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At present, vitreoretinal surgery has gradually entered the era of minimally invasive surgery, and high-speed vitrectomy, micro-incision approach and wide-angle illumination have also pushed it to a higher level. Minimally invasive vitreoretinal surgery has become one of the key and difficult points in the teaching of ophthalmic microsurgery. The three-dimensional (3D) surgical display system can provide high-resolution, multi-level magnification, and stereoscopic images for surgery teachers and multiple observers at the same time, breaking the traditional "one-to-one" teaching of the main surgeon and assistant mirrors, realizing "one-to-more" and "head-up" surgery teaching, and thereby improving the teaching effect of vitreoretinal surgery.
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Purpose@#NUF2 has been implicated in multiple cancers recently, suggesting NUF2 may play a role in the common tumorigenesis process. In this study, we aim to perform comprehensive meta-analysis of NUF2 expression in the cancer types included in the Cancer Genome Atlas (TCGA). @*Materials and Methods@#RNA-sequencing data in 31 cancer types in the TCGA data and 11 independent datasets were used to examine NUF2 expression. Silencing NUF2 using targeting shRNAs in hepatocellular carcinoma (HCC) cell lines was used to evaluate NUF2’s role in HCC in vitro and in vivo. @*Results@#NUF2 up-regulation is significantly observed in 23 out of the 31 cancer types in the TCGA datasets and validated in 13 major cancer types using 11 independent datasets. NUF2 overexpression was clinically important as high NUF2 was significantly associated with tumor stages in eight different cancers. High NUF2 was also associated with significantly poorer patient overall survival and disease-free survival in eight and six cancers, respectively. We proceeded to validate NUF2 overexpression and its negative association with overall survival at the protein level in an independent cohort of 40 HCC patients. Compared to the non-targeting controls, NUF2 knockdown cells showed significantly reduced ability to grow, migrate into a scratch wound and invade the 8 μm porous membrane in vitro. Moreover, NUF2 knockdown cells also formed significantly smaller tumors than control cells in mouse xenograft assays in vivo. @*Conclusion@#NUF2 up-regulation is a common feature of many cancers. The prognostic potential and functional impact of NUF2 up-regulation warrant further studies.
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In view of the problems and shortcomings of the domestic ophthalmic microsurgery training system, drawing lessons from the training programs of famous ophthalmic centers abroad, our hospital has explored a set of hierarchical comprehensive training system for ophthalmic microsurgery. Through the four levels-eight scales microsurgery training, the hierarchical comprehensive training system organically integrates the multimedia theoretical teaching, the microscopic practice of Wet-Lab laboratory, microscopic training of surgical simulator and the clinical practice to achieve a better teaching effect in clinical practice, being widely praised by teachers and students.
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Proper wound healing and scaring are the key factor to the success of trabeculectomy in glaucoma patients.The inadequate wound healing will lead to bleb leakage and ocular hypotension after surgery;however,excessive wound healing and scaring will cause the failure of the surgery and eventually increase the intraocular pressure.The applying of antimetabolic drugs such as mitomycin C (MMC) and 5-fluorouracil (5-FU) are able to relieve the excessive wound healing in some degree;however,the side effects like ocular hypotension,dysesthesia endophthalmitis can never be ignored.What is worse,some patients are not sensitive to such drugs.Subconjunctival injection of CAT-152 (monoclonal antibody against transforming growth factor-β) was able to control wound healing in animal trabeculectomy model,while failed in multi-center clinical trial.Recent studies have focused on the role of vascular endothelial growth factor (VEGF) and VEGF inhibitors on the wound healing after trabeculectomy.This paper aims to review the mechanism of wound healing after trabeculectomy,as well as the role of anti-VEGF on this kind of wound healing and scaring.
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Pharyngeal foreign body is a common disease. The diagnosis and treatment are easy. However, in a few cases, pharyngeal foreign bodies migrated to other part of body, which often causing missed diagnosis or misdiagnose to delaythe treatment, and even lead to fatal complications. Here we present a case report of a 52-year-old female patient.who was found to have cervical mass 20 days before. Contrast-enhanced computed tomography showd a foreign body and foreign body granuloma on the left side of the neck. To look back on the history, the patient swallowed a fish bone in mistake one month ago.
