Characterization, isolation, and culture of spermatogonial stem cells in / 亚洲男科学杂志(英文版)

Spermatogonial stem cells (SSCs) have great applications in both reproductive and regenerative medicine. Primates including monkeys are very similar to humans with regard to physiology and pathology. Nevertheless, little is known about the isolation, the characteristics, and the culture of primate SSCs. This study was designed to identify, isolate, and culture monkey SSCs. Immunocytochemistry was used to identify markers for monkey SSCs. Glial cell line-derived neurotrophic factor family receptor alpha-1 (GFRA1)-enriched spermatogonia were isolated from monkeys, namely Macaca fascicularis (M. fascicularis), by two-step enzymatic digestion and magnetic-activated cell sorting, and they were cultured on precoated plates in the conditioned medium. Reverse transcription-polymerase chain reaction (RT-PCR), immunocytochemistry, and RNA sequencing were used to compare phenotype and transcriptomes in GFRA1-enriched spermatogonia between 0 day and 14 days of culture, and xenotransplantation was performed to evaluate the function of GFRA1-enriched spermatogonia. SSCs shared some phenotypes with rodent and human SSCs. GFRA1-enriched spermatogonia with high purity and viability were isolated from M. fascicularis testes. The freshly isolated cells expressed numerous markers for rodent SSCs, and they were cultured for 14 days. The expression of numerous SSC markers was maintained during the cultivation of GFRA1-enriched spermatogonia. RNA sequencing reflected a 97.3% similarity in global gene profiles between 0 day and 14 days of culture. The xenotransplantation assay indicated that the GFRA1-enriched spermatogonia formed colonies and proliferated in vivo in the recipient c-Kit

Do males bond? A study of male-male relationships in Nicobar long-tailed macaques Macaca fascicularis umbrosus

Due to their unique properties, carbon nanotubes (CNTs) are being widely explored for industrial and medicalapplications. This has necessitated a thorough assessment of the effect of CNTs on human and animalphysiology and health. Impact of CNTs on epithelial tight junctions has not been evaluated in the context oftheir toxic effects in many biological systems. In the present study, we examined the effect of acid functionalized single-walled carbon nanotubes (AF-SWCNTs) on the function and expression of two tight junctionproteins (ZO-1 and occludin) in the Madin-Darby canine kidney (MDCK) cell line. Treatment of MDCK cellswith AF-SWCNT resulted in a downregulation of tight junction proteins, decreased trans-epithelial electricalresistance (TER), increased paracellular permeability, and disruption of tight junctions. Taken together, ourdata demonstrate that AF-SWCNT disrupts tight junction barrier by downregulating tight junction proteins inMDCK epithelial cells.

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.

Clinical effectiveness of live preparation of lactobacillus in treatment of bacterial vaginosis in pregnancy: A meta-analysis

Objective: To evaluate the clinical effectiveness of live preparation of lactobacillus in treatment of bacterial vaginosis in pregnancy. Methods: Randomized controlled trials of live preparation of lactobacillus in the treatment of bacterial vaginosis in pregnancy were collect by searching PubMed, Web of Science, OVID, CNKI, WanFang, VIP, CBM and Elsevier databases. Quality of the included trials were evaluated by two researchers independently, and data were extracted according to Cochrane systematic evaluation. RevMan 5.3 software was used for meta-analysis. Results: Twenty-one randomized controlled trials involving 2 930 patients were included, which showed that there was significant difference in the clinical effectiveness between vaginal medication of live preparation of lactobacillus and vaginal medication of metronidazole [total effective rate (RR=1.05, 95% CI: 1.02-1.07, P=0.000 4]; significant differences were found in premature delivery rate (RR=0.49, 95% CI: 0.32-0.73, P=0.000 4), premature rupture of membrane rate (RR=0.54, 95% CI: 0.38-0.77, P=0.000 7), infant of low-birth weight rate (RR=0.45, 95% CI: 0.22-0.94, P=0.03), puerperal infection rate (RR=0.60, 95% CI: 0.39-0.94, P=0.03) between the two groups. Conclusions: Vaginal medication of live preparation of lactobacillus was more clinically effective than vaginal medication of metronidazole for bacterial vaginosis in pregnancy. Live preparation of lactobacillus is associated with a lower premature delivery rate, a lower premature rupture of membrane rate, a lower low-birth weight rate and a lower puerperal infection rate.

