Table of Content

2012, Volume 43 Issue 5
06 October 2012 Previous Issue    Next Issue
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神经生物学
细胞和分子生物学
肿瘤生物学
解剖学
组织学胚胎学发育生物学
人类学
技术方法


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神经生物学

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Variability of directed differentiation potential into neurons between neural stem cells from embryonic rats and immortalized neural stem cell line C17.2 2012, 43 (5):  581-587.  doi: 10.3969/j.issn.0529-1356.2012.05.001 Abstract ( )   Objective To compare directed differentiation potential into neurons between neural stem cells(NSCs) from early embryonic rats and immortalized neural stem cell line C17.2(C17.2-NSC), and to establish a screening assay on the level of NSCs for high content screening in order to discover drugs that induce directed neurons differentiation of NSCs. Methods Three types of cells, the C17.2-NSC, 17 days embryonic hippocampus NSCs and 11 days embryonic cerebral cortex NSCs were used in this study. Cells from each type were divided into the control and experimental groups, and induced to differentiate by 0 μmol/L and 1 μmol/L retinoic acid (RA) for 5 days under DMEM/F12 medium containing 2%(V/V) B27 and 37℃, 5% CO2 normal culture condition. The variability of expression level of nestin and βⅢ tubulin (Tuj1), which are the specific markers of neural stem and progenitor cells and neurons, respectively, in both control group and experimental group were detected throughimmunohistochemistry. BR>Results Compared with the control group, C17.2-NSC did not differentiate into neurons under the induction of 1 μmol/L RA, but 17-day embryonic hippocampus NSCs and 11-day embryonic cerebral cortex NSCs effectively differentiated into neurons under the same treatment.Conclusion The directed differentiation capacity of early embryonic NSCs into neurons is higher than that of C17.2-NSC, thus early embryonic NSCs are suitable for high content drug screening which aims to discover new types Related Articles | Metrics

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Screening and identification interaction proteins of dynamin-1 PRD functional domain in rat brain synaptosome 2012, 43 (5):  588-593.  doi: 10.3969/j.issn.0529-1356.2012.05.002 Abstract ( )   Objective Synaptic vesicles complete of synaptic vesicles cycle by exocytosis and endocytosis. The retrieval of synaptic vesicle membrane after exocytosis is essential for the maintenance of synaptic transmission in central nervous system synapses. Dynamin-1 is a 96kD multidomain GTPase enzyme that is crucial for the fission stage of synaptic vesicle recycling and vesicle traffick. Several models range from viewing Dynamin-1 strictly as a mechanochemical enzyme to considering it as a regulatory protein for the recruitment of the downstream binding partners responsible for scission. To address the role of Dynamin-1 and its interaction proteins in synaptic vesicle endocytosis, we screened and identified interaction proteins of Dynamin-1 PRD functional domain in rat brain synaptosomes. Methods pGEX-4T-2-PRD, a prokaryotic expression plasmid of PRD functional domain, was constructed into Dynamin-1. GST-PRD fusion proteins were obtained by Escherichia coli (E.coli) expression system combined glutathione agarose purified column. Rat brain synaptosomal fractions were isolated by ultracentrifugation. Glutathione S-transferase(GST) pull-down assay was employed to screen interaction proteins between rat brain synaptosome and GST-PRD fusion proteins. Subsequently, these proteins were identified using liquid chromatography spectroscopy (LC-MS). Result We successfully purified the GST-PRD fusion proteins and extracted rat brain synaptosome. Thirty-five interaction proteins of Dynamin-1 PRD functional domain in rat synaptosome were isolated and identified, consisting of synaptic vesicle-associated proteins, cytoskeletal proteins, metabolic enzymes and other proteins. Conclusion Here we reported a comprehensive set of candidate proteins that are closely related to synaptic vesicle recycling. The study has laid the foundation for clarifying the function, regulatory mechanism of Dynamin-1 and cracking the pathway/protein network of synaptic vesicle recycling. Related Articles | Metrics

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Role of apoE-mimetic peptide1410 in the regulatory efficacy of metabtropic glutamale receptor/extracellular signal regulated kinase pathway during cerebral vasospasm in mice 2012, 43 (5):  594-600.  doi: 10.3969/j.issn.0529-1356.2012.05.003 Abstract ( )   Objective To investigate mechanisms of the extracellular signal-regulated kinase signaling pathway, and the protective effect of apoE-mimetic peptide1410 after severe cerebral vasospasm (CVS)in mice. Methods A SAH model was established by endovascular perforation without opening cranium. 134 male ICR mice were randomly divided into four groups: sham operation group, model control group, LY367385 group and apoE group. Ten minutes after SAH, 5μl of LY367385 were microinjected into the lateral cerebral ventricle in LY367385 group. Before 30 min of operation mice were injected with apoE-mimetic peptide1410(0.6 mg/kg)via the tail vein and repeated every 12 hours for three days in apoE group. The neurological behavioral function was evaluated.At different time points (6, 24, 48 hours) after operation, changes in brain tissue were observed with light and electron microscopy. The expression of mGluR1 mRNA was detected using RT-PCR, Western blotting assay was used to examine mGluR1 and p-ERK1/2 expression. The number of apoptotic cells was determined by TUNEL method. Results In model control group, the neurological function scores were significantly lower, and the mGluR1 mRNA, mGluR1, p-ERK1/2 level and the number of apoptosis cells were significantly enhanced. Some neurons displayed histopathologie changes of necrosis, and organelles decreased. Mice treated with LY367385 or apoE-mimetic peptide1410 had reduction in mGluR1 mRNA, mGluR1, ERK1/2 activity, and neuronal apoptosis. apoE-mimetic peptide1410 notably relieved the neurological scores, and improved the ultrastructural changes. Conclusion The increased expression of mGluR1 in hippocampus may induce apoptosis by activation of ERK1/2 signaling after CVS. LY367385 has a good therapeutic effect on Related Articles | Metrics