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Animals , Female , Humans , Middle Aged , Contrast Media , Deglutition , Foreign Bodies , Diagnosis , Granuloma , Diagnosis , Neck , Pathology , Pharynx , Pathology , Seafood , Tomography, X-Ray ComputedABSTRACT
Objective To investigate the effects of endogenous sulfur dioxide(SO2) on pulmonary vascular inflammation in rats with monocrotaline (MCT)-induced pulmonary hypertension.Methods Thirty-two Wistar rats were randomly divided into 4 groups(n =8 for each group):control group,MCT group,MCT + L-aspartic acid-β-hydroxamate(HDX) group,and MCT + SO2 group.Rats in the MCT group,MCT + HDX group,and MCT + SO2 group were subcutaneously injected with MCT(60 mg/kg) on the first day.For rats in MCT + HDX group,HDX(25 mg/kg,on day 0,7 and 14) was given orally after injection of MCT; and rats in MCT + SO2 group were subcutaneously injected with the SO2 donor sodium sulfite/sodium bisulfate(Na2SO3/NaHSO3,and mole ratio was adjusted to approximately 3:1) each day.Rats in the control group received only the same volume of solvent vehicle only.After 3 weeks,mean pulmonary artery pressure(mPAP) of each rat was evaluated by using a right cardiac catheterization procedure.Immunohistochemistry was used to detect the expression of inflammatory related factor intercellular adhesion molecule-1 (ICAM-1) and the key molecules of nuclear factor-κB (NF-κB) signal transduction pathway,including p65 and inhibitor of NF-κB (IκBα) in the small pulmonary artery endothelial cells.Results The differences in mPAP,expression of ICAM-1,IκBα and p65 in the small pulmonary artery endothelial cells were found among the 4 groups (mPAP:F =53.334,P ＜ 0.01 ; ICAM-1:F =183.82,P ＜ 0.01 ; IκBα:F =142.89,P ＜ 0.01 ; p65:F =105.46,P ＜0.01).The mean pulmonary artery pressure(mPAP) was significantly raised in MCT group rats as compared with that of the control group along with upregulated expressions of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells,while the expression of IκBα protein in small pulmonary artery endothelial cells was significantly low.After administration of HDX,the mPAP and the expression of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells further increased compared with those of MCT group,while the expression of IκBα protein in small pulmonary artery endothelial cells was significantly lower than that of MCT group.Whereas with treatment of SO2 derivatives,the mPAP,the expression of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells were significantly lower than those of MCT group,while the expression of IκBα protein in small pulmonary artery endothelial cells increased significantly compared with that of MCT group.Conclusions Endogenous SO2 might inhibit the activation of NF-κB pathway in the small pulmonary artery endothelial cells,attenuate the pulmonary vascular inflammation and prevent the MCT-induced pulmonary hypertension in rats.
