1.Influence of oxidative/antioxidative biomarkers and inflammatory cytokines on rats after sub-acute orally administration of titanium dioxide nanoparticles.
Di ZHOU ; Zhang Jian CHEN ; Gui Ping HU ; Teng Long YAN ; Chang Mao LONG ; Hui Min FENG ; Guang JIA
Journal of Peking University(Health Sciences) 2020;52(5):821-827
OBJECTIVE:
To evaluate the sub-acute oral effect of titanium dioxide (TiO2) nanoparticles on the oxidation/antioxidation biomarkers and inflammatory cytokines in blood, liver, intestine, and colon in rats.
METHODS:
Twenty four 4-week-old clean-grade Sprague Dawley (SD) rats were randomly devided into 4 groups by body weight (n=6, control, low, middle, and high), in which the rats were orally exposed to TiO2 nanoparticles at doses of 0, 2, 10 and 50 mg/kg body weight/day for 28 consecutive days separately. Food intake, body weight and abnormal behaviors during the experiment were recorded. The rats were euthanized on the 29th day. The blood was collected via abdominal aortic method and centrifuged to collect the serum. Tissues from liver, intestine and colon were collected and homogenated. Then enzyme-linked immunosorbent assay (ELISA) and microwell plate methods were used to detect oxidation/antioxidation biomarkers including superoxide dismutase (SOD), glutathione (GSH), glutathione peroxidase (GSH-Px), total mercapto (T-SH), glutathione disulfide (GSSG), malomdialdehvde (MDA) and inflammatory cytokines including interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the serum, liver, intestine and colon in the rats.
RESULTS:
Compared with the control group, no significant differences in body weight, food intake and organ coefficients were observed in all the three groups after TiO2 gavage. No significant changes in GSH, GSH-Px, T-SH, and IL-6 were observed. Compared with the control group, significant increase of SOD activity in serum in high dose group, signi-ficant increase of GSSG concentration in intestine in middle and high dose group and significant increase of MDA concentration in liver in low and high dose group were observed. Compared with the control group, a significant increase of TNF-α in liver in middle and high dose group was observed.
CONCLUSION
TiO2 nanoparticle can increase antioxidant enzymes activities in blood, increase oxidative biomarkers in liver and intestine, increase inflammatory cytokines in liver in rats after a 28-day sub-acute orally administration. Among blood, liver, intestine, and colon, liver is most sensitive to the toxicity induced by TiO2 nanoparticles, followed by intestine, blood, and colon in sequence.
Animals
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Antioxidants
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Biomarkers
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Cytokines
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Nanoparticles
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Oxidative Stress
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Rats
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Rats, Sprague-Dawley
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Titanium
2.hVEGF siRNA inhibits initial growth of A549 tumor cells in nude mice and angiogenesis on CAM assay.
Rong LIU ; Wen-jiang MA ; Jian-ya ZHOU ; Guang-di YANG ; Jian-ying ZHOU
Journal of Zhejiang University. Medical sciences 2009;38(2):136-144
OBJECTIVETo investigate the effect of small interfering RNA (siRNA) targeting human vascular endothelial growth factor (hVEGF) on A549 cell growth in nude mice and angiogenesis on chorioallantoic membrane (CAM) assay.
METHODSThree pairs of hVEGF siRNA-plasmid and non-silencing-plasmid were constructed, and transfected into A549 cells through lipofectamine 2000, respectively. The most effective pair of hVEGF siRNA-plasmid was selected by ELISA and real-time RT-PCR. A549 cells transfected with selected hVEGF siRNA- plasmid, A549 cells transfected with non-silencing-plasmid and A549 cells without transfection were inoculated into nude mice, respectively. Chick embryos were randomly divided into four groups and CAM was treated by different solutions for 48 h: culture media DMEM as negative control group,un-transfected A549 cell culture supernatants as positive control group, hVEGF siRNA A549 cell culture supernatants as hVEGF siRNA group and nonsilencing siRNA A549 cell culture supernatants as non-silencing siRNA group. The CAMs were harvested on d12 for microscopic assays.