细胞和分子生物学

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Generation of induced pluripotent stem cells from human foreskin fibroblasts 2012, 43 (5):  601-607.  doi: 10.3969/j.issn.0529-1356.2012.05.004 Abstract ( )   Objective To optimize the method for establishment of induced pluripotent stem cells from human foreskin fibroblasts. Methods We used GFP as an indicator to determine the optimized dose of the retrovirus during the reprogramming of human foreskin fibroblasts into human induced pluripotent stem cells (iPSCs) by Yamanaka’s classical strategy with the combination of the small-molecule compounds VPA, a histone deacetylase inhibitor. The characteristics of established iPSCs were tested via alkaline phophatase (AP) staining, immunofluorescent detection of human embryonic stem (ES) cells surface antigens, and RT-PCR analysis of three germ layers specific gene expression in cell during differentiation in vitro. Results Using GFP expression as an indicator of the retrovirus transduction, we established a high efficient system to successfully reprogramme human foreskin fibroblasts into induced pluripotent stem cells. The iPSCs showed positive for alkaline phosphatase, expressed high levels of human ES cells pluripotency markers such as OCT4,TRA-1-60 and TRA-1-81. RT-PCR analysis indicated that the germ layers differentiation related gene expression was detected in the embryonic body when derived from these hiPS cells. BR>Conclusion Our results showed that we successfully generated hiPS cells from adult foreskin fibroblasts in the indication of GFP with the combination of the small-molecule compoun Related Articles | Metrics

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Relationship between cytotoxicity of neural cell and CYP1A1 expression induced by benzo(a)pyrene 2012, 43 (5):  608-613.  doi: 10.3969/j.issn.0529-1356.2012.05.005 Abstract ( )   Objective To investigate the relationship between neural cell cytotoxicity, apoptosis and cytochrome P4501A1(CYP1A1) mRNA exposed to benzo (a) pyrene ［B(a)P］. Methods Primary neural cells were dissociated from the cerebral cortex of 1-3 days old SD rats and cultured in a DMEM incubator at 37℃. After cultivation for 5 days, the well-grown neural cells were selected and treated with the concentrations 0, 10, 20 and 40μmol/L of B(a) P respectively. The neural cells were further cultivated for 40 hours. Neural cell vitality was detected using cck8 kit. Apoptosis rate was measured by flow cytometry using annexinV -FITC and propidium iodide (PI) staining. CYP1A1 mRNA was tested with RT-PCR technique. CYP1A1 mRNA protein expression was determined by immunohistochemistry SABC. Results With the increase of B(a) P concentration, the neural cell vitality declined in both the medium dose group and the high dose group compared with controls, and early apoptotic rate increased steadily only in the high dose group compared with controls. Trend test indicated that cell vitality declined and apoptotic rate increased in a dose dependent manner. Also, there was a dose-reaction relationship between B(a) P and CYP1A1 mRNA. The apoptotic rate of neural cell was positively correlated with CYP1A1 mRNA ( r =0.831, P <0.01) and CYP1A1 protein expression ( r =0.780, P <0.01). Conclusion B(a) P may induce apoptosis of neural cell, and CYP1A1 induction may be Related Articles | Metrics

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Effect of endothelial microparticles on monocytes to promote endothelial cell proliferation in inflammation 2012, 43 (5):  614-618.  doi: 10.3969/j.issn.0529-1356.2012.05.006 Abstract ( )   Objective To investigate the effect of endothelial microparticles (EMPs) on monocytes in promoting inflammatory angiogenesis and in expressing endothelial markers. Methods EMPs were isolated from the supernatants of cultured human umbilical vein endothelial cells (HUVECs) treated with TNF-αby ultracentrifugation, identified under an electron microscope and applied to cultured THP-1 cells. The level of VEGF-A and VEGF-C expressed by THP-1 cells was examined with ELISA. The expression of von Willebrand factor (vWF) and vascular endothelial growth factor receptor 2 (VEGFR-2) in THP-1 cells were studied with RT-PCR and immunofluorescence. HUVECs were co-cultured with THP-1 cells either treated with EMPs not. The proliferation of HUVECs was assessed. Results TNF-α enhanced HUVECs to release EMPs, which promoted THP-1 cells to secrete vascular endothelial growth factor A (VEGF-A) and vascular endothelial growth factor C (VEGF-C). THP-1 cells did not express vWF or VEGFR-2 genetically after either 3 days or 8 days of culture. When cultured with THP-1 cells, HUVECs were more active in proliferation. Conclusion EMPs are able to activate monocytes which in turn enhanced endothelial cells growth by secreting Related Articles | Metrics

