Echocardiographic reference ranges for sedated healthy cynomolgus monkeys (Macaca fascicularis).

Echocardiographic imaging has become the primary tool to evaluate cardiac structure and function in human and veterinary medicine. The cynomolgus monkey (Macaca fascicularis) is a nonhuman primate species frequently used in biomedical research, particularly for the study of human cardiovascular disease and toxicology, yet echocardiographic reference ranges are not available for this species. Using standard 2-dimensional and M-mode imaging, we performed echocardiographic evaluation of 118 female and 119 male cynomolgus monkeys under sedation with either ketamine hydrochloride (10 mg/ kg IM) alone or with a combination of tiletamine hydrochloride and zolazepam (4.0 mg/kg IM) and atropine sulfate (0.015 mg/kg IM). Reference ranges were developed (tolerance interval methodology) separately for each gender for heart rate, left ventricular (LV) size (interventricular septum in diastole, LV internal diameter in diastole and systole, LV free wall in diastole), left atrial diameter, and aortic diameter. LV functional parameters (fractional shortening, aortic peak flow velocity, LV ejection time, and LV preejection period) and mitral valve E point to septal separation were also measured. After normalization for body weight (1.7 to 6.3 kg), the data were analyzed for gender- and sedation-associated differences. Using a large number of healthy subjects (118 of each gender), we have developed gender-specific echocardiographic reference ranges for cynomolgus monkeys.     

Multidetector computed tomographic morphology of ovaries in cynomolgus macaques (Macaca fascicularis).

Macaques are important models for menopause and associated diseases in women. A sensitive, noninvasive technique for quantifying changes in ovarian morphology would facilitate longitudinal studies focused on the health-related sequelae of naturally occurring or experimentally induced alterations in ovarian structure and function. Multidetector computed tomography (MDCT) is a fast, non-invasive imaging technique that uses X-rays, multiple rows of detectors, and computers to generate detailed slice images of structures. The purpose of this study was to describe the utility of MDCT for reliably characterizing ovarian morphology in macaques. Five macaques were scanned using contrast-enhanced MDCT. The following characteristics were described: 1) appearance of ovaries and adjacent landmarks, 2) effects of varying technical protocols on ovarian image quality, 3) radiation doses delivered to the pelvic region during scanning, and 4) MDCT estimates of ovarian volume and antral follicle counts versus those measured directly in ovarian tissue. Ovaries were distinguishable in all MDCT scans and exhibited heterogeneous contrast enhancement. Antral follicles appeared as focal areas of nonenhancement. Ovarian image quality with 5 pediatric scanning protocols was sufficient for discriminating ovarian margins. Pelvic region radiation doses ranged from 0.5 to 0.7 rad. Antral follicles counted using MDCT ranged from 3 to 5 compared with 3 to 4 counted using histology. Ovarian volumes measured using MDCT ranged from 0.41 to 0.67 ml compared with 0.40 to 0.65 ml by water displacement. MDCT is a promising technique for measuring longitudinal changes in macaque ovarian morphology reliably and noninvasively.     

Echocardiographic diagnosis of muscular ventricular septal defect in a cynomolgus monkey (Macaca fascicularis).

There have been only a few case reports of heart disease in monkeys. In the case we present, a cardiac murmur was auscultated in a 26-month-old male cynomolgus monkey during a routine physical examination. Echocardiography of this monkey revealed features indicating blood flow from the left ventricle to the right ventricle through the ventricular septum. These findings clarified that the animal had a muscular interventricular septal defect.     

Idiopathic new bone formation in the femoral shafts of a cynomolgus monkey (Macaca fascicularis).