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<p><b>OBJECTIVE</b>To assess the effect of CatWalk automated gait analysis system for evaluation of motor function of rats with traumatic brain injury (TBI) after umbilical cord mesenchymal stromal cell (UC-MSC) treatment.</p><p><b>METHODS</b>Eighteen Wistar rats were randomized equally into normal control group, TBI ∓ saline group, and TBI ∓ UC-MSCs group. The rats in the latter two groups were subjected to weight-drop impact to induce TBI followed by injection UC-MSCs or saline into the lesion 7 days after TBI. The neurological function was assessed using CatWalk system and modified neurological severity scores (mNSS) before and 3 days after TBI and 7 days after UC-MSC transplantation. The rats were sacrificed 14 days after the cell transplantation and the brain sections were stained for immunohistochemical analyses.</p><p><b>RESULTS</b>Three days after TBI, mNSS test showed moderate injury of the rats. Seven days after the cell transplantation, the rats showed significant motor function improvement and CatWalk analysis indicated partial recovery of the gait parameters of the 4 limbs compared to the rats with saline treatment. Histological analyses showed that DiO-labeled UC-MSCs were present in the lesion boundary and expressed glial fibrillary acidic protein and β-tubulin III.</p><p><b>CONCLUSION</b>UC-MSC transplantation can promote functional improvement of the brain after TBI in rats. Compared with mNSS test, CatWalk analysis is more sensitive and objective for assessing neurological function and also provides more detailed information on specific gait parameters.</p>
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Animals , Male , Rats , Brain Injuries , General Surgery , Disease Models, Animal , Gait , Mesenchymal Stem Cell Transplantation , Mesenchymal Stem Cells , Cell Biology , Rats, Wistar , Recovery of Function , Umbilical Cord , Cell BiologyABSTRACT
<p><b>OBJECTIVE</b>To construct a prokaryotic expression vector of human tau multiepitope peptide for examining the immunogenicity of a TauP1/P2 DNA vaccine in mice using the expressed product.</p><p><b>METHODS</b>The coding sequence of Tau multiepitope peptide gene was amplified from the plasmid pVAX1-Tau by PCR and inserted into the prokaryotic expression vector pGEX-4T-2 to construct the recombinant plasmid pGEX-4T-2-TauP1/P2. The positive recombinants were transformed into E.coli BL21 cells, and the expression of fusion protein GST-TauP1/P2 was induced by IPTG and identified by SDS-PAGE. Mice was immunized with TauP1/P2 DNA vaccine and the production of the specific antibodies was detected by Dot-blot analysis using the purified fusion protein.</p><p><b>RESULTS</b>A gene fragment 300 bp in length was amplified. Enzyme digestion and DNA sequencing verified correct construction of the prokaryotic expression plasmid pGEX-4T-2-TauP1/P2. The expression of target fusion protein GST-TauP1/P2 was detected by SDS-PAGE. Specific antibodies against TauP1/P2 were detected in the serum of mice immunized with the DNA vaccine using GST-TauP1/P2 fusion protein.</p><p><b>CONCLUSION</b>The constructed prokaryotic expression plasmid of human Tau multiepitope peptide is capable of expressing the target fusion protein, which specifically recognizes the specific antibodies against TauP1/P2 in mice immunized with TauP1/P2 DNA vaccine.</p>
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Animals , Humans , Mice , Epitopes , Allergy and Immunology , Escherichia coli , Genetics , Metabolism , Genetic Vectors , Genetics , Mice, Inbred C57BL , Peptides , Genetics , Metabolism , Recombinant Proteins , Genetics , Allergy and Immunology , Vaccines, DNA , Allergy and Immunology , tau Proteins , Genetics , Allergy and ImmunologyABSTRACT
Objective To study prognosis and grading of somatosensory evoked potential(SEP)in long-term unconscious patients after severe traumatic brain injury(TBI).Methods Five prognostic factors including age,sex,injury mechanism,history of temporal craniotomy and SEP grading were selected and analyzed in 47 patients after severe TBI with a duration of unconsciousness longer than two weeks.The prognosis was judged by Glasgow Outcome Scale.Results Prognosis was closely associated with SEP grading(P=0.024).The accuracy of SEP in assessing the prognosis was 91.5%.About 95%-100% of patients with SEP at grade Ⅲ-Ⅲ ended up with severe disability,persistent vegetative state or death.However,43.75% of patients with SEP at grade Ⅰ had good prognoses.Conclusions The SEP grading can objectively and accurately evaluate patients' prognosis and demonstrate the brain function.
