Increased CD68/TGFβ Co-expressing Microglia/Macrophages after Transient Middle Cerebral Artery Occlusion in Rhesus Monkeys

The function of microglia/macrophages after ischemic stroke is poorly understood. This study examines the role of microglia/macrophages in the focal infarct area after transient middle cerebral artery occlusion (MCAO) in rhesus monkeys. We measured infarct volume and neurological function by magnetic resonance imaging (MRI) and non-human primate stroke scale (NHPSS), respectively, to assess temporal changes following MCAO. Activated phagocytic microglia/macrophages were examined by immunohistochemistry in post-mortem brains (n=6 MCAO, n=2 controls) at 3 and 24 hours (acute stage), 2 and 4 weeks (subacute stage), and 4, and 20 months (chronic stage) following MCAO. We found that the infarct volume progressively decreased between 1 and 4 weeks following MCAO, in parallel with the neurological recovery. Greater presence of cluster of differentiation 68 (CD68)-expressing microglia/macrophages was detected in the infarct lesion in the subacute and chronic stage, compared to the acute stage. Surprisingly, 98~99% of transforming growth factor beta (TGFβ) was found colocalized with CD68-expressing cells. CD68-expressing microglia/macrophages, rather than CD206⁺ cells, may exert anti-inflammatory effects by secreting TGFβ after the subacute stage of ischemic stroke. CD68⁺ microglia/macrophages can therefore be used as a potential therapeutic target.

Prevalência de Campylobacter spp. termofílicos em primatas não humanos da espécie Macaca mulatta provenientes de centro criatório para uso como biomodelos / Prevalence of thermophilic Campylobacter spp. in non-human primates of the Macaca mulatta species from a breeding center for use as biomodels

Objetivo: A campilobacteriose é uma das principais doenças entéricas do mundo. Ocorre não só no homem mas também em primatas não humanos, sendo desta forma importante no monitoramento sanitário de colônias de animais provenientes de criatórios científicos. É causada por bactérias do gênero Campylobacter, cuja detecção em biotérios possibilita não só controlar a doença e prevenir sua disseminação, mas assegurar a qualidade das pesquisas que serão realizadas nestes biomodelos. Com base na importância deste isolamento, nosso objetivo foi a verificação do status sanitário de uma colônia de criação de Macaca mulatta, usando como referência a presença de Campylobacter spp. Métodos: Durante o manejo médico anual coletaram-se 52 swabs fecais de primatas não humanos adultos, o que representou um percentual de 10% da colônia total. Esse material foi submetido ao esquema de semeadura/incubação e identificação de Campylobacter sp. seguindo as recomendações de cultivo microbiológico, incluindo o isolamento, prova de Gram e testes bioquímicos. Todo o processo levou de cinco a sete dias e foi realizado em atmosfera de microaerofilia. Resultados: Em 14 indivíduos foram isoladas bactérias do gênero Campylobacter. Destes, sete eram portadores de Campylobacter coli, seis portadores de Campylobacter jejuni e em um indivíduo não foi possível definir a espécie de Campylobacter isolada. Conclusão: Apesar da baixa prevalência (27%), esses resultados reforçam a necessidade de constante monitoramento microbiológico dos primatas pertencentes à colônia, visando não só a qualidade dos animais fornecidos, mas minimizando o risco de contaminação dentro da colônia e de contágio pelos profissionais que lidam com os animais, já que o Campylobacter possui importante potencial zoonótico.

Effects of recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immta ure dendritic cells on T cell proliferation / 医学研究生学报

