西藏小型猪与人血液流变学指标的比较分析

目的 探讨西藏小型猪与人血液流变学指标间的变化规律.方法 采集成年西藏小型猪和人的血液样品,分别检测其WBV(高、中、低切变率)、PV、HCT、ESR及Fi等5项血液流变学指标.结果 (1)雄性西藏小型猪的150s-1、30s-1、5s-1、1s-1WBV值和Fi含量均显著低于雌性(P<0.05, P<0.01, P<0.05, P<0.01, P<0.01);(2)在人的血液流变学指标中男性的WBV值(1s-1)和HCT值显著高于女性(P<0.05,P<0.01);(3)西藏小型猪的HCT值和Fi含量与人相比存在显著差异(P<0.01),其余指标均无显著性差异(P>0.05).结论 西藏小型猪除HCT和Fi两指标与人有差异外,其余指标均与人基本接近.     

实验用西藏小型猪线粒体DNA控制区变异类型及其血液指标比较研究

目的 分析实验用西藏小型猪线粒体DNA控制区碱基序列变异分化类型,比较血液生理生化指标的差异,为实验动物新品种的培育奠定分子遗传学基础.方法 采集59头西藏小型猪血液,分别进行全基因组DNA提取、线粒体DNA控制区的扩增、测序并分类,同时进行血液生理生化指标的测定和比较研究.结果 根据串联重复区重复片段的变化,西藏小型猪线粒体DNA控制区分为A、B两种类型,分别占57.6%和42.4%,几乎与5'端序列在305、500和691的三个主要转换位点的变化相对应.A、B两类西藏小型猪18项血液生理生化指标统计结果显示,红细胞数(RBC)差异(P<0.01)有统计学意义,其他指标差异均无统计学意义.结论 根据线粒体DNA控制区序列碱基变异,西藏小型猪分为两个类群,两个群体之间红细胞数存在明显差异.

Abstract：

Objective To analyze the mitochondrial DNA (mtDNA) D-loop region sequence variation in Tibet Mini-Pigs in relation to the blood parameters and provide the molecular genetic basis for developing new species of laboratory animals.Methods The genomic DNA was extracted from the whole blood samples of 59 Tibet mini-pigs to amplifying the mtDNA D-loop for sequence analysis. Nine physiological and nine biochemical blood parameters of Tibet mini-pigs were measured .Results Based on the variation of the tandem repeat motif, the mtDNA D-loop region of Tibet mini-pigs was classified into two types, namely type A and B with the percentage of 57.6% and 42.4%, respectively, roughly matching the 3 transform sites (305, 500, 691) at the 5' end. In the 18 blood parameters, only red blood cell count showed significant differences between types A and (P<0.01). Conclusion Based on the sequence variation of the mtDNA D-loop region, Tibet mini-pigs can be divided into two types that show a significant difference in red blood cell count.     

西藏小型猪与人血液流变学指标的比较分析

目的探讨西藏小型猪与人血液流变学指标间的变化规律。方法采集成年西藏小型猪和人的血液样品,分别检测其WBV（高、中、低切变率）、PV、HCT、ESR及Fi等5项血液流变学指标。结果（1）雄性西藏小型猪的150s-13、0s-15、s-11、s-1 WBV值和Fi含量均显著低于雌性（P〈0.05,P〈0.01,P〈0.05,P〈0.01,P〈0.01）;（2）在人的血液流变学指标中男性的WBV值（1s-1）和HCT值显著高于女性（P〈0.05,P〈0.01）;（3）西藏小型猪的HCT值和Fi含量与人相比存在显著差异（P〈0.01）,其余指标均差异无显著性（P〉0.05）。     

Experimental Study on Observation of Hemorheologic Parameters of Hydrocortisone Caused Shen-Yang Deficiency in Rat Models

Objective:To explore the relationship between Shen deficiency and blood stasis. Methods: Using consecutive intramuscular injection of hydrocortisone to build Shen-yang deficiency model rats, and detect their hemorheological and biochemical parameters. Results: Comparison between the model group and control group showed that the in vitro thrombus significantly lengthened in the model group (P<0.05), the dry weight increased (P<0.01), hematocrit and whole blood viscosity under high sheared change lowered (P<0.05), blood glucose elevated (P<0.01), and RBC infiltration index, aggregation index increased (P close to 0.05). Conclusion: Under the condition of Shen-yang deficiency, although blood does not show such properties of blood stasis as being "viscous, concentrated, coagulated and aggregated", partial change of hemorheology occurs, and it is liable to form thrombus, which proves that the Shen deficiency status is accompanied with the tendency of blood stasis.     