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Expresssion of Nanog protein in mouse CD73+ adipose tissue derived mesenchymal stem cells 2012, 43 (5):  619-623.  doi: 10.3969/j.issn.0529-1356.2012.05.007 Abstract ( )   Objective To investigate expression of Nanog protein in mouse adipose-derived mesenchymal stem cells （ADMSCs). Methods ADMSCs were isolated and cultured. The ADMSCs subpopulations for CD73+ and CD73- were selected by flow cytometry and then were cultured and passaged.Immunocytochemistry and immunofluorescence were used to detect the expresssion of nanog protein and compared expression difference between two subpopulations. Results The percentage of CD73+ cell in all ADMSCs was about 5%. According to their morphology,these cells were divided into big cells and small cell. The strong positive expression rate of Nanog protein for CD73+ ADMSCs was（16.85±6.83）%, the weak positive rate was（81.78±7.19）%, and the negative positive rate was（1.63±3.59）%; The strong positive expression rate of Nanog protein for CD73- ADMSCs was（5.74±2.79）%, the weak positive rate was（85.84±37.31）%， the negative rate was（8.42±4.12）%. The different mitotic division phase of the Nanog positive cells was detected. The dividing cells of CD73+ ADMSCs were more than those Related Articles | Metrics

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RNAi-mediated Bip knockdown improves secretion of FVIII activity by protein splicing-based two-chain gene transduced cells 2012, 43 (5):  624-628.  doi: 10.3969/j.issn.0529-1356.2012.05.008 Abstract ( )   Objective To investigate the effect of RNA Interfering-mediated down -regulation ER chaperon protein, immunoglobulin heavy-chain binding protein (Bip) on secretion of spliced FVIII and bioactivity from protein splicing based two-chain co-transgenic HEK293 cell. Methods After treatment with Bip-siRNA, the HEK293 cells were co-transfected with intein-contained B-domain-deleted FVIII(BDD-FVIII) heavy and light chain genes. The expression of Bip was observed by Western blotting with the cell growth atatus assayed by MTT. The quantitative secretion of spliced BDD-FVIII protein and bioactivity were measured by ELISA and coatest chromogenic method respectively. Results Bip was obviously down-regulated by RNA interference technonogy but with no effect on the cell proliferation. It showed a great higher level of spliced BDD-FVIII and heavy chain in terms of secretion by Bip-downregulated co-transgenic cell ［(142±33)μg/L and (197±43)μg/L］ compared to control［(89±23)μg/L and (120±27)μg/L］. The FVIII bioactivity in cell culture supernatant showed an elevated levels in Bip-downregulated co-transgenic cell ［(1.05±0.16)IU/mL］, greater than that of control ［(0.64±0.17)IU/mL］. Co Related Articles | Metrics

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Change of myocardial cells and collagen Ⅰ/Ⅲ protein expression of the left ventricular induced by isoprenaline hydrochloride in rats 2012, 43 (5):  629-634.  doi: 10.3969/j.issn.0529-1356.2012.05.009 Abstract ( )   Objective To probe the change of myocardial cells and collagen I/III protein expression of the left ventricular induced by isoprenaline hydrochloride (ISO) in rats. Methods Adult rats were treated with intraperitoneal injection of ISO by 4mg/(kgI>&#/I>8226;d) for 1, 4 and 8 weeks to prepare SD rats model of myocardial ischemia. The control group were treated with the same volume of the physiologic saline. The tissue structure of the left ventricular was observed with the electron and light microscopes. The expression of collagen I/III protein was detected with immunohistochemistry and Western blotting. The expressive intension and the area percentage were statistically treated with the HAPIS-2000 image analysis system and SPSS10.0 software. Results Under the light microscope, the necrotic area of myocardial ischemia appeared near the endocardium when induced by ISO. In the necrotic area of ischemia collagen I/III was of high expression, and the percentage of the necrotic area was of the decreasing tendency.The difference between groups of 1 week and 4 weeks was of no statistical significance( EM>P /EM>>0.05), and the difference between the groups of 4 weeks and 8 weeks was of statistical significance ( EM>P/EM> <0.05). The difference of the expression intensity when compared among groups was of no statistical significance( EM>P/EM> >0.05). The early period of myocardial cells in the non-necrotic area of ischemia was the mass collection of glycogenosome, and the characteristic of the later period was the increase of the mitochondria and the loose myofibrilla. There was no obvious change in the area percentage of collagen I protein when detected with immunochemistry, and the difference among groups was of no statistical difference (EM> P/EM> >0.05). The difference of the expression intensity when compared among the experimental groups was of no statistical significance ( EM>P /EM>>0.05), but compared with the control group, the expression intensity decreased and the difference was of statistical difference ( P <0.05). The area percentage of collagen III protein was of the increasing tendency when detected with immunochemistry, and the difference between the groups of 1 week and 4 weeks was of statistical significance ( EM>P/EM> <0.05). The expression intensity was stable and the difference among groups was of no statistical significance ( P <0.05). The result of Western blotting showed that the collagen I/III protein among the experimental groups was of a tendency from increasing to decreasing, and the difference among the groups was of statistical significance ( EM>P /EM><0.05). Conclusion Long-time inducement may lead to the ischemic necrosis of the myocardial cells near the endocardium. The necrotic area of ischemia is generally of a decreasing tendency and fibrous change appears in this area, which is associated with the increase of stiffness of the left ventricular wall. The hypertrophy of the myocardial cells in the nonnecrotic area of ischemiais associated with the energy metabolism. Collagen I/III protein does not involve in the stiffness and the compliance of left ventr Related Articles | Metrics