Comparative safety assessments of the biosimilar APZ001 and Erbitux in pre-clinical animal models

Abstract Purpose: To evaluate the toxicity of Erbitux as well as its biosimilar APZ001 antibody (APZ001) in pre-clinical animal models including mice, rabbits and cynomolgus monkeys. Methods: We performed analysis of normal behavior activity, autonomic and non-autonomic nervous functions, nervous-muscle functions, nervous excitability and sensorimotor functions on CD-1 mice. Subsequently, we studied that effects of APZ001 and Erbitux on respiratory system, cardiovascular system and kidney in Cynomolgus monkey models and performed local tolerance experiments on New Zealand rabbits. Results: The comparisons between APZ001 and Erbitux showed no significant differences in mice autonomic nervous system, nervous muscle functions, non-autonomic nervous functions, nervous excitability and sensorimotor functions between treated and untreated group (p>0.05). APZ001 and Erbitux showed negative effect on CD-1 mice in the present of pentobarbital sodium anesthesia (p>0.05). Single administrations of high, medium or low doses of APZ001 did not lead to monkey urine volume alterations (p>0.05). In human tissues, APZ001 and Erbitux showed positive signals in endocardium, lung type II alveolar epithelial cell and surrounding vessels, but showed negative results in kidney and liver tissues. No hemolysis phenomenon and serious side-effects in vessels and muscles were observed in rabbits when administrated with APZ001 and Erbitux respectively. Conclusion: The safety comparisons between APZ001 antibody and Erbitux showed that these two antibodies showed highly similarities in mice, rabbits and cynomolgus monkey animal models in consideration of pharmaceutical effects, indicating APZ001 might be a suitable substitute for Erbitux.

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.

Investigation of natural depression model under the attack-yield behavior of caged cynomolgus monkeys / 上海交通大学学报（医学版）

Objective • To investigate the natural depression model of Macaca fascicularis induced by submissive-aggressive behaviors. Methods • In a simulated wild environment, two Macaca fascicularis groups (G1 and G2) were respectively established. The two groups consisted of 19 and 14 Macaca fascicularis respectively. The effective frequency of each group's submissive-aggressive behaviors was observed and recorded, the matrix of submissive-aggressive behaviors was analyzed, each individual of the group was calculated separately by David's score(DS), according to the level of individual DS, each group was divided into attack group and yield group; the behavior changes of each individual in 7 periods were recorded by the focus observation and the correlation analysis of 11 different behaviors was carried out. Results • After the formation of these groups, there was fierce conflict between the cynomolgus monkeys. 1 122 and 1 409 submissive-aggressive behaviors were recorded in the two groups respectively. In the submissive-aggressive behavior matrix, the differences between high and low DS in anxiety behavior (t1=-4.053, P1=0.005; t2=-3.041, P2=0.012), conflict behavior (t1=8.478, P1=0.018; t2=7.651, P2=0.002), depression behavior (t1=-3.691, P1=0.006; t2=-2.431, P2=0.045) and exercise behavior (t1=9.639, P1=0.007; t2=3.568; P2=0.002) were statistically significant. Conclusion • The natural depression model of caged Macaca fascicularis is a social defeat model induced by yield-attack behavior.