Objective HLA-G widely participates in immune tolerance by its combination with immunoglobulin-like tran-scripts IL-2 and IL-4 on the surface of dendritic cells (DCs).The aim of the article was to explore the effects of recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immature dendritic cells on T cell proliferation . Methods Marrow blood was collected from macaca mulattas by the puncture needle after anesthesia .Density gradient centrifugation method was applied in separating mononuclear from the extracted blood on which CD 34+cells were collected and pu-rified by means of immunomagnetic separation .Small doses of cyto-kines were added to get the immature dendritic cells after induced dif-ferentiation of CD34+cells.After the recombinant adnovirus-mediated HLA-G transfection in macaca mulatta immature dendritic cells , observation was done on the viral infection efficiency and western blot was used in detecting the expression of HLA -G in immature den-dritic cells.Taking T cells in macaca mulatta as responders and DCs transfected by recombinant adnovirus -mediated HLA-G as stimu-lators, mixed lymphocyte test was conducted .T cells were divided into 5 groups: mDC group ( mature DCs ) , imDC group ( immature DCs), imDC(L) group(addition of 100 ng/mL lipopolysaccharide after getting imDC at 7th day) , imDC(V) group (imDCs infected by recombinant adnovirus-mediated HLA-G) , imDC( L+V) group ( imDCs infected by recombinant adnovirus-mediated HLA-G along with the addition of 100 ng/mL lipopolysaccharide in culture process ) . Results We obtained the immature dendritic cells and recom-binant adenovirus of HLA-G expressed in these cells .Flow cytometry showed DC purity was up to 92.3 %, imDC purity was up to 72.39%and positive percentage of CD 4+T was greater than 80%.In comparison with imDC group ,the proliferation of stimulated T cells in mDC and imDC(L) groups was obviously intensified (P<0.01).In comparison with imDC(V) group, the proliferation of stim-ulated T cells in imDC, mDC, imDC(L), and imDC(L+V) groups was obviously intensified (P <0.01).In comparison with imDC(L+V) group, the proliferation of stimulated T cells in mDC and imDC(L) groups was obviously intensified(P<0.01). Conclu sion Im-mature DCs infected by recombinant adnovirus can inhibit the proliferation of T cells effectively .

Study on the expression level of 8-Hydroxy-2'-deoxyguanine in rhesus monkeys of different ages / 中华老年医学杂志

Objectives To investigate the expression of 8-Hydroxy-2'-deoxyguanine(8-oxodG)in white blood cell,plasma and urine of rhesus monkey of different age group.Methods 30 female rhesus macaques at different age(1y,5y,10y,15y,20y,25y)were selected and grouped(n=5,each).10 mL of morning urine and 5 mL of fasting venous blood were collected.The level of 8-oxodG expression in plasma,leukocyte and urine was measured by high-performance liquid chromatography-mass spectrometry(HPLC-MS) method.Results The level of 8-oxodG in leukocytes,plasma and urine was increased along with aging.The level of 8-oxodG was 1.8,1.6 and 1.4 times higher in 25 year group than in 1 year group in plasma,white blood cell and urine,respectively(P＜0.05).The 8-oxodG level was more than 40 times higher in urine than in plasma.Conclusions The expression level of 8-oxodG is increased along with aging.It may be one of the experimental evidence of the aging markers.

Expression and identification of VEGF165 in bone marrow mesenchymal stem cells of rhesus / 天津医药

Objective To detect the transferred vascular endothelial growth factor (VEGF)165 gene expression in rhesus autologous bone marrow mesenchymal stem cells (MSCs), and to explore the functional viability of transgenic MSCs. Methods MSCs from rhesus bone were isolated by Ficoll, which were used to detect the phenotype. After the culturing, the expression vector pcDNA-eGFP-VEGF165 was transfected into bone marrow MSCs. Fluorescence microscope and flow cytometry were used to detect the enhanced green fluorescent protein (eGFP) expression. At the same time, the phenotype in transfected MSCs was also indentified. The VEGF165 expression level was detected by RT-PCR. Results The highly purified MSCs were collected successfully. The transfected MSCs and daughter cells showed expressions of eGFP and VEGF165, which also remained the characteristics of MSCs. Conclusion The VEGF165 gene that is transfected into MSCs can maintain characteristics of MSCs, and stably express foreign genes.

Variation of T cell subset during acute rejection after liver transplantation in rhesus monkeys / 中国组织工程研究