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Effects of ischemic acute kidney injury on morphology of hepatocytes in rats 2012, 43 (5):  635-640.  doi: 10.3969/j.issn.0529-1356.2012.05.010 Abstract ( )   Objective To observe morphological effects of ischemic acute kidney injury (IAKI) on hepatocytes in rats. Methods Western blotting, immunohistochemical staining, light and electron microscopy were used in this observation. Results Necrotic lesions, including pale and vacuolization necrosis lesions, in varied sizes were found in livers of IAKI animals. Cytologically, hepatocyte with either pale necrosis or vacuolization necrosis, hepatocytes with pyknotic cell death and undergoing apoptosis constituted the hepatic damages induced by IAKI but necrosis was more evident. Western blotting assays provided that both PARP-1 and Caspase-3 expressed much stronger in livers of IAKI animals. In addition immunohistochemical staining of both PARP-1 and Caspase-3 revealed that positive hepatocytes were more evident during the acute hepatic damage induced by IAKI. Conclusion Necrosis, PARP-1 mediated cell death and apoptosis, Caspase-dependent cell death are involved in hepatic damages induced by IAKI, but necrosis is more predominated. Related Articles | Metrics

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Comparison of characteristics between rib perichondrium-derived and rid cartilage-derived mesenchymal stem cells of the rabbit 2012, 43 (5):  641-646.  doi: 10.3969/j.issn.0529-1356.2012.05.011 Abstract ( )   Objective To separate rabbit-rib perichondrium cells and cartilage cell, and compare their characteristics. Methods Rib perichondrium cells and rid cartilage cells were isolated from New Zealand white rabbits,aged 3 and 4 weeks by the enzyme digest and adherent culture method and then cultured in vitro. The growth, phenotype and potention of dipogenic and osteogenic differentiation in both types of the cells were observed. Results In the primary generation of rib perichondrium cells were round or polygon and assemble as conglobation; The cells from two-passage began to show a spindle-shape and whirlpool arrangement; and the cells grew to the 15th generation. The primary generation of rib cartilage cells was in cobblestone appearance，and then gradually changed as spindle-shape cells from two-passage to there-passagee cells. But the cells appeared senescence in the five-passage, and then were dead gradually. The results of immunocytochemical stain showed that CD29，CD90 and CD34 were negative in primary rid perichondrium and cartilage cells. From the two passage cells of rib perichondrium and rid cartilage began expressing CD29 and CD90, but not CD34. The collogen type Ⅱwas expressed by the primary rid cartilage cells, and then vanishied in the three-passage cells. The adipogenic and osteogenic differentiation were induced in three-passage rib perichondrium and rid cartilage cells EM>in vitro/EM>. Conclusion The stem cells can be isolated from rib perichondrium and cartilage of rabbit. The cartilage cell may become the stem cell through dedifferentiation EM>in vitro/EM>. The growth of the stem cell derived from rib perichondrium was surp Related Articles | Metrics