A 6.5-y-old cynomolgus monkey was referred to the Veterinary Medical Teaching Hospital at Chungnam National University for suspected bone fracture. The monkey had been reared singly in a cage at a laboratory facility. An animal caretaker incidentally found a bone fragment protruding through the skin of the right leg. Radiographic examination revealed 2 new bone fragments clearly distinguishable from the original femurs; the fragments seemed to be inserted into both femurs. One of the new bone fragments was easily separated surgically from the right femur. Although the bone fragment consisted of a medullary cavity and bone cortex, the periosteal structure was incomplete. New bone formation in nonhuman primates, as manifested in this case, has been reported previously. However, growth of an additional long bone from the original, penetrating the skin without motional disturbance, has not been reported previously.     

Aspirin inhibits development of coronary atherosclerosis in cynomolgus monkeys (Macaca fascicularis) fed an atherogenic diet.

The effect of aspirin in the primary prevention of diet-induced atherogenesis in cynomolgus monkeys was studied. The diet consisted of 2% cholesterol and 10% butter by weight for 24 wk. Six monkeys received only the atherogenic diet and five monkeys received the diet plus aspirin, 81 mg/monkey per day. Aspirin did not affect plasma cholesterol levels or aortic atherosclerosis. Platelet aggregation to arachidonic acid was almost completely suppressed. Aspirin decreased significantly the number of coronary vessels with atherosclerotic involvement, and the number of coronary vessels narrowed by 20% or more. Thus, aspirin appears to exert a protective effect in the primary prevention of diet-induced coronary atherosclerosis in a primate model.     

Use of operant performance to guide and evaluate medical treatment in an adult male cynomolgus macaque (Macaca fascicularis)

A 6-y-old male cynomolgus macaque presented with noticeable swelling of the left forearm and signs of discomfort, as indicated by nonuse of the arm even in a behavioral task that he previously had been well-motivated to perform. Examination under anesthesia revealed lacerations to the arm. Radiography of the forearm showed no fractures, indicating that the damage was limited to soft tissue. The daily operant behavioral session assessed the amount of force the monkey emitted when touching the screen with the affected arm and how long each touch was sustained. We then used these parameters (force and duration of touch) as objective measures of putative pain relief and recovery of function to guide the medical treatment. The affected monkey received ketoprofen, buprenorphine, or their combination but continued to perform poorly during daily operant behavioral sessions. Only after treatment with dexamethasone did performance return to preinjury levels, suggesting inflammation near the radial or ulnar nerve. These findings indicate that performance of a trained operant task performance can be useful in guiding medical treatment, evaluating pain relief, and objectively monitoring health in laboratory animals.     

Disseminated Zygomycosis in a Cynomolgus Monkey (Macaca fascicularis)

Here we describe a case of zygomycosis in a 4-y-old male cynomolgus monkey (Macaca fascicularis) newly received into our facility. Results of physical exam, clinical chemistry analysis, hematology, and tuberculin skin testing and routine thoracic radiographs performed during the quarantine period are reported. Routine chest radiographs taken during the quarantine period revealed enlargement of the cranial mediastinum. During quarantine, the macaque developed severe respiratory distress and was euthanized. Microscopic examination of tissue collected at the postmortem exam revealed nonseptated, branching hyphae in sections of the stomach and mediastinal lymph nodes consistent with a zygomycete. This is the first reported case of zygomycosis in a cynomolgus monkey.Zygomycosis refers to the angiotropic (blood-vessel–invading) infection produced by fungi in the class Zygomycetes (formerly Phycomycetes). Two orders in the Zygomycetes class are responsible for the majority of clinical cases in humans. Organisms in the order Entomophthorales affect humans in Asia, Africa, and South America, causing subcutaneous lesions in otherwise healthy patients. Fungi of order Mucorales occur worldwide in several clinical forms, including cutaneous, systemic, and rhinocerebral, and typically occur in patients predisposed by other conditions.Zygomycetes are well distributed in nature and survive in organic matter, with infection generally the result of exposure to spores via aerosol or ingestion. In humans, the disease is associated with preexisting conditions such as diabetes, nutritional deficiencies, severe burns, or other immunocompromising conditions. Infection typically involves the rhinofacial area, lungs, gastrointestinal tract, and skin and less commonly other organs. Pulmonary zygomycosis is acquired through inhalation, and antemortem diagnosis is uncommon. These organisms show a predilection for blood vessel (arterial) invasion, resulting in necrosis and embolization, but dissemination to other organs is rare. Despite treatment, the prognosis for complete resolution in humans is poor, and if left untreated, often results in death.     