RESULTCompared with control group, hVEGF siRNA-plasmid induced 48% reduction in hVEGF secretion by A549 cells accompanied by 70% reduction in hVEGF mRNA. Compared with non-silencing siRNA group, the mean tumor volume of murine xenograft was reduced by 58% in hVEGF siRNA group; time for xenografts growing to 50 mm(3)was delayed by 5.4 d. hVEGF contents in xenograft were reduced by 54%; but mean doubling time of tumors and the growth rate of tumors were not significantly reduced. In CAM assays, hVEGF content was zero in negative group, and in hVEGF siRNA group that was 40%-44% of non-silencing siRNA group or positive group; vessels branch points of CAM in hVEGF siRNA group or non-silencing siRNA group or positive group were increased by 45%-55% compared with negative group; total vessel length of CAM in hVEGF siRNA group was increased by 53% compared with negative group, while in non-silencing siRNA group or positive group that was increased by 97% or 99%. Compared with negative control group, the proliferation of microvessels was increased when cell culture supernatant with hVEGF was added in hVEGF siRNA group, significant proliferated vessels were observed in non-silencing siRNA group or positive group.
CONCLUSIONA plasmid-mediated hVEGF siRNA has been constructed and verified, which can effectively downregulate the expression of hVEGF in human A549 cells, resulting in the inhibition of angiogenesis. hVEGF siRNA can delay initial growth of A549 tumor xenograft but not reduce the growth rate.
Adenocarcinoma ; pathology ; Animals ; Cell Line, Tumor ; Cell Proliferation ; Chick Embryo ; Chorioallantoic Membrane ; blood supply ; metabolism ; Female ; Humans ; Lung Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Physiologic ; RNA Interference ; RNA, Messenger ; genetics ; RNA, Small Interfering ; genetics ; Vascular Endothelial Growth Factor A ; antagonists & inhibitors ; genetics
3.Genetic association of apoE and apoCI gene polymorphisms with coronary heart disease.
Chun-Hong WANG ; Xin ZHOU ; Guang-di ZHOU ; Xiao-dong TAN ; Ding-fen HAN ; Fang ZHENG ; Fang LIU
Chinese Journal of Epidemiology 2004;25(11):982-985
OBJECTIVETo study the genetic association of apolipoprotein (apo) E and apoCI gene polymorphisms with coronary heart disease (CHD) in China.
METHODSapoE genotypes were identified by multiplex amplification refractory mutation system (multi-ARMS) and the apoCI promoter polymorphisms were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 186 cases with CHD (age: 65.0 +/- 10.5 years) and 350 controls (age: 63.6 +/- 8.3 years). The haplotype frequencies were estimated.
RESULTSThe frequencies of apoE E4/3 genotype (26.9%) and epsilon4 (14.5%) in CHD group were significantly higher than that in the control group (12.6%, 7.0%), P <0.05. The significant difference was also found for the apoCI locus and the CHD group showed higher rate of both for the H2 allele and genotypes, carrying this allele. Estimation of the haplotype frequencies indicated that the association between the apoE-CI haplotype and CHD was significantly strong. The apoE-epsilon4/apoCI-H2 was estimated to be responsible for 9.86% of CHD.
CONCLUSIONWhen the subjects carrying both epsilon4 and H2 alleles, they would have higher risk of suffering from CHD than controls.
Adult ; Aged ; Aged, 80 and over ; Alleles ; Apolipoproteins C ; genetics ; Apolipoproteins E ; genetics ; China ; epidemiology ; Coronary Disease ; blood ; epidemiology ; genetics ; Female ; Genotype ; Haplotypes ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Risk Factors
4.Modulation of lianbizi injection (andrographolide) on some immune functions.
Guang-yong PENG ; Feng ZHOU ; Ru-ling DING ; Huan-di LI ; Kun YAO
China Journal of Chinese Materia Medica 2002;27(2):147-150
OBJECTIVETo study the effects of andrographolide on immune functions and the immune mechanism of its clinical application.
