Construction of corneal neovascularization rabbit model by suture / 解剖学报

Objective To compare the effect of different sutures and suture method on corneal neovascularization ( CNV) in rabbit models. Methods NV was induced by placing sutures at the corneal periphery of rabbits (n = 45). To observe the NV status, 45 rabbits were randomly divided into 5 equal groups. Group A applied 8-0 absorbable suture (A1 single loop parallel suture, A2 single loop vertical suture). In group B, 10-0 nylon suture was used (B1 double loop parallel suture, B2 double loop vertical suture, B3 three loop radial suture). The development of CNV was observed with slit lamp microscope and photographed. Therefore the effective model for neovascularization induction was selected. Histological examination, immunofluorescent staining and ELISA analysis for the vascular endothelial growth factor( VEGF) were performed before suture, 7 and 14 days after suture. Results Sutures fell off and CNV gradually atrophied in group Al and A2; At the 14th day after suture, Sparse or short cluster CNV grew into the corneal margin in group B1 and B2, while CNV was vigorous and grew in bundles in group B3. The expression of VEGF in aqueous humor increased in B3 group after suturing, and increased in 14 days as compared with 7 days after suture. Corneal edema, neovascularization and little immunofluorescence staining for VEGF were detected in group B3 after 7 days suture. More neovascularization and immunofluorescence staining for VEGF were detected in group B3 after 14 days suture. Conclusion Corneal NV can be induced successfully in rabbit model by suturing. The method of 10-0 thread with three sets of circular seams (B3) is stable and effective.

Screening the differentially expressed genes after spinal cord injury by RNA-seq and subsequent functional analysis for partial candidate genes / 解剖学报

Objective: We apply RNA-Seq technology to characterize the temporal changes in global gene expression after spinal cord injury (SCI) in rats. Methods: Spinal cord contusion injury was produced with the Infinite Horizon Device. A total of 48 rats were randomly divided into sham control, and contusion injury for 1 day, 4 days and 7days. RNA-Seq technology was carried out to screen the differentially expressed genes (DE genes) after SCI. We also performed expression pattern and pathway analysis for the DE genes, and selected the candidates to further expression variation validation. Results: Compared with sham group, there were 944DE genes at the first day, 1362 DE genes at the 4th day and 1421 DE genes at the 7th day. The expression variation patterns were roughly divided into 8 kinds of forms. In addition, Real-time PCR results showed that the expression patterns of heme-oxygenases 1 (Hmoxl), Plau, Serpinel and Ncf2 were consistent with RNA-seq analysis. The result of immunohistochemistry showed that Hmoxl was highly expressed in spinal cord neurons after injury. Conclusion: RNA-Seq analysis is useful to screen the DE genes after SCI, and the validated genes could partially explain the molecular mechanism of SCI.

Human bone marrow-derived mesenchymal stem cells differentiate into retinal cells in vitro / 中国药理学通报

Aim To study the differentiation of human bone marrow-derived mesenchymal stem cells ( HM-SCs) into retinal cells in vitro. Methods HMSCs were isolated from human bone marrow after Ficoll den-sity gradient centrifugation. The adherent cells after at least 5 passages were used for study. Immunopheno-type of the cells was analysed by flow cytometer, and cellular differentiation was identified by immunofluores-cence labeling technique. Results The target cells derived from human bone marrow adhered to the plate with fibroblastic-like morphology, whose surface mark-ers were similar to mesenchymal stem cells. Major cells were positive for CD90 , CD44 , CD147 , while they were all negative for CD34, CD45, HLA-DR. In the differentiation study, HMSCs cultured in induced me-dium can differentiate into nestin ( neural stem cell ) -positive cell, GFAP ( glial fibrillary acidic protein ) -positive glial cells and retina-specific neurons express-ing Rhodopsin with CD90 ( mesenchymal stem cells )-negative. Conclusion HMSCs have the ability to dif-ferentiate into retinal neural cells in vitro.

Expression of adhesion molecules in polymyositis / 医学研究生学报

Objective:To study the role of adhesion molecules in the pathogenesis of polymyositis. Methods:The abnormal expression of adhesion molecules on T cells in peripheral blood and muscle fibers from patients with myositis was analyzed by two colour immunofluoresence and RT PCR methods respectively. Results:The expression of adhesion molecules including lymphocyte function associated antigen 1(LFA 1 ),very late antigen 4(VLA 4) on T cells in peripheral blood and intercellular adhesion molecule l(ICAM 1) on muscle fibers from patients with myositis was markedly higher than that in the healthy control group. Conclusion: These findings suggested that adhesion molecules may be responsible for the migration of T cells and destraction of muscle fibers.

