Table of Content

2009, Volume 40 Issue 3
06 June 2009 Previous Issue    Next Issue
---
论著
技术方法


For Selected:

Download Citations EndNote Reference Manager ProCite BibTeX RefWorks

Toggle Thumbnails

论著

Select

Changes of Aβ antibody and cytokines in peripheral blood from patients with Alzheimer’s disease 2009, 40 (3):  345-349.  doi: 10.3969/j.issn.0529-1356.2009.03.001 Abstract ( )   Objective To observe changes of peripheral immune function in patients with Alzheimer’s disease and to investigate their clinical meanings. Methods A total of 48 cases in Guangzhou Welfare Center from June 2006 to April 2007 were consecutively selected as the observed subjects, including 19 mild to moderately severe AD patients, 10 severe AD patients and 19 healthy age-matched controls.Level of the anti-β-amyloid peptide antibody was detected by ELISA. Distribution of the T, B-cell subsets was determined by flow cytometry (FACS). The production of IFN-γ and IL-4 by peripheral blood mononuclear cell(PBMC) after stimulation with 1-42 beta-amyloid (Aβ) fragments was also evaluated by ELISA and ELISPOT. Results Compared to the controls, the levels of AβSUB>42/SUB>antibody in serum in the patients with different course of AD were significant decreased (EM>P/EM><0.05 and EM>P/EM><0.01, respectively). The levels of CDSUB>8/SUB> in peripheral blood from patients with different degree of AD were upregulated to some extent（EM>P/EM><0.05)；In conurast, CDSUB>4/SUB> were significant downregulated（EM>P/EM><0.05）；After stimulated with antigen AβSUB>42/SUB>, the level of IFN-γ in the cultured medium of PBMC in HC was significant higher than that in both AD groups （EM>P/EM><0.05）. After stimulated with antigen AβSUB>42/SUB>, in every 4 Related Articles | Metrics

Select

Hair follicle neural stem cells promote dedifferentiation of macroglial cells in injured optic nerve of rats 2009, 40 (3):  350-357.  doi: 10.3969/j.issn.0529-1356.2009.03.002 Abstract ( )   Objective To investigate the effect of cultured hair follicle neural stem cells （HFNSCs） on dedifferentiation of macroglial cells in injured optic nerve of rats. Methods Follicle bulge cells isolated from vibrissa pad of male SD rats weighing about 90g were cultured in DMEM/F12 without serum or containing 15% FBS, identified as HFNSCs by immunofluorescence cytochemistry and PCR. The cells were stably transfected with rAAV2EGFP. Adult male SD rats were randomly divided into 3 groups as normal control group, injury group and transplantation group in which HFNSCs were transplanted into injured optic nerves. At 7 days, 14 days and 30 days after EGFPHFNSCs transplantation, optic nerves were observed under fluorescence microscope. At 7 days post operation, optic nerves from injury group and transplantation group were detected by gene chip of Affymetrix and realtime PCR. At 7 days, 14 days post operation, optic nerves were harvested and detected by HE staining and immunohistochemistry. Results Bulge cell in primary culture were labeled by neural progenitor cell markers, however the cells induced by 15% FBS expressed some makers of mature neural cells. EGFPHFNSCs could survive and migrate in the injured optic nerve 30 days after transplantation. With the injury group, there were 240 differentially expressed genes including genes related stem cell, apoptosis, proliferation, transcription, differentiation and development, cell adhesion, signal transduction and so on in the transplantation group. The result of real time PCR was consistent with that of gene chip. There were more cells, more immunoreactivity of Nestin, MBP, Erk1/2, and less immunoreactivity of GFAP in the distal optic nerves and more immunoreactivity of NF in the proximal optic nerves in the transplantation group than that in the injured group. Conclusion HFNSCs regulate some genes expression of glial cells in the injured optic nerve to Related Articles | Metrics

Select

Effect of glucocorticoid on the expression of Huntingtin-associated protein 1 in the rat hypothalamus 2009, 40 (3):  358-362.  doi: 10.3969/j.issn.0529-1356.2009.03.003 Abstract ( )   Objective To investigate the effect of glucocorticoid on the expression of Huntingtinassociated protein 1（HAP1）in rat hypothalamus. Methods Eighty male Wistar rats were divided into 4 groups: control group (injection of 0.9% saline, n=24）, corticosterone group［(injection of corticosterone, 5mg/(kg.12h), EM>n/EM>=40］, corticosterone+ RU38486 group［con-injection of corticosterone with its receptor antagonist RU38486, 10mg/(kg.12h), EM>n/EM>=8］ and Hydrocortisone group［orally given hydrocortisone (0.01g/L）in drink water, EM>n/EM>=8］. Corticosterone group was further divided into corticosterone 1day, corticosterone 3days, corticosterone 5days and corticosterone 7days group according to the difference in time after corticosterone -injection. The expression of HAP1 was studied 3days after co-injection of corticosterone and RU38486 in corticosterone +RU38486 group. Hydrocortisone was consecutively applied in the drink water for 1 month in hydrocortisone group. RT-PCR and Western blotting were used to explore the expression of HAP1 in rat hypothalamus. Results The expression of HAP1 and the level of its mRNA were decreased significantly in hypothalamus 1day and 3days after injection of corticosterone. The expression of HAP1 began to increase 5days after injection of corticosterone and recovered to the control level 7days after injection. The mRNA of HAP1 increased markedly 5days after injection and the level was higher than that of control group. The expression of HAP1 in hypothalamus significantly decreased in hydrocortisone group. Co-injection of corticosterone with RU38486 blocked corticosterone-induced action on HAP1 expression in hypothalamus. Conclusion Related Articles | Metrics