肿瘤生物学

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Effects of the gap junctional connexin on the Ca SUP>2+/SUP> -mediated migration and invasion of breast cancer cells 2012, 43 (5):  647-653.  doi: 10.3969/j.issn.0529-1356.2012.05.012 Abstract ( )   Objective To explore effects of the connexin on the Ca SUP>2+/SUP> -mediated migration and invasion of breast cancer cells. Methods MDA-MB-231 and MCF-7 cells, with high and low metastatic potentials, respectively, were treated with a safe concentration of octanol (100μmol/L) for 24 hours. The morphology change was observed under an inverted phase contrast microscope.The location ofconnexin43(Cx43), arrangement of microfilament(MF)and concentration of intracellular Ca SUP>2+/SUP> were determined by confocal microscopy, and cell migration was checked by wound healing and Transwell chamber assays. Results Functional blockade of gap junctions during the formation of a cellular monolayer resulted in discordance of actin stress fibers between neighboring cells, even though whole-cell morphology of these cells did not change. Confocal microscopy revealed that immunoreactivity of Cx 43 was in the cell membrane，particularly at the region of cell-to-cell apposition regardless of the presence of gap junction inhibition. The number of MDA-MB-231 cells was significantly increased in migration assays, as compared with the control. The concentration of intracellular Ca SUP>2+/SUP> was significantly decreased. EGTA treatment enhanced the inhibition of cell migration and invasion which induced by octanol alone. However, inhibition had no effects on the migration and invasion of low metastatic potential MCF-7cells. Conclusion These data imply that gap junctional inhibitor does not interrupt the formation but rather the function of gap junctions, and the underlying mechanism may be related to the decrease of intracel Related Articles | Metrics

解剖学

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Distribution of intramuscular nerve and muscle spindle in gluteus maximus 2012, 43 (5):  654-657.  doi: 10.3969/j.issn.0529-1356.2012.05.013 Abstract ( )   Objective To study the distribution of intramuscular nerve branches and muscle spindles of the gluteus maximus. Methods Dissection, modified Sihler’s neural staining technique ( EM>n/EM> =6) and hematoxylin-eosin staining ( EM>n/EM> =6) were used to study the gluteus maximus from 12 cadavers. Restults The number of secondary branches of the inferior gluteal nerve were 11-14. They ran along the direction of muscle bundles. Spindle density and spindle index of human gluteus maximus were （14.91±3.05）number/cm3 and （14.12±4.34）number/g, respectively. The highest spindle density and spindle index area of gluteus maximus were in the lower part of the muscle. BR>Conclusions There are many secondary branches of the inferior gluteal nerve. They travel along the direction of muscle bundles. The highest spindle density and spindle Related Articles | Metrics

组织学胚胎学发育生物学

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Expression of Bcl-2 during the development of mouse kidney 2012, 43 (5):  658-661.  doi: 10.3969/j.issn.0529-1356.2012.05.014 Abstract ( )   Objective To explore the expression of Bcl-2 during mouse kidney development. Methods Fifty-five Kunming mice from embryonic day 16(E16d), 18 and postnatal day 1(P1d), 3, 5, 7, 14, 21 were chosen to be used to investigate the expression of Bcl-2 on epoxy sections with immunohistochemistry technique and stereological methods. Results During mouse kidney development, the positive staining of Bcl-2 was mainly observed in proximal tubules. The increase with numerical area density(N A) of Bcl-2 was the fastest from E18d to P1d, while not much changed after P7 d. Conclusion During the pristine kidney development, the expression of Bcl-2 influences survival or death of proximal tubule. Related Articles | Metrics

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Toxic effects of lead on embryonic development of zebrafish 2012, 43 (5):  662-666.  doi: 10.3969/j.issn.0529-1356.2012.05.015 Abstract ( )   Objective To explore toxic effects of lead on embryonic development of zebrafish ( EM>Danio rerio/EM>).Methods Zebrafishes (wild type, AB line)were exposed to lead acetate (PbAc) at the doses of 0, 0.1, 0.5, 2.5 and 12.5μmol/L, respectively, from 1 hour post fertilization (hpf) to embryonic hatching or death. There were 50 embryos in each group, and 0 dose group was used as the control. Morphology of the developing embryo was observed with a phase contrast microscope, and the visible toxic effects were recorded. The embryos were photographed at time points of 4, 8, 16, 24, 48 and 72 hpf. The ultrastructure of diencephalon neurons at 36 hpf was observed with a transmission electron microscope. Results The embryonic mortalities of exposure groups were significantly increased compared to the control group ( EM>P/EM> 0.05) with a higher mortality at the early stage of embryonic development. In general, with increasing concentration of PbAc, the mortality increased ( EM>P/EM> 0.01). The aberration rates of the treated embryos were slightly higher than the control group though there was no statistical significance. With the exception of the 12.5 μmol/L lead acetate group, the average hatching times of the exposed groups were longer than that of the control group ( EM>P/EM> <0.05). The results of ultrastructure analysis showed that pathological changes, such as karyopyknosis, cellular edema, and damaged organelles, occurred in the embryonic neuron in the exposed groups.Conclusion Lead may result in considerable toxic effects on zebrafish embryos, especially at the early stage. Low level Pb-exposure may also lead to a Related Articles | Metrics