Assessment of effects of 7,8-dihydroxyflavone on the striatum in normal cynomolgus monkeys with 99TCm-TRODAT-1 SPECT/CT imaging / 中华核医学与分子影像杂志

Objective To assess the effects of 7,8-dihydroxyflavone (7,8-DHF) on the striatum (ST) in normal cynomolgus monkeys using 99Tcm-TRODAT-1 imaging.Methods A total of six healthy female cynomolgus monkeys were included in this study.Three of them were fed with normal food (control group),and the other three were given oral administration of 7,8-DHF in addition to normal food (experimental group).The SPECT/CT imaging was performed at different time after 99Tcm-TRODAT-1 injection.The ROI of ST was drawn on images of 3 consecutive transverse slices that could be visualized best.The cerebellum (CB) was taken as the background reference area.The radioactivity uptake ratios of ST/CB at 1,3,4 and 5 h were calculated respectively.Paired-t test was used to analyze the data.Results ST radioactive uptake ratios showed continuing increase on the delay images.ST/CB uptake ratios of the control group at 1,3,4 and 5 h were 1.43±0.04,1.82±0.06,2.04±0.12,2.42±0.23,respectively,and those of the experimental group were 1.35±0.08,2.40±0.09,2.74±0.13 and 3.25±0.15 respectively.There was no significant difference between the two groups at 1 h (t =2.57,P＞0.05),while ST/CB uptake ratios of the experimental group at 3,4 and 5 h were significantly higher (t values:2.77,2.87 and 2.92,all P＜0.05).Conclusion 99Tcm-TRODAT-1 SPECT/CT imaging can be used to assess the DAT activation effect by 7,8-DHF on ST of cynomolgus monkeys.

Study on SCARB1 mediated coagulation dysregulation in recipients after liver xenotransplantation / 器官移植

To investigate the changes in the expression levels of scavenger receptor class B member 1 (SCARB 1) in the liver tissues before and after liver xenotransplantation and analyze the relationship between the variations in the SCARB1 expression and coagulation regulating dysfunction in the recipients.Methods The Wuzhishan miniature pig with α-1,3-galactosyltransferase gene-knockout(GTKO) was utilized as the donor and Macaca thibetana was chosen as the recipient.Heterotopic auxiliary liver xenotransplantation models were established.The liver tissue specimen was collected before and after liver xenotransplantation.Primary hepatocytes were extracted from the pig using collagenase digestion method.Human peripheral blood mononuclear cells were obtained by immunomagnetic bead sorting.These two types of cells were co-cultured and supplemented with human plasma to establish cell models with coagulation regulating dysfunction following liver xenotransplantation.Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western blot were performed to quantitatively measure and statistically compared the expression levels of messenger ribonucleic acid (mRNA) and protein of SCARB1 in the tissue and cell samples.At the cellular level,the expression of SCARB 1 was interfered by lentiviral vector.The coagulation time was detected to validate the effect upon coagulation function.Results The expression levels of SCARB1 mRNA and protein were significantly down-regulated after liver xenotransplantation (both P＜0.05).In the cell models,the expression levels of SCARB1 mRNA and protein in the porcine hepatocytes co-cultured with human monocytes were significantly down-regulated compared with those in porcine hepatocytes without intervention (both P＜0.05).Compared with the non-intervention group,the coagulation time was significantly prolonged after the expression of SCARB1 was interfered by lentiviral vector (P＜0.05).Conclusions The down-regulated expression of SCARB1 in the liver graft is one of the main causes of mediating coagulation regulating dysfunction.Intervention of SCARB1 expression contributes to resolve the coagulation regulating dysfunction in the recipients after liver xenotransplantation.

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.

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 .

Safty evaluation, migration and distribution of human bone marrow derived-mesenchymal stem cells in the cns of young macaca fascicularis / 中国比较医学杂志

Objective To evalutate the safty of hBMSCs transpalntation and to observe their migration and distribution in the brain of young macaca fascicularis. To establish a new technology platform and theoretical basis for the treatment of central nervous system diseases in children. Methods Labelled hBMSCs were transplanted into the striatum of young macaca fascicularis. Brain sections were examined to evalutate the inflammatory reaction and immunological rejection of local injection sites by HE observation and immunohistochemical staining. Migration and distribution of transplanted?hBMSCs was observed by real?time fluorescence quantitative PCR of male DNA and fluorescence microscope. Results The results showed that the direct intracerebral injection of hBMSCs did not cause systemic symptoms in animals. There is no inflammatory reaction and immunological rejection was detected, and degeneration and necrosis of neural cells and proliferation of glial cells were absent in the local injection sites. The transplanted hBMSCs survived, and migrated into the brain after 4 weeks transplantation. Its migration and distribution have certain regularity and were overlapping between transplant recipients. In addtion, hBMSCs tended to extend rostrally into the forebrain and showed preference of migrating toward the blood vessels and below the ependyma. Conculsions Intracerebral transplantation of hBMSCs is safe. And hBMSCs can survive and migrate into the brain.