Select

Changes of Connexin 43 and 32 expression in the dorsal horn of spinal cord of rat with neuropathic pain and analgesic effect of carbenoxolone by intrathecal injection 2009, 40 (3):  363-368.  doi: 10.3969/j.issn.0529-1356.2009.03.004 Abstract ( )   Objective To study the changes of connexin 32 (Cx32) and Cx43 signals in the dorsal horn of spinal cord of rat with neuropathic pain induced by spared nerve injury (SNI) to sciatic nerve, and the analgeaic effect of 10μl carbenoxolone (CBX, a gap junction blocker, 1mmol/L）on neuropathic pain via intrathecal administration. Methods Adult SD rats were randomly divided into 3 groups, normal group (the animals did not receive any treatment), sham group (incised the skin and only exposed the sciatic nerve) and the SNI group, exposed three branches of the sciatic nerve, i.e. the tibial nerve, the common peroneal nerve and the sural nerve, and cut the tibial nerve and the common peroneal nerve but remained sural nerve. The paw withdrawal mechanical threshold (PWMT) of normal, sham and SNI groups was measured at 1day before SNI and at 3, 5, 10, 20 and 30 days after SNI, then the animals were killed and the spinal cords (L4-S1 segments) were removed and were used for Western blotting and anti-Cx32 and anti-Cx43 immunofluorescent staining. CBX was intrathecally injected at 20 days after SNI, and measured PWMT induced with an innocuous mechanical von Frey filaments stimulation. Results The PWMT of left behind leg in normal and sham groups did not reduce, while the left behind leg in SNI animals developed a conspicuous hypersensitivity to innocuous mechanical von Frey filaments stimulation. The PWMT of SNI rat decreased significantly as compared with normal and sham groups, and peaked at 20 days after SNI, it could maintain for as long as the animals were monitored. AntiCx32 and antiCx43 immunofluorescent reaction in the dorsal horn of spinal cord of SNI side increased significantly as compared with that in normal and sham group or the nonSNI side. After intrathecal injection of CBX 3hours, the PWMT of SNI side increased from the(2.5±1.0)g to (20.0±3.2)g, CBX inhibited the mechanical allodynia. In NS intrathecal injection group did not affect PWMT on SNI rats. Conclusion The gap junction Related Articles | Metrics

Select

Triptolide inhibits the Aβ deposition and senile plaques formation in the hippocampus of APP/PS1 double transgenic mice 2009, 40 (3):  369-373.  doi: 10.3969/j.issn.0529-1356.2009.03.005 Abstract ( )   Objective To examine the effects of Triptolide (T10) on the Aβ deposition and senile plaques formation in the hippocampus by using the APP/PS1 double transgenic mice model of AD (APP/PS1dtg). Methods Eighteen Male APP/PS1dtg mice aged at 45 months were used in this study. The mice were divided into 3 groups randomly, High-5μg/(kgI>&#/I>8226;d) T10 (T10 H,EM>n/EM>=6), Low-1μg/g(kgI>&#/I>8226;d) T10 (T10 L,EM>n/EM>=6) and placebo (PLC,EM>n/EM>=6). Intraperitoneal administration of T10 and capacity solvents (PLC group) was given in a period of a total of 45days. After 45days of treatment, the mice were sacrificed and the brains removed for processing. The brains were separated along the middle sagittal sulcus. The left side was used for the histological study staining 6E10 immunohistochemical and Congo red methods, and quantitative analysis by unbiased stereological counting. The right hippocampus was dissected, Western blotting was performed to assess the change of protein level for Aβ. Results Compared with the PLC group, the total area of 6E10 positive Aβ plaques in the hippocampus of T10 H was reduced by 35% (P<0.001), and that of T10 L reduced by 18% (P<0.05); and the total area of SP in the hippocampus of T10 H group was reduced by 32% (P<0.001), and in the T10 L reduced by 27% (EM>P/EM><0.05). Western blotting protein analysis also displayed that T10 H group Aβ protein levels in the hippocampus was lowest, Aβ protein levels in PLC group Related Articles | Metrics

Select

Effect of EPO-activated astrocyte conditioned medium on differention of neural stem cells and its protective effect on differentiated neural stem cells after injury EM>in vitro/EM> 2009, 40 (3):  374-378.  doi: 10.3969/j.issn.0529-1356.2009.03.006 Abstract ( )   Objective To investigate whether erythropoietin (EPO) activated astrocytes could release some bioactive factors which affect differentiation of neural stem cells and protect these differentiated cells after hydroxyl free radical induced injury. Methods Primarily cultured astrocytes were prepared and purified, and the astrocytes conditioned medium was collected and used to culture neural stem cells. After 5 days in vitro the morphological changes of the neural stem cells were observed under a phase contrast microscope, NF-200 immunocytochemical staining was conducted to evaluate the differentiation of neural stem cells. In the mean time, FeSO4 and H2O2 were added to neural stem cells for 20 min, and these cells were cultured for 48 hours with or without EACM, then the viability of cells was determined with MTT assays, and survived cells were counted as well. Results The expression of NF-200 in the neural stem cells showed strong positive whereas control group exhibited less morphological changes and NF-200 immunocytochemical staining was almost negative in corresponding period. As for EACM protective effects on differentiated neural stem cells after hydroxyl freeinduced injuries generated by adding FeSO4 and H2O2, the absorbance value of the experimental group was 0.381±0.027 while the data was only 0.27±0.013 in control group (EM>P/EM><0.01). Conclusion These results suggest that EPO activated astrocytes can secrete some factors to either promote differentiation of neural stem cells into neuron or protect differentiated neu Related Articles | Metrics

Select

Comparison of the effects of acute administration and chronic exposure of βamyloidpeptide 25-35 on the Ca SUP>2+/SUP>-independent KSUP>+/SUP> currents in rat  BR>hippocampal and cerebral cortical neurons BR> 2009, 40 (3):  379-384.  doi: 10.3969/j.issn.0529-1356.2009.03.007 Abstract ( )   P> Objective To study the different effects between the acute administration and chronic exposure of βamyloidpeptide 2535(Aβ25-35) on the Ca2+independent K+ currents in rat hippocampal and cerebral cortical neurons. Methods After different applications (acute administration and chronic exposure) of Aβ25-35 in rat hippocampal or cerebral cortical neurons, the Ca2+independent K+ currents were recorded by wholecell patch clamp technique and the cell viability was detected by Calcein-AM staining. BR> Results The amplitudes of the Ca SUP>2+/SUP>independent K+ currents were significantly decreased by acute administration to AβSUB>25-35/SUB> in acute isolation of hippocampal neurons (EM>n/EM>=11) while the amplitudes were marke Related Articles | Metrics

Select

Expression of Caspase-3 and NF-κB in the APPSWE transgenic mouse 2009, 40 (3):  385-389.  doi: 10.3969/j.issn.0529-1356.2009.03.008 Abstract ( )   Objective To investigate the roles between the neuroapoptosis and the onset, development of Alzheimer’s disease, and the NF-κB’s regulation in neuroapoptosis. Methods APPSWE transgenic mice from postnatal day 0 to postnatal day 180 were used for Nissl staining, Caspase-3 and NF-κB immunohistochemistry and RT-PCR analysis. Results The neuroapoptosis and NF-κB immunostaining in CA3 areas decreased gradually with age increasing. In the meantime, the Caspase-3 positive pyramidal cells and NF-κB positive pyramidal cells were measured. The densities of both Caspase-3 and NF-κB positive cells in APPSWE transgenic mice were higher than that of the agematched controls, and there were statistical differences after P14 (P<0.01 or P<0.05). In addition, the result of RT-PCR was coincident with that of immunohistochemistry. Conclusion The onset and development of Alzheimer’s Related Articles | Metrics