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Morphological observation on the development of the airway smooth muscle in mouse embryos 2012, 43 (5):  667-672.  doi: 10.3969/j.issn.0529-1356.2012.05.016 Abstract ( )   Objective To investigate the developmental pattern of the airway smooth muscle in mice and the expression characteristics of different muscle-specific proteins. Methods Serial sections of mouse embryos from embryonic day 10 (ED10) to embryonic day 18 (ED18) were stained with monoclonal antibodies against α-SMA, α-SCA and Desmin. Results With the foregut gradually separating into trachea and esophagus, α-SMA positive cells were detected in the posterior wall of the initial segment of the trachea at ED12 and the expression intensity was tapered off towards the caudal segment. A few α-SMA and α-SCA positive cells were observed in the mesenchyme surrounding the developing pulmonary vein. At ED13, the expression intensity of α-SMA in the posterior wall of the trachea became stronger, while the very weaker expression of α-SCA and Desmin was only initiated. At ED14, strong α-SMA expression showed C-shaped patterns in the wall of the left and right bronchi. However, staining intensity of α-SCA and Desmin at the same segment was weaker than that of α-SMA. In addition, the muscle cells of the pulmonary vein showed strong expression of α-SMA. At ED15, α-SMA positive cells were found in the wall of small bronchioles. Between ED17 and ED18, with progression towards the terminal bronchioles, expression intensity of α-SCA and Desmin became weaker in the airway smooth muscle while the expression of α-SMA became stronger. Conclusion The development of the airway smooth muscle begins from the upper segment of the trachea at ED12 and gradually extends distally. At ED18, airway smooth muscle cells have extended to the terminal bronchioles. The expression of Desmin marks the formation of smooth muscle cytoskeleton structures and implies the further maturation of the airway smooth muscle. Besides, it may help airway smooth muscle Related Articles | Metrics

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Network remodeling of collagenⅠand Ⅲ protein in the heart of renal hypertension rats 2012, 43 (5):  673-678.  doi: 10.3969/j.issn.0529-1356.2012.05.017 Abstract ( )   Objective To explore the network remodeling of collagenⅠand Ⅲ protein in the heart of renal hypertension rats. Methods The animal model was prepared by bilateral renal artery stenosis. Thirty-two SD rats were randomly divided into sham operation group, two weeks group, five weeks group and seven weeks group after operation. The model quality of hypertensive rats was assessed with noninvasive blood pressure analysis. The variation characteristics of collagenⅠand Ⅲ in left ventricular wall were detected by immunohistochemistry and Western blotting, and the quantitive analysis was performed with the microscopic image analysis system. Results BR>The collagenⅠand Ⅲ networks had no noticeable change at the second week after operation, but at the fifth week after operation,the thick fibers of collagenⅠaround cardiomyocyte bundle were seen markedly broken and gathered together, or absent. The broken or accumulated fibers with irregular arrangement in collagenⅠsheath in tunica adventitia of arteriole were also detected. The thick and thin fiber bundles of the collagen Ⅲ around cardimyocyte became loose and disorder at the fifth week after operation, and the collagen Ⅲ sheath of tunica adventitia of arteriole turned to be loose. The results of microscopic image analysis showed that the difference of the mean optical density value and the average area ratio of collagen I had both significance between the fifth week and the seventh week after operation ( EM>P /EM><0.05). The difference of mean optical density value of collagen Ⅲ had no significance between groups ( EM>P/EM> >0.05), and the difference of the average area ratio had significance between the second week and the fifth week group after operation (EM>P /EM><0.05). The results of Western boltting showed that the expression level of collagen I was gradually increased, and collagen Ⅲ was gradually decreased from the fifth week after operatio Related Articles | Metrics

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Protective effects and mechanisms of resveratrol on the rats suffering with osteoporosis 2012, 43 (5):  679-684.  doi: 10.3969/j.issn.0529-1356.2012.05.018 Abstract ( )   Objective To investigate the protective effect and its mechanism of resveratrol (RES) on rats with osteoporosis. Methods A total of 72 clean grade female SD rats were randomly divided into six cohorts: control cohort, osteoporosis model cohort, estrogen pretreated cohort, low, median and high dosage RES pretreated cohorts. Except for rats in the control cohort, rats in other cohorts were treated by bilateral oophorectomy and pretreated with 17β-E2 and different dosage of resveratrol respectively, ranging from 1 to 12 weeks after operation. The blood parameters associating with osteoporosis were tested. Morphological observation of bone tissue was performed by HE staining under a microscope. Bone mineral density was measured by a dual-energy X-ray absorptiometry. The mRNA expression level of antioxidant enzymes (gpx1, trx1 and γ-gt) were assayed by RT-PCR. Results Serum calcium, serum phosphate, ALP and TRAP increased obviously, but estrogen and calcium level in serum decreased sharply in the animal model. The bone tissue of the model manifested typical osteoporosis characteristics, and the expression of antioxidant enzymes was at a low level. After treated with estrogen or different dosages of resveratrol, the parameters and phenotypes mentioned above were almost close to the control cohort.Conclusion Resveratrol, likely estrogen, can maintain a balance of bone formation and absorbance, which is critical to the homeostasis of bone, antagonizing the osteoporosis pathology progression. This protective mechanism is associated with the expression and activity of antioxidant Related Articles | Metrics