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.

Establishment and evaluation of a rhesus monkey model of experimental type 2 diabetes mellitus / 中国组织工程研究

BACKGROUND:At present, there are few reports about the non-human primate models of type 2 diabetes mel itus in domestic and abroad, so it lacks of standardized production methods and evaluation criteria. OBJECTIVE:To establish a safe and effective type 2 diabetes mel itus model of rhesus monkey and evaluation method. METHODS:Twelve rhesus monkeys were randomly assigned to experimental group (n=9) and control group (n=3). Rhesus monkeys in the experimental group were fed with high-glucose and high-fat diet for 4 weeks, and intraperitoneal y injected with 30 mg/kg streptozotocin to establish models of type 2 diabetes mel itus. Rhesus monkeys in the control group were fed with an equal volume of physiological saline. At 12 weeks after injection, peripheral blood serum was col ected to measure fasting blood glucose, lipids, insulin, and C-peptide levels. Intravenous glucose tolerance test and C-peptide release test were used to detect pancreatic gland and pancreatic islet function. Histopathological examination was performed in pancreas, kidney and liver. RESULTS AND CONCLUSION:(1) 12 weeks after injection, fasting blood glucose, triglycerides, and total cholesterol levels were significantly higher in the experimental group than in the control group (P<0.05). Insulin and C-peptide levels were significantly lower in the experimental group than in the control group (P<0.05). (2) The area under the curve for intravenous glucose tolerance test was increased in the experimental group than in the control group (P<0.05). The area under the curve for C-peptide response test was significantly reduced in the experimental group than in the control group (P<0.05). (3) The pathological sections of pancreas, kidney and liver showed typical pathological changes of diabetes in the experimental group. (4) It is confirmed that we got high achievement about rhesus monkey models of type 2 diabetes mel itus made by high-glucose and high-fat diet combined with low-dose streptozotocin. It is a feasible, safe and effective method.

Small hairpin RNA targeting inhibition of NF-κB gene in endometriosis therapy of Macaca fascicularis / 中华妇产科杂志

Objective To observe the therapeutic effect of NF-κB gene short hairpin RNA (shRNA) on endometriosis and identify the function of NF-κB on the maintenance and development of endometriosis in Macaca fascicularis.Methods The Macaca fascicularis model of endometriosis was developed,which divided into experimental group,negative control group and simple model group.The high specificity adenovirus vector mediated shRNA targeting NF-κB gene and negative control shRNA adenovirus with no-load NF-κB gene were synthesised.The experimental group injected the adenovirus which carried the NF-κB shRNA into the endometriosis lesions under laparoscopy surgery,the negative control group with no-load shRNA adenovirus and the simple models group injected with normal saline.Four weeks later after the injection,an observed operation was performed through laparoscopy and some lesions were collected.The CD34 immunohistochemistry of these lesions were done to detect the microvessel density,then the variation of the microvessel density among each group were observed.The expression of the NF-κB and proliferating cell nuclear antigen (PCNA) were detected through western blot.Results First,the Macaca fascicularis model of endometriosis was successful developed,and the experimental group has an evident atrophy in ectopic lesions compared with the previous.The lesions' microvessel density in experimental group decreased evidently compared with the negative control group and simple model group (0.002 0±0.000 3 versus 0.021 9±0.002 6 versus 0.024 5±0.003 3),and the differences was statistically significant (P＜0.01).The expression of PCNA (0.37±0.17 versus 0.57±0.26 versus 0.57±0.28) and NF-κB (0.338 ± 0.174 versus 0.678 ± 0.021 versus 0.645 ±0.098) in experiment group was lower than the negative control group and simple model group,the differences were statistically significant (all P＜0.01).Conclusion Through targeting suppressed the NF-κB gene expression by NF-κB shRNA,we can inhibit the development of endometriosis through reducing the ability of angiogenesis and cell proliferation of ectopic endometrial cells.