Select

Effects of taurine on the survival and differentiation of neuroepithelial cells treated with hyperthermia 2009, 40 (3):  390-394.  doi: 10.3969/j.issn.0529-1356.2009.03.009 Abstract ( )   Objective To study the effects of taurine on the survival and differentiation of neuroepithelial cells treated with hyperthermia. Methods Neuroepithelial cells were dissociated from embryonic neural tube of E12.5d mice. They were divided into the control group, the hyperthermia group and the taurine groupⅠ,Ⅱ. The morphological changes were observed under the light microscope. MTT was used to examine the proliferative activitiy of various groups. Apopotosis percentage was determined by DAPI staining. The immunofluorescence staining was used to identify the expression of the Caspase-3, MAP-2 and GFAP in cultured cells. The immunostaining positive cells were counted at high fields in different groups. Results In the hyperthermia group, compared to the control group,the proliferative activitiy of neuroepithelial cells decreased significantly, the apoptosis percentage and the immunostaining positive cells of Caspase-3 increased, the immunostaining positive cells of MAP-2 decreased. However, these changes were significantly reversed by the taurine preconditioning. Conclusion Taurine may protect neuroepithelial cells from hyperthermia, and may promote these cells to differentiate i Related Articles | Metrics

Select

Effect of ethanol on the serotonergic systems in rat brain 2009, 40 (3):  395-400.  doi: 10.3969/j.issn.0529-1356.2009.03.010 Abstract ( )   Objective To study the effect of ethonal on the serotonergic systems by analyzing the altered expression of tryptophan hydroxylase (TPH), serotonin (5-HT) and serotonin transporter (SERT) in the brain of ethanoltreated rats. Methods Wistar rats were treated with 20% ethanol for 6 months. Immunocytochemistry, flow cytometry and Western blotting were used to analyze the altered expression of the serotonergic markers in different brain regions of the ethanoltreated rats. Results 1. Immunohistochemistry showed the number of TPH or 5-HT positive neurons in dorsal raphe nucleus (DRN) is less in ethanoltreated rats than that in control (EM>P/EM>0.01), the diameter of TPH neurons in DRN is smaller than that in control (EM>P/EM>0.01) and the mean gray value of TPH, 5-HT and SERT in observed regions of the ethanoltreated rats is larger than that in control (EM>P/EM>0.05). 2. Flow cytometry showed the expression of TPH, 5-HT and SERT in the studied regions of the ethanoltreated rats decreased significantly compared with control (EM>P/EM>0.05). 3. Western blotting showed the ratio of SERT/β-actinin or TPH/β-actinin in analyzed regions of the ethanol-treated Related Articles | Metrics

Select

Effects of ferric ammonium citrate on the expression of proteins related to iron release in primary cultured rat cardiac myocytes 2009, 40 (3):  401-404.  doi: 10.3969/j.issn.0529-1356.2009.03.011 Abstract ( )   Objective To observe the effect of ferric ammonium citrate (FAC) on the pulsatile frequency and the expression of proteins related to iron releas in primary cultured rat cardiac myocytes. Methods Newborn rat cardiac myocytes were isolated and cultured with the medium containing FAC. The cells were divided into control group, 20mg/L FAC group, 40mg/L FAC group and 80mg/L FAC group. Every group had 6 repeats. The cells’ livability, beat rhythmically and beat amplitude of cardiac myocytes were detected. The expressions of ceruloplasmin (CP), hephaestin(HP) and ferroportion 1(FP1) were also examined by immunohistochemistry. Results Compared with control, FAC had no clearly influence on cells’ livability, but it could decrease cells’ beat rhythmically and beat amplitude. CP, HP and FP1 expression increased after the incubation of FAC. Conclusion FAC has no influence on cardiac myocytes’ livability, but it could affect the cells’ living state. IRP-IRE may be involved in the regulation of FP1 expression, the expression Related Articles | Metrics

Select

Effect of corticosterone on antizyme mrna and protein expression in early regenerating rat liver 2009, 40 (3):  405-408.  doi: 10.3969/j.issn.0529-1356.2009.03.012 Abstract ( )   P>Objective The effect of corticosterone on both antizyme mRNA and protein levels in early regenerating rat liver induced by partial hepatectomy (PH) was investigated. Methods Bilateral adrenalectomies (ADX) were performed 3 days before PH. Corticosterone in sesame oil was injected subcutaneously to ADX rats. Antizyme mRNA and its protein level were examined by RT-PCR and Western blotting respectively. Results When compared with control (n=6, the same below) group, there were no significant changes in antizyme mRNA levels in ADX, 10 and 20 mg/kg body weight corticosterone-treated groups, whereas 40 mg/kg body weight corticosterone treatment markedly induced antizyme mRNA expression. Antizyme protein content in ADX group decreased significantly at 5, 7 and 9 hours after PH, however, it could be stimulated in a dosedependent manner following corticosterone administration during the whole experiment observed, particularly, at 5 hours post-PH antizyme protein levels in 10, 20 and 40 mg/kg body weight corticosterone-treated rats increased by 43%, 79% and 93% respectively, as compared with control group at the same time point. Conclusion Corticosterone may dose-dependently induce antizyme protein synthesis in early regenerating rat liver; 10 or 20 mg/kg body weight corticosterone treatment has little effect on antizyme mRNA level, but 40 mg/kg body weight corticosterone stimulates antizyme gene transcription./P> Related Articles | Metrics

Select

Production of transgenic cloned mouse embryos by somatic cell nuclear transfer with different donor cells 2009, 40 (3):  409-413.  doi: 10.3969/j.issn.0529-1356.2009.03.013 Abstract ( )   Objective To generate transgenic mouse embryos by somatic cell nuclear transfer (SCNT), and compare effects of different donor cells on developmental potential of cloned embryos. Methods Mouse embryo fibroblasts and mouse embryonic stem (ES) cells were transfected with a plasmid (pCE-EGFP-IRES-NEO) containing the enhanced green fluorescent protein (GFP) and neomycinresistant (Neor) genes by electroporation. Transgenic cell lines were obtained for SCNT after 14 days selection with G418. At the same time, SCNT was performed using cumulus cells and normal fibroblasts as control. Results There was no significant difference in blastocyst rates between reconstructed embryos derived from transgenic fibroblasts and nontransgenic ones (154/160,19.23% vs 22.91%, EM>P/EM>＞0.05). Reconstructed embryos from transgenic ES cells showed higher blastocyst developmental rates than that from transgenic fibroblasts donor (152/154,41.54% vs 19.23%, P<0.05). Cumulus cloned embryos had better developmental potential than that of fibroblasts cloned ones (171/160, 41.17% vs 22.91%, P<0.05). Conclusion Our results showed that transgenic donor cells did not affect EM>in vitro/EM> developmental potential of mouse SCNT embryos. Using EGFP maker, mouse transgenic blastocysts could be produced effectively Related Articles | Metrics