METHODThe amounts of IFN-alpha, IFN-gamma, TNF-alpha, IL-8 in the peripheral blood mononuclear cells(PBMCs) culture supernatants deal with by different concentrations of LianBiZhi (LBZ) injection made of andrographolide were detected by biological activity test or ELISA in vitro. The effects of LBZ injection on macrophage phagocytotic function and natural killer cells cytotoxicity were examined by means of macrophage to phagocytize cock erythrocyte and measurement of lactate dehydrogenase(LDH) activity released from the damaged cells, respectively.
RESULTThe LBZ injection could promote IFN-alpha, IFN-gamma, TNF-alpha inductions of PBMCs, but had no effect on IL-8. At the same time, the LBZ injection could not only enhance the phagocytosis activity of peritoneal macrophage from guinea pig to phagocytosis cock erythrocyte, but also augment the cytotoxicity mediated by natural killer cells from PBMCs to damage the K562 cell lines.
CONCLUSIONAndrographolide is an immunostimulant agent which can modulate both antigen specific and nonspecific immune function by means of its natural killer cells and macrophage and cytokines induction.
Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Chickens ; Diterpenes ; pharmacology ; Guinea Pigs ; Humans ; Injections ; Interferon-alpha ; metabolism ; Interferon-gamma ; metabolism ; Killer Cells, Natural ; drug effects ; Macrophages, Peritoneal ; drug effects ; Neutrophils ; metabolism ; Phagocytosis ; drug effects ; Tumor Necrosis Factor-alpha ; metabolism
5.Analysis of closed abdominal injury in pregnant women.
Chang-di TU ; Shao-juan WANG ; Ri-guang ZHOU ; Yang-yi WEI ; Qiang TAN
Chinese Journal of Traumatology 2005;8(4):205-208
OBJECTIVETo explore the characteristics of closed abdominal injury in pregnancy women and its treatment.
METHODSThe clinical data of 37 pregnancy patients with closed abdominal injury treated in our hospital from June 1993 to June 2003 were collected and analyzed.
RESULTSAll the 37 patients were treated with operation. Among them 2 early pregnancy patients with intestinal rupture and 1 patient with retroperitoneal hematoma were treated under laparoscope; in other 34 pregnancy patients laparotomy was performed. Of the 34 patients 8 used cesarean section because premature separation of placenta and enlarged womb interrupted the management of intra-abdominal organ injury. In the 37 patients 33 (89.1%) were cured, 4 (10.8%) die, postoperative complication rate was 16.2% (6/37). Two patients (5.4%) suffered from abdominal cavity infection, 3 (8.1%) from pulmonary infection, and 1 (2.7%) had multi-organ failure.
CONCLUSIONSFor pregnancy patients with closed abdominal injury, besides obsteric diseases intra-abdominal injury should be given much attention. Accurate diagnosis and timely treatment can gain the time to save the life of both mother and fetus.
Abdominal Injuries ; diagnosis ; surgery ; Adult ; Female ; Humans ; Laparoscopy ; Pregnancy ; Pregnancy Complications ; diagnosis ; surgery
6.Preliminary investigation of relationship between sperm apoptosis and male infertility.
Guang-Rong WANG ; Zeng-Di ZHOU ; Zheng-Min GE ; Ming-Jun ZHAO
National Journal of Andrology 2002;8(1):25-27
OBJECTIVESTo evaluate the relationship between sperm apoptosis and male infertility.
METHODSPercentage of apoptotic sperm (PAS) in spermatozoa of fertile and infertile men were tested by flow cytometry (FCM).
RESULTSSperm apoptosis had happened in all different people. PAS in fertile and infertile group was (4.28 +/- 1.66)% and (18.67 +/- 8.55)% respectively, and difference was significant between two groups (P < 0.01). There was negative correlation between PAS and semen volume, sperm density, percentage of forward motility, percentage of normal morphology (P < 0.01).
CONCLUSIONSThere was very close relationship between sperm apoptosis and male infertility. FCM used to test sperm apoptosis is rapid, accurate, objective and reliable to analyse sperm functions and male fertility.
Apoptosis ; Flow Cytometry ; Humans ; Infertility, Male ; pathology ; Male ; Spermatozoa ; pathology
7.Studies on triterpenoid constituents in leaf of Ilex pernyi.