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Influence of the ambient calcium level on stanniocalcin-1 gene expression and plasma membrane Ca2+-ATPase activity in kidney of EM>Bufo bufo gargarizans/EM> 2012, 43 (5):  685-689.  doi: 10.3969/j.issn.0529-1356.2012.05.019 Abstract ( )   Objective To investigate the influence of the plasma calcium level on stanniocalcin-1 (STC1) gene expression and on plasma membrane Ca2+-ATPase (PMCA) activity in kidney. Methods Seventy-two adult toads, EM>Bufo bufo gargarizans/EM> , were randomly divided into 3 groups, which were exposed to the tap water only, the tap water with 0.1mol/L CaCl2, and the tap water with 0.03mol/L ethylene diamine tetraacetic acid(EDTA), respectively. The toad’s plasma was prepared. Kidneys were excised at pre-exposure and post-exposure 12, 24, 48, 72, 96, 120, and 144 hours, respectively. Both of plasma Ca2+ level and PMCA activity in kidney were detected with colorimetry. STC1 gene expression in kidney were analyzed with semi-quantitative RT-PCR. STC1 immunohistochemical staining were showed by using rabbit antibody for mouse STC1 in conjunction with streptavidin/HRP-conjugated second antibodies. Results Compared with the un-treated group, 0.1mol/L CaCl2 exposure induced the plasma calcium level to increase at the 12th and 24th hour, and to restore to the normal level at the 48th hour, and to increase after 72 hours. Furthermore, 0.1mol/L CaCl2 exposure enhanced PMCA activity, and up-regulated STC1 gene expression during 12-24 hours, and restored them to the normal level at the 72th hour. Compared with un-treated group, the plasma calcium level decreased, but both of PMCA activity and STC1 gene expression did not change after 0.03mol/L EDTA exposure. The immunohistochemistry staining showed that STC1 immunoreactive cells were found in distal convoluted tubules, and in medullary collecting ducts. Moreover, 0.1mol/L CaCl2 exposur enhanced STC1 immunoreactivity. Conclusion High plasma calcium level stimulates PMCA activity and up-regulation of STC1 gene expression in kidney. The up-regulation of STC1 decreases the plasma calcium level. However, substained high plasma calcium level results in inhibition of PMCA activity and down-regulation of Related Articles | Metrics

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Expression and significance of JNK/c-Jun pathway in rat thoracic aortic aneurysm 2012, 43 (5):  690-693.  doi: 10.3969/j.issn.0529-1356.2012.05.020 Abstract ( )   Objective To investigate the expression and significance of c-Jun N-terminal kinase (JNK) and c-Jun in the experimental rat thoracic aortic aneurysm. BR>Methods Twenty adult male Wistar rats were randomly divided into two groups: sham group and operated group. The thoracic aortic aneurysm models were induced by CaCl2 The thoracic aortic segments were collected at the 4th week after surgery. Immunohistochemistry and Western blotting were performed to detect the expression of JNK and c-Jun in the wall of the aneurysm. Results Immunohistochemical staining results showed that p-JNK and p-c-Jun were located extensively in smooth muscle cells of the tunica media in the aortic aneurysm, while there was only a little positive staining in the control group. Western blotting results indicated that the expression levels of p-JNK and p-c-Jun in the aortic aneurysm were higher than those in the control. Conclusion The expressions of p-JNK and p-c-Jun in thoracic aortic aneurysm are stronger than that in the control. The JNK/c-Jun pathway may play a significant role in the development of aortic aneurysm. BR> Related Articles | Metrics

人类学

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Physical characteristics of Xiang languages group 2012, 43 (5):  694-702.  doi: 10.3969/j.issn.0529-1356.2012.05.021 Abstract ( )   Related Articles | Metrics

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A study on the physical characteristics of Hakka in Jiangxi 2012, 43 (5):  703-711.  doi: 10.3969/j.issn.0529-1356.2012.05.022 Abstract ( )   P>Objective To find the physical characteristics and Mongoloid type position of Hakka in Jiangxi province. Methods Investigation was undertaken in accordance with the international academic methods.Eighty-six physical characteristics of 337 males (154 urban adults and 183 rural adults)and 346 females (150 urban adults and 196 rural adults) of Hakka were investigated in Ganzhou county of Jiangxi province. Twenty-three physical indices were calculated and the distributions of them were analyzed accordingly. The physical characteristics of Hakka of Jiangxi were preliminarily analyzed. Results (1) The percentages of eyefold of upper eyelid of Hakka were lower than others.The percentages of Mongoloid fold were higher. Opening height of eyeslits was narrow. External angles were higher than internal angles in direction of eyeslits. The nasal root height was of middle-type, the nasal profile was straight and the nasal base was upturned. The height of alae nasi was of middle-type. The breadth of alae nasi was of wide-type. The upper lip skin height was of middle-type. Thin lips were slightly higher than medium type in males. Thickness of lips was mainly thin in females. Hair color was black and skin color was yellow. The eye color was brown. The indices of head and face observation of Jiangxi Hakka showed the characteristics of head and face in the ethnic groups North Asian type of Mongoloid, but the nose was wider. It also showed some physical characteristics of the ethnic groups South Asian type of Mongoloid.(2)Typical physical characteristics of Jiangxi Hakka belonged to brachycephaly, hypsicephalic type, metriocephalic type and mesorrhiny. Males were of euryprosopy, but females were of leptoprosopy. Jiangxi Hakka anthropometry indices of head and face were relatively close to the North of China type ethnic groups, but the indices of head and face were between the North of China ethnic group type and the South of China ethnic group type of Mongoloid. (3) Typical physical characteristics of Jiangxi Hakka were middle-sized stature, long trunk and subbrachyskelic type. In addition, males and urban females had middle chest circumference, but rural females had broad chest circumference. Males and rural females had medium shoulder breadth and broad shoulder breadth. Males were of a medium distance between iliac crests. Conclusion The results of cluster analysis show that the physical characteristics of Hakka are between the ethnic group types of the North and the South of China of Mongoloid./P> Related Articles | Metrics