Select

Recombinant adeno-associated virus containing antisense nucleotide against cyclin b1 inhibits osteosarcoma cell U2OS growth and induces apoptosis 2009, 40 (3):  414-418.  doi: 10.3969/j.issn.0529-1356.2009.03.014 Abstract ( )   To investigate the effect of recombinant adenoassociated virus (rAAV) containing antisense nucleotide against cyclin B1 (ASCCNB1) on osteosarcoma cells in vitro. Methods The rAAV was produced by Cotransfecting HEK293 cell line with the rAAV plasmid pd16-95-AS-CCNB1/neo and AAV/Adenoviral helper plasmid pSH3. The rAAV particles expressing ASCCNB1 were purified and concentrated. Subsequently the virus titer was assayed by dotbloting analysis. Then, human osteosarcoma U2OS cells were infected by rAAV at the pfu of 10SUP>5/SUP>-10SUP>6/SUP>/cell titer. CCNB1 expression was detected by Western blotting and RT-PCR methods, human osteosarcoma U2OS cells growth curve was assayed by MTT, cell cycle and apoptosis were detected by FACS. Results The rAAV-AS-CCNB1 plasmid was successfully constructed and the virus titer is up to 1.0×10SUP>11/SUP> v.g./ml. After infected with rAAV EM>in vitro/EM>, U2OS cells growth was inhibited significantly. Both Western blotting and RT-PCR assay showed that CCNB1 expression of U2OS cells was downregulated by rAAV-AS-CCNB1,and cell apoptosis and GSUB>1/SUB> arrest were observed. Conclusion The recombinant adenoassociated virus Related Articles | Metrics

Select

Effect of different concentrations of endothelin on intracellular calcium level in cultured human pericardial mesothelial cells 2009, 40 (3):  419-422.  doi: 10.3969/j.issn.0529-1356.2009.03.015 Abstract ( )   Objective To investigate the effect of different concentrations of endothelin on intracellular calcium level in cultured human pericardial mesothelial cells. Methods Pericardial mesothelial cells from patients undergoing open heart surgery were cultured in vitro. Cells of passage 8-10 were used in this experiment. When confluence was reached, the mesothelial cells were loaded with Fluo3/AM. The intracellular calcium concentrations in theses cultured cells were determined by confocal laser scanning microscopy after 10SUP>-7/SUP>mol/L, 10SUP>-8/SUP>mol/L, 10SUP>-9/SUP>mol/L ET being added respectively. Results Calcium concentrations in cultured mesothelial cells increased remarkably after adding three kinds of ET concentration(EM>P/EM><0.001). There was interaction between endothelin concentration and observation time (P<0.001). Conclusion Calcium in cultured mesothelial cells changed distinctly after the stimulation of different concentration of ET; Human pericardial mesothelial cells may possess important biological activity and endothelin may play an important role in the pericardium in an autocrine and/or paracrin Related Articles | Metrics

Select

Study on the anticancer effect of three new selenium compounds in K562 cells 2009, 40 (3):  423-427.  doi: 10.3969/j.issn.0529-1356.2009.03.016 Abstract ( )   Objective To investigate the antitumor activities of three selenium compounds in EM>vitro/EM> as well as EM>in vivo/EM>, and to explore the mechanism. Methods Assay of MTT and growth inhibition of S180 and H22 on tumorbearing mice were used to study the antitumor effect, morphological observation, flow cytometry and confocal laser scanning microscopy assay were used to study the mechanism of three selenium compounds. Results They could significantly inhibit proliferation of K562 cells in vitro（EM>P/EM><0.05）, and the growth of S180 and H22 (EM>P/EM><0.01,EM>n/EM>=10). Electron microscopic observation revealed typical apoptotic features, including shrinkage of cellular and nuclear membranes, condensed heterochromatin around the nuclear periphery, and cytoplasmic vacuolation in the K562 cells treated with NaSeVO, SeMoV and SeWV 5 mg/L for 24 hours. The cell cycle was redistributed by selenium compounds and a significantly subG showed at high dosage. The fluorescence intensity of intracellular CaSUP>2+/SUP>,MgEM>SUP>2+/SUP>/EM> and ROS was greatly increased after treatment with selenium compounds as compared with control group（EM>P/EM><0.01）. However, the fluorescence intensity of intracellular pH value and MMP decreased（EM>P/EM><0.01）. Conclusion Three selenium compounds have antitumor activity EM>in vivo/E Related Articles | Metrics

Select

Effect of heat shock protein 70 on the expression of Bax and Bcl-2 and the conformational change of Bax in glucose deprived HeLa cells 2009, 40 (3):  428-432.  doi: 10.3969/j.issn.0529-1356.2009.03.017 Abstract ( )   Objective To observe the effect of heat shock protein 70 (Hsp70) on apoptosis of HeLa cells induced by glucose deprivation and investigate the relationship between Hsp70 and the key proteins of apoptosis: Bcl-2 family members. Methods HeLa cells were stably transfected with the plasmid of pcDNA31(+)Hsp70 to establish the Hsp70 overexpressed cell model; Hsp70 normalexpression and overexpression cells cultured with glucose free medium to create stress models. Every detection had three parallel samples. MTT assay was applied to evaluate the cell viability; Hoechst 33258 and Giemsa stain were used to examine the rate of cell apoptosis. RT-PCR was used to determine the expression level of Bax and Bcl-2 and to calculate the ratio of Bax/Bcl-2; Immunofluorescence and immunocytochemistry were applied to examine the conformational change of Bax. Results Cell viability was increased and the rate of cell apoptosis was decreased in Hsp70 overexpression cells cultured with glucose free medium. At the same time, the expression of Bax and Bcl-2 and the conformational change of Bax were also inhibited by Hsp70. Conclusion Hsp7 Related Articles | Metrics

