2.Advances in the study of aldehyde oxidases.
Acta Pharmaceutica Sinica 2014;49(5):582-589
Aldehyde oxidase (AOX), a highly conserved molybdoflavoenzyme in mammal cytoplasm, has broad substrate specificity and ability to catalyze the oxidation of aldehydes and nitrogen, oxygen-containing heterocyclic rings. AOX was found to widely distribute with the individual differences in vivo and plays an important role in phase I metabolism of drugs and xenobiotics. The biological characteristics of AOX and its contributions in drug metabolism are introduced briefly in this review.
Aldehyde Oxidase
;
antagonists & inhibitors
;
chemistry
;
metabolism
;
Animals
;
Drug Discovery
;
Humans
;
Liver
;
enzymology
;
Oxidation-Reduction
;
Pharmaceutical Preparations
;
metabolism
;
Raloxifene Hydrochloride
;
pharmacology
;
Substrate Specificity
;
Xenobiotics
;
chemistry
;
metabolism
3.Overexpression of FKS1 to improve yeast autolysis-stress.
Jia LI ; Jinjing WANG ; Qi LI
Chinese Journal of Biotechnology 2015;31(9):1344-1354
With the development of high gravity brewing, yeast cells are exposed to multiple brewing-associated stresses, such as increased osmotic pressure, enhanced alcohol concentration and nutritional imbalance. These will speed up yeast autolysis, which seriously influence beer flavor and quality. To increase yeast anti-autolytic ability, FKS1 overexpression strain was constructed by 18S rDNA. The concentration of β-1,3-glucan of overexpression strain was 62% higher than that of wild type strain. Meantime, FKS1 overexpression strain increased anti-stress ability at 8% ethanol, 0.4 mol/L NaCl and starvation stress. Under simulated autolysis, FKS1 showed good anti-autolytic ability by slower autolysis. These results confirms the potential of FKS1 overexpression to tackle yeast autolysis in high-gravity brewing.
Autolysis
;
Beer
;
Echinocandins
;
genetics
;
Glucosyltransferases
;
genetics
;
Hypergravity
;
Membrane Proteins
;
genetics
;
Saccharomyces cerevisiae
;
cytology
;
Saccharomyces cerevisiae Proteins
;
genetics
4.Effect of different drugs for ocular surface healing in patients with preoperative dry eye after phacoemulsification
International Eye Science 2017;17(9):1700-1704
AIM:To evaluate changes in tear film stability and meibomian gland function and the clinic efficacy of three different artificial tears in patients with preoperative dry eye after phacoemulsification.METHODS:All 90 cataract patients (90 eyes) with preoperative dry eye who underwent phacoemulsification randomly divided into three groups (Group A treated with protein-free calf blood extract ophthalmic gel;Group B treated with sodium hyaluronate eye drops;Group C treated with Vitamin A palmitate eye gel).Ocular Surface Disease Index (OSDI), Schimer's I test(SⅠt), tear film break time (BUT), corneal fluorescein staining (FL) and meibography were performed for all patients preoperatively and 7, 30 and 90d postoperatively.RESULTS:No statistical differences existed among the three preoperative groups (P>0.05).Except meibomian gland score, there was no statistical significance among preoperatively and 7, 30, 90d postoperatively of the three groups (P>0.05).At 7d postoperatively, SⅠt and BUT of every group were lower than those before treatment, FL scores and OSDI scores were higher than those preoperative (P<0.05);there were no statistical differences among the three groups(P>0.05).At 30d postoperatively, SIt, BUT, OSDI scores in group A and C were better than in group B, which displayed statistical differences (P<0.05);FL scores in group A were significantly better than in group B and C (P<0.05).At 30, 90d postoperatively, SIt, BUT, FL scores, OSDI scores were better than preoperative results, which displayed statistical differences (P<0.05).There were no statistical differences among the three groups at 90d postoperatively (P>0.05).CONCLUSION:Tear film stability and meibomian gland function were affected by phacoemulsification.Topical application of deproteinised calf blood extract eye gel, sodium hyaluronate eye drops and Vitamin A palmitate eye gel all hase a clearly beneficial effect on subjective symptoms.Deproteinised calf blood extract eye gel and Vitamin A Palmitate Eye Gel had more powerful effect on BUT than sodium hyaluronate, however deproteinised calf blood extract eye gel is more effective in superficial punctuate keratopathy.
5.Assessing drug targeting of Yougui Pill, Zuogui Pill, and their disassembled prescriptions using infrared thermography.
Xia ZHENG ; Yan-Li DENG ; Qi-Jia LI ; Hua LU
Chinese Journal of Integrated Traditional and Western Medicine 2014;34(4):446-449
OBJECTIVETo dynamically assess drug targeting of Yougui Pill (YP) and Zuogui Pill (ZP) using infrared thermography.