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Objective To study the potential antidepressive-like effect of tetramethylpyrazine (TMP). Methods Forced-swimming and chronic mild stress (CMS) tests were performed to assess the antidepressant-like activity of TMP. Male Sprague-Dawley (SD) strain rats were divided into six matched groups (n= 13 or 14 in each group) based on their sucrose consumption:control, CMS, CMS + fluoxetine, and CMS + TMP groups. The rats except control were housed separately in different rooms, and the rat model of depression was established by exposing to an unpredictable sequence of stressors for 28 days; the rats in CMS + fluoxetine were exposed to CMS and received administration of FLU (2.0 mg· kg-1·d-1 ,ig) for 28 days; the rats in CMS + TMP groups were exposed to CMS and received administration of TMP (10,20,40 mg·kg-1·d-1 ,ig) , respectively, for 28 days. The rats in control group were given ordinary daily care and received ig administration of normal saline simultaneously. The body weight, food intake and fluid consumption were measured, and the behaviors were examined by open field during the duration of the stress procedure, and forced-swimming test was performed 1 day after last unpredictable stressor. Results Acute administration of TMP markedly decreased the duration of immobility during forced-swimming test((89.0 ±37.0)s vs (117.1 ±32. 1)s, P<0.05) . Chronic administration of TMP partially countered the effects of CMS on consumption of sucrose solution and locomotion and exploration behavior, and potently shortened the immobility time during forced-swimming test following CMS in rats. The results showed that long-term administration of TMP partially reversed the effects of CMS on the body weight gain,the consumption of sucrose solution,the squares crossing in open field test and the immobility time during forced-swimming test in rats ((91.9 ±31.5) vs (124.4±27.0)s,P<0.05).Conclusion TMP shows obvious antidepressant-like activity.
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Objective To explore the protecting mechanism of erythropoietin (EPO) on learning and memory of Alzheimer disease (AD) rats. Methods The AD model was made by injected into rat hippocampal CA1 subregion with amyloid beta-protein(Aβ). Male Spraque-Dawley rats were as the experimental objects, which were randomly separated into 3 groups including Sham, Saline control and EPO treatment. After Aβ was injected into rats hippocampal CA1 subregion ,saline or EPO was respectively injected into the lateral ventricle of rats,with help of stereotaxic coordinates, upon the designed conditions. Hippocampal CA1 subregion Bcl-xl expression changes were observed 24 hours after the operation, and learning and memory abilities were checked 4 weeks after the operation. Results 24 hours after the operation Bcl-xl expression in the EPO group and the Saline group was less than the Sham control ,while Bcl-xl positive cells( 100.42 ± 12.43/field) in the EPO group were more than in the Saline group( 82.06 ± 19.68/field ) (P ＜ 0. 05 ). 4 weeks after the operation learning ability in the EPO group ( 20.38 ± 5.88 ) was better than Saline group ( 25.50 - 3.25 ) (P ＜ 0. 05 ), and memory ability in the EPO group (4.75 ± 1.75 ) was better than the Saline group(2.88 ± 1.55 )(P ＜ 0.05 ). Conclusion EPO could improve the learning and memory abilities in the model rats,and it could be related with EPO restraining Bcl-xl expression decreasing.
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BACKGROUND: It is a new tendency to treat central nervous system injury or tumor therapy using the combination of seed cells and gene therapy.OBJECTIVE: To observe the dose-relationship between transfection and expression of rabbit bone marrow mesenchymal stem cell (BMSCs) with adenovirus vector carrying green fluorescent protein (Ad-GFP), and to study its effects on cell biological properties, in addition, to explore the feasibility of using Ad-GFP vector to construct gene modified BMSCs.DESIGN, TIME AND SETTING: A randomized grouping, contrast observation. The experiment was performed at the Southern Medical University between August 2008 and March 2009.MATERIALS: New Zealand white rabbits, irrespective of genders, weighing 2.0-3.0 kg, were selected.METHODS: BMSCs were separated and cultured in vitro, and then the cell immunophenotypes were detected by flow cytometry.The adenovirus was obtained by packaging 293 cells and was used to transfect BMSCs with various liters (1 ×-10~3-1×10~(10) PFU/mL).Cytometry was used to analyze the transfection efficiency.MAIN OUTCOME MEASURES: Cell morphological changes were detected under an invert microscope. The cell proliferation was detected by CCK8 kits. BMSCs transfected with Ad-GFP were induced differentiating into neuron-like cells by adding of β-mercaptoethanol.RESULTS: The surface markers of 3-6-generation BMSCs were negative to CD34 and CD45, but positive for CD29 and CD44.When the virus titers were 1 ×10~7 PFU/mL, the transfection rate was 55%, which were 85% when the virus titers were 1 ×10~9 and1×10~(10) PFU/mL. However, cell pathology phenomenon occurred when the virus titer was 1 ×10~(10) PFU/mL. The fluorescence was strongest expressed at day 7, and it still can be seen at day 28. The BMSCs trasfected with Ad-GFP could differentiate into neuron-like cells under induction of p-mercaptoethanol, with positive neuron-specific enolase.CONCLUSION: Ad-GFP with suitable titers can infect BMSCs effectively with little influence on the biology property or differentiation function. BMSCs can serve as seeds cell in gene therapy field when utilizing ad-GFP vector system.