Ischemic brain damage after ketamine and xylazine treatment in a young laboratory monkey (Macaca fascicularis).

After a 4-year-old female laboratory cynomolgus monkey manifested neurological abnormalities, including tetanic spasm, after intramuscular injection of 20 mg/kg ketamine, we administered 2 mg/kg xylazine in an attempt to control the seizure. However, the animal continued to display opisthotonus, nystagmus, and symptomatic epilepsia. Analysis of blood chemistry revealed a dramatically increased creatine phosphokinase level. Abnormal histopathological findings included acute neuronal necrosis or glial reaction or both in the cerebral cortex, nucleus lentiformis, hippocampus, cerebellar cortex and nucleus, and medulla oblongata; severe myocardial hemorrhagic necrosis; and hepatic subcapsular hematoma. Although the mechanism of this neuronal damage has not been clarified, it may be attributable to an ischemic condition in the brain, probably due to temporal cardiac arrest or hemorrhagic change in the liver and heart, with subsequent decreased blood pressure, after ketamine and/or xylazine treatment. Because both drugs often are used as general anesthetics in veterinary medicine, attention should be paid to this rare case with neural damage.     

超声引导穿刺活检对胸膜病变的诊断价值分析

目的探讨和分析超声引导穿刺活检对胸膜病变的诊断价值。 方法回顾性选取浙江省肿瘤医院2010年1月至2017年12月行超声引导胸膜病变穿刺活检、影像及病理资料完整的病例共118例。所有患者均行超声引导下穿刺活检术。以外科手术病理组织学及临床随访结果为"金标准"，应用四格表计算超声引导穿刺活检诊断胸膜病变的敏感度、特异度、阳性预测值、阴性预测值和准确性。 结果依据诊断"金标准"，118例患者中良性病变17例，恶性病变99例，不确定2例。超声引导胸膜病变穿刺活检病理诊断恶性病变87例，良性病变29例，不确定2例。超声引导穿刺活检诊断胸膜病变的敏感度、特异度、阳性预测值、阴性预测值和准确性分别为87.9%、100.0%、100.0%、58.6%和89.7%。118例患者中4例出现并发症。 结论超声引导胸膜病变穿刺活检术具有实时、操作简便、无放射线危害，以及并发症发生率低等特点，对于胸膜病变具有较高的诊断价值，值得临床推广应用。     

Nonsurgical Repair of a Pseudoaneurysm in a Cynomolgus Macaque (Macaca fascicularis)