Select

Ezrin, MMP-2 and TIMP-2 might exert a synergistic action in invasion and metastasis of human cervical carcinoma 2009, 40 (3):  433-436.  doi: 10.3969/j.issn.0529-1356.2009.03.018 Abstract ( )   Objective To identify their expressions of Ezrin, atrix metalloproteinases2(MMP-2) and tissue inhibitor of metalloproteinase 2 (TIMP-2)in human cervical carcinoma, to clarify their relationships between these expressions and invasion, metastasis of human cervical carcinoma. Methods Ezrin, MMP-2 and TIMP-2 were detected immunohistochemically in 40 cases of squamous cell carcinoma of the uterine cervix, in 20 cases of adenocarcinoma of the uterine cervix, in 29 cases of cervical intraepithelial neoplasia (CIN) and 16 cases of normal cervices. Results Immunohistochemical staining of tumor cells for Ezrin, MMP-2 and TIMP-2 were noted in intracytoplasm. There were significant differences for Ezrin, MMP-2 and TIMP-2 in squamous cell carcinoma and adenocarcinoma of the uterine cervix, CIN and normal cervices respectively (P<0.01). The positive rates of Ezrin and MMP-2 in lymph node metastasis were obviously higher than that in without lymph node metastasis(P<0.01)，but for TIMP-2 it was contrary. The expressions of Ezrin between MMP-2 and TIMP-2 protein were positively correlated in cervical lesions respectively（EM>rSUB>S/SUB>/EM>=0422, EM>P/EM>=0.000; EM>rSUB>S/SUB>/EM>=0294, EM>P/EM>=0.002）. Conclusion The expressions for Ezrin, MMP-2 and TIMP-2 in squamous cell carcinoma and adenocarcinoma of the uterine cervix are higher than that in control group. The overexpressions of three markers possibly play a key role in invasion and lymphnode metastasis of carcinoma of the cervix Related Articles | Metrics

Select

Impairment of osteogenesis potential of bone marrow mesenchymal stem cells and bone injury caused by irradiation 2009, 40 (3):  437-440.  doi: 10.3969/j.issn.0529-1356.2009.03.019 Abstract ( )   Objective To investigate the effects of irradiation on the osteogenesis potential of bone marrow mesenchymal stem cells (BMMSCs) and EM>in vivo/EM> bone system. Methods At 28 days after 4Gy total body irradiation (TBI), BMMSCs were isolated from irradiated or normal male C57BL/6 mice and cultured. Their osteogenesis potential was evaluated by alkaline phosphatase (ALP) and Von Kossa staining after 1 and 3 weeks osteogenic differentiation EM>in vitro/EM> respectively. Bone histomorphometric analysis including hematoxylin and eosin (HE) and toluidine blue staining, bone mineral density (BMD) measurment were performed to detect the changes in bone systemEM>in vivo/EM>. Results Significant impairment of osteogenic differentiation of BMMSCs at 28 days post irradiation was observed in both ALP and Von Kossa staining with less ALPpositive cells after 1 week of culture in osteogenic culture and a calciumless mineralized matrix in the induction dish after 3 weeks of induction. Meanwhile, the trabecular structure was gradually fragmented after irradiation and at 15 weeks after TBI, the irradiated mice had decreased total body bone mineral density. Conclusion Irradiation resulted in decrease in osteogenesis potential of BMMSCs accompanied by bone injury, and the damage in BMMSCs may be, at the stem cell level, invo Related Articles | Metrics

Select

Expression of α-smooth muscle actin in the early development of the outflow tract of the human embryonic heart 2009, 40 (3):  441-445.  doi: 10.3969/j.issn.0529-1356.2009.03.020 Abstract ( )   Objective To explore the expression pattern and significance of the α-smooth muscle actin (α-SMA) in the myocardium and endocardial cushion of the outflow tract in the early human embryonic heart. Methods Serial sections of thirtytwo human embryonic hearts from Carnegie stage 10 to Carnegie stage 16 were stained immunohistochemically with antibodies against α-SMA, α-sarcomeric actin (α-SCA) and myosin heavy chain ( MHC ) to observe the α-SMA expression pattern in the myocardium and endocardial cushion during the remodeling of the outflow tract. Results During C10 to C15, the new cardiomyocytes differentiating from the pericardial splanchnic epithelium were added to the distal pole of the outflow tract. The expression of α-SMA of these cells was earlier than the expression of α-SCA and MHC. At C16, α-SMA positive cells were observed in the ridges neighboring with the myocardial cells of the outflow tract wall. The latter linked with these α-SMA positive cells by their protrusion. From C12 to C15, α-SMA positive cells gradually migrated into the endocardial cushion. At the same time, the endothelial cells of the outflow tract began to express α-SMA and differentiated toward mesenchymal cells. Mesenchymal cells of different origins aggregated to form two opposite spiral ridges. Conclusion α-SMA could be regarded as an early marker Related Articles | Metrics

Select

Effects of recombinant human endostatin on the expression of p21, cyclin D1 and CDK4 in synovial tissue in rats with adjuvant arthritis 2009, 40 (3):  446-450.  doi: 10.3969/j.issn.0529-1356.2009.03.021 Abstract ( )   Objective To observe the effect of recombinant human endostatin(rhEndostatin) on the expression of p21,cyclin D1 and CDK4 genes in synovial tissue in rats with adjuvant arthritis(AA) and explore molecular mechanisms of the inhibitory effect of rhEndostatin on proliferation of fibroblast-like synoviocytes(FLS) in AA rats. Methods Thirty-six male SD rats were used in the study. They were divided at random into normal, AA model and rhEndostatin (2.5mg/kg)treated groups. Twelve rats were taken from each group. Adjuvant arthritis was induced in rats in AA model and rhEndostatin(2.5mg/kg)treated group. The effect of rhEndostatin on the expression of p21,cyclin D1,CDK4 mRNA and cyclin D1 protein in synovial tissue in AA rats was examined quantitatively by realtime fluorescent quantitative PCR and Western blotting assays,respectively. Results The expression levels of p21, cyclinD1 mRNA and cyclinD1 protein were significantly decreased by rhEndostatin, compared with the levels in AA synovial tissues (P<0.01). However, CDK4 mRNA expression levels in synovial tissue treated with rhEndostatin were significantly higher than those in AA control samples (P<0.01). Conclusion Recombinant human endostatin decreases cyclinD1 expression levels in AA synovial tissues, which may be one of the molecular mechanisms of the inhibitory effect of rhEndostatin on AA FLS pro Related Articles | Metrics