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Background To provide clinical evidence for ablative application by comparison of the analgesic effect following different thalamotomy in rats.Methods Thirty rats were randomly assigned into sham and 4 thalamotomies groups: central medial thalamic nucleus ( CM), parafascicular thalamic nucleus ( PF), ventral posterolateral thalamic nucleus (VPL), and CM +Cigulum (cg). Two μL 10% phenol dissolved in glycerin were used for stereotactic thalamotomy. The thermal pain thresholds before and after procedures were evaluated with the tail stimulate test. The formalin test was carried out in an open field apparatus where the animal formalin-induced responses (licking duration, flexing duration, and flinching frequency of the injected paw) were recorded for 60 min.Results Changes of pain thresholds in all ablative groups were significantly higher than that in the sham group, especially it was higher in VPL group. Differences of the factor thalamotomy were found to be due to the shorter licking in the ablative groups than that in the sham group (P ＜0.01 ), whereas flexing duration and flinching frequency were only slightly affected by thalamotomy. Moreover, licking duration was lower in VPL group than in CM and CM + cg groups ( P ＜0.05), whereas nociceptive responses did not differ between the CM and CM+cg groups (P ＞0.05).Conclusions In acute period, CM, PF, VPL, CM + cg neurolysis all showed to elevate the thermal pain threshold and to reduce the pain-induced behavioral responses related to supraspinal neural circuits (licking of the injected paw). Among them, the damage of VPL might be the most active one. CM + cg damage did not get better antinociceptive effect than single CM ablation.
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Our previous study showed that transmembrane TNF-alpha (TM-TNF-alpha) had broader tumoricidal spectrum than secretory TNF-alpha (s-TNF-alpha). This study examined the difference between the two kinds of TNF-alpha in inducing cells and the relationship between the apoptosis induced by TM-TNF-alpha and the cell cycle. Bioassay was employed to compare the cytotoxic effect of two kinds of TNF-alpha on cell lines L-929 and HepG2. TUNEL was used to detect apoptosis and the TdT and PI co-staining were used for determining the phase of apoptotic cells. Our results showed that TM-TNF-alpha could kill not only s-TNF-sensitive L929 cells but also s-TNF-tolerant HepG2 cells. TM-TNF-alpha predominantly induced apoptosis while s-TNF could induce both apoptosis and necrosis. The apoptosis of L-929 cells induced by TM-TNF-alpha mainly occurred in S phase and the apoptosis of HepG2 predominantly took place in G(1) phase. It is concluded that the cytotoxic effects of the two TNF differ substantially. Since TM-TNF-alpha works locally, mainly induces apoptosis and has broader anti-tumor spectrum, it may be more effective for the treatment of tumor than s-TNF.
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Our previous study showed that transmembrane TNF-α (TM-TNF-α) had broader tumoricidal spectrum than secretory TNF-α (s-TNF-α). This study examined the difference between the two kinds of TNF-α in inducing cells and the relationship between the apoptosis induced by TM-TNF-α and the cell cycle. Bioassay was employed to compare the cytotoxic effect of two kinds of TNF-α on cell lines L-929 and HepG2. TUNEL was used to detect apoptosis and the TdT and PI co-stainin gwere used for determining the phase of apoptotic cells. Our results showed that TM-TNF-α could kill not only s-TNF-sensitive L929 cells but also s-TNF-tolerant HepG2 cells. TM-TNF-α predominantly induced apoptosis while s-TNF could induce both apoptosis and necrosis. The apoptosis of L-929 cells induced by TM-TNF-α mainly occurred in S phase and the apoptosis of HepG2 predominantly took place in G1 phase. It is concluded that the cytotoxic effects of the two TNF differ substantially.Since TM-TNF-α works locally, mainly induces apoptosis and has broader anti-tumor spectrum, it may be more effective for the treatment of tumor than s-TNF.
