1.Flow cytometry analysis of unusual peripheral monocyte population
Lixin ZHANG ; Jun YE ; Mei LIN ; Taohong LU ; Zulong XU ; Xuejun ZHU ; Yabao CHEN
Chinese Journal of Laboratory Medicine 2008;31(5):504-508
Objective To analyze a population of cells on the right lower lateral of monocyte population in forward scatter/side scatter(FSC/SSC)(X-axis/Y-axis)scatterplot of peripheral blood leucocyte by flow cytometry(FCM)and its influencing factors.Methods The type of cells were identified based on cluster of differentiation antigen(CD)by FCM.The impact of temperature,hemolysin concentration,and incubation time was evaluated.Blood lipid tests were performed to observe the relation between them by statistical methods.Results (1) Phenotypo of this population of cells on the right lower lateral of monocytes in FSc/SSC scatterplot is CD+45 CD+13 CD+14 CD3- CD-19 ,which was the same as monocyte cells:(2)The monocytes in FSC/SSC scatterplot shifted to left side after using haemolysin;(3)The monocytes showed less resistance to antihemolysin in 37℃ than that in 220C:There were more monocytes shifting to left side with the increase of haemolysis time:(4)The swarming ratio of monocytes in patients (31.5%,40/126) Was higher than it in normal controls (5.1%,5/98)(x2=22.74,P<0.01);(5)The levels of total serum cholesterol(TC),triglyeride(TG),low density lipoprotein cholesterol(LDL-C), apoprotein B100(Apo B100) in patients with swarming monocytes were lower than that in the patients without swarming monocytes,(P<0.05).There was no statistical significance between the two groups with respect to levels of total bilirubin(TBIL),albumin(Alb),hish density lipoprotein cholesterol(HDL-C),apoprotein A I(Apo A I),lipoprotein(Lpa).Conclusions Peripheral blood monocytes can be divided in two groups in FSC/SSC scatterplot when analyzed with FCM.The presence of this population of cell Was related to resistance to hemolysin.It can be influenced by haemolysis time and incubation temperature.Therefore,the effect of swarming monocytes and abnormal cell membrane should be taken into consideration when the markers and function of monocytes are detected by FCM.
2.Identification of melatonin receptor in human embryonic peripheral tissue
Zhi-Min LIU ; Jun-Jie ZOU ; Yu-mei SHEN ; Zu-Qian LU
Academic Journal of Second Military Medical University 2001;22(1):8-11
Objective: To determine protein binding characteri stic and signal transmission pathway of melatonin(Mel) receptor(MR) in human e mbryonic peripheral organ tissues. Methods: MR was measured by radio ligand-binding assay and the effect of GTPγS on melatonin specific bindi ng was studied. Results: Mel specific binding sites were det ermined in 16 kinds of human embryonic tissue and this binding could be inhibit ed by GTPγS, supporting the theory that MR is coupled to inhibitory G-proteins system. Conclusion: MR is measured in human embryo tissue, the se results provide experimental data for elucidating the mechanism of the effect of Mel.
3.Preliminary report of sunitinib as first line treatment in patients with metastatic renal cell carcinoma
Xinan SHENG ; Siming LI ; Zhihong CHI ; Lu SI ; Chuanliang CUI ; Mei HAN ; Jun GUO
Chinese Journal of Urology 2011;32(2):134-137
Objective To evaluate the efficacy and safety of sunitinib as first line treatment in patients with metastatic renal cell carcinoma (RCC). Methods This study included 46 Chinese patients who were diagnosed with metastatic RCC after radical nephrectomy. The patients received oral sunitinib (50 mg once daily on a 4 weeks on, 2 weeks off) on a 6 weeks cycle dose schedule until disease progression or intolerable toxicities occurred. Results The overall objective response rate was 32.6% (95% confidence interval [CI, 19.1% to 46. 1%]), and the disease control rate was 86.9%,with complete response (CR) 0 (0%), partial responses (PRs) 15 (32.6%), stable disease (SD) 25(54.3 %), and progression disease (PD) 6 ( 13. 1%). The median progression-free survival was 11 months, and the 1-year survival rate was 65.2%, while the median overall survival (mOS) has not been reached. The main adverse events included fatigue 33 (71.7%), skin discoloration 29 (63.0 %),anorexia 28 (60.9%), hand-foot syndrome 26 (56.5%), oral mucositis 25 (54.3%), hypertension 19 (41.3%), facial edema 18 (39.1%), diarrhea 17 (37.0%), hemorrhage 17 (37.0%), nausea 15 (32.6%), and hematological toxicity: leukopenia 32 (69.6%), neutropenia 30 (65.2%), thrombocytopenia 28 (60.9%), anemia 21 (45.7%). Most of grade 3/4 serious adverse events were thrombocytopenia in 15 (32. 6%) patients. Conclusions Sunitinib has a prominent effect in metastatic renal cell cancer in a Chinese population with mostly mild to moderate adverse reactions. More attention should be paid to grade 3/4 adverse reaction of thrombocytopenia.