Select

Expressions of TGF-β,AQP-2 and Bax during kidney development of EM>Bufo raddei/EM> 2009, 40 (3):  451-457.  doi: 10.3969/j.issn.0529-1356.2009.03.022 Abstract ( )   Objective To observe the expressions of transforming growth factorβ(TGF-β)，aquaporin2(AQP-2) and Bax during development of renal tubules in Bufo raddei， and explore the role and regulating significance of them. Methods The expressions of TGF-β(β2，β3)， AQP-2 and Bax were observed by immunohistochemistry combined with stereological methods in Bufo raddei kidney at 1838th， young and adult incubation stages. There were 5 samples at each stage.The expression intensity of them was analyzed by IPP software. Results At 2632th stages the epithelial cells of proximal tubules and distal tubules of pronephron showed TGF-βSUB>2/SUB> immunoreactivity， at 34=38th stages the epithelial cells of proximal tubules of mesonephron showed TGF-βSUB>2/SUB> immunoreactivity and there was the same in a young and an adult Bufo raddei; At 26-30th stages the epithelial cells of proximal tubules and distal tubules of pronephron showed TGF-βSUB>3/SUB> immunoreactivity， at 32-38th stages the epithelial cells of proximal tubules of mesonephron showed TGF-βSUB>3/SUB> immunoreactivity and there was the same in a young and an adult EM>Bufo raddei/EM>; The epithelial cells of collecting tubules always appeared AQP-2 immunoreactivity at 26-38th stages and there was the same in a young and an adult EM>Bufo raddei/EM>; at 2634th stages the epithelial cells of proximal tubules and distal tubules appeared Bax immunoreac Related Articles | Metrics

Select

Immunohistochemical observation and quantitative analysis of IL-6, IFN-β and IP-10 in human spleens with severe acute respiratory syndrome 2009, 40 (3):  458-462.  doi: 10.3969/j.issn.0529-1356.2009.03.023 Abstract ( )   Objective To investigate the pathological changes and pathogeneses of spleen in severe acute respiratory syndrome(SARS) with immunohistochemical observation and quantitative analysis of the expressions of interleukin6 (IL-6)，interferon β (IFN-β) and IFN-γ inducible protein10 (IP-10). Methods Three specific antibodies were used to detect the expression of IL-6，IFN-β and IP-10 in the spleens of six cases who died of SARS and six normal cases as the controls with immunohistochemistry. The results were analyzed with image analysis system. Results In the splenic red pulp of SARS patients, lots of cells expressed IL-6 averagely, as bolus in cytoplasm and nucleus, and compared with normal cases, average absorbance (AEM>A/EM>) in IL-6 positive cells had significant difference (EM>P/EM><0.05). The IFN-β positive cells dispersed in splenic red pulp of SARS patients, yellow bolus was in cytoplasm and nucleus, compared with normal cases, AEM>A/EM> in IFN-β positive cells had significant difference (EM>P/EM><0.05). There were lots of IP-10 positive cells in splenic red pulp of SARS patients, the positive productions as bolus were in cytoplasm and nucleus, and compared with normal cases, AEM>A/EM> in IP-10 positive cells had significant difference (EM>P/EM><0.05). In the relic splenic corpuscle and periarterial lymphatic sheath, immunostainings of these three cytokines were all negative. Conclusion Immunohistochemical intensities of IL-6, IFN-β and IP-10 in spleen of SARS patient Related Articles | Metrics

Select

Expression and localization of 5-HT, somatostatin and substance P of rectum in heroin dependent rats 2009, 40 (3):  463-468.  doi: 10.3969/j.issn.0529-1356.2009.03.024 Abstract ( )   Objective To explore the morphological changes of 5-HT, Somatostatin（SS）, Substance P（SP） immunoreactive(IR) cells in rat rectum during heroin dependence period. Methods Normal SD rats were divided into normal control group (NCG), saline control group(SCG),and heroin dependence group(HDG).To make the heroin dependence model of rats by subcutaneous injection of herion, and excise the rectum. Immunohistochemical SABC method, counting the number of cells and image analysis were used in the study. Results Compare with the NCG and SCG, All the number and immunohistochemical reaction of 5-HT, SS, SPIR cells increased(EM>P/EM><0.05) in rectum during heroin dependence. The result of image analysis showed that the mean of grey degree of the rectum 5-HT, SS, SPIR cells decreased in HCG as compared with the NCG and SCG(EM>P/EM><0.05).And in 5-HT, SS-IR cells, the mean grey scale were the lowest on 31st day(EM>P/EM><0.05). But in SP-IR cells, the mean grey scale was the lowest on 38th day(P<0.01). Conclusion The changes manifested that the function of 5-HT, SS and SP-IR ce Related Articles | Metrics

Select

Expression of Bcl-2/Bax in postnatal rat ovaries at different developmental days 2009, 40 (3):  469-473.  doi: 10.3969/j.issn.0529-1356.2009.03.025 Abstract ( )   P>Objective To investigate the relationship between the expression of Bcl-2/Bax protein and follicular development and apoptosis. Methods Immunohistochemical technology was utilized to study the distribution and positive reaction intensity of apoptosis protein Bcl-2/Bax in 95 rat ovaries from different postnatal developmental days. Value of absorbance of Bcl-2/Bax positive reaction in ovary sections was measured by the computer image analysis system. Results Positive staining for Bcl-2/Bax existed in the oocytes of different class follicles from 0 dayold to 360 dayold rat ovaries. Bcl-2 positive reaction in health oocytes was stronger than Bax,whereas Bax positive reaction in degenerative oocytes was stronger than Bcl-2 The expression pattern of Bcl-2/Bax was different in follicular cells, in which Bcl-2 was positive reaction from the postnatal first 21days rat, negative reaction from 21 dayold to 150 dayold rat,and negative in health follicles and positive in atretic follicles from older rat than 180 dayold. Bax reaction in follicular cells was all negative from younger rat than 180 dayold and liked Bcl-2 from older rat than 180 dayold. Expression of Bcl-2/Bax in theca cells was similar to follicular cells. Expression of Bcl-2/Bax in corpora luteum was observed only from older rat than 180 dayold. Conclusion Bcl-2/Bax may play an Related Articles | Metrics

Select

Long-term ovarian function in rats after heterotopic autologous transplantation of cryopreserved ovarian tissues 2009, 40 (3):  474-479.  doi: 10.3969/j.issn.0529-1356.2009.03.026 Abstract ( )   Objective To assess the longterm efficacy of cryopreserved ovarian tissues after heterotopic autologous transplanted in rats. Methods Seventyeight adult Wistar rats were divided into four groups randomly as follows: sham-operated group (group 1,EM>n/EM>=15), freshtransplanted group (group 2,EM>n/EM>=24), frozen-thawed-transplanted group (group 3,EM>n/EM>=24) and ovariectomized group (group 4,EM>n/EM>=15). Fresh and frozen-thawed ovarian tissues of adult rats were autologous transplanted under renal capsule. One third animals of each group were slaughtered and their ovaries and uteruses were removed for morphometric analysis at 5 months after transplantation; one third additional animals were slaughtered at 8 months; and the remaining animals were slaughtered at 10 months. The measurement of serum estradiol (ESUB>2/SUB>) and progesterone (P) concentrations during the estrus stage and vaginal cytology were performed to assess the secretary function of implanted ovarian tissues. Results Both fresh and frozen ovarian grafts survived in all the animal models. In groups 2 and 3, the serum ESUB>2/SUB> and progesterone concentrations during estrus remained comparable to the shamoperated rats’ hormone levels (group1), and significantly higher than the concentrations in group 4 at 5, 8, 10 months respectively (EM>P/EM><0.01). Morphologically, no significant differences were observed in the proportion of each stage of follicles between frozenthawed tissues and fresh ovarian tissues (EM>P/EM>> 0. 05). And the proportion of each stage follicles in both types of grafts was comparable to that in group 1 (P>0.05). The number of primordial follicles in fresh grafts and frozen grafts were significantly less than that in group 1 Grafts in group 2 contained 59.1% of the primordial follicles in sham-operated controls while frozen grafts contained 54.5% at 5 months after implantation. Conclusion Cryopreservation of small pieces of ovarian tissues is feasible for keeping cell viability. After autologous implanted under the renal capsule, the cryopreserved ovarian tissues have no significant morphological changes. The follicles survive and develop well, though th Related Articles | Metrics