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0.05). The movement and invasion ability were evidently inhibited in AQP1-siRNA groups compared with those in blank control groups and negtive control groups [(36? 3), (23?4)/HP vs(70?5),(65?4)/HP and(72 ?4),(69?4)/HP, respectively, P0.05).CONCLUSION The knock down of AQP1 by RNAi have no effect on the adhesion ability of Hep-2 cells, whereas could prevent the movement and invasion ability of laryngocarcinoma cells, which may be related with the metastasis of laryngeal carcinoma.
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BACKGROUND: Researches indicated that bone marrow stem cells (BMSCs) could differentiated into neural stem cells in vitro, but what was the role of neural stem cells(NSCs) in the recovery of cortical injury,whether the NSC is capable of growing and migration in injured still remained unknown.OBJECTIVE: To explore the growing state of autograft NSC derived from crab-eating macaque BMSC transplanted in brain.DESIGN: Prospective case control study based on experimental animals.SETTING: Department of neurosurgery in a hospital of a military medical university.MATERIALS: This study was carried out at Center Laboratory of Neurological Research Institute, Zhujiang Hospital affiliated to the First Military Medical University of Chinese PLA. Six healthy adult crab-eating macaques were purchased from the South China Primate Animal Center.INTERVENTIONS: BMSCs harvested from six crab-eating macaques were cultured in vitro and induced to differentiate into neural stem cells, which then labeled by bromodeoxyuridine(BrdU) and autografted into brains.MAIN OUTCOME MEASURES: Brain tissues underwent hematoxylin and eosin(HE) staining and immunohistochemical staining before observed under optical microscope.RESULTS: The results of HE staining showed that the cell number in injured brain vas obviously higher in both instant and delayed transplanting groups than sham-transplanting group; moreover cells were proved reacting to BrdU by immunohistochemical staining in cortical injuries of both groups at 1-6 months following stem cells autograft, as well as at neighboring white matters at half year later, but no BrdU positive cells could be found in traumatic controls, sham-transplanting group and normal brains.CONCLUSION: NSCs derived from in vitro cultured BMSCs were proved capable of surviving, proliferating, differentiating and migrating in cortex after autograft, so that BMSCs is considered as replacing cells or the source of NSCs; moreover autograft stem cells could survive, proliferate and migrate in old cortical traumatic focus.
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BACKGROUND: Lipopolysaccharide(LPS), as a polyclonal immune exciter, can simulate immune excitation status, which is useful in the observation of whether the catecholaminergic neurons in paraventricular nucleus of hypothalamus(PVN) projected from medullary visceral zone(MVZ) react towards LPS stimulation that is to provide a theoretical gist for the researches on the protection of brain function.OBJECTIVE: To observe whether PVN catecholaminergic neurons projected from MVZ react towards LPS stimulation for the exploration of the impacts of MVZ-PVN catecholaminergic access in "immune-to-brain communication".DESIGN: A randomized controlled study using experimental animals as subjects.SETTING: An Institute of Neurosurgery and Neurology of one Military University of Chinese PLA.MATERIALS: The study was conducted in the Department of Neurosurgery of Zhujiang Hospital affiliated to Southern Medical University and the Institute of Neurology of the Fourth Military Medical University of Chinese PLA from January to December in 2002. Ten healthy adult SD rats in cleanness grade were obtained from the experimental animal center of the Fourth Military Medical University of Chinese PLA.METHODS: WGA-HRP was injected into PVN of one side of the rat, and the immune exciter LPS was injected into the abdominal cavity after 48 hours of survival to induce immune response. Samples were stained by triple labels of WGA-HRP method and double immunohistochemical staining of anti-Fos and anti-TH antibodies.MAIN OUTCOME MEASURES: To observe the distributions and expressions of WGA-HRP labeled cells, Fos protein, and catecholaminergic neuron(labeled by TH) in MVZ.RESULTS: Seven immune-reactive(IR) positive neurons were found in MVZ, i. e., HRP, Fos or TH single labeled cells, Fos/HRP, Fos/TH or HRP/TH double labeled cells, and Fos/HRP/TH triple labeled cells. Fos/HRP double labeled neurons and Fos/HRP/TH triple labeled neurons accounted for 12. 5% and 39.6% of HRP labeled cells respectively.CONCLUSION: MVZ reacts to LPS immune stimulation, which could upload the immune message to PVN through Catecholaminergic neurons.MVZ might be a relay station in "immune-to-brain communication", which exerts immune modulatory impact through "MVZ→PVN" access.