4.Association Study of the Disrupted in Schizophrenia 1(DISC1)Gene Polymorphisms with Schizophrenia
Wei SUN ; Jun YAN ; Lifang WANG ; Mei QU ; Yanyan RUAN ; Tianlan LU ; Weihua YUE
Chinese Mental Health Journal 2002;0(08):-
T,P=0.008; Genotype:P=0.031)of DISC1 gene were significantly associated with schizophrenia.The haplotypes constructed by these two markers were significantly associated with schizophrenia,such as AT(?2=7.065,P=0.008,OR=1.42,95%CI=1.10~1.83)and GA(?2=6.009,P=0.014,OR=0.80,95%CI=0.68~0.96).When the subjects examined with the positive and negative syndrome scale(PANSS),the risk haplotype AT was not significantly correlated with positive,negative,excitement,depression and cognitive impairment factors of PANSS. Conclusion:These findings provide further evidence for DISC1 as a predisposing gene involved in schizophrenia in the Chinese Han Population.However,no positive association is found between DISC1 polymorphisms with schizophrenia clinical symptoms.
5.Effects of cholesterol-lowering probiotics on the metabolism of bile acid in a rat model of non-alcoholic fatty liver disease and the possible mechanism
Limei BAI ; Pengyuan ZHENG ; Jun ZHANG ; Fuguang LI ; Lu MEI ; Huang HUANG ; Yirui DING ; Simeng LIU
Chinese Journal of Microbiology and Immunology 2016;36(2):110-116
Objective To investigate the effects of two cholesterol-lowering probiotics, DM9054 (Lac-tobacillus Rhamnosus GG, LGG) in combination with 86066 (Lactobacillus plantarum WCFS1, LP), on the metabolism of bile acid via a rat model of non-alcoholic fatty liver disease (NAFLD) and the possible mecha-nism. Methods Twenty-one SD male rats were randomly divided into three groups including control group, NAFLD model group and probiotics intervention group. Rats in the control group received normal diet. The rat model of NAFLD was established by feeding rats with chronic high fat diet (45% of calories derived from fat di-et) for 20 weeks. Rats in the probiotics intervention group were given high fat diet together with cholesterol-low-ering probiotics through oral gavage. General indexes of each group including body weight and the levels of tri-glyceride, cholesterol and CK18-M30 in serums samples were detected. The expression of cholesterol 7-alpha hydroxy-lase (CYP7A1), fibroblast growth factor receptor 4 (FGFR4), farnesoid X receptor (FXR), fibroblast grwoth factor 15 (FGF15) and apical sodium-dependent bile acid transparter(ASBT) at mRNA level were de-tected by using real-time polymerase chain reaction (real-time PCR). Western blot assay was used to detect the protein expression of CYP7A1, FXR in liver tissues and ASBT in ileum tissues. The expression of FXR in liver and ileum tissues were analyzed by immunohistochemistry. Results Rats with NAFLD showed loss of body weight and decreased levels of the serological markers after treating with the probiotics (P<0. 05). Compared with the rats in model group, enhanced expression of CYP7A1 and inhibited expression of FXR in liver tissues, activated FXR-FGF15 pathway in ileum tissues as well as down-regulated expression of ASBT in ileum tissues were detected in rats receiving probiotics intervention (P<0. 05). No significant difference in the expression of FGFR4 at mRNA level was observed between NAFLD rats with or without probiotics intervention (P>0. 05). Conclusion Probiotics intervention might up-regulate the expression of CYP7A1 by suppressing the FXR path-way in liver tissues and inhibiting the expression of ASBT in ileum tissues. Treating NAFLD rats with cholester-ol-lowering probiotics could activate the FXR-FGF15 pathway in ileum tissues and enhance the metabolism of bile acid, which contributed to the alleviation of NAFLD.