A cynomolgus macaque presented with an ecchymotic and edematous left leg approximately 1 wk after a blood sample had been collected from the left femoral vein. Ecchymosis was noted in the femoral triangle, prepuce, and scrotum. The animal was not febrile or exhibiting signs of pain or distress. Duplex Doppler ultrasound imaging was used to evaluate the area. An arteriovenous fistula between the femoral artery and vein, accompanied by a pseudoaneurysm arising from the femoral artery, was identified. Various invasive and noninvasive treatment options for the pseudoaneurysm, including surgical repair, thrombin injection, stent placement, and ultrasound-guided compression repair (UGCR), were considered. UGCR was chosen as the first option for treatment. After a total of 20 min of UGCR at the neck of the pseudoaneurysm, complete thrombosis was achieved. Subsequent imaging of the lesion revealed resolution of the pseudoaneurysm. Because of the risks involved with invasive management techniques for this vascular lesion, UGCR is a valuable noninvasive treatment option for the repair of pseudoaneurysms.Abbreviation: CDI, color Doppler imaging; UGCR, ultrasound-guided compression repairVascular malformations have been reported to occur in a wide variety of species.^{2,3,5-7,10,11,13,15-22,24,25,27-35} These abnormalities include arteriovenous fistulas, vascular shunts, hemangiomas, vascular atresias, aneurysms, pseudoaneurysms, telangectasia, and lymphangiomas, with the overwhelming majority of reports describing arteriovenous fistulas.^{2,3,5,6,11,15-19,25,27,30,33,35} Aneurysms have occurred in several primates including chimpanzees, gorillas, squirrel monkeys, howler monkeys, owl monkeys, African green monkeys, spider monkeys, patas monkeys, and capuchin monkeys.²⁴ Pseudoaneurysms are reported infrequently. Pseudoaneurysms (or false) aneurysms are the result of leakage of blood from an artery into a defined space. A pseudoaneurysm associated with the femoral artery in a black and white colobus monkey was detected 6 d after manual restraint for routine blood collection from the femoral artery.³² The pseudoaneurysm was excised, and the resulting vascular defect was closed with an autologous graft. Another report¹⁰ describes a pseudoaneurysm associated with the femoral artery in a rhesus monkey. The diagnosis was obtained interoperatively, and excision of the defect was successful. Because surgical treatment of pseudoaneurysm has inherent risks and requires considerable surgical skill for a successful outcome, a noninvasive approach would be a valuable and cost-effective treatment option. This report is the first to describe the clinical presentation, diagnosis, and nonsurgical treatment of a pseudoaneurysm in a cynomolgus macaque.     

Pharmacokinetics of 2 Formulations of Buprenorphine in Macaques (Macaca mulatta and Macaca fascicularis)

Buprenorphine is the cornerstone of pain management in nonhuman primates, but the pharmacokinetics of this widely used drug are unknown. The purpose of this study was to evaluate the pharmacokinetic profiles of buprenorphine (0.01 and 0.03 mg/kg IM) and sustained-release buprenorphine (0.2 mg/kg SC) in 2 macaque species (M. mulatta and M. fascicularis) by using mass spectrometry. The pharmacokinetics did not differ significantly between species, and buprenorphine was dose-proportional at the tested doses. The low and high doses of buprenorphine had elimination half-lives of 2.6 ± 0.7 and 5.3 ± 2.0 h, respectively, but the low-dose data were constrained by the sensitivity of the analytical method. Sustained-release buprenorphine had an elimination half-life of 42.6 ± 26.2 h. The AUC_0-Tlast of buprenorphine were 9.1 ± 4.3 and 39.0 ± 25.1 ng×h/mL for the low and high doses, respectively, and sustained-release buprenorphine had an AUC_0-Tlast of 177 ± 74 ng×h/mL. Assuming a hypothesized therapeutic buprenorphine plasma concentration threshold of 0.1 ng/mL in macaques, these results suggest that buprenorphine doses of 0.01 mg/kg IM should be administered every 6 to 8 h, whereas doses of 0.03 mg/kg IM can be administered every 12 h. These results further demonstrate that a single 0.2-mg/kg SC injection of sustained-release buprenorphine maintains plasma concentrations above 0.1 ng/mL for 5 d in macaques. These findings support a new dosing strategy using sustained-release buprenorphine to improve pain management, decrease animal stress, improve animal welfare, and simplify the postoperative management of nonhuman primates in laboratory animal and zoological settings.Abbreviation: λ_z, elimination constant; C_max, maximal observed plasma concentration; HDB, high-dose buprenorphine; LDB, low-dose buprenorphine; MRT, mean residence time; SRB, sustained-release buprenorphine; T_last, time of last quantifiable plasma analyte concentration; T_max, time to C_max; V, volume of distributionBuprenorphine is a key component of veterinary multimodal pain management, especially in nonhuman primates. The long duration of action, low risk of respiratory depression, and negligible cardiovascular effects in healthy animals make it an advantageous opioid analgesic agent.⁴³ The widely accepted dosage range for buprenorphine in nonhuman primates is 0.01 to 0.03 mg/kg IM twice daily.^13,14,20 This dosage is based on the canine dose and anecdotal evidence, because few studies in the primary literature address therapeutic dosages in laboratory animal species.^24,38,42 The premise that the current recommended dosing regimen of buprenorphine provides appropriate analgesia is unsubstantiated, introducing the possibility that nonhuman primates do not gain sufficient pain control from the opioid component of the pain management plan.A new formulation of buprenorphine is reported to have analgesic activity for up to 72 h in cats and rats.^7,15 The manufacturer reports that, when administered at 0.27 mg/kg SC, this sustained-release buprenorphine (SRB; ZooPharm, Fort Collins, CO) reaches maximal plasma concentration within 1 h and remains above 1.0 ng/mL for 72 h after injection in dogs. In light of the prolonged duration attained in dogs, this new formulation warrants further evaluation in nonhuman primates.Drugs with prolonged durations of action are preferred in veterinary medicine and are typically developed for either production or companion animals rather than laboratory animal species. Recently, 2 studies evaluated cefovecin sodium in nonhuman primates, with the expectation that this third-generation cephalosporin antibiotic would have an extended duration of activity in nonhuman primates as it does in dogs and cats. Unfortunately, both studies concluded that the plasma clearance of the antibiotic was 20-fold higher in nonhuman primates than in dogs, providing only 12 to 24 h of antibiotic activity and therefore no dosing advantage over other cephalosporin antibiotics in nonhuman primates.^37,39 One study further demonstrated differences in the metabolism of the drug between nonhuman primates species.³⁹ Collectively, these findings highlight the importance of determining optimal dosing strategies for any drug in targeted nonhuman primates species, rather than simply using a published dose for a different species without further evaluation.The purpose of this study was to evaluate the plasma concentrations and elimination kinetics of buprenorphine and SRB at clinically relevant dosages and administration routes in the 2 most common Old World nonhuman primate species used in research. Specifically, we used liquid chromatography–electrospray ionization–tandem mass spectrometry to confirm that buprenorphine and SRB achieved quantifiable plasma concentrations after injection and to verify how long buprenorphine and individual metabolites remained detectable in the plasma.     