Pharmacological Investigation of Voltage-dependent Ca^2＋ Channels in Human Ejaculatory Sperm in vitro

The types of the voltage-dependent calcium channels (VDCCs) in human ejaculatory sperm and the effects of calcium channel blocker (CCB) on human sperm motility parameters in vitro were investigated. The human sperm motility parameters in vitro in response to the pharmacological agents nifedipine (NIF, inhibitor of L-type VDCC) andω-conotoxin (GVIA, inhibitor of N-type VDCC) were compared and analyzed statistically. The results showed that NIF (1, 5, 10μmol/L) could not only significantly affect human sperm's shape but also spermatozoa motility after incubated at least 10 min in vitro (P<0.001). GVIA (0.1, 0.5 and 1μmol/L) could just only significantly affect human sperm's progressive motility (a % b %) after incubated for 20 min in vitro (P<0.01), but they both could not significantly affect spermic abnormality rate. It is suggested that L-type VDCC, non L-type VDCCs and isoform of L-type VDCC exist in the cell membrane of human sperm solely or together, and they participate in the spermic physiological processes especially the spermic motility.     

DNA content and morphological parameters in synovial sarcoma and synovioma.

By using image analysis technique with 6 normal synovial tissue specimens as controls,nuclear DNA content and morphological parameters in specimens from 51 synovial sarcomas and 15 synovioma were examined quantitatively.The DNA content in synovial sarcoma and synovioma was significantly different (P<0.01), and that of synovioma and normal synovial tissue was also different (P<0.05).Morphological parameters including nuclear areas and perimeters differed significantly among the above three groups(P<0.01).There was no correlation between the DNA content of synovial sarcoma and its histologic type (P>0.05), but the DNA content greatly differed between its higher and lower differentiated grade in each type (P<0.01). The DNA content in synoviai sarcoma IIA stage, according to Enneking's staging system, was lower than that in ⅡB, ⅢA and ⅢB (P<0.05).The 5-year survival rate of synovial sarcoma with diploid/nearly diploid (D/ND) pattern was higher than that of aneuploid (AN) pattern (P=0.028). The study suggests that analysis of the DNA content and morphological parameters of tumor cells is helpful in the diagnosis and pathologic grading of synovial sarcoma.It also provides a guide to clinical operation and estimation of the prognosis. Although synovioma is usually a well-differentiated and benign lesion,this investigation also suggested that increased DNA content may be an indication of poor prognosis for this neoplasm.     

1种实验用小型猪的血液参数

作者测定了30头1～6月龄实验用小型猪仔猪(雌雄各半)的血液参数。该实验用小型猪为北京农业大学实验动物研究所引进育成之品系。作者并将测定结果(附表)与大鼠、Beagle犬、猕猴及人的有关数据作了比较。     

