Neuroprotective effect of novel Rho kinase inhibitor FSD-C10 on a mouse model of Alzheimer disease

AIM: To explore the neuroprotective effect of novel Rho kinase inhibitor FSD-C10 on Alzheimer disease (AD) model of APP/PS1 double transgenic (Tg) mice. METHODS: Male APP/PS1 Tg mice (n=20) at 8 months of age were randomly divided into 2 groups:model group and FSD-C10 treatment group. The mice were treated with normal saline or FSD-C10 (25 mg·kg^-1·d^-1) by intraperitoneal injection, once daily for 2 months. Age-and sex-matched wild-type (WT) mice without treatment were used as the control. The Morris water maze (MWM) test and SMART 3.0 behavioral record system were applied to examine and analyze the spatial cognitive function of the mice. The protein levels and distribution of Aβ, p-tau and synapse-associated proteins such as synaptophysin, α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) and postsynaptic density protein 95 (PSD-95) were determined by immunofluorescence staining. The protein levels of phosphorylated amyloid precursor protein at Thr668[p-APP(Thr668)], beta-site APP-cleaving enzyme 1 (BACE1) and synapse-associated proteins in the brain were analyzed by Western blot. RESULTS: Compared with model group, FSD-C10 treatment significantly improved the cognitive function of the APP/PS1 Tg mice, accompanied by reduced Aβ deposition and p-tau level, increased protein level of p-APP (Thr668) in the central nervous system, decreased expression of BACE1, and increased expression of synapse-associated proteins in the brain of the mice (P<0.05). CONCLUSION: FSD-C10 has neuroprotective potential in the APP/PS1 Tg mice. The mechanism may be related to enhancing the non-amyloidogenic APP cleavage pathway, reducing the production of Aβ oligomers, the deposition of senile plaques and the amount of tau protein, up-regulating synapse-associated proteins, and increasing synaptic plasticity.     

Characterisation of adiponectin and its receptors in the bovine mammary gland and in milk

Adiponectin is an adipocyte-derived hormone, which circulates in the form of homo-multimers. The individual oligomers have a distinct profile of activity, playing crucial roles in several biological processes, including metabolism and inflammation. Adiponectin exerts many of its effects by interacting with the receptors, AdipoR1 and AdipoR2. In the present study, mRNA expression of adiponectin, AdipoR1 and AdipoR2 was evaluated by quantitative PCR in different areas of the mammary gland in healthy lactating cows. The adiponectin isoforms in milk and blood were investigated by Western blotting and 2D-electrophoresis, and the presence of adiponectin protein was determined by immunohistochemistry.Low level expression of adiponectin mRNA was found in all areas of bovine mammary gland tissues examined. AdipoR1 and AdipoR2 mRNAs were also detected in mammary tissues and their expression was particularly prominent in the parenchyma and cistern. Western blotting revealed a heterogeneous electrophoretic pattern, indicating that different adiponectin isoforms exist in milk, compared with blood. In particular, milk shows a low molecular weight isoform of adiponectin, corresponding to the globular domain. Adiponectin in milk is characterised by a more complex 2D electrophoretic pattern, compared with blood, as illustrated by the presence of proteins of different molecular weights and isoelectric points. Adiponectin protein was detected by immunohistochemistry in epithelial cells lining the secretory alveoli, in secretum within the alveolar lumen and in small peripheral nerves. The study findings support a role for adiponectin in regulating metabolism and immunity of the bovine mammary gland and potentially the calf intestine, following ingestion of milk.     