Guang-Bo XIE ; Si-Xiang ZHOU ; Lian-Di LEI ; Peng-Fei TU
China Journal of Chinese Materia Medica 2007;32(18):1890-1892
OBJECTIVETo investigate the chemical constituents of Ilex pernyi.
METHODThe chemical constituents were isolated by various column chromatographic methods. The structures were identified by spectral data.
RESULTEight triterpenoid compounds were isolated and identified as ursolic acid (1), lupeol (2), alpha-amyrin (3), uvaol (4), 3beta-hydroxyurs-11-ene-13beta-olide (5), pomolic acid (6), lup-20 (29)-ene-3beta, 24-diol (7), 3beta, 23-dihydroxy-urs-12-en-28-oic acid (8).
CONCLUSIONThe eight compounds were obtained from this plant for the first time.
Ilex ; chemistry ; Oleanolic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Triterpenes ; chemistry ; isolation & purification
8.Effect of different hemopoietic microenvironment on the differentiation of hemopoietic cells from human embryonic stem cells.
Hui-ping ZHAO ; Hai-jun ZHAO ; Ge LIN ; Di ZHOU ; Tian-cheng LIU ; Guang-xiu LU
Journal of Central South University(Medical Sciences) 2007;32(6):992-996
OBJECTIVE:
To observe the inductive efficiency of deriving hematopoietic cells from human embryonic stem (hES) cells co-cultured with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells,in order to discuss the effect of the different hemopoietic microenvironment on hemopoietic cytogenesis.
METHODS:
We used two-step method to induce the hES cells into the hematopoietic cells. In the first step the hES cells were co-cultured with cytokines by formation of the day 5 embryoid bodies (5d EBs). In the second step the 5d EB cells were induced into the hematopoietic cells by co-culturing with human yolk sac stromal cells, fetal liver stromal cells or fetal bone marrow stromal cells for 10 days. The inductive efficiencies of deriving hematopoietic cells from hES cells co-cultured with the different hemopoietic microenvironment were reflected by the expression levels of flk, CD34 and CD45 antigen.
RESULTS:
Flow cytometry analysis demonstrated that the population of the cells co-cultured with human yolk sac stromal cells contained flk (1.80%+/-0.56%), CD34 (1.30%+/-0.14%) or CD45 (1.05%+/-0.63%) positive cells; the population of the cells co-cultured with human fetal liver stromal cells contained flk (34.00%+/-25.45%), CD34 (38.40%+/-24.80%) or CD45 (72.60%+/-25.70%) positive cells; the population of the cells co-cultured with human fetal bone marrow stromal cells contained flk (2.50%+/-1.48%), CD34 (3.20%+/-0.56%) or CD45 (1.65%+/-0.21%) positive cells. Compared with spontaneous differentiation of EBs, all of the three stromal cells could induce EBs into the hematopoietic cells (P<0.05).
CONCLUSION
The inductive efficiency of deriving hematopoietic cells from EBs co-cultured with human fetal liver stromal cells was higher than EBs co-cultured with human yolk sac stromal cells and fetal bone marrow stromal cells.
Antigens, CD34
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Cell Differentiation
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Cells, Cultured
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Cellular Microenvironment
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Coculture Techniques
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Embryonic Stem Cells
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cytology
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Fetus
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cytology
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Hematopoietic Stem Cells
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cytology
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Humans
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Leukocyte Common Antigens
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Mesenchymal Stem Cells
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cytology
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Stromal Cells
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cytology
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Yolk Sac
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cytology
9.CT axial imaging of the iliolumbar ligament and its significance on locating lumbosacral vertebral segments.
Ke-Chang LIU ; Guang-Zhang XIANG ; Guang-Hua CHEN ; Yao ZHOU ; Xiao-Qing CAO ; Jian-Di XIA ; Ke LI
China Journal of Orthopaedics and Traumatology 2010;23(11):854-858
OBJECTIVETo study the CT axial manifestations of iliolumbar ligament(ILL) and discusses its clinical effects on locating lumbosacral vertebral segments.