高脂环境下五指山小型猪和西藏小型猪冠心病相关血脂和炎症易感基因的表达

目的观察在高脂环境下五指山小型猪、西藏小型猪血脂和炎症细胞的变化,分析冠心病相关血脂和炎症易感基因的表达。方法取雄性五指山小型猪、西藏小型猪各5只,高脂饲喂连续6个月,每月检测血脂和血常规,并检测饲喂前和饲喂6个月时外周白细胞中LDLR、LPL、PCSK9、ApoAV、5-LO、FLAP、MMP-9、ICAM-1、VCAM-1基因和蛋白的表达,及血清中CRP、TNF-α水平;并在饲喂6个月时取腹主动脉组织进行HE染色病理学检查。结果高脂饲喂后,两种小型猪TC、HDL-C、LDL-C水平均显著升高(P0.01),西藏小型猪WBC、NEUT%、MONO%和五指山小型猪MONO%亦呈升高趋势(P0.05);五指山小型猪TC、LDL-C水平高于西藏小型猪,而西藏小型猪的WBC、NEUT%和MONO%显著高于五指山小型猪(P0.05,P0.01),且西藏小型猪血清CRP和TNF-α水平显著升高(P0.05),而五指山小型猪未见显著差异(P0.05)。高脂饲喂6个月后,两种小型猪均出现明显的AS斑块,五指山小型猪LDLR、ApoAV mRNA和蛋白表达均显著降低(P0.05,P0.01),PCSK9、LPL mRNA表达显著升高(P0.05,P0.01);西藏小型猪LPL mRNA和蛋白表达均显著升高(P0.01);五指山小型猪5-LO、FLAP、ICAM-1、VCAM-1 mRNA表达和5-LO蛋白表达均显著升高(P0.05,P0.01),西藏小型猪FLAP、MMP-9、ICAM-1、VCAM-1 mRNA和蛋白表达均显著升高(P0.05,P0.01),且西藏小型猪MMP-9、ICAM-1、VCAM-1 mRNA和蛋白表达均高于五指山小型猪,而5-LO mRNA和蛋白表达则五指山小型猪高于西藏小型猪。结论高脂环境下五指山小型猪、西藏小型猪均出现不同程度的血脂异常和炎症反应,两种小型猪的冠心病相关血脂和炎症易感基因存在明显异质性,其中,高脂诱导下五指山小型猪外周血白细胞对LDLR、PCSK9、5-LO表达较敏感,而西藏小型猪外周血白细胞中对LPL、ICAM-1、VCAM-1和MMP-9表达较敏感。     

Experimental study of chuanxiong on cerebrovascular hemodynamic parameters.

In the experimental rabbit arteriosclerosis, the change of the cerebrovascular hemodynamic parameters (CVHP) and the effect of Chuanxiong (CX) on CVHP were observed. In arteriosclerosis group (AS group), the mean flow (Qmean), mean velocity (Vmean), maximal velocity (V max) and minimal velocity (Vmin) of the carotid blood flow and cerebrovascular compliance for zero pressure (CO) were significantly decreased (P < 0.05, 0.01), but the values of cerebrovascular peripheral resistance (R) and characteristic impedance (Zc) were significantly increased (P<0.05 and 0.01), and the value of R showed significant positive correlation with the extent of carotid lesions (P < 0.01). All indexes of CVHP of the Chuanxiong group (CX group) were close to and had no significant difference from those of the normal group (N group) but Qmean, Vmean, Vmin, CO and R were significantly better than those of the AS group (P<0.05, 0.01). These results showed that CX can effectively improve cerebrovascular hemodynamics.     

ROLE OF B LYMPHOCYTE AND ITS SUBPOPULATIONS IN PATHOGENESIS OF IMMUNORELATED PANCYTOPENIA

Objective To measure the quantities and apoptosis-related protein levels of B lymphocyte in the patients with immunorelated pancytopenia(IRP)and explore the action of B lymphocyte in the pathogenic mechanism of IRP. Methods Quantities of whole B lymphocytes and CD5 B lymphocytes as well as the expressions of Fas and Bcl-2 in B lymphocytes in 35 patients with untreated IRP, 15 IRP patients in complete remission (CR), and 10 normal controls were assayed by flow cytometry. Results The percentages of B lymphocyte and CD5 B lymphocyte were significantly higher in untreated IRP patients than in CR IRP patients and normal controls (P<0.05), and there was no significant difference between the latter two groups (P>0.05). There was no significant difference of Fas expression in B lymphocyte among three groups (P>0.05). The expression of Bcl-2 in B lymphocyte was significantly higher in untreated patients than in CR patients or normal controls (P<0.01), and significantly higher in CR patients than in normal controls (P<0.01). The apoptosis-related index was significantly lower in untreated patients than in CR patients or normal controls (P<0.05), and significantly lower in CR patients than in normal controls (P<0.05). The percentage of B lymphocyte was positively correlated with post-treated response time(r=0.53, P<0.01). Conclusion The production of auto-antibodies in IRP patients probably has some relationship with the abnormal quantities of B lymphocyte and its subpopulations as well as with the inhibition of B lymphocyte apoptosis.     