6.Study on the Characteristics of Nicotine Degradation by Strain DN2 and its Application
Yong-Jun YUAN ; Zhao-Xin LU ; Xiao-Mei BIE ; Feng-Xia LV ;
China Biotechnology 2006;0(03):-
The kinetics of nicotine degradation by O.intermedium DN2 and its application in tobacco waste were investigated. The results showed that the optimum temperature of nicotine degradation by O.intermedium was 30 ℃, the pH value was 6.5 and a mount of inoculum was 5 %. Under above conditions, the kinetics of nicotine degradation of initial concentration 500 mg/L was studied. The results indicated that the degradation process of nicotine with no-induced strain DN2 followed inverse S-shaped curve, and degradation process of nicotine with induced cells of DN2 followed Eckenfelder mode. The half life of nicotine degradation was 17.43 h and 4.10 h, respectively. And the results also showed that tolerance of O.intermedium DN2 to nicotine was up to 5000 mg/L when 0.1 % of glucose was added. Nicotine (2 220 mg/L) in extract of tobacco wastes degraded about 95.22 % by strain DN2 in 60 h incubation, indicating that strain DN2 was of application value in treatment nicotine pollution.
7.Comparison of Three ELISA Kits for the Differentiation of Foot-and-mouth Disease Virus-infected from Vaccinated Animals
Yi-mei, CAO ; Zeng-jun, LU ; Zai-xin, LIU ; Qing-ge, XIE
Virologica Sinica 2007;22(1):74-79
A study was performed to validate 3 FMDV 3ABC-I-ELISA kits developed in China for the differentiation of FMDV infected and vaccinated animals.Sets of sera from naive and vaccinated cattle as well as from cattle that had been infected were tested for antibodies against nonstructural proteins (NSPs) of FMDV by commercial diagnosis kits,Ceditest(R)FMDV-NS (Ceditest(R) kit),UBI(R) FMDV NONSTRUCTURAL PROTEIN ELISA DIRECTION INSERT (UBI(R) kit) and a FMDV 3ABC-I-ELISA kitdeveloped at the Lanzhou Veterinary Research Institute.The test parameters (sensitivity and specificity) of the three kits were determined,and the result obtained from FMD 3ABC-I-ELISA kit was compared with that obtained from two foreign kits.The results indicated that the coincidence rate between the FMDV 3ABC-I-ELISA and Ceditest(R) kits was 98.05%,and the coincidence rate between the FMDV 3ABC-I-ELISA and UBI(R) kits was 94.4%; the sensitivity of both Ceditest(R) and FMDV 3ABC-I-ELISA kit was 100%.However,the sensitivity of the UBI(R) kit was only 81.8%.With sera from naive or vaccinated non-infected animals,the specificity of all tests exceeded 90%.
8.Effects of arsenic trioxide combined with bortezomib on proliferation and apoptosis of K562 cells and their mechanism.
Hong-Ying WU ; Song-Mei WANG ; Jun-Qian HUANG ; Lu-Mei HAO ; Xue-Mei LI ; Li-Yan WEI
Journal of Experimental Hematology 2012;20(6):1361-1364
This study was aimed to investigate the effect of arsenic trioxide (As(2)O(3)) alone and in combination with bortezomib (Bor) on proliferation and apoptosis of leukemia cell line K562, and to analyze the potential mechanism. K562 cells were treated with different concentrations of As(2)O(3) or Bor (alone or combination) for 24, 48 h. MTT method was used to detect the cell proliferation. After K562 cells were treated with 0.5 µmol/L As(2)O(3) alone or in combination with 10 nmoL/L Bor, the apoptosis rate and cell cycle were measured by flow cytometry, and the activity of NF-κB was analyzed by SP immunohistochemistry. The results indicated that the different concentrations of As(2)O(3) and Bor could inhibit the K562 cell proliferation in a time- and dose-dependent manners (P < 0.05). The IC(50) of Bor and As(2)O(3) in 48 h were 20 nmol/L and 0.6 µmol/L respectively. When K562 cells were treated with As(2)O(3) or Bor alone for 24 h, the apoptotic rate of K562 cells increased, and the apoptotic rate in combination group was higher than that in As(2)O(3) or Bor group. The cells were apparently arrested in G(2)/M phase in Bor group and G(0)/G(1) phase in As(2)O(3) group. The activity of NF-κB decreased significantly in As(2)O(3) or Bor group (P < 0.05), this effect was most significant in the combination group (P < 0.01). It is concluded that both As(2)O(3) and Bor can inhibit the proliferation and induce apoptosis of K562 cells, a synergistic effect can be observed when a low dose of As(2)O(3) combined with low dose of Bor. The different cell cycle block site and the decrease of activity of NF-κB may be one of the mechanisms underlying their synergic effect.