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Objective To approach the feasibility of transfecting the DNA plasmid of encoding red fluorescent protein directly into the nucleus of rabbit primary bone marrow stromal cell with recently developed nucleofection technique. Methods Rabbit primary bone marrow stromal cells (BMSCs) were harvested by means of density gradient centrifugation following a thighbone puncture. The primary BMSCs were cultured and either transfected to pDsRed1-N1 by nucleofectorTM technique (as DsRed group) or left uninfected(as control group) in vitro. The cellular viability, adhesive rate, the growth curves and the efficiency of transfection of both DsRed and control groups were analyzed. Result DsRed were successfully expressed at 48h after nucleofection. Similar morphology evolvement, adhesive rates and growth curves were obtained from the two groups. The positive DsRed expression enhanced gradually alone with a prolonged culturing time, and reached its peak value at the 10th day after marked, with about 54.2% of DsRed-positive cells in the total BMSCs. The DsRed did not attenuate even until 1 month following the mark. Conclusion Neuclofection of pDsRed1-N1 showed no significant effect on the proliferation of rabbit BMSCs. DsRed worked efficiently for the purpose of stable gene marking of rabbit BMSCs, and nucleofection is an efficient method for transferring genes into primary rabbit BMSCs.
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BACKGROUND: It is considered traditionally that epilepsy is a kind of complicated nervous conduct disorder caused by abnormally excited neuron in different area in brain. While the research on the function of astrocyte in epileptic attack is very rare.OBJECTIVE: To study the reaction of neuron and astrocyte in medullary visceral zone after epilepsy induced by pentetrazole in rats.DESIGN: A randomized controlled experimental research.SETTING and MATERIALS: The experiment was done in the Neurosurgery Laboratory of Zhujiang Hospital Affiliated to Southern Medical University and Neuroscience Institute of Fourth Military Medical University of Chinese PLA. Fourteen healthy adult SD rats, weighing 180 - 220 g, clean grade, were provided by the Experimental Animal Center of Fourth Military Medical University of Chinese PLA.INTERVENTIONS: Distribution of neuron and astrocyte in MVZ 1 hour after epileptic attack was shown by laserconfocal microscopic technique combined with triplication immunofluorescence histochemistry of anti-Fos protein, anti-tyrosine hydroxylase(TH) and anti-glial fibrillary acidic protein(GFAP).MAIN OUTCOME MEASURES: Observation of distribution of positive cells of Fos, GFAP and TH in MVZ and relationship between GFAP positive astrocyte and neuron.RESULTS: Fos positive neurons and GFAP positive astrocytes in MVZ increased significantly. Triplication immunofluorescence histochemistry showed reaction neuron(Fos positive) closely related with reaction astrocyte(GFAP positive) . Three kinds of N-ASC compounds with different labels were found, which were TH +/Fos +/GFAP + three labeled compound, TH + /GFAP +/Fos- and Fos+/GFAP +/TH- two labeled compound.CONCLUSION: Neuron and astrocyte in MVZ reacted strongly when epilepsy attacks. N-ASC as a functional unit may regulate onset of epilepsy.