亚热带气候环境下西藏小型猪部分血液生理生化指标的比较

目的比较研究西藏小型猪在亚热带气候环境下的部分血液生理生化指标。方法常规方法测定原代和F1代西藏小型猪血液的9项生理和9项生化指标,比较两代之间以及雌、雄之间的不同,并与其他实验动物及人类的同类指标进行对比分析。结果在生化指标中,原代雌雄之间AST差异显著;F1代雌雄之间TG、ALB有差异,BUN、CHOL有显著性差异。在生理指标中,原代雌、雄间无差异,F1代雌雄间MONO有显著性差异。结论西藏小型猪已经基本适应亚热带地区的环境。与其他实验动物相比西藏小型猪许多生理生化指标更接近人类,非常适合替代犬、猴用于生物医学研究。     

F1代西藏小型猪早期生长发育特点

目的研究西藏小型猪早期生长发育的规律。方法对本基地F1代西藏小型猪的体重、体长、胸围、体高、体温、呼吸频率、心率及血液生理生化指标进行了测定。结果西藏小型猪体型小,生长性能稳定,生长速度与原产地西藏小型猪基本一致;西藏小型猪的体温、呼吸次数和心率与原产地西藏小型猪相近,且雌雄之间差异不显著(P>0.05);F1代雌雄间除嗜碱性粒细胞、单核细胞数和尿素氮外其他指标也基本一致,大部分指标与人接近。结论由于西藏小型猪体型小、生长性能稳定,血液生理生化值与人类相近,因而是理想的医学研究用实验动物。     

西藏小型猪线粒体D-loop区及微卫星多态性的遗传学分析

目的通过分析西藏小型猪线粒体控制区(D-loop区)及微卫星位点的遗传多态性,检测西藏小型猪的遗传背景,从而为其作为实验动物提供分子生物学方面的可靠依据。方法利用特异性引物对西藏小型猪的线粒体D-loop区及10个具有多态性的微卫星位点进行扩增,割胶纯化并对线粒体D-loop区进行测序,另外采用聚丙烯酰胺凝胶电泳的方法分离微卫星位点的等位基因。结果西藏小型猪线粒体D-loop区全序列没有多态性,微卫星位点则具有高度的遗传多态性和杂合度,分别为0.584和0.573。结论西藏小型猪线粒体基因组无多态性,证明其在母系进化和遗传上与其他猪种较为一致,本实验所用的西藏小型猪生长于一个封闭的环境,导致其微卫星位点遗传多态性的中低度水平。     

Beagle犬的血液学、血液生化生理值测定

用微量细胞计数仪、全自动生化仪测定５２ 只Ｂｅａｇｌｅ 犬的血液学、血液生化值,并比较不同性别间的差异。结果表明除ＡＰＴＴ 值雌雄间比较有显著性差异外（Ｐ＜ ０－０５） ,其他指标均无统计学差异,与国外研究结果基本相同。     

泛酸钠对Beagle犬心血管指标的影响

目的 观察泛酸钠对Beagle犬心血管指标的影响.方法 选取健康Beagle犬16只,雌雄各半,随机分为泛酸钠低剂量(L,0.26 mg/kg)、中剂量(M,0.52 mg/kg)、高剂量(H,1.04 mg/kg)组及生理盐水(NS)对照组,每组4只.静脉给予质量分数3％戊巴比妥钠麻醉,分离一侧颈动脉,插入充满肝素-NS溶液的动脉插管,测定收缩压、舒张压;分离一侧后肢股静脉,按1 mL/kg静脉单次给药;将针状电极插入四肢皮下,待动物各项观察指标稳定后,分别记录药前、药后5 min、15 min、30 min、45 min和60 min的血压、心率及心电图(ECG)指标.结果 L、M、H三个剂量组与NS组比较,泛酸钠对Beagle犬颈动脉平均动脉压、心率及心电等指标的差异均无统计学意义(P＞0.05).结论 泛酸钠对麻醉状态下Beagle犬心血管指标无明显影响.     

