一项小鼠采血新技术——颌下静脉丛采血法

目的介绍小鼠颌下静脉丛采血方法。方法选用2月龄昆明小鼠,固定小鼠后,将采血注射针头刺入颌下静脉丛血管取血。结果单人操作约1 min内可完成小鼠颌下静脉丛采血,采血量达到0.3-0.5 mL。结论此方法无需麻醉、创伤小、采血量大并可重复采血,是一种简便、安全、快速、采血量多的采血方法。     

Long-term blood-sampling technique in piglets

A technique to enable long-term blood sampling from piglets aged 2-3 months is described. Piglets were housed individually in expandable cages and a heparinized polyurethane catheter was inserted into the external jugular vein. A technique was used which prevented the catheter from pulling out of the vein with growth of the animals. Blood samples could be obtained for more than 1 month, and levels of cortisol, glucose, white blood cell count, haematocrit, rectal temperature and heart rate were compared for samples obtained from simulated conventional venopunctures and from the cannula using this technique. It was shown that restraint and needle pricks raised these levels considerably.     

A rapid and non-surgical procedure for jugular catheterization of pigs

A rapid and non-surgical method for jugular catheterization in pigs was set up in 30 piglets of 6.2 kg, 23 pigs of 46 kg and 84 kg and two lactating multiparous sows. The animal was restrained on a V-shaped table (piglets) or with a rope around the mandible (slaughter pigs and sows). The vein was located with the Vacutainer system and a wire guide was inserted into the Vacutainer needle up to the vein lumen. When the needle was removed, the catheter was inserted over the wire guide and advanced until it penetrated the skin and thereafter, the vein wall. The catheter was fixed outside by a large tape and coiled inside a patch just behind the ears. The technique utilizes readily available material and is no more risky for the animal than a single blood sampling. Moreover, it can be performed within 15 to 20 min (including animal restraint) within pens. This new approach might have important implications not only for research purposes by facilitating repeated blood samplings but also for projects which require a rapid and easy method for testing of any kind of pharmaceutical or other type of products under husbandry conditions.     

Non-surgical cannulation of the vena cava for chronic blood collection in mature swine.

A nonsurgical cannulation technique for blood collections from mature swine was evaluated. Primiparous Yorkshire-Landrace sows (n = 6) received an indwelling jugular vein cannulae for 7 days duration. Recannulation was performed at monthly intervals for a total of 14 months. During cannulation, sows were restrained in a standing position using a rope snout snare. A 12-gauge by 10 cm needle was inserted into the jugular vein. Sterilized polyvinyl chloride tubing was advanced through the needle into the vein and a blunted 18-gauge needle and attached intermittent injection hub was inserted into the free end of the tubing. Surgical tape was used to form a butterfly on the tubing by suturing the tape to the animals' skin. Foam padding, livestock cement, and elastic tape helped to keep the tubing in position. Problems with cannulae patency and maintenance were few. No behavioral problems or systemic signs of illness were noted and necropsy examinations performed after the final cannulation revealed few abnormalities associated with chronic intermittent cannulation. This technique provides a safe, quick, effective means for multiple and repeated cannulae placement for blood collection from mature swine with minimal effects on the animal and without the risks associated with surgical techniques.     

The Circulatory System of Amphioxus (Branchiostoma lanceolatum (Pallas))

The morphology of the circulatory system of Amphioxus ( Branchiostoma lanceolatum (Pallas)) has been investigated using a new intravascular injection technique. A survey of the vessels of Amphioxus using this technique is given. The dorsal arteries and their ramifications are described in detail. The new injection technique brought to light myoseptal plexi, supplied from the dorsal arteries, between every two myomeres. Also the ventral parietal arteries have a much more complicated course than hitherto accepted. They are connected with an atrial plexus which is a continuous net of small vessels in the whole length of the dorso-lateral wall of the atrial cavity. It is postulated that this plexus has a supplementary function in respiration. Plexi of minute vessels in the gonads and a real blood circulation with afferent and efferent gonadal vessels have been demonstrated. Two vessels connecting the liver plexus with the cardinal vein (or the atrial plexi) have been noticed, the v. communicans accessoria anterior mentioned in 1900 by Burchardt, and a so-called oblique vessel never described before. The vessels of the caudal region are analyzed completely and also here a real blood circulation appears possible.     