Abstract BACKGROUND: Looking for the early diagnosis of acute rejection indicators after liver transplantation can assess the risk after liver transplantation quickly and effectively, and T lymphocytes play the significant role in acute rejection. OBJECTIVE:To observe the relationship between acute rejection and variation of expression of T cel subset in blood after liver transplantation in rhesus monkey. METHODS: The sixteen liver transplant models in rhesus monkey which were constructed successfuly by the method of “double-cuff and one support tube” were divided into two groups randomly: experiment group (no treated by immunosuppressant in perioperative period) and control group (treated by immunosuppressant in perioperative period). Then the blood specimen and liver tissue respectively were colected at 6, 12, 24 and 72 hours after operation. The levels of alanine transferase, aspartate aminotransferase, and total bilirubin were detected with the fuly automatic biochemical analyser. The levels of CD4+/CD8+were tested by flow cytometry. The liver tissue in rhesus monkey after liver transplantation was detected by hematoxylin-eosin staining. The degree of acute rejection was evaluated by Banff Score System. RESULTS AND CONCLUSION: Acute rejection appeared in the experiment group at 12, 24, and 72 hours after liver transplantation. Levels of alanine transferase, aspartate aminotransferase, and total bilirubin were significantly higher in the experimental group than in the control group at 24 and 72 hours after transplantation (P < 0.05). The expression of CD4+/CD8+of the experiment group and control group began to rise at 6 hours after surgery, but the experiment group increased the most obvious. CD4+/CD8+ expression was significantly greater in the experimental group than in the control group at 24 and 72 hours after transplantation (P < 0.05). Morphological pathology was severer, and Banff score was higher in the experiment group than in the control group at 72 hours (P < 0.05). These data suggested that the variation of expression of CD4+/CD8+was earlier than the change of liver tissue pathology and the change of liver function in the early acute rejection after liver transplantation. The rise of level of CD4+/CD8+ after liver transplantation indicated the increase of celular immunity in body, which had an important role in the early diagnosis of acute rejection after liver transplantation.

Interleukin-6 expression in rhesus monkey models with acute rejection after liver transplantation / 中国组织工程研究

BACKGROUND:Interleukin-6 is an important cytokine in the immune inflammatory response, strongly links with graft rejection reaction, and plays an important role in diagnosis of graft rejection and evaluation of anti-rejection. OBJECTIVE:To measure the expression of interleukin-6 in acute rejection of the liver transplantation in the rhesus monkey, and to evaluate the value as an early diagnosis of acute rejection after liver transplantation. METHODS:A total of 16 rhesus monkeys were used as the object and randomly divided into experimental group (no treated by immunosuppressant in perioperative period), and control group (treated by immunosuppressant in perioperative period). The al ograft orthotopic liver transplantation models were established in those monkeys. Then serum and liver tissue were col ected at 6, 12, 24, and 72 hours after surgery. Al ograft rejection was monitored by liver function tests, and hematoxylin-eosin staining of liver and Banff score. Final y, the expression levels of interleukin-6 were detected by enzyme linked immunosorbent assay and immunohistochemistry.RESULTS AND CONCLUSION:Acute graft rejection reaction appeared at 12, 24 and 72 hours after liver transplantation in the experimental group. The expressions of alanine aminotransferase, aspartate aminotransferase and total bilirubin were significantly higher in the experimental group than in the control group at 24 and 72 hours (P<0.05). Histological manifestations were severer and Banff score was higher in the experimental group at 72 hours than in the control group (P<0.05). Interleukin-6 levels were significantly higher in the serum and liver tissue of experimental group than in the control group at 12, 24 and 72 hours after liver transplantation (P<0.05), especial y at 72 hours. Results suggested that interleukin-6 possibly participated in rejection after liver transplantation. The expression of interleukin-6 was probably of significance in the early diagnosis of acute rejection after orthotopic liver transplantation in rhesus monkeys.

Anesthetic management in intrauterine surgery to evaluate an experimental model of myelomeningocele in non human primates (Macaca mulatta) / Anestesia em cirurgia intra-uterina para avaliar um modelo experimental de mielomeningocele em primatas não humanos (Macaca mulatta)

PURPOSE: Evaluate the anesthetic management in intrauterine surgery to induce myelomeningocele in non human primates Macaca mulatta. METHODS: A total of nine fetuses had intrauterine surgery; laminectomy was performed on them in L5 and L6. The studied variables were: maternal death, fetus death, cardiac frequency, respiratory frequency, arterial pressure, temperature, and oxygen saturation. RESULTS: No maternal or fetal deaths occurred; the only variable that was reported below the normal ranges was temperature. CONCLUSION: No maternal or fetal deaths occurred; the only variable that was reported below the normal ranges was temperature.