METHODSIn this self-control experiment, five healthy volunteers were recruited. By using infrared thermography 10 to 11 thermal images of different body locations were taken from each participant after they took warm water, YP, ZP, and their dissembled prescriptions at 30, 70, 100, 130, and 160 min, respectively. The heat values in the lower quadrant abdomen, uterus, Du channel, and Shenque (CV8) were statistically analyzed after scanning for 125 times.
RESULTSAdministration of YP and its disassembled prescriptions enhanced the heat value of the locations of the Du channel and Shenque (CV8), but did no enhance the heat value of the lower quadrant abdomen at 30 min. Administration of ZP and its disassembled prescriptions reduced the heat value in the locations of the lower quadrant abdomen, uterus, Du channel, and Shenque (CV8) at each time point.
CONCLUSIONThe drug targeting of ZP and YP focused on the locations of the Du channel and Shenque (CV8), not on the locations of the lower quadrant abdomen or uterus.
Adult ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Infrared Rays ; Thermography ; methods ; Young Adult
6.Comparison of early developmental differences of hair follicles in different skin areas of neonatal mice
Yuchen JIA ; Qi CHEN ; Shaowei LI ; Yuanqiang ZHENG ; Li WANG
Acta Laboratorium Animalis Scientia Sinica 2014;(3):84-87
Objective The aim of this study was to observe the growth difference and expression of cytochrome C of skin hair follicles in neonatal mice .Methods The morphology of different skin hair follicles of neonatal mice ( postnatal day 1-9)were observed by HE staining histology and cytochrome C was detected by immunohistochemistry .Results The skin hair follicles in different parts of neonatal mice showed differences not only in morphology but also in developmental pe -riods.Hair follicle growth in the back and tail skin had a nonlinear and growing period .After the nonlinear and growing pe-riod they began to grow rapidly .The tail development was slightly slower than that on the back .The hair follicles of vibris-sae were very special , and started to develop without a stable period .Conclusions The results of morphological observa-tion and cytochrome C immunohistochemistry demonstrate that differences exist in the hair follicle morphology and develop -mental times in the skin of different parts of the body in neonatal mice .
7.CRD-BP and β-TrCP in colorectal cancer
Li CHENG ; Li CHENG ; Dandan JIA ; Wenjuan QI ; Changjiang ZHOU
Journal of International Oncology 2013;40(12):930-932
Coding region instability determinant (CRD) is one of the influence factors of oncogene c-myc.Coding region instability determinant-binding protein (CRD-BP) can connect with CRD in order to protect CRD from nuclease attack,prevent rapid degradation of c-myc mRNA,and increase c-myc protein content.Beta-transducin repeats-containing protein (β-TrCP) can affect cell growth,differentiation,apoptosis and oncogenesis by regulating multiple signaling pathways and cell cycle.The overexpression of CRD-BP can upregulate the expression of β-TrCP and both of them play important roles in the tumorigenesis,progression,metastasis and invasion of colorectal cancer.
8.Significance of anti-cell membrane associated DNA antibodies in diagnosis of systemic lupus erythematosus lacking of specific autoantibodies
Jia-Long GUO ; Li-Qi BI ; Zhan-Guo LI ;
Chinese Journal of Laboratory Medicine 2003;0(09):-
Objective To investigate the effects of anti-cell membrane associated DNA (mDNA) antibodies in the diagnosis of systemic lupus erythematosus (SLE) lacking of specific autoantibodies including anti Sm,anti ds-DNA,and anti-nucleosome antibodies.Methods Indirect immunofluorescence assay was used to measure anti-mDNA antibodies in serum of 145 SLE patients,and indirect immunofluorescence,Western-blot and ELISA were used to detect the anti-dsDNA ,anti-Sm and anti- nueleosome antibodies respectively to analysis the value of anti-mDNA antibodies on the specific autoantibodies negative patients with SLE.Results The sensitivity for anti-mDNA antibodies (69.7%) in SLE was significantly higher than anti-Sm (19.7%),anti-dsDNA ( 31.9% ) and anti-nucleosome (45.8% ).The incidences of anti-mDNA antibodies in SLE lacking of anti-dsDNA,Sm and anti-nueleosome antibodies (AnuA) were 64.3% ,70.2% and 60.3% respectively.Conclusion Anti-mDNA antibodies are serologic marker of SLE and important in diagnosis of SLE lacking of anti-dsDNA,Sm and nucleosome antibodies.