An alternative, less invasive blood sample collection technique for serologic studies utilizing triatomine bugs (Heteroptera; Insecta)

The collection of blood samples for serological studies is often stressful for the focus animal. Recently, the use of bloodsucking bugs, such as Dipetalogaster maximus or Triatoma infestans (Reduviidae; Triatominae; Heteroptera), has been suggested as a new and less invasive method for blood collection. To evaluate this technique, we collected paired blood samples from 20 domestic rabbits (Oryctolagus cuniculus) during a study of rabbit hemorrhagic disease virus (RHDV). For each rabbit, blood samples were collected by the conventional method (needle and syringe from the vena auricularis) and through feeding by D. maximus. Samples were tested for RHDV antibodies using standard test kits at three different dilutions. Antibody titers were identical for 56 paired samples and differed in only four cases. The simple matching indices were 1 for the 1:10 dilution and 0.9 for the 1:100 and 1:1000 dilutions. The major advantages of the new technique are 1) the possibility to obtain blood from animals where veins are inaccessible and 2) the fact that anesthesia of focus animals may not be necessary.     

磁共振血管造影对颈部静脉系统疾病诊断的应用价值

目的：探讨三维动态增强磁共振血管成像（30 DCE MRA）对于颈部静脉系统疾病的临床应用价值。方法：对7例疑有颈部静脉病变的病人进行了3D DCE MRA检查,观察对病变血管及正常血管显示情况。结果：7例检查图像都能清晰显示正常血管结构及病变情况。其中2例正常,2例颈静脉血栓,1例颈静脉癌栓,1例颈静脉扩张,2例动静脉瘘。所有病例均经手术及（或）DSA证实。结论：3D DCE MRA是一种简便,易行,有效的血管检查方法,分辨率高,假阳性率低,在临床上已广泛应用,但临床上多限于动脉系统疾病的诊断。只要选择合适的序列、扫描时间及对比剂的剂量就能较好显示静脉血管情况。此项技术的应用,在颈部静脉病变诊断中有很大的实用价值。     

A new method for isolation of leukocytes from the peripheral blood of amphibians [Bufo himalayanus, (Gunther)] and study of their surface morphology by scanning electron microscopy

Using percoll as the density gradient, a new single step method to isolate leukocytes from the peripheral blood of amphibians (B. himalayanus) has been described. Isolated leukocytes were photographed under the scanning electron microscope and an attempt has been made to characterize the leukocyte population on the basis of surface morphology. Apart from regular blood cell types, B. himalayanus have slender, elongated and slightly curved leukocyte type cells in their peripheral blood. Such slender elongated cells were absent in the blood of a related species B. stomnaticus and hence could not be categorized under the known blood cell types.     

Comparisons of blood viscosity between amphibians and mammals at 3°C and 38°C

(1) The range of temperature exposure of endotherms is narrow compared to ectotherms that can experience daily and seasonal temperature fluxes. (2) Comparison of the blood viscosity of amphibians (bullfrog, Woodhouse's toad, and marine toad) and mammals (horse, dog, and rat) at 3°C and 38°C was undertaken to determine if the effect of temperature on blood viscosity was diminished in amphibians relative to mammals. (3) Mammals did not consistently show greater changes in blood viscosity, plasma viscosity, or relative viscosity with decreasing temperatures relative to the amphibians in this study. (4) These data do not support our hypothesis that blood viscosity of amphibians is less affected by temperature than mammalian blood.     

Improved method of studying the blood for sterility

The technique used for the inoculation and subculturing of blood samples in testing them for sterility is described. This technique eliminates the possibility of contaminating the culture medium and the blood sample under test with extraneous bacterial flora. Blood samples were inoculated without opening the containers with the culture medium. Inoculation was made with the syringe and the needle used for taking the blood sample through the punctured rubber stopper closing the container. Subculturing on solid culture media was also carried out without opening the containers: the rubber stopper was punctured and the contents of the container withdrawn with a pipette needle. The use of this new technique made it possible to detect bacteremia in 12.8% of cases, only in persons with purulent and septic diseases, whereas by using the existing technique bacteremia was detected both in sick and healthy persons, in 38.6% and 26.6% of cases, respectively.     

Rat liver-targeted naked plasmid DNA transfer by tail vein injection

High levels of foreign gene expression in mouse hepatocytes can be achieved by "hydrodynamics-based transfection," the rapid injection of a large volume of a naked deoxyribonucleic acid (DNA) solution into the tail vein. Rats are more tolerant of the frequent phlebotomies required for monitoring blood parameters than mice and, thus, are more suitable for some biomedical research. Recently, we demonstrated that hydrodynamics-based transfection can also be used to deliver naked plasmid DNA into the normal rat, which is more than 10 times larger than the mouse. We performed the tail vein injection using a syringe with a winged needle equipped with an external tube. Injection of a lac Z expression plasmid, pCAGGS-lac Z by this technique resulted in the exclusive detection of beta-galactosidase in the liver. We also injected a rat erythropoietin (Epo) expression plasmid, pCAGGS-Epo (800 microg). Maximal Epo gene expression was achieved when a 25-mL injection volume (approx 100 mL/kg body wt) was transferred within 15 s.     