OBJETIVO: Avaliar o manejo anestésico em cirurgia intra-uterina para induzir mielomeningocelo em primatas não humanos, Macaca mulatta. MÉTODOS: Operaram-se um total de nove fetos in útero que foram submetidos à laminectomia em L5 e L6. As variáveis a estudar foram mortes maternas ou fetais, freqüência cardíaca e respiratória, pressão arterial, temperatura e saturação de oxigênio. RESULTADOS: Não se apresentaram mortes maternas ou fetais, a temperatura se manteve abaixo dos 36°C, não tendo repercussões no bem-estar dos macacos. CONCLUSÃO: Não ocorreu nenhum óbito materno ou fetal, sendo que a única variável abaixo do normal foi a temperatura.

Sex differences in the length ratios of metapodials in Macaca mulatta from the Taihang mountains / 解剖学报

Objective Sex difference of the length ratios of metacarpals and metatarsals in Macaca mulatta from the Taihang mountains was studied in our laboratory. Methods The lengths of 27 metacarpals (10 males, 17 females) and 30 metatarsals(12 males, 18 females) were measured from the skeletons of 30 adult Macaca mulatta. Length ratios were constructed for all possible pairings of the five bones in each individual hand and foot. One-Way ANOVA adopting SPSS13.0 for windows was used to study the sex differences of length ratios of metacarpals and metatarsals. Results For Macaca mulatta, several of these lengths ratios exhibited substantial differences between the sexes. The metacarpal(Mc) length ratios showing the largest sex differences were 2Mc∶5Mc and 4Mc∶5Mc in both hands (P<0.01), and the metatarsal(Mt) length ratios showing the largest sex difference was 1Mt:3Mt in both feet (P<0.05). Conclusion The sex differences of metacarpals and metatarsals remained when specimens of similar size were compared. It showed that body size was not the basis for these sex differences. Various facts suggested that the sex difference of length ratios in primate metapodials was associated with sex hormones exposure, possibly during prenatal development.

Validation of different gradients orientation diffusion tensor tractography of macaque monkeys with manganese-enhanced MR imaging / 中华放射学杂志

Objective Using manganese enhanced MR imaging(ME-MRI)as a standard to validate the accuracy of white matter diffusion tenor tractography(DTT)with different gradients orientations in macaque monkeys.Methods Seven adult male macaque monkeys were scanned for DTY images with different number of gradients orientations.When all data were acquired.3 μl of 0.8 mol aqueous solution of MnCl2 was injected into the primary motor cortex representing forelimb for ME-MRI scanning 48 hours later.On the images of fractional anisotropy(FA),seed region for fiber tracking was chosen according to the subcortical enhanced range on ME-MRI,then the DTT fiber tracts proiecting on axial images of FA were compared with enhancement region on T1WI of ME-MRI.Results Under the calculation method for fiber tracking in the study,different motion-probing gradients orientations led to results consistent with ME-MRI for the brain above the midbrain level.All group's concordance score of fibertracking results were good.Among them,30 and 64 orientation fiber tracking results were optimal.Below the midbrain level,there was significant difference between DTT fiber tracking and ME-MRI fiber tracking. Conclusion Different motion-probing gradients orientations do not influence DTT fiber tracking.But below the midbrain level.the results of DTT fiber tracking were not reliable according to the standard results of ME-MRI fibe tracking.

IgG immune response of Macaca mulatta monkeys after immunization with human rotavirus strains that were used for vaccine production

Background: Rotavirus acute diarrhea is a common disease in children aged 6 to 24 months, accounting for 50-70% of hospitalizations in Vietnam. Vaccines recommended by the WHO are quite expensive, so vaccination for this disease isn\u2019t widely used in Vietnam. Objectives: To evaluate the immune responses of 3 human rotavirus strains in Macaca mulatta monkeys. Subjects and method: 32 healthy monkeys aged 6-12 monthswere divided into 4 groups that received orally the G1P8 strains (Master seed- MS and Working seed- WS), G1P4 strains (MS and WS), G4P6 strains (MS and WS),and placebo. All monkeys were evaluated on general status, gastrointestinal symptoms and blood samples taken for immune analysis. Results: By ELISA technique, the Master Seed (MS) and Working Seed (WS) of Rotavirus, including G1P8 (KHI008), G1P4 (2001019210) and G4P6 (2001019203) strains showed high titer of IgG antibody in monkey at least four-fold after 3 doses of immunization. Conclusion: These 3 rotavirus strains produced by the Center for Research and Production of Vaccines and Biologicals could be candidates for vaccine production.