Serological survey of Toxoplasma gondii in captive nonhuman primates in zoos in Spain

Toxoplasma gondii is a widely distributed zoonotic protozoan parasite, which can affect most warm-blooded species. Some species of non-human primates (NHPs) are highly susceptible to T. gondii infection. The aim of the study was to determine the seroprevalence and risk factors associated with T. gondii infection in NHPs housed in zoos in Spain. Sera from 189 NHPs belonging to 33 species were collected in eight zoos. Additionally, 10 of the 189 animals were longitudinally sampled. Anti-T. gondii antibodies were detected in 48 NHPs (25.4%; confidence interval of 95% (CI_95%): 19.2–31.6) using a modified agglutination test (MAT; cut-off = 25). Seropositive animals had titers of 25 (6.3%), 50 (8.3%), 100 (8.3%) and ≥500 (68.8%). Seropositivity was detected in 15 of the 33 species (45.5%). Of the 10 NHPs sampled more than once, two animals (one Barbary macaque [Macaca sylvanus] and one common chimpanzee [Pan toglodytes]) seroconverted along the study period, while one seropositive chimpanzee increased antibody titers over time. The Hominidae family (OR = 5.9; CI_95%: 2.7–12.8) and sex (females) (OR = 2.1; CI_95%: 1.1–4.1) were risk factors potentially associated with seropositivity to T. gondii. Our results evince a widespread circulation of T. gondii in NHPs in zoos in Spain, which may be of conservation concern. Control measures should be implemented to minimize the risk of exposure of these species to T. gondii.     

Neuroprotective effect of fasudil combined with bone marrow-derived neural stem cells on mice with experimental autoimmune encephalomyelitis

AIM: To explore the neuroprotective effect of fasudil combined with bone marrow-derived neural stem cells (BM-NSCs) on the mice with experimental autoimmune encephalomyelitis (EAE). METHODS: Female C57BL/6 mice (8~10 weeks old, n=32) were immunized with myelin oligodendrocyte glycoprotein 35-55 (MOG_35-55) to establish chronic EAE model. The mice were randomly divided into control (ddH₂O) group, fasudil group, BM-NSCs group, and fasudil+BM-NSCs group. The clinical score and body weight were recorded every other day. The expression of neurotrophic factors was determined by immunofluorescence staining. RESULTS: In comparison with ddH₂O group, fasudil combined with BM-NSCs delayed onset and ameliorated severity of EAE. The numbers of brain-derived neurotrophic factor, glial cell-derived neurotrophic factor, nerve growth factor, neurotrophin-3 and ciliary neurotrophic factor positive cells in fasudil group, BM-NSCs group and fasudil+BM-NSCs group were all increased in various extents. In particularly, the expression of these neurotrophic factors in fasudil+BM-NSCs group was significantly higher than that in the mice treated with fasudil or BM-NSCs alone (P<0.01). CONCLUSION: Fasudil combined with BM-NSCs promotes the expression of neurotrophic factors and improves microenvironment of central nervous system, thus playing a positive role in neural restoration and regeneration through a synergistic and superimposed effect.     

哺乳动物原始卵泡生长启动的调控

哺乳动物卵泡发育是一个被众多内分泌、旁分泌和自分泌因素精确调控的生理过程。原始卵泡生长启动可能受卵巢内在因子，如生长因子等的调控，而不是通常认为的FSH(促卵泡素)；原始卵泡细胞连接和原始卵泡的细胞(卵母细胞和颗粒细胞)之间相互作用也参与调节原始卵泡的生长启动     

用冻干精子生产ICSI小鼠

前期研究中，我们采用小鼠卵母细胞胞浆内精子注射（ICSI）技术，成功地用新鲜精子获得了生理健康的ICSI小鼠。在此基础上，本文结合细胞冻干技术，进行了冻干精子ICSI生产试管小鼠的尝试。B6D2F1成年公鼠的精子在Tris-HCl-EDTA（pH8.2）溶液中冻干4 h后解冻，将其精子头注入到KM小鼠成熟卵母细胞的胞质中。6 h后，83.0%的卵子受精。用CZB溶液体外培养发现，冻干精子生产的胚胎，体外发育至2-C (92.0% vs 99.5%)、4-C（52.7% vs 97.2%）、桑椹胚（36.6% vs 86.3%）和囊胚的比例（21.4% vs 68.7%）极显著地（p<0.01）低于新鲜精子生产的ICSI胚胎。移植2-C期冻干精子ICSI胚胎检测体内发育，结果发现，D10的着床率为63.0%（17/27）；胎鼠的出生比例为21.7%（13/60），这些ICSI小鼠成年后的生理和生殖能力正常。试验结果说明，精子冻干后，仍能生产ICSI动物，冻干技术可以用于哺乳动物精子的保存。     