Chronic vascular catheterization in the rat. Adaptation to injections and semi-continous blood sampling for hormonal studies]

A method for frequent sampling of blood and injections of fluids in undisturbed rats is described. The right external jugular vein and the left carotid artery are cannulated without completely blood stream interruption in the vessel. Injection or perfusion can be performed during several weeks. Blood was collected for at least 15 post-operative days. The technique appears suitable for pituitary kinetics studies which can be carried out several times in rats bearing indwelling catheters.     

Ultrastructure of the epidermis of the lizard (Lacerta vivipara) at the resting stage of the sloughing cycle

The ultrastructure of the epidermis of the lizard ( Lacerta vivipara ) one day after sloughing is described. The non-keratinized layers of the epidermis are essentially similar in structure to those of amphibians and mammals. The cells of the basal layer are not however separated from each other by the large spaces described in the amphibian (Farquhar & Palade, 1965). The middle layers of the epidermis at this stage of the sloughing cycle produce neither the characteristic mucous granules found in amphibians nor the keratohyalin granules of mammals. A small number of granules corresponding in size and location to the "Odland bodies" of both mammalian and amphibian epidermis are, however, present. The intermediate layer cells also contain a number of bodies similar in appearance to those described by Farquhar & Palade as lysosomes in amphibian skin. These structures are both osmium iodide and acid phosphatase positive. Unlike the condition in amphibians and mammals, the cytoplasm of cells in the layer immediately beneath the keratinized strata is honeycombed with small vesicles, and contains large irregular vacuoles of uncertain content. Certain nonkeratinizing elements within the epidermis are tentatively interpreted as nerve terminations. Two morphologically distinct keratinized strata can be distinguished, the inner stratum consisting of flattened cells similar to those of the stratum corneum of mammalian epidermis; individual cell outlines cannot be distinguished in the outer stratum, which has a structure similar to that of avian feather keratin. A shallow surface zone of the outer keratinized stratum has been identified as the Oberhautchen. This consists of longitudinally disposed leaflets or laminae which are responsible for the sculptured pattern of the epidermal surface. The observations reported here provide a basis for analysis of changes occurring at other stages of the sloughing cycle.     

Fetal blood sampling in baboons (Papio spp.): Important procedural aspects and literature review

Background  The baboons ( Papio cynocephalus ) have similarities with human placentation and fetal development. Fetal blood sampling allows investigators to assess fetal condition at a specific point in gestation as well as transplacental transfer of medications. Unfortunately, assessing fetal status during gestation has been difficult and fetal instrumentation associated with high rate of pregnancy loss. Our objectives are to describe the technique of ultrasound guided cordocentesis (UGC) in baboons, report post-procedural outcomes, and review existing publications.
Methods  This is a procedural paper describing the technique of UGC in baboons. After confirming pregnancy and gestational age via ultrasound, animals participating in approved research protocols that required fetal assessment underwent UGC.
Results  We successfully performed UGC in four animals (five samples) using this technique. Animals were sampled in the second and third trimesters with fetal blood sampling achieved by sampling a free cord loop, placental cord insertion site or the intrahepatic umbilical vein. All procedures were without complication and these animals delivered at term.
Conclusions  Ultrasound guided fetal umbilical cord venipuncture is a useful and safe technique to sample the fetal circulation with minimal risk to the fetus or mother. We believe this technique could be used for repeated fetal venous blood sampling in the baboons.     

Effect of lead acetate on the in vitro engulfment and killing capability of toad (Bufo arenarum) neutrophils

Lead is an element of risk for the environment and human health and has harmful effects that may exceed those of other inorganic toxicants. The immune system is one of the targets of lead. Its immunomodulatory actions depend on the level of exposure, and it has been demonstrated that environmental amounts of the metal alter immune function. Very little information is available regarding the effect of the metal on different aspects of the immune system of lower vertebrates, in particular of amphibians. The aim of this study was to investigate the effect of sublethal lead (as acetate) on the function of polymorphonuclear cells of Bufo arenarum. The results revealed that phagocytic and lytic functions of the adherent blood cells collected from sublethal lead-injected toads and incubated with suspensions of Candida pseudotropicalis were affected negatively. The decrease of the phagocytic activity was correlated with increased blood lead levels (P < 0.0001). Additional information referred to the total and differential leukocyte counts was presented; the only difference found was in the number of blast-like cells that resulted augmented in the samples of lead-injected toads. It was concluded that the evaluation of these parameters might be a reliable tool for the biological monitoring of the immune status of amphibians.     