Determination of adventitous agent in monkeys and monkey kidney cells culture using for OPV production in 2006

Background: In 2006, Center for research and production of vaccines and biologicals \ufffd?Ha Noi produced 10 million doses of poliomyelitis vaccine met quality standard which given by World Health Organization (WHO). The monkeys for vaccine production were selected and monkey kidney cells were tested arcording to the WHO requirements (the strong moykeys with no adventitous agents). Objectives: To assess the rates of mokeys (at the Reu island in Quang Ninh province) with no adventitous agent infection and the cells using to produce type 1 poliomyelitis vaccine in 2006. Subjects and method: The study was performed by using microneutralization technique on 39 Macaca mulatta mokeys with weights were 1.5 to 2.5 kg. Results: 11 out of 39 monkeys (28,21%) are negative to SIV, SV40, poliomyelitis viruses, foamy virus as well as tuberculin test. 100% lots of primary monkey kidney cells culture produced in 2006 are free from adventitous agents arcording to the WHO requirements. Conclusion: Center for research and production of vaccines and biologicals - Ha Noi implemented strictly under the WHO guidelines on selection of monkeys and monkey kidney cells for OPV production from Sabin strains.

Changes of Circulating Immune Complex in Monkeys Infected by Simian Immunodeficiency Virus / 广州中医药大学学报

[Objective] To observe the dynamic changes of circulating immune complex (CIC) in monkeys infected by simian immunodeficiency virus (SIV). [Methods] Agglutination test of complement-sensitized yeast cell was used to determine the serum CIC level in 30 cases of monkeys, which were infected with SIVmac251 and sampled in different time-points after infection. Sixty-eight cases of normal monkeys were also examined as controls. [Results] After SIV infection, CIC can't be detected in all 30 monkeys until the 4th week, the total positive rate being 30% . In the 8th week, CIC were detected in 46.7% of these monkeys and then declined gradually in the following 12 weeks. Since the 20th week, the CIC in these monkeys maintained lower liter and lower positive rate which was close to that of the normal monkeys (about 10%). [Conclusion] CIC appeared and increased during the primary SIV infection and declined accompanying with the virus clearance from the circulalion. The formation of CIC may not benefit to the control of virus replication and the induction of anti-virus immunity; CIC has a role in the pathogenesis after SIV infection.

Experimental Study on ATA Prescription in Treating Monkey with Chronic Infection of Simian Immunodeficiency Virus / 广州中医药大学学报

[Objective] To investigate the effect of ATA, a herbal medicine compound, for the treatment of the chronic infection of simian immunodeficiency virus ( SIV) in monkeys. [ Methods ] Eight Rhesus monkeys were infected with SIVmac251 to establish monkey models with chronic infection of SIV. After 18 months, the monkeys were randomized into model group and ATA group, administered with saline and ATA 2 g?kg-1?d-1 for 56 days respectively. The changes of signs were observed, plasma levels of T-lymphocyte subsets CD4+ and CD8+ were detected by flow cytometry and the lymph node biopsy were observed under light microscope before and after treatment. [Results] After treatment, the body weight increased and the incidence of infective diarrhea decreased in ATA group, the difference being insignificant. ATA had no obvious effect on white blood cells counting and the replication number of plasma SIV, but increased the plasma levels of T-lymphocyte subsets CD4+ and CD8+(P

The effects of Nd:YAG laser on dental pulp of rhesus monkeys / 实用口腔医学杂志

Objective: To observe the effects of Nd:YAG laser on dental pulp of rhesus monkeys and compare the effects between methylene blue and silver nitrate on laser irradiation. Methods: 27 teeth of 3 monkeys were irradiated respectively by 1 592 mJ/mm 2 ,3 185 mJ/mm 2 or 4 777 mJ/mm 2 Nd:YAG laser. Befor irridiation 9 teeth were treated with methylene blue on the surfaces and 9 treated with silver nitrate.The other 3 teeth were used as controll.One hour after irradiation,pulp samples were obtained and observed with histological examination. Results: No harmful response was observed in the dental pulp following laser irradiation at 1 592 mJ/mm 3 no matter what teatment of occlusional surfaces. Mild change were observed in the pulp irradiated with the laser at 3 185 mJ/mm 2. Severe damages were found in the 4 777 mJ/mm 2 laser treated group.Methylene blue could enhance pulp damage but silver nitrate could minish the damage by isolating tooth surface from laser beam. Conclusion: Great output energy of Nd:YAG laser may cause damaging effects on dental pulp and the use of absorption dye onto the surface of teeth may enhance the effect of laser irradiation.