大鼠胰腺炎相关性急性肺损伤模型的探讨

目的研究重组纳豆激酶(recombinant nattokinase,r-NK)大剂量重复给药对Beagle犬和小型猪凝血和纤溶系统的不同影响。方法用血球计数仪测定血小板相关参数;用血凝仪采取凝固法依次测定纤维蛋白原(Fib)含量、活化部分凝血酶时间(APTT)、凝血酶原时间(PT)、凝血酶时间 (TT);以ELISA法分别测定纤溶酶原(PLG)、纤溶酶原激活因子(t-PA)活性。结果药物对Beagle 犬和小型猪有相似的血管刺激作用。给药1周后Beagle犬血小板计数下降,PT明显延长,Fib含量明显降低,PLG活性升高。而药物对小型猪的各项参数均无明显影响。结论不同动物对相同剂量r-NK毒性反应存在较大的差异性,r-NK大剂量重复给药可破坏Beagle犬的凝血/纤溶系统平衡,而不影响小型猪凝血/纤溶系统功能。     

低频超声抗兔早孕的形态学研究

目的探讨低频超声终止妊娠的可能性及其对胎盘形态学的影响。方法利用低频聚焦声束超声(353 kHZ“超声终止早孕治疗机”样机)6种声强各60 s体内直接照射妊娠第10天兔胚胎,妊娠20 d时观察各组胚胎死亡率和胎盘、子宫的形态学变化。结果对照组胚胎总死亡率为12.65%(32/253);照射组中声强30 W/cm2、35W/cm2及≥40 W/cm2组的胚胎死亡率分别为25.00%(12/48)、72.88%(43/59)和100%(64/64、43/43、40/40、35/35)。超声辐照后的即刻改变:光镜下主要为胎盘(绒毛及子宫蜕膜)出血、坏死;电镜下胎盘细胞正常结构消失,子宫平滑肌、内膜上皮细胞中线粒体肿胀。变化随剂量的增加而明显。照射后10 d观察:光镜下见绒毛、蜕膜细胞仍有变性或坏死;电镜下胎盘组织细胞坏死呈致密团块状,子宫内膜上皮、平滑肌及血管内皮细胞均未见异常。结论胎盘较子宫对低频超声更敏感,更易受损,子宫照射后变化轻微且可恢复,低频超声抗早孕是有效而可行的。     

激光脱毛动物模型的建立

目的 采用黑毛西藏小型香猪为动物模型,探讨翠绿宝石激光脱毛术后的毛囊组织学和超微结构的改变.方法 对12只黑毛白皮两藏小型香猪用翠绿宝石激光脱毛,术前、术后即刻、1 h及1、3、5、10、15、30、60d分别取材,在光镜和透射电镜下观察皮肤标本.结果 激光治疗术后毛囊中毛干凝固坏死、碳化及脱落,部分外根鞘及毛球细胞变性坏死;术后1 h毛囊外根鞘、毛球细胞出现核固缩、核碎裂或核溶解等凋亡表现,胞浆空泡化明显,以术后1、3 d达到高峰,并伴有毛乳头血管内皮变性、真皮组织充血水肿和淋巴细胞浸润.术后5 d毛囊的损伤性炎症减轻,术后10、15 d表皮、真皮及毛囊结构基本恢复正常,术后60d毛囊数目减少,残余毛囊结构正常.结论 翠绿宝石激光对毛囊有选择性损伤作用,通过直接热凝同作用和诱导细胞凋亡造成毛囊不可逆的损伤,达到永久性脱毛的目的 .黑毛西藏小型香猪可作为临床激光脱毛的动物模型.     

米非司酮对恒河猴促性腺激素分泌水平的抑制作用

目的测定西藏小型猪以及巴马小型猪、贵州香猪和五指山猪的mtDNA控制区5’端序列,比较研究其遗传变异和与国内家猪的亲缘关系。方法扩增四种小型猪的mtDNA控制区5’端,测序并进行所测序列多重比对,确定变异位点、单倍型,并建立亲缘关系树。结果与荣昌猪相比,西藏小型猪mtDNA控制区5’端侧翼区704bp,有20个变异位点,由此归纳出26个单倍型;其中有两种主体单倍型,三个转换位点（305,500,691）的碱基分别为t,a,a和c,g,g。巴马小型猪、贵州香猪和五指山猪mtDNA控制区5’端变异位点较少,分别只有4、4、3种单倍型。亲缘关系树的分析表明,西藏小型猪与其它三种小型猪有较近的亲缘关系。结论西藏小型猪群体存在遗传分化,与巴马小型猪、贵州香猪和五指山猪亲缘关系较近;巴马小型猪、贵州香猪和五指山猪遗传均一性较高,可能与长期的人工选择有关。