9.Biological effect of down-regulating of MTRR gene on cisplatin-resistant ovarian cancer SKOV3 cells in vitro and in vivo studies
Jia CHEN ; Qi WANG ; Wei ZHANG ; Li LI
Chinese Journal of Obstetrics and Gynecology 2016;(2):126-134
Objective To study the biological effects of down-regulatingof methionine synthase reductase (MTRR) gene on cisplatin resistant ovarian cancer SKOV3/DDP cell in vitro and in vivo. Methods (1) Establishing the cell line of MTRR down-regulated. Four short hairpin RNA (shRNA) for MTRR gene (U6-GFP-Neo-homo-1106, U6-GFP-Neo-homo-1931, U6-GFP-Neo-homo-419, U6-GFP-Neo-homo-1460) were designed respectively. Western blot was used to detect the interference efficiency and selected the most efficient shRNA. The MTRR 1106 was selected as the best silencing effect of interference fragment and then therecombinant plasmid vector pSicoR-1106 was constructed and transfected into SKOV3/DDP cells.The stably transfected cells was obtained by screening of flow cytometry(FCM).Fluorescence quantitative reverse transcription (RT)-PCR and western blot were used to detect the expression of MTRR mRNA and protein. (2) Study in vitro: recombinant plasmid expression vector pSicoR-1106, pSicoR-NC and packaging plasmid were respectively transfected into 293T cell. SKOV3/DDP cells were transfected by viral supernatant. The experiment was divided into three groups, namely SKOV3/DDP-MTRRi (down-regulated MTRR group), SKOV3/DDP-NC (negative control group), and the SKOV3/DDP (blank control group). The cell growth curves and half maximal inhibitory concentration (IC50) of cisplatin were made by methyl thiazolyl tetrazolium (MTT) method. Three groups cells were treated with different concentration of cisplatin (0, 1, 2 and 4 μg/ml). The clonogenicity efficiency was observed by clony formation test. The cell cycles were measured by FCM . (3) Study in vivo: three groups cells were subcutaneously inoculated into the nude mice to develop a tumor model. Mice were injected intraperitoneally with cisplatin at 2.5 mg/kg (once every 2 days, in 21 rounds), then the tumor growth was observed. The expression of MTRR and proliferation-related Ki-67 antigen by immunohistochemistry in xenograft tumors were measured. Results (1) Results showed that U6-GFP-Neo-homo-1106 was the best shRNA with interference effect to MTRR. The recombinant plasmid pSicoR-1106 was constructed and transfected into SKOV3/DDP. The MTRR mRNA and protein were down-regulated after transfected. This result showed that MTRR down-regulated SKOV3/DDP cell line was constructed successfully. (2) The cell growth curves showed that the growth of SKOV3/DDP-MTRRi cells were significantly decreased compared with that in the SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). The IC50 of SKOV3/DDP-MTRRi, SKOV3/DDP-NC and SKOV3/DDP were 4.01, 7.90, and 8.91 μg/ml, respectively. The IC50 of SKOV3/DDP-MTRRi was significantly lower than that in control cell groups (P<0.05).Clony formation tests showed that the clony numbers of varied concentration of cisplatin of SKOV3/DDP-MTRRi were significantly less than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). FCM showed that when the cisplatin concentration rose to 4 μg/ml, the G0/G1 phase cell ratio in SKOV3/DDP-MTRRi cells group was (72.8±5.0)%, which was significantly higher than those in the SKOV3/DDP-NC cells group and SKOV3/DDP cells group [(64.4±2.5)%and (64.3±3.0)%], respectively (all P<0.05).(3) Six weeks after nude mice intraperitoneal injection with cisplatin, the tumor volume of SKOV3/DDP-MTRRi, SKOV3/DDP-NC and SKOV3/DDP were respectively (97 ± 32), (168 ± 45), and (173 ± 32) mm3, the tumor weight were (0.36±0.17), (1.08±0.17), and (1.11±0.20) g, in which tumor volume and weight of SKOV3/DDP-MTRRi were significantly less than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (all P<0.05). In three groups tumor tissue, positive rates of MTRR were respectively 2/8, 5/8, and 7/8, the positive rates of Ki-67 were respectively1/8, 6/8, and 7/8, in which SKOV3/DDP-MTRRi was significantly lower those SKOV3/DDP-NC cells and SKOV3/DDP cells (all P<0.05). Conclusion The growth and cisplatin resistance of ovarian cancer cells could be decreased by down-expressing of MTRR gene in vitro and in vivo.