Isolation and culture of rhesus putative epidermal stem cells / 中国病理生理杂志

AIM: Our study was designed to look for an easy and feasible approach to isolate and culture rhesus epidermal stem cells.METHODS: Skin specimens were cut into strips and immersed into 0.25% trypsin overnight.Then transparent cuticular layeres were striped off with ophthalmic microscopic forceps.The epithelial layers were scratched,harvested and transferred to culture in skin epithelium media.The cells in 2nd-4th passages were harvested by 0.25% trypsin and relayed in IV collagen plate at 37 ℃,5% CO_2 for 20 min to harvest rapid attaching cells.Flow cytometric analysis,immunohistology and RT-PCR were conducted to identify rapid attaching cells.RESULTS: The specific protein and mRNA of epidermal stem cells(?6 integrin,K15 and ?1 integrin) were identified in rapid attaching cells.No K1/K10(marker of terminally differentiated cells) and CD71 expression were found.CONCLUSION: Our method of isolation and culture can apply for rhesus epidermal stem cells.

Specific humoral immune responses in adult rhesus monkeys vac cinated with A?_(1-15) peptide vaccine / 中国病理生理杂志

AIM: To observe the humoral immune response in adult rhesus monkey induced by A? 1-15 vaccine. METHODS: 5 adult male rhesus monkeys were injected intramuscularly with A? 1-15 vac cine at baseline and at week 2, 6, 10, 14, 18, 22. The titer and IgG isotypes of the antibody against A? 1-42 in the serum were measured with ELISA. The specificity of the antibody against A? 1-42 was determined by Wester n blotting. The A? plaques in Tg2576 transgenic mouse brain were stained with t he antisera using immunohistochemistry method. RESULTS: At the eighth week after the vaccination, antibody against A? 1-42 bega n to develop significantly i n the serum. The titers of the antibody increased following vaccine boosted and reached 1: 3 840 at the twenty-fourth week, then decreased after the terminat ion o f inocunation. The IgG1 was accounted for the highest level in the antisera pool . The antibody against A? 1-42 showed high specificity. The A? plaques in Tg2576 transgenic mouse brain were labeled with the antisera. CONCLUSION: A? 1-15 vacci ne could induce vigorously specific humoral immune responses in adult rhesus mon key.

THE COMPARATIVE STUDY ON THE DISTAL ARTICULATION OF HUMERUS AMONG Macaca mulatta, M. thibetana and M.assamensis / 解剖学报

M. mulatta, M. thibetana and M. assamensis have a part of sympatry in distribution areas and closed relationships in phylogeny, but there also exist certain differences among them in the morphology and function of skeleton. In recent years, on their locomotion and adaptive behaviors, the different viewpoints were suggested by many authors. The comparison of distal articular surface of humerus in these species were carried out in this paper. The major results based on 24 humerus including 48 sides are as follows:1. The results of the discriminant analysis suggest that the features and structures of humeral distal articulation of M. thibetana is similar to that of M. assamensis than that of M. mulatta.2. The relative width of articulation (AFT), olecranon fossa (OFW) and internal epicondyle (MEW), and the relative length of trochlea (THL) play more important rule to distinguish the three species of macaques. The. first discriminate function is: F1=13.64MEW-15.57ATF+23.500FW+14.09THL-9.63.3. The characters on distal articulation of humerus of the three species of monkeys differ evidently from the apes.4. The results of principal component analysis show that the locomotion of M. mulatta is similar to Presbytes obscurus, Hylobates and M. nemestrina, and it seems that they are more adaptive to arboreal movement than that in M. thibetana and M. assamensis based on the osteological features and functional morphology of the distal articular surface of humerus.