Long-term toxicity of fully humanized anti-human tumor necrosis factor-αmonoclonal antibody for injection in cynomolgus monkeys / 中国药理学与毒理学杂志

OBJECTIVE To evaluate the long-term toxicity of fully human anti-human tumor necrosis factor-α monoclonal antibody(anti-hTNF-α FHMA)for injection in cynomolgus monkeys. METHODS Forty cynomolgus monkeys were randomly divided into 5 groups (4 males and 4 females in each group):negative control group,adalimumab 10 mg·kg-1 group,anti-hTNF-αFHMA 2,10 and 50 mg·kg-1 groups. Cynomolgus monkeys in each group were injected sc once a week for 5 consecutive times, followed by 4 weeks of recovery. During the test,general clinical observation,body mass,body temperature,electrocardiogram(ECG),hematology,coagulation function,blood biochemistry,urine, ophthalmology,immune index,and pathological changes in organs and tissues were observed. At the same time,plasma drug concentrations were detected and the toxicokinetics parameters were analyzed. RESULTS No significant toxicological changes related to drugs were observed in general clinical observation,body mass,body temperature,ECG,ophthalmic examination,blood cell counts,coagu?lation function,blood biochemistry,urine analysis,lymphocyte subsets,cytokines,serum immuno?globulin,serum complement. Neutralizing anti-drug antibody(ADA)could be detected in adalimumab group and anti-hTNF-αFHMA groups. Anti-hTNF-αFHMA showed linear dynamic characteristics in cyno?molgus monkeys. At the same dose(10 mg·kg-1),anti-hTNF-αFHMA had similar immunogenicity and kinetics characteristics to adalimumab. CONCLUSION The level of anti-hTNF-α FHMA at which no adverse effect was observed was 50 mg · kg-1,which is equivalent to 75 times clinical dosage of quasi (0.67 mg·kg-1),which suggests that anti-hTNF-αFHMA be safe in clinical use.

Comparative Morphological and Anatomical Study on Thymus Gland of Human and Primate.

Context: The comparative morphological and anatomical study on thymus was carried out in human and primate. The prenatal stage of Macaca radiata was selected for the present study. Study Design: Cross sectional analytical type of study. Place and Period of study: Department of Anatomy, Dr. A.L.M. PG Institute of Basic Medical Sciences, Chennai from July 1999 to June 2000. Materials: The comparative morphology and anatomy of thymus of human embryonic, 10 weeks, 15 weeks and prenatal foetuses, and monkey foetus was carried out. Methods: Comparative micro-anatomical study was done by paraffin processing method. The sections were stained as per the method published by Culling (1974). Results: In monkey foetus, the thymus gland is slightly elongated, whereas in human foetuses it is not elongated and oval in shape. The size of the thymus is larger in human foetuses than monkey foetus. In both cases cells are parenchymal in nature. Due to spatial organization in human foetuses, the lymphocytes aggregation is more in cortex than in medulla. In monkey foetus the lymphocyte aggregation is simpler in arrangement through spatial organization is much less.

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.

The establishing of renal transplantation model in non-human primate / 中国基层医药

Objective To explore renal transplantation model in non-human primate cynomolgus monkeys.Methods 50 non-human primates' kidneys were transplanted into the lower part of the abdomen with end-to-side anastomosis of renal artery to aorta and renal vein to inferior vena eava,and with end-to-end anastomosis of ureter to bladder.Results In the 50 cases,1 case death as accident of anesthesia;7 cases with postoperative complications,and all with creatinine sudden rise,after ultrasonic examinations showed that 2 cases with renal vein thrombosis,and 5 cases appeared urinary leakage.All animal models were without surgical infections,and with normal serum creatinine,urine output.Conclusion Non-human primate animal kidney transplantation model establishment method is reliable,but should pay attention to the the surgical technique training,complications prevention.The model is valuable for application in the research of immune tolerance,heterogeneous transplant.