男性不育症的睾丸细针穿刺与活检对比分析

目的　探讨睾丸细针吸液在诊断男性不育症中的实用价值。方法　采用三指固定睾丸法。０ ．６ｍ ｍ 空针头注射器抽吸睾丸、计算各种细胞及精子所占的百分比。普通钳取法做活检对比。结果　精子密度在正常范围８例（１７ ％ ） ;少精症患者６ 例（１３ ％ ） ;无精症患者１４ 例（３０ ％ ） ;非阻塞性无精症６ 例（１３ ％ ） ,以上各组均吸出睾丸液体。结论　睾丸细针吸法在诊断男性不育症中较其他诊断方法先进、快捷、准确并有较高的临床价值。     

Use of animal-operated folding perches by rhesus macaques (Macaca mulatta).

Providing captive or laboratory animals with the best possible living conditions has led to many ideas about how caging environments can be enhanced and the animals' lives can be enriched. This study focused primarily on 2 issues: more efficient use of existing caging and providing animals with a measure of control over their environments. We designed a new springloaded folding perching apparatus that, when modified for size, could be added to almost any caging system. Experiment 1 measured usage by animals in standard laboratory caging for rhesus macaque monkeys (Macaca mulatta). Experiment 2 measured usage by this same species in social groups in a 5-acre outdoor-indoor field setting, where several other forms of enrichment were available to the animals. Results indicated that the folding perches were used in both environments. Animals quickly learned to fold down the devices to use as a place to perch, even in the presence of permanent fixed perches. The folding perches did not significantly affect existing behavioral repertoires, but they altered how the animal used the cage. Increased animal presence near folding perches during experiment 2 suggests that these devices actually were preferred. The preference results can only partially be explained by novelty. The folding perches afforded animals a measure of control over their immediate environment without interfering in research or animal care efforts. Including at least 1 folding perch per cage satisfies both the letter and the spirit of regulations on environmental enhancement for captive primates.     