Select

Effect of heparin on cardiac hypertrophy mediated by the activity of RhoA/Rho kinase signaling pathway 2009, 40 (3):  480-484.  doi: 10.3969/j.issn.0529-1356.2009.03.027 Abstract ( )   Objective Cardiac hypertrophy may be regulated primarily by hypertrophystimulating factors.However，little is known about the cardiac growthinhibitory factors that negatively regulate the formation of cardiac Hypertrophy. Recently，a RhoA/Rho kinase signaling pathway was found to be involved in cardiac hypertrophy.The inositol 1，4，5 trisphosphate receptor(IP-3R)is an intracellular Ca SUP>2+/SUP> release channel and specially inhibited by heparin.Heparin was shown to prevent the growth and proliferation of cardiovascular cells besides its wellcharacterized anticoagulant action. We postulated that activation of IP-3R mediated RhoA/Rho kinase pathway had a potential rote in regulation of cardiomyocyte hypertrophy and heparin may antagonize this signaling pathway. Methods Primary neonatal rat cardiomyocytes were cultured in fetal calf serum. Western blotting and RT-PCR were used to evaluate the protein and gene expression. Results IP-3(10SUP>-7/SUP>mol/L)caused a time-dependent increase in c-fos，c-myc， α-actin and βmajor histocompatibility complex (β-MHC) expression of cardiomyocytes(EM>P/EM>0.05).Stimulation of cardiomyocytes with lP3 revealed a timedependent increase in the expressions of RhoA and Rho kinase gene (EM>P/EM>0.05), and heparin could inhibit the effects above mentioned (EM>P/EM>0.05). Concl Related Articles | Metrics

Select

Effect of simvastatin on the fusion process of cervical intervertebral cortical bone allograft transplantation 2009, 40 (3):  485-490.  doi: 10.3969/j.issn.0529-1356.2009.03.028 Abstract ( )   Objective To investigate the effect of simvastatin on fusion process after cortical bone allograft transplantation for anterior cervical intervertebral fusion. Methods Forty--four male Wistar rats，undergone anterior cervical discectomy and fusion with allogenous cortical bone transplantation, were divided into 4 groups: control group, simvastatin group, recombinant human bone morphogenetic protein2(rhBMP-2) group and bland group. The animals of the first 3 groups were sacrificed to obtain cervical spine specimen and performed pathological examination at 2, 4, 8, and 12week intervals. The bone mineral density of cervical vertebral body was also measured. Results The bone mineral density of control group decreased markedly compared with that of blank group. No difference was found among that of the simvastatin, BMP2 and blank groups. Inflammatory fibrous capsule existed for a prolonged period and little endochondral calcification in the soft tissue was observed in control group. In both simvastatin and rhBMP-2 groups, bony fusion between the trabecula and the surface of allograft bone was achieved earlier. But in the rhBMP-2 group, fusion was observed only at the surface of the implant. Invasion of soft tissue and new blood vessels into the core region of the implants were apparent in the simvastatin group. And scattered neonate osteons were found within allograft. Conclusion Simvastatin demonstrated the potency of promotin Related Articles | Metrics

Select

Influence of combination regimen of low-dose gossypol acetic acid with steroid hormones on apoptosis of germs and iNOS expression 2009, 40 (3):  491-495.  doi: 10.3969/j.issn.0529-1356.2009.03.029 Abstract ( )   Objective To investigate the influence on apoptosis of germs and expression of inducible nitric oxide synthase (iNOS) expression in male rats testis by administered with combination regimen of low-dose gossypol acetic acid(GA)with steroid hormones(desogestre/ethinylestradiol/testosterone undecanote(DSG/E/TU). Methods Adult male rats were randomly divided into four groups. Group GH：rats were fed orally with GA［125mg(kg.day)］and DSG［125μg/(kg.day)］/E［25μg/(kg.day)］/TU［100mg/(kg.day)］; Group G: rats were administered with a single dose of GA［125mg/(kg.day)］; Group H: rats were administered with DSG［125μg//(kg.day)］, E［25μg/(kg.day)］and TU［100mg/(kg.day)］; Group C: rats only received vehicle(1％ methyl cellulose). The testes were removed at 4,6,8 weeks respectively. TUNEL staining was observed. The expressions of iNOS were detected by immunohistochemical staining and Western blotting. Results The TUNEL positive stainings were found in the residual bodies in the methylcelluosetreated group(C) and in the pachytene spermatocytes nuclei in gossypol combined with steroid hormones group(GH). The amount of apoptosis cells were much larger in the GH group than that in the control group with treatment time increasing(EM>P/EM>0.01).iNOS was expressed constitutively in Leydig cells and in a stagede Related Articles | Metrics

Select

Expression and significance of nNOS, VEGF and Bcl-2 proteins in the developing anterior horn of human embryonic spinal cord 2009, 40 (3):  496-499.  doi: 10.3969/j.issn.0529-1356.2009.03.030 Abstract ( )   Objective To explore the rule of distribution and expression significance of neuronal nitric oxide synthase（nNOS）, vascular endothelial growth factor （VEGF） and Bcell lymphoma/leukemia2（Bcl-2）proteins in the developing spinal cord anterior horn of human embryos. Methods In the fourth month of gestation, the expression productions of nNOS, VEGF and Bcl-2 proteins were investigated in the anterior horn of human embryonic spinal cord using the immunohistochemical methods. Results The intensities of the immunohistochemical signals of both nNOS and Bcl-2 proteins were progressively enhanced from the second to the fourth month of gestation. There was negative immunohistochemical signals of VEGF in the anterior horn of human embryonic spinal cord in the second month of gestation. At the third to fourth month of gestation, the expression of VEGF protein was from weak to strong in the anterior horn of human embryonic spinal cord. Conclusion nNOS, VEGF and Bcl-2 proteins a Related Articles | Metrics

