Pharmacokinetic study of baicalein after oral administration in monkeys

Baicalein, a flavonoid originally isolated from the root of Scutellaria baicalensis Georgi, has numerous pharmacological activities. Up to now, several studies regarding the pharmacokinetic profiles of baicalein have been described, while there is no such study reported in monkey, the species which is more similar to human. The purpose of this study was to investigate the pharmacokinetic profiles of baicalein after oral administration in monkeys. After orally administrating three doses of baicalein in monkeys, multi-peaks of the plasma concentration-time curves were observed and the non-linear pharmacokinetics for baicalein and its metabolite baicalin were found at doses of 50-500mg/kg. In order to calculate the absolute bioavailability, the intravenous pharmacokinetic study was also carried out after intravenous administration of 10mg/kg baicalein. The absolute bioavailability of baicalein in different doses was ranged from 13.1% to 23.0%. In this study, baicalein and baicalin were determined by LC-MS method. The chromatographic separation was performed on Agilent Poroshell 120 SB-C18 column (2.7μm, 2.1×50mm). Baicalein and baicalin were detected by single quadrupole mass spectrometer equipment with electrospray ionization interface with the selected ion monitoring mode. The assay was linear for both baicalein and baicalin with the correlation coefficients>0.99. The intra- and inter-day precisions for baicalein and baicalin were all less than 15% by relative standard deviation. The analytes were stable during samples storage and handling, and no matrix effects were observed. The method we developed in this study was sensitive, precise, stable and producible.     

生物反应器悬浮培养Vero细胞制备水貂犬瘟热活疫苗

为实现利用生物反应器制备规模化、自动化生产水貂犬瘟热Vero活疫苗,在7 L生物反应器中悬浮培养Vero细胞,并考察培养基、细胞初始接种密度、培养方式、病毒感染复数和感染时间等参数对细胞增殖、病毒滴度以及细胞代谢的影响。结果显示,在含有5 g/L微载体的DMEM培养基中接种Vero细胞(30～40 cells/球),设定p H、温度、溶氧值和转速分别为7.2、37℃、50%和55 r/min,培养方式为批次培养(0～48 h)+灌流培养(48～96 h)组合方式,培养Vero细胞3～4 d或细胞密度达到200 cells/球以上时,按照MOI=0.0001～0.001吸附接毒,调低温度(33℃)继续培养100～120 h,即可获得高滴度病毒液(10~(6.5)～10~(7.2)TCID_(50)/0.1 m L),经无菌检验合格后,配制水貂犬瘟热Vero细胞活疫苗(CDV3-CL株,悬浮培养),疫苗符合《中国兽药典》三部(2015版)的规定。试验建立了7 L生物反应器悬浮培养Vero细胞制备水貂犬瘟热活疫苗的新工艺,为进一步规模化生产奠定了基础。     

Isolation and identification of a South China strain of Plasmodium inui from Macaca fascicularis

Southeast Asian macaques are hosts of a number of Plasmodium infections, some of which are transmittable to humans. During examination of blood films of five wild-caught long-tailed macaques Macaca fascicularis from South China, malaria infection was detected in one of the monkeys. In order to isolate this parasite for identification and characterization, we experimentally passed this parasite through both Assamese (M. assamensis) and rhesus (M. mulatta) monkeys by intravenous injection of infected blood. This parasite morphologically resembled Plasmodium inui, and had a typical 72 h quartan periodicity. This parasite was infective to Anopheles dirus mosquitoes, and salivary gland sporozoites appeared 13 days post feeding. Feeding by 20 infected An. dirus mosquitoes on another Assamese monkey produced infection with a prepatent period of 8 days. Molecular analysis of the small subunit rRNA genes and the mitochondrial genome confirmed this parasite as an isolate of P. inui. In spleen-intact macaques, the infection had a protracted duration with parasites being detected during the rearing of the infected monkeys for over two years. In summary, this study identified a P. inui isolate and successfully passed this parasite through Assamese monkeys by both intravenous inoculation and mosquito transmission.     

恒河猴多部位不脱钙骨组织学观察

观察恒河猴骨不同部位不脱钙骨制片的骨组织，方法采用健康成年雄性恒河猴，进行骨荧光双标记后取材，对桡骨远端，中段和跟骨、肋骨及腰椎行 骨包埋，分别行切片，磨片及染色和观察。结论本实验对恒河猴动物模型的骨组织学作了补充。     

Molecular characterization of Blastocystis isolates from children and rhesus monkeys in Kathmandu, Nepal

To investigate the possible transmission of Blastocystis organisms between local rhesus monkeys and children in Kathmandu, Nepal, we compared the subtype (ST) and sequence of Blastocystis isolates from children with gastrointestinal symptoms and local rhesus monkeys. Twenty and 10 Blastocystis isolates were established from 82 and 10 fecal samples obtained from children and monkeys, respectively. Subtype analysis with seven sequence-tagged site (STS) primers indicated that the prevalence of Blastocystis sp. ST1, ST2 and ST3 was 20%, 20% and 60% in the child isolates, respectively. In contrast to human isolates, ST3 was not found in monkey isolates and the prevalence of ST1 and ST2 was 50% and 70%, respectively, including three mixed STs1 and 2 and one isolate not amplified by any STS primers, respectively. Since Blastocystis sp. ST2 has been reported as the most dominant genotype in the survey of Blastocystis infection among the various monkey species, sequence comparison of the 150 bp variable region of the small subunit rRNA (SSU rRNA) gene was conducted among ST2 isolates of humans and monkeys. Sequence alignment of 24 clones developed from ST2 isolates of 4 humans and 4 monkeys showed three distinct subgroups, defined as ST2A, ST2B and ST2C. These three subgroups were shared between the child and monkey isolates. These results suggest that the local rhesus monkeys are a possible source of Blastocystis sp. ST2 infection of humans in Kathmandu.     