Comparison of Methods for Determining ABO Blood Type in Cynomolgus Macaques (Macaca fascicularis)

Thorough examination of ABO blood type in cynomolgus monkeys is an essential experimental step to prevent humoral rejection during transplantation research. In the present study, we evaluated current methods of ABO blood-antigen typing in cynomolgus monkeys by comparing the outcomes obtained by reverse hemagglutination, single-nucleotide polymorphism (SNP) analysis, and buccal mucosal immunohistochemistry. Among 21 animals, 5 were type A regardless of the method. However, of 8 serologically type B animals, 3 had a heterozygous type AB SNP profile, among which 2 failed to express A antigen, as shown by immunohistochemical analysis. Among 8 serologically type AB animals, 2 appeared to be type A by SNP analysis and immunohistochemistry. None of the methods identified any type O subjects. We conclude that the expression of ABO blood-group antigens is regulated by an incompletely understood process and that using both SNP and immunohistochemistry might minimize the risk of incorrect results obtained from the conventional hemagglutination assay.Abbreviation: SNP, single nucleotide polymorphismCynomolgus macaques (Macaca fascicularis) have been recognized as an appropriate model for transplantation research because of their similarity to humans in immunologic, genetic, and physiologic aspects.^8,9 Identification of ABO blood-group antigens is a prerequisite not only for donor–recipient matching to prevent rejection but also for transfusion during emergency animal care. In humans, the forward hemagglutination test, in which blood type is determined by using the antigen–antibody reaction between antigens on RBC and antisera, is recognized as the ‘gold standard’ for ABO blood-antigen determination. However, such a system is infeasible in macaques because they lack ABO antigens on their RBC.⁷ Therefore, hemagglutination assays using surrogate human blood cells and the antisera of the test animal is used.Although the hemagglutination assay is simple, cost-effective, and reliable on many occasions, its results are prone to subjectivity, and it has several technical limitations. The largest drawback of the agglutination assay arises from age-associated differences in the levels of serum antibody secreted in each animal.²⁰ The gel test method, in which hemagglutination is examined by assessing the differential movement of nonagglutinated and agglutinated RBC in dextran acrylamide gel after centrifugation, was introduced as an alternative to conventional hemagglutination in cynomolgus monkeys.⁴ However, interpreting the grade (0 to +4) can be subjective; therefore other validation data, obtained by using methods, such as DNA analysis and immunohistochemistry, are required to determine the blood type of cynomolgus monkeys.It is well established in humans that the A and B allele encode distinct glycosyltransferases that convert the H antigen to the respective A and B antigens, whereas an O-type person expresses an unmodified H antigen.²² Although a specific single-nucleotide polymorphism (SNP) in the ABO locus controls this activity, the molecular mechanism of blood-group antigen expression in cynomolgus macaques is not fully delineated. A proposed new method for ABO typing in cynomolgus macaques is based on the results of sequence comparison between various nonhuman primates and uses quantitative real-time PCR coupled with the TaqMan probe assay.^16,17 Such genetic analysis can clearly show the antigenic characteristics on the DNA level with the strength of high-throughput analysis of multiple samples.⁶ Despite the tremendous potential of that approach in immunohematologic applications, the SNP that govern ABO antigen expression in cynomolgus monkeys remain unclear. For example, the null allele (O) in cynomolgus monkeys lacks the stop codon mutation found in the human ABO locus,^16,22 prompting questions regarding whether O-type animals exist and how the A- or B-antigen moiety is generated from H antigen in cynomolgus monkeys. Therefore, discrepant outcomes between hemagglutination and modern DNA analysis methods are still possible.ABO antigens in Old World monkeys are expressed in a variety of tissues, including the endothelium of the vasculature of most organs, the epithelium of the gastrointestinal tract, saliva, and exocrine secretions.^15,19 If appropriately set up, immunohistochemistry provides unequivocal results regarding blood-group antigen expression.² However, the level of antigen expression on the epithelium differs among subjects,²⁰ and the quality of cells collected from epithelial tissues (for example, buccal mucosa) can vary widely, depending on the salivation status and general health conditions (for example, hydration) of the tested animals.Given those viewpoints, we evaluated current ABO typing methods for cynomolgus macaques by comparing the results from hemagglutination assays with those of SNP analysis and immunohistochemistry.     