High-frequency Ultrasound Imaging of Mouse Cervical Lymph Nodes

High-frequency ultrasound (HFUS) is widely employed as a non-invasive method for imaging internal anatomic structures in experimental small animal systems. HFUS has the ability to detect structures as small as 30 µm, a property that has been utilized for visualizing superficial lymph nodes in rodents in brightness (B)-mode. Combining power Doppler with B-mode imaging allows for measuring circulatory blood flow within lymph nodes and other organs. While HFUS has been utilized for lymph node imaging in a number of mouse  model systems, a detailed protocol describing HFUS imaging and characterization of the cervical lymph nodes in mice has not been reported. Here, we show that HFUS can be adapted to detect and characterize cervical lymph nodes in mice. Combined B-mode and power Doppler imaging can be used to detect increases in blood flow in immunologically-enlarged cervical nodes. We also describe the use of B-mode imaging to conduct fine needle biopsies of cervical lymph nodes to retrieve lymph tissue for histological  analysis. Finally, software-aided steps are described to calculate changes in lymph node volume and to visualize changes in lymph node morphology following image reconstruction. The ability to visually monitor changes in cervical lymph node biology over time provides a simple and powerful technique for the non-invasive monitoring of cervical lymph node alterations in preclinical mouse models of oral cavity disease.     

Operations on large arteries; application of recent advances

With the use of anticoagulants to prevent recurrence of thrombosis it is now possible to open a major artery which has been occluded by a thrombus, remove the thickened lining with the clot and restore the continuity of the vessel. Free vein grafts have been used successfully to replace arterial segments in the extremities. Vein inlay grafts are preferred in dealing with aneurysms of the abdominal aorta. The aneurysmal wall can be used to support the venous segments. It has been possible to by-pass aneurysms of the abdominal aorta by anastomosing the splenic artery to the left iliac artery. The use of the left superficial femoral artery as a by-pass to supply blood to the right leg is described.     

OPERATIONS ON LARGE ARTERIES—Application of Recent Advances

With the use of anticoagulants to prevent recurrence of thrombosis it is now possible to open a major artery which has been occluded by a thrombus, remove the thickened lining with the clot and restore the continuity of the vessel. Free vein grafts have been used successfully to replace arterial segments in the extremities. Vein inlay grafts are preferred in dealing with aneurysms of the abdominal aorta. The aneurysmal wall can be used to support the venous segments.It has been possible to by-pass aneurysms of the abdominal aorta by anastomosing the splenic artery to the left iliac artery. The use of the left superficial femoral artery as a by-pass to supply blood to the right leg is described.     

The technique of subclavian vein cannulation

A modification of the technique for subclavian vein cannulation, which avoids the risk of fatal air embolism, is described. Other essential precautions designed to avoid the usual complications are emphasized. The technique as described has been used successfully in over 50 consecutive cases without complications.     

A new method to gently place biopsy needles or treatment electrodes into tissues with high target precision

We present a new core needle biopsy and treatment electrode precision placement technique which, regardless of needle size, target lesion hardness and elasticity, makes it possible to precisely place an image guided device inside the abnormal tissue. Once inside the abnormal lesion, multiple tissue samples can be collected using a dedicated trocar and collecting system. Our unique “Fourier” driver substitutes the commonly used spring-loaded device or complements the jerky insertion technique used by experienced interventional physicians. It enables the physician to precisely and with extreme tactility maneuver even large diameter core needles or treatment-electrodes into the lesion using only a diminutive external force. This is achieved by applying supporting servo-controlled mechanical high-acceleration micro-pulses, proportional to the average vector directed by the physician. The Fourier-needle or Fourier-electrode stands completely non-moving when the system automatically goes into full idling. This means that the angle of attack successively and arbitrary can be aligned to hit the target, becoming successively symmetrically inserted into even small tumors to be treated as well as exactly hit any point outlined by real time ultrasound guiding. This kind of biopsy needle or treatment electrode placement results in a uniquely accurate and less traumatic procedure. Due to the risk of disseminating viable tumor cells the precision placement device can be combined with a computer controlled anti-seeding system, denaturizing tumor cells detached during penetration of the biopsy needle or treatment electrode.