real-time PCR in detection for the ctxA and the tdh genes in the strains tested were both 100.0%, compared to the results by routine PCRs. In the detection for V. parahaemolyticus specific gyrB using the multiplex real-time PCR, all of 116 V. parahaemolyticus strains were positive, and 9 other strains and 72 V. cholerae strains were all negative. The multiplex real-time PCR is a sensitive, specific and quick assay not only for detecting virulence genes of V. cholerae and V. parahaemolyticus but also for identifying V. parahaemolyticus at species level. In addition, two real-time PCRs for detection of V. parahaemolyticus virulence genes trh1 and trh2 were also developed.