METHODSFrom May 2008 to March 2010, 706 adult patients diagnosed lumbar disc disease were performed with axial scans by single slice helical CT. Among the patients, 436 patients were male and 270 patients were female, ranging in age from 25 to 82 years, the median age was 44 years, 78 cases with lumbosacral transitional vertebrae (LSTV) were verified by X-radiography or fluoroscopy. The morphology, origin and insertion, courses of ILL and the relationship of ligament and spinal segments on axial plane images were used to study. The location method of spinal segments by ILL was compared with the other four location methods on CT.
RESULTSOf the 628 cases with normal lumbosacral segmentations sides of ligament, the main part of ILL originated from L5 transverse processes and terminated at the iliac crest, the morphological characters were divided into two types: double band (71.8%, 451/628) and single band (28.2%, 177/628). The tiny branches from posterior and outside edge of L4, lumbar disc were seen simultaneity in 3 cases. The ILL of 78 cases with LSTV all also originated from L5 transverse processes. Using ILL as a marker of the L5 vertebral level, 78 cases with LSTV were correctly numbered, the accuracy rate was higher than the other location methods, there was statistical significance between the location method by ILL and the location method by iliac crest (P < 0.05).
CONCLUSIONThe main part of ILL originates from L5 transverse processes, the anatomic location is relatively steady and can be clearly displayed on axial CT, which can be used as a measure in the idenlification of LSTV in clinical practice, it is worthy to be applied widely in basic-level hospitals.
Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Ilium ; diagnostic imaging ; Ligaments ; diagnostic imaging ; Lumbar Vertebrae ; diagnostic imaging ; Lumbosacral Region ; Male ; Middle Aged ; Tomography, X-Ray Computed
10.Serum testosterone reduction and metabolism in aging male rats: correlation and mechanism.
Di GUAN ; Fang ZHOU ; Zheng-Yan GE ; Wen-Hong LU ; Dong YUAN ; Xiao-Wei LIANG ; Shan-Jie ZHOU ; Zhi-Guang LI ; Yi-Qun GU
National Journal of Andrology 2011;17(7):579-585
OBJECTIVETo investigate the mechanism of serum testosterone reduction, its relationship with metabolism, changes in the number and morphology of Leydig cells and endocrine function in aging male rats.
METHODSThe levels of serum total testosterone (tT), LH, FSH, HDL, LDL, TG, TC, Glu, INS, IRG and LP were determined in young (9 mo) and aging rats (12, 15, 18 and 21 mo), with 6 in each group. The morphological changes of Leydig cells were observed under the microscope. The concentrations of testosterone secreted from the cultured Leydig cells with the stimulation of hCG and Forskolin were assayed. The apoptosis rates of Leydig cells were detected by TUNEL. The visceral fat was isolated and weighed, and the Lee's index calculated. All the above indexes were recorded and compared among different age groups.
RESULTSThe aging rats showed a significant decrease in the levels of serum tT and TSI ([1.26 +/- 0.65] ng/ml and [0.07 +/- 0.65] ng/mIU) as compared with the young rats ([3.24 +/- 0.38] ng/ml and [0.21 +/- 0.01] ng/mIU) (P < 0.01). Obvious differences were found in the morphology of Leydig cells among different age groups. The T secretion of Leydig cells at 24, 48 and 72 h in aging rats was markedly decreased (P < 0.05) while their TUNEL positive rate remarkably increased in the aging rats (17.36% +/- 1.31%) compared with the young ones (7.02% +/- 1.05%) (P < 0.05). There were statistically significant differences between the young and aging rats in all the biochemical parameters including IRG, HDL, LDL, TG, TC and visceral fat content (P < 0.05), except the levels of serum Glu, INS and LP (P > 0.05).
CONCLUSIONThe serum T level and secreting capacity of Leydig cells are significantly lower in aging rats than in young ones, and the metabolic parameters undergo regular changes with the decreasing level of serum T. The reduction of testosterone in aging male rats may be associated with the decreased secreting capacity and number of Leydig cells and declined function of the pituitary.
Aging ; Animals ; Leydig Cells ; cytology ; metabolism ; secretion ; Male ; Rats ; Rats, Sprague-Dawley ; Testosterone ; blood ; metabolism