THE STUDY OF DIRECT DIFFERENTIATION OF MICE EMBRYONIC STEM CELLS INTO CARDIOMYOCYTES INDUCED BY TGF-?_1 AND CO-CULTURE WITH VISCERAL ENDODERM LIKE END-2 CELLS / 解剖学报

Objective To study the differentiation of embryonic stem cells(ESCs) induced by transforming growth factor-?_1(TGF-?_1) and co_cultured with visceral endoderm like END_2 cells,and explore cardiomyocytes induction effects of the combined techniques Methods Day 2-3 embryoid bodies(EBs) were derived from ESCs,and then TGF-?_1 was added or/and co_cultured with END_2 cells or END_2 cells conditioned medium.Spontaneous differentiation was as a control.The expression of cardiac specific ?-sarcmeric actin(?-actin) and cardiac troponin_T(TnT) was detected by immunofluoresence staining.The ultrastructural analysis for ESCs_derived cardiomyocytes was scanned by transmission electron micrograph. Results The total percentage of beating EBs treated with TGF-?_1,co_cultured with END_2 cells,or END_2 cell conditioned medium was(43?2.08)%,(69?3.61)%,(65?3.06)%,respectively.All the beating cardiomyocytes derived from ESCs expressed cardiac_specific proteins for ?-actin and TnT,and could be observed the cardiac_specific ultrastructure.Interestingly,the total percentage of beating EBs treated with the combined method was(91?1.52)%.(P

CONNECTIONS BET WEEN VESICULAR GLUTAMATE TRANSPORTERS-,GAD-LIKE IMMUNOREACTIVE TERMINALS AND GABA_A RECEPTOR ?3 SUBUNIT- LIKE POSITIVE NEURONS IN MESENCEPHALIC TRIGEMINAL NUCLEUS OF THE RAT / 解剖学报

Objective To observe the association of vesicular glutamate transporter of typeⅠ (VGluT1 ) like immunoreactive(LI) ,the differentiation-associated Na+ dependent inorganic phosphate cotransporter(DNPI) LI and glutami cacid decarboxy lase(GAD) LI terminals with GABAA receptor ?3 subunit(GABAAR ?3) LI neurons in mesencephalic trige minal nucleus of the rat.Methods Triple immun of luorescencehis to chemical staining technique and confo call aser scanning micros copy were used.Results Alargenumber of neuronal cell bodies showed GABAA R?3 LI immunoreactivity atallrostrocaudal level softhe Vme ,and most of GABAAR ?3 LI cells were large (2 5 5 0 ?m)pseudouni polarneurons.The dense VGluT1 LI,DNPL LI and GAD LIterminal sdistri buted widelyin Vme ,some VGlu T1 DNPI LI and GAD LIterminals surrounded the somata of the GABAAR ?3 LI Vmeneurons ,and made close contacts with them .Conclusion Proprioceptive sensory signals from the or of acialregionmight be modulated at the level of the primary afferent cell bodies in the Vme both by glutamatergic and GABAergic axonal terminals from other brain areas,and the effect of GABAergic terminals might be mediated by post synaptical GABAA receptors .

THE EFFECT OF TRANSPLANTATION OF MOUSE PRIMORDIAL GERM CELLS ON THE ACUTE DAMAGED LIVER / 解剖学报

Objective To investigate the milieu-dependent differentiation of primordial germ cells(PEGs) in the acute damaged liver microenviroment. Methods After PGCs were cultured and proliferated,these cells were labelled with 5-bromo-2-deoxyuridine(BrdU),then transplanted into the acute damaged liver by CCl_4 through tail vein.Two and four weeks later,the liver was extracted and 10?m-cryostat continuous sections were obtained.The existing and differentiation of the transplanted cells were identified by immunohistochemistry,immunofluorescence double staining and histochemistry for BrdU and hepatic-specific ALB,and the glycogen. Results Transplanted PGCs were found to be incorporated into the acute damaged liver and differentiated into hepatocytes,compensating for acute liver failure.Conclusion PGCs can be induced to differentiate into hepatocytes in the acute damaged liver microenvironment,and can be used for cellular hepatoplasty to treat severe liver disease.

CONNECTIONS BETWEEN GAD-LIKE IMMUNOREACTIVE TERMINALS AND COEXISTENT POSITIVE NEURONS OF PAG-AND GABA-A-RECEPTOR ?3-LIKE IN THE MESENCEPHALIC TRIGEMINAL NUCLEUS OF THE RAT / 解剖学报

Objective To observe the connections between the glutamic acid decarboxylase\|like immunoreactive(GAD\|LI) axonal terminals and the positive neurons showing phosphate actived glutaminase(PAG)\|LI or GABA\-A\|receptor ?3 subunit (GABA\-AR?3)\|LI and parvalbumin(PV)\|LI in the mesencephalic trigeminal nucleus(Vme) of the rat. Methods The triple\|immunofluoresence histochemical staining techniques were used in the present study,and stained sections were observed under a confocal laser\|scanning microscope. Results Many neuronal cell bodies through the whole rostrocaudal extent of Vme were showed dense GABA\-AR?3\|,PAG\| and PV\|like immunoreactivities,respectively.The majority of them were large pseudounipolar neurons(diameter 25\|50?m).About 90% of PAG\|LI neurons expressed both GABA\-AR?3\|LI and PV\|LI.Confocal laser\|scanning microscope further revealed that dense GAD\|LI axonal varicosities were found to surround the somata of Vme neurons showing both GABA\-AR?3\| and PAG\|like immunoreactivities,and formed the close contact each other.Conclusion\ The present results suggested that the GABAergic axonal terminals projecting to Vme might exert inhibitory regulation and control effect through GABA\-AR?3 subunit localizated in somata of Vme neurons on the transmission of orofacial region proprioceptive sensory information mediated by glutamate.