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Association study on MRC1 gene polymorphism with pulmonary tuberculosis in Chinese Han and Uygur populations 2012, 43 (5):  712-716.  doi: 10.3969/j.issn.0529-1356.2012.05.023 Abstract ( )   Objective The present study is aimed to investigate potential association of SNPs in exon 7 of the MRC1 gene with pulmonary tuberculosis (TB) in Chinese Han and Kazak populations. Methods Six SNPs (G1186A, G1195A, T1212C, C1221G, C1303T and C1323T) of the MRC1 gene were genotyped using the PCR and DNA sequencing methods in 454 Chinese Han and 595 Uygur subjects. Linkage disequilibrium analysis was performed to determine any linkage between the polymorphic sites. Results In a Chinese Han population, we found that the allele frequency of G1186A had a significant difference in frequency distribution between the two groups ( P =0.037; OR=0.76; 95% CI, 0.58-0.98). The AG genotypes were significantly correlated with pulmonary TB ( EM>P /EM><0.01; OR=0.57; 95% CI, 0.37-0.87). After adjustment for age and gender, G1186A site was found to be dominant ( EM>P /EM><0.01; OR=0.59; 95% CI, 0.40-0.87), over-dominant ( EM>P/EM> =0.045; OR=0.69; 95% CI, 0.47-0.99) and additive models (EM> P/EM> =0.041; OR=0.76; 95% CI, 0.59-0.99) in association with pulmonary TB. In the Chinese Uygur population, we found that the allele frequency of G1186A was a significant difference between the two groups ( EM>P/EM> =0.031; OR=1.29; 95% CI=1.02-1.62). The AA genotype was significantly correlated with pulmonary TB ( EM>P /EM>=0.033; OR=1.64; 95% CI=1.04-2.60). After adjustment for age and gender, G1186A site was found to be additive models ( EM>P/EM> =0.033; OR=1.28; 95% CI=1.02-1.61) in association with pulmonary TB. By calculating the pairwise LD between 6 SNPs in the Chinese Uygur population, we found that the frequency of the haplotype GGTCCT ( EM>P /EM>=0.032; OR=0.75; 95% CI=0.57-0.97) and GGTCCC (EM> P/EM> =0.044; OR=0.57; 95% CI=0.33-0.99) was significantly associated with pulmonary TB. No association was found between the other 5 SNPs and TB ( EM>P /EM>>0.05) in the Chinese population. Conclusion This study reports that genetic variants in the MRC1 gene may be associated with pulmonary TB in a Chinese population. Related Articles | Metrics

技术方法

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Preparation and identification of the pancreatic decellularized Bio-derived scaffold in isolated rats 2012, 43 (5):  717-722.  doi: 10.3969/j.issn.0529-1356.2012.05.024 Abstract ( )   Objective To prepare a pancreatic decellularized bio-derived scaffold(PDBC) in vitro, providing a novel natural bio-derived scaffold for islet and pancreatic tissue engineering. Methods The pancreas from adult SD rats was obained. To prepare the PDBC, we used physical freeze thawing, socking and perfusing that based on retrograde pancreatic and bile ducts’ perfusion. After decellularization, normal pancreatic tissue and PDBC were examined by genomic DNA quantification, and through HE staining,transmission electron microscope and immunohistochemistry.The conservation of collagen type Ⅳ, fibronectin and laminin were identified. Results BR>The DNA gelelectrophoresis showed that no bands in the experimental group while a 100bp band appeared in the control group. Compared with the DNA in the control group, BR>the DNA in PDBC was less than 5%. Histochemical staining and transmission electron microscopy indicated that PDBC preserved the complete extracellular matrix and no cells attached to PDBC surface.Immunofluorescence results demonstrated that collagen, elastin and other extracellular components were well preserved, without obvious nuclear components. Conclusion The PDBC prepared by freeze thawing, socking and perfusing was decellularized completely and preserved the ext Related Articles | Metrics