Select

Expression and significance of histone deacetylase 1 in endometriosis 2009, 40 (3):  500-502.  doi: 10.3969/j.issn.0529-1356.2009.03.031 Abstract ( )   Objective To investigate the expression of histone deacetylase1 (HDAC1) in the eutopic and ectopic endometrium of women with endometriosis. Methods The expression of HDAC1 in the eutopic and ectopic endometrium of 20 cases of women with endometriosis, and the expression of HDAC1 in the endometrium of 20 cases with hysteromyoma were mesured by immunohistochemical technique and Western blotting. Results HDAC1 was located in the nuclei of epithelial and stromal cells. The expression level of HDAC1 in eutopic endometrium was significantly higher than that in the control group (EM>P/EM><0.01). In Western blotting, semiquantitative analysis showed that the relative expression of HDAC1 protein in eutopic and ectopic endometrium were significantly higher than that in the control group (2.67±0.69；2.55±1.36 vs 1.63±0.93, EM>P/EM><0.01，EM>P/EM><0.05). But there was no significant difference between eutopic and ectopic endometrium (P>0.05). Conclusion The higher expression of HDAC1 in eutopic and ectopic endometrium with endometriosis indicates that it may play an import Related Articles | Metrics

Select

Gene frequencies of 16 genetic traits in Yi and Bai nationalities in Guizhou province 2009, 40 (3):  503-506.  doi: 10.3969/j.issn.0529-1356.2009.03.032 Abstract ( )   Objective To investigate gene frequencies and it’s distributive characteristic on some genetical traits of Yi and Bai nationalities in Guizhou province. Methods Using the methods of population genetics, we investigated 16 genetic characters of 879 cases (Yi nationality 472, Bai nationality 407) of Yi and Bai nationalities in Guizhou province. Gene frequencies of 16 genetical traits were calculated. Using the EM>U/EM>test, the significant differences were checked between Yi and Bai nationalities, and then compared with other populations. Results Gene frequencies on 16 genetical traits (curly top of tongue,curly sides of tongue, thickness of lips, chin type, hair matter, hair vortex, eyelid, front form of hair, eyelash, cerumen, darwinian point, handedness, thumb style, hair of middle finger, style of fore and ring fingers（males）, style of fore and ring fingers(females) and curl of little finger) in Yi nationality in Guizhou province are 0.0424, 0.3427, 0.5539, 0.3527, 0.1894, 0.3860, 0.4960, 0.1089, 0.5132, 0.1363, 0.3652, 0.6285, 0.2190, 0.3397, 0.0673, 0.0797 and 0.3144 respectively; In Bai nationality, they are 0.0450, 0.3868, 0.4940, 0.2993, 0.2210, 0.3188, 0.4075, 0.0901, 0.6285, 0.0808, 0.4238, 0.7034, 0.2550, 0.3707, 0.0678, 0.0950 and 0.2901 respectively. Conclusion There are very low positions on 5 genetical traits(curly top of tongue, hair of middle finger, hair vortex, thumb style and curl of little finger), and very higher level on 2 genetical traits (chin type and eyelash), and the middle positions on other 9 genetical traits(curly sides of tongue, thickness of lips, hair matter, style of fore and ring fingers（males）, style of fore and ring fingers(females), cerumen, darwinian point, handedness, eyelid, front form of hair) of Yi and Bai n Related Articles | Metrics

Select

Biofilm maturation is correlated to drug resistance of EM>Candida albicans/EM> 2009, 40 (3):  507-509.  doi: 10.3969/j.issn.0529-1356.2009.03.033 Abstract ( )   ［Abstract］ Objective To observe the drug resistance at different stages of EM>Candida(C.) albicans /EM>biofilm formation. Methods To form the biofilm in glass slide, then observe the biofilm morphology by inverted microscope at different development stages(2, 4， 8, 24, 48 hours).Using National Committee for Clinical Laboratory Standards(NCCLs) microdilution to detect drug resistance of dissociative EM>C.albicans/EM> and 2,3 bis(2methoxy 4 nitro 5 sulfophenyl) 5(phenylamino) carbonyl 2H tetrazolium hydroxide(XTT) reduction method to test the change of EM>C. albicans/EM> drug resistance in biofilm. Results Antifungal resistance of biofilmgrown cells increased as the biofilm maturation. Compared with dissociative C.albicans strains, the drug resistance was stronger more than 100 times. Conclusion There is positive cor Related Articles | Metrics

技术方法

Select

Isolation，purification and identification of macrophages from mouse uterus 2009, 40 (3):  510-512.  doi: 10.3969/j.issn.0529-1356.2009.03.034 Abstract ( )   Objective To establish an efficient and reliable method of isolating，purifying and identifying macrophages from mouse uterus. Methods Five female mice were killed and their uteri were obtained. Mechanical dissection and digestive enzymes were used to prepare cell suspension of mouse uterus. Uterine macrophages were separated by density gradient centrifugation. And these cells were purified by anchoring cultivation. Then the cell viability was assessed by trypan blue exclusion staining. The phagocytosis of cell was evaluated by phagocytosis of chicken erythrocytes. The purity of cell was determined by acid phosphatase staining，αacetic acid naphthol esterase staining and surface specific antigen CD14 staining. Results By using this new method，the cell viability was（85.35±0.43）％. And the purity of uterine macrophages was （86.12±1.19）%,（ Related Articles | Metrics

Select

Comparision of three modified Ranson’s methods on displaying rats’ spinal cord tissue 2009, 40 (3):  513-515.  doi: 10.3969/j.issn.0529-1356.2009.03.035 Abstract ( )   Objective The aim of this study was to compare effects of three modified Ranson’s methods on the staining of spinal cord of rats. Methods In the control group, following fixed with alcohol and ammonia, the tissue of spinal cord was embedded with wax, sectioned and stained with traditional Ranson’s method, then, observed under the light microscope. For experimental groups, the tissue of spinal cord was treated with three modified methods namely, replaced the ammonia with sodium hydroxide、sodium carbonate or 95% alcohol respectively, other staining processes were as same as those in the traditional Ranson’s staining method. Results On slices of the experimental group, the soma and neuron filaments were showed with color of yellowbrown, and those nerve fibers were stained with deep brown. Compared with the control group, no significant difference was found in three experimental groups. However, the best staining result was found in the group which the ammonia was replaced with 95% alcohol. Conclusion Three modified Ranson’s staining m Related Articles | Metrics