Apoptosis
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drug effects
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Arsenicals
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pharmacology
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Boronic Acids
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pharmacology
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Bortezomib
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Cell Proliferation
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drug effects
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Drug Synergism
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Humans
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K562 Cells
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NF-kappa B
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metabolism
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Oxides
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pharmacology
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Pyrazines
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pharmacology
9.Effects of hyperbaric oxygenation combined with As(2)O(3) on proliferation of K562 cells and associated mechanism.
Song-Mei WANG ; Hong-Ying WU ; Jun-Qian HUANG ; Lu-Mei HAO ; Li-Yan WEI ; Xue-Mei LI
Journal of Experimental Hematology 2012;20(4):863-866
This study was aimed to explore the effects of hyperbaric oxygenation (HBO) alone or combined with As(2)O(3) on proliferation, apoptosis and expression of HIF-1a, VEGF, caspase-3 mRNA of K562 cells, and the molecular mechanism of As(2)O(3) enhancing the anti-leukemic effect of HBO so as to provide a scientific basis for clinical treatment of chronic myeloid leukemia. The effects of drugs on proliferation of K562 cells was assayed by MTT method, the apoptosis rate of K562 cells was detected by flow cytometry with Annexin V/PI double staining, the expressions of HIF-1a, VEGF, caspase-3 mRNA of K562 cells were determined by real-time quantitative PCR. The results showed that as compared with As(2)O(3) alone, HBO combined with As(2)O(3) could increase inhibitory rate of K562 cell proliferation, and enhance apoptotic effect, obviously down-regulate expressions of HIF-1a and VEGF mRNA, up-regulate expression of caspase-3 mRNA. The effect of HBO combined with As(2)O(3) was higher then effect of As(2)O(3) alone, and their effects were synergistic (P < 0.05). It is concluded that HBO combined with As(2)O(3) can increase the expression of caspase 3 mRNA and decrease the expression of HIF-1a and VEGF mRNA, which may be one of molecular mechanisms underlying their synergistic antileukemia efficacy.
Apoptosis
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drug effects
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Arsenicals
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pharmacology
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Caspase 3
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metabolism
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Cell Proliferation
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drug effects
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Humans
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Hyperbaric Oxygenation
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Hypoxia-Inducible Factor 1, alpha Subunit
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metabolism
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K562 Cells
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Vascular Endothelial Growth Factor A
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metabolism
10.Morphology and immunological function of immature dendritic cells induced by sodium butyrate in human monocytes
Jun MIN ; Lu LIU ; Jie WANG ; Changzhen SHANG ; Yunle WAN ; Qiang YU ; Mei YANG ; Jun CAO ; Jisheng CHEN
Chinese Journal of Tissue Engineering Research 2007;11(37):7485-7488
BACKGROUND: The immature dendritic cell (imDC) can induce immunological tolerance and has widely application in the field of organ transplant. At present, the methods of inducing imDC are insufficient, so the new induction method is demanding.OBJECTIVE: To investigate the effect of sodium butyrate (SB) on the maturation and immunological function of human peripheral blood-derived imDC.DESIGN: Controlled observation and in vitro cytological trial.SETTING: Department of Hepatobiliary Surgery in the Second Affiliated Hospital of Sun Yat-sen University.MATERIALS: Five samples of human peripheral blood were obtained from the healthy volunteers (aged 20-23 years) of Sun Yat-sen University, totally 500 mL. Then peripheral blood mononuclear cells (PBMCs) and lymphocytes were isolated within 2 hours.METHODS: The experiment was carried out in the Medical Research Center of the Second Hospital Affiliated to Sun (1 mmol/L) was added for induction, while those supplemented with maturation promoting factor lipopolysaccharide (LPS)the beginning of induction, while LPS was added on the sixth day for second stimulation.MAIN OUTCOME MEASURES: Cell morphological change, flow cytometry was used to detect DC phenotype,FITC-labeled Dextran was used to detect the endocytosis of DC, the production of IL-12 was determined by means of enzyme-linked immunosorbent assay, and the proliferation of lymphocyte induced by DC was assayed with mixed lymphocyte reaction.expressions of CD80, CD83 and HLA-DR were significantly lower in the imDC of routine induction group following SB maturity promoting, compared with LPS group (P<0.01). On the sixth day, LPS was added into the SB-induced imDC,Endocytosis of DC: The imDC of routine induction group possessed a significantly lower endocytic activity after induced by LPS, and there were extremely significant differences compared with blank control group and SB maturation Production of IL-12: The production of IL-12 in the mDC induced by LPS was significantly higher than that in control group, SB maturation promoting group and SB induction group, the mDC induced by LPS in routine induction group stimulated significantly stronger proliferation of lymphocyte (P<0.01).