Effect of prolonged ketamine exposure on cardiovascular physiology in pregnant and infant rhesus monkeys (Macaca mulatta).

Physiologic measurements in nonhuman primates usually are collected from animals that are chemically or physically restrained. Both types of restraint may affect the parameters measured, and those effects can vary with age. Heart rate, respiratory rate, oxygen saturation, expired CO2, blood pressure, temperature, blood glucose, hematocrit, and venous blood gasses were measured in rhesus monkeys that were either infused intravenously with ketamine for 24 h or were cage-housed and physically restrained for sample collection. The subjects were pregnant monkeys at gestational day 120 to 123, infants 5 to 6 d old, and infants 35 to 37 d old. Heart rate and blood pressure were lower in ketamine-treated monkeys than physically restrained monkeys. Heart rate was higher in infants than adults, whereas blood pressure was lower in infants. Respiratory rate was higher in infants than adults and higher in physically restrained infants than ketamine-sedated infants but was not affected by ketamine in pregnant adults. Hematocrit was decreased in older infants. In summary, both physical restraint and ketamine sedation altered several physiologic parameters in pregnant and infant rhesus macaques. Investigators should consider these effects when designing experiments and evaluating experimental outcomes in monkeys.     

超声及微波辐射技术在制备穿刺活检组织切片中的应用

肝、肾、肺等穿刺活检组织体积微小、质软易碎，送检时常被预先固定。对其做快速病理诊断时，冷冻制片法常造成切片不全、细胞肿胀、冰晶裂隙等假象；传统快速石蜡制片法安全性差，易出现组织收缩、染色不清晰。我们将超声、微波辐射技术用于制备穿刺活检组织切片，既克服了上述不足，又缩短了制片时间，基本达到常规石蜡切片的效果。     

Elimination of chronically consumed caffeine in the pregnant monkey (Macaca fascicularis)

Characterization of alterations in caffeine elimination during pregnancy is essential in assessing the potential exposure of the fetus to caffeine and its metabolites. Female monkeys (Macaca fascicularis) were exposed to caffeine in their drinking water 7 days/week before, during and after pregnancy. The low exposure (0.15 mg/ml) corresponded to a level sometimes consumed by pregnant women (10-15 mg/kg/day) whereas the high exposure (0.35 mg/ml) was above average human consumption (25-30 mg/kg/day). Blood samples and 24-hr urine samples were collected every 2 weeks throughout dosing. Caffeine and metabolite concentrations in serum and urine were determined by high-performance liquid chromatography. Before pregnancy, geometric mean serum caffeine concentrations were approximately 1.6 and 4.9 micrograms/ml and serum theophylline concentrations were 6.6 and 13.3 micrograms/ml for the low and high dose groups, respectively. During pregnancy, serum caffeine concentrations increased by approximately 100% for both dose groups and, after parturition, declined to prepregnancy concentrations. Serum theophylline concentrations were usually greater than serum caffeine concentrations and did not change during pregnancy. The amount of caffeine and theophylline excreted in the urine over 24 hr increased during pregnancy and returned to prepregnancy levels after parturition. The results of this study indicate that pregnancy decreased caffeine elimination, resulting in a significant increase in serum caffeine levels. The changes in caffeine elimination may be related to alterations in serum estrogen and progesterone levels.