10.Effect of MTRR gene on apoptosis and autophagy pathways in multi-resistant epithelial ovarian cancer
Jia CHEN ; Qi WANG ; Wei ZHANG ; Li LI
Chinese Journal of Obstetrics and Gynecology 2016;51(4):285-292
Objective To explore the effect of down-regulated methionine synthase reductase (MTRR) gene on the apoptosis and autophagy pathway, and offer a possible approach for the MTRR to reverse the multi-resistant ovarian cancer. Methods (1) The experiment was divided into 3 groups, SKOV3/DDP-MTRRi (down-regulated MTRR group), SKOV3/DDP-NC (negative control group), and SKOV3/DDP (blank control group). Different concentration of cisplatin (0, 1, 2, and 4 μg/ml) treated on 3 groups cells. The apoptosis rate was measured by flow cytometry (FCM). Autophagy was detected by immunofluorescence. Autophagy microtubule associated protein light chain 3β(LC3B) and p62 were detected by western blot. The formation of autophagosome of cells was observed by transmission electron microscope. (2) Detection of autophagy and apoptosis of SKOV3/DDP-MTRRi induced by rapamycin. The experiment was divided into 4 groups included rapamycin group (5 nmol/L rapamycin), rapamycin+cisplatin group (5 nmol/L rapamycin+4μg/ml cisplatin), cisplatin group (4μg/ml cisplatin) and blank control group. LC3B and p62 protein were detected by western blot. The survival rate cells were detected by methyl thiazolyl tetrazolium (MTT) method. The apoptosis rate was measured by FCM. (3) The 3 groups cells (SKOV3/DDP, SKOV3/DDP-NC and SKOV3/DDP-MTRRi) induced by a certain concentration of cisplatin (4 μg/ml) after 48 hours, then detecting the protein expression of caspase, Bcl-2 family in apoptosis pathway and the key proteins in phosphatidylinositol-3 kinase (PI3K)/protein kinase B (Akt) autophagy pathways by western blot, getting the time when the proteins′expression changed. Results (1) The 3 groups cells (SKOV3/DDP, SKOV3/DDP-NC, and SKOV3/DDP-MTRRi) induced by a certain concentration of cisplatin (4 μg/ml) after 48 hours, apoptosis and autophagy of 3 groups of cells were gradually increased with the increased concentration of cisplatin. The apoptosis rate of SKOV3/DDP-MTRRi cells [(26.2 ± 1.4)%] were significantly increased compared with the SKOV3/DDP-NC cells or SKOV3/DDP cells [(14.8 ± 2.4)%, (14.2 ± 2.4)%;all P<0.05] at 2μg/ml cisplatin. Immunofluorescence tests revealed that the aggregates of LC3B in SKOV3/DDP-MTRRi cells were more than that of SKOV3/DDP-NC cells and SKOV3/DDP cells. The expression of LC3B of SKOV3/DDP-MTRRi cells was lower than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). The expression of p62 of SKOV3/DDP-MTRRi cells was higher than those of SKOV3/DDP-NC cells and SKOV3/DDP cells (P<0.05). The structure of chloroplast was integrity and autophagosome was dispersing in plastids of SKOV3/DDP-NC cells and SKOV3/DDP cells. Organelles disappear and vacuoles increased obviously in SKOV3/DDP-MTRRi cells, no autophagosome was observed. (2) The expression of LC3B of rapamycin+cisplatin group was higher than those of other 3 group cells (1.72±0.08,1.43±0.04, 1.37±0.11, and 1.11 ± 0.09;P<0.05). The expression of p62 of rapamycin + cisplatin group was significant decreased (0.58 ± 0.10,0.94 ± 0.12, 1.21 ± 0.11, and 1.57 ± 0.10; P<0.05). The survival rate of rapamycin + cisplatin group was higher than that of cisplatin group [(0.78±0.03)%vs (0.62±0.03)%;P=0.018], the apoptosis rate was significant decreased in rapamycin+cisplatin group [(59.0 ± 3.9)% vs (40.4 ± 3.0)%, P=0.019]. (3) The 3 groups cells (SKOV3/DDP, SKOV3/DDP-NC, and SKOV3/DDP-MTRRi) induced by a certain concentration of cisplatin (4μg/ml) after 48 hours, the expression of Bax in 3 groups cell were not evidently changed (P=0.661). The expression of Bcl-2 was significantly decreased in SKOV3/DDP-MTRRi cells (P=0.030). The expression of caspase-3, caspase-7, caspase-9, and poly (ADP-ribose) polymerase (PARP) were not evidently changed (P>0.05), but cleaved caspase-3, cleaved caspase-7, cleaved caspase-9, and cleaved PARP were significantly increased in SKOV3/DDP-MTRRi cells (P<0.05). For the autophagy pathway, the expression of phosphorylated Akt (p-Akt) and phosphorylated mammalian target of rapamycin (p-mTOR) were significantly increased (P<0.05), but Akt and mTOR had no significant variation. The expression of phosphatase and tensin homologue deleted on chromosome ten (PTEN) was significantly decreased (P<0.05). Conclusions MTRR silencing significantly increase cisplatin-induced apoptosis and reduce the autophagy induced by cisplatin in SKOV3/DDP cells. Down-regulation of MTRR enhanced the chemosensitivity of cisplatin-resistant ovarian cancer cells may be by activating caspase and Bcl-2 apoptosis family and inhibiting the PI3K/Akt autophagy pathway.