猴卡氏肺孢子虫肺炎模型的建立及蒿甲醚的疗效试验

从住家附近诱捕到 13只野鼠 ,经皮下注射醋酸可的松 10周 ,建立野鼠卡氏肺孢子虫肺炎模型 ,取野鼠肺组织制成接种物。 1～ 6号实验猴经气管接种 3 6× 10 6 卡氏肺孢子虫包囊 ,接种前 5d至接种后 35d注射醋酸可的松。 7～ 12号实验猴经气管接种 7 2× 10 6 卡氏肺孢子虫包囊。接种后 ,1～ 6号实验猴出现严重的临床症状 ,病理变化呈典型的卡氏肺孢子虫肺炎 ;7～ 12号实验猴为亚临床症状 ,经注射醋酸可的松后 ,则出现类似于 1～ 6号实验猴的严重症状和病理变化。实验结果表明实验猴免疫力低下时 ,接种外源性卡氏肺孢子虫包囊可导致典型的卡氏肺孢子虫肺炎 ;呈隐性感染的实验猴经免疫抑制处理后 ,亦可导致卡氏肺孢子虫肺炎。用蒿甲醚对呈卡氏肺孢子虫肺炎的实验猴进行治疗 ,取得明显的治疗效果     

A survey investigating the current practice of French health professionals regarding infection risk after monkey bites

International tourism is steadily increasing, with 15% of travellers reporting health problems when they come back. Animal bites represent 2% of consulting causes, of which 20% are due to monkey bites. The Monkey B virus (Macacine alphaherpesvirus 1) is an alphaherpesvirus (Herpesviridae, genus Simplexvirus) enzootic in macaques (Genus Macaca). Zoonotic infections with the Monkey B virus following exposure to macaques are exceptionally rare, but can cause fatal encephalomyelitis in humans. An observational survey was undertaken in 2018 to assess the practice of French health professionals regarding infection risk after monkey bites. French health professionals practicing in vaccination and rabies centres were specifically targeted for this study. Standardized questionnaires were sent by email to a sample of French health professionals. They were asked to participate on a voluntary and anonymous basis. The questionnaires requested epidemiological details and included multiple‐choice questions about the infection management of monkey bites. The response rate was 33.5%. The frequency of monkey bites in 2017 was variable with a minority of centres managing more than 6 per year (12%), 46% managing 1–5 monkey bites and 42% none. Most of the monkey bites were described as occurring in South Asia at tourist sites, on naked upper limbs, shortly after the travellers arrived at their destination. Tetanus status verification, rabies post‐exposure prophylaxis and antibiotic therapy were said to be prescribed in most cases. Knowledge about the Monkey B virus was reported as scarce for 38% of the participants. The number of monkey bites managed per year per centre varied greatly but practices regarding infectious risk after monkey bites were generally homogeneous. The risk of Monkey B virus transmission did not readily come to mind in the differential diagnosis of infection risk for many French health professionals.     

川金丝猴解剖一头骨

本研究为川金丝猴骨骼系统的一部分,标本来源于上海动物园,通过解剖进行观察金丝猴头骨各部分以及与猕猴头骨间的差异,现将结果报导如下:     

Vaccination of vervet monkeys against cutaneous leishmaniosis using recombinant Leishmania ‘major surface glycoprotein’ (gp63)

Vervet monkeys (Cercopithicus aethiops) were shown to give a positive delayed-type hypersensitivity (DHT) reaction to gp63, a major surface glycoprotein of Leishmania parasites, and also produce antibodies to the molecule following a triple vaccination with a total dose of 150 μg of recombinant gp63 mixed with Bacille Calmette Guerin (BCG). However, peripheral blood leucocytes (PBL) from these animals neither proliferated nor produced any interferon-gamma (IFN-γ) following in vitro stimulation with the antigen. Analysis of lymphocyte subsets following vaccination did not reveal any striking phenotypic alteration of cellular sub-populations in PBL. When vaccinated animals were rechallenged, via the needle, with virulent Leishmania major promastigotes containing salivary gland extracts from vector sandflies, only partial protection was achieved. We conclude from these studies that gp63 produced in Escherichia coli is a safe vaccine molecule which gives only partial protection following vaccination in the vervet monkey host. The molecule requires further improvement for vaccine and/or immunodiagnosis application.     

Occurrence of hemotropic mycoplasmas in non-human primates (Alouatta caraya,Sapajus nigritus and Callithrix jacchus) of southern Brazil

Hemoplasmas, the erythrocyte-associated mycoplasmas, have been detected in several primates, causing mostly subclinical infection. This study aimed to determine the prevalence of hemoplasma infection in captive and free-ranging monkeys from southern Brazil, as well as factors and hematological abnormalities associated with infection. Blood samples from 40 non-human primates (NHP) were tested for hemoplasmas and coinfections. An overall of 10/40 (25.0%) NHP tested positive for hemoplasmas using PCR-based assays, including 9/14 (64.3%) black howler monkeys (Alouatta caraya) and 1/24 (4.2%) black-horned capuchin (Sapajus nigritus). Infection was not statistically associated with anemia, but wild-born monkeys and male black howler monkeys were more likely to be positive when compared with captive-born animals and female black howler monkeys, respectively. The sequences from the black howler monkey hemoplasma were similar (94% identity) to the squirrel monkey hemoplasma (“Candidatus Mycoplasma kahanei”) and were phylogenetically located in a different cluster when compared to the human hemoplasma (“Candidatus Mycoplasma haemohominis”).