卵泡大小和颗粒细胞对山羊卵泡卵母细胞体外成熟和体外受精的影响

用切割法采集卵泡液,收集卵丘一卵母细胞复合体（Cumulus oocytes comlexs,COCs）和自然裸卵,将部分COCs去除卵丘细胞获得机械裸卵,COCs放入体外成熟培养液中培养为成熟卵母细胞,加入获能的精子液,进行体外受精。结果表明：卵母细胞的体外成熟率和卵裂率与卵泡直径密切相关,大卵泡（80．95％,P〈0．01）和中等卵泡（75．50％,P〈0．05）的卵母细胞成熟率高于小卵泡（50．27％）;犬卯泡（53．53％）和中等卵泡（47．13％）的卵裂率显著高于小卵泡的32．26％（P〈0．05）。COCs、机械裸卵和自然裸卵的体外成熟率分别为75．0％、54．2％和10．5％,差异极显著（P〈0．01）,卵裂率分别为53．8％、10．8％和0％,差异极显著（P〈0．01）。对照组和1×10^5、1×10^6个／mL颗粒细胞组卵母细胞体外成熟率分别为68．6％、69．6％和67．8％,无显著差异（P〉0．05）,但均显著高于1×10^7个／mL（51．5％,P〈0．05）和1×10^10个／mL（35．5％,P〈0．05）颗粒细胞组,但各组间的体外受精率无显著差异（P〉0．05）。结果提示,大卵泡和中卵泡的卵母细胞的体外成熟率和卵裂率显著高于小卵泡,体外成熟培养液中添加高浓度的颗粒细胞能显著抑制卵母细胞的体外成熟。     

Inflammation and coagulation activation are associated with gallstone formation in golden hamsters

AIM: To elucidate the relationship of inflammation, coagulation and cholesterol gallstone formation. METHODS: Hamsters were divided into four groups: control group (feeding normal diet), lithogenic diet (LD) 2 weeks group, LD 6 weeks group and LD+aspirin 6 weeks group. Gallstone incidence, antithrombin antigen (AT-Ⅲ:Ag), antithrombin activity (AT-Ⅲ:Ac), thrombin (F-Ⅱa:Ac), plasminogen activator inhibitor activity (PAI:Ac), plasmin activity (Plm:Ac), D-dimer:Ag and C-reactive protein (CRP) in gallbladder bile were observed as read-out parameters. RESULTS: The incidence of gallstones in control group, LD 2 weeks group, LD 6 weeks group and LD+aspirin group were 0%, 20%, 73% and 25%, respectively. AT-Ⅲ:Ag, F-Ⅱa:Ac, D-dimer and CRP in LD 2 weeks group and LD 6 weeks group were significantly higher than those in the control group (P<0.01). AT-Ⅲ:Ag, PAI:Ac, Plm:Ac, D-dimer and CRP in LD 6 weeks group were significantly higher than those in LD 2 weeks group (P<0.01). AT-Ⅲ:Ag, F-Ⅱa:Ac, PAI:Ac, D-dimer and CRP were strikingly decreased in LD+aspirin group compared with those in LD 6 weeks group (P<0.01). The level of CRP was closely correlated with the concentration of D-dimer (γ=0.752, P<0.01). CONCLUSIONS: Coagulation cascade is activated and crosslinked fibrin forms in gallbladder bile before gallstone formation. “Lithogenic bile” might play a central role in the inflammation of gallbladder mucosa, which activates extrinsic coagulation pathway and leads cholesterol gallstone formation.     

家庭宠物对弓形虫病流行的影响

随着经济的发展,家庭盛行豢养宠物已经成为一种时尚,与宠物有关的寄生虫病亦随之而来,人群感染弓形虫病的比率逐年上升,弓形虫的潜在危害直接影响着人民的身体健康。如何正确认识弓形虫对人类的危害,对疾病预防和控制有着重要的意义。