NOD/SCID雌性小鼠动情周期的观察

目的：观察NOD/SCID小鼠动情周期及卵巢切除术后阴道涂片的变化。方法：连续观察9天NOD/SCID鼠,每日进行两次阴道涂片,计算动情周期时间及发生率。卵巢切除后,阴道涂片并观察阴道涂片变化。结果：NOD/SCID小鼠动情周期为4-6天。有规律动情的小鼠占80%。阴道口状态与动情周期无明显相关。卵巢切除术后,阴道涂片呈动情后期或动情间期改变。结论：采用阴道脱落细胞涂片法可判断NOD/SCID雌性小鼠的动情周期及特点,NOD/SCID雌性小鼠卵巢切除手术有效。     

不同方法确定动情周期及动情期生殖器官结构

目的 为确定最佳合笼时间,对两种常见鉴定小鼠动情周期方法进行比较,并确定处于动情期的子宫内膜和输卯管的形态结构,为动物生殖发育及胚胎研究提供基础技术指导.方法 对43只昆明小鼠分别应用阴道脱落细胞检查法、宫颈黏液结晶法,确认小鼠动情周期,与子宫、输卵管和卵巢做成石蜡切片,HE染色后镜下观察的结果进行统计分析比较,同时对处于动情期的小鼠子宫内膜和输卵管进行HE染色.结果 阴道脱落细胞检查法所确定小鼠动情周期与子宫镜检所见确定的动情周期中,在动情期的判定上吻合率较高;小鼠的阴道脱落细胞在动情周期各个时期有不同的形态特点,处于动情期的小鼠的子宫内膜和输卵管的形态与处于非动情期的小鼠有明显变化.结论 阴道脱落细胞染色检查确定动情期比宫颈黏液结晶法更为可靠;小鼠激素水平的变化使阴道上皮细胞发生周期性变化,动情期小鼠输卵管和子宫内膜发生相应改变分别为受精和着床做准备.     

宫腔注射无水乙醇对小鼠动情周期的影响

目的探讨宫腔内注射无水乙醇对小鼠动情周期的影响。方法利用阴道脱落细胞染色法选择动情期规律的小鼠20只,宫腔注射无水乙醇,术后第一个动情周期内每日测定其动情周期,随机择10只在第二个动情周期每2 h测一次,观察动情周期的改变程度。结果实验小鼠宫腔注射无水乙醇后动情周期整体发生大幅度延长,且动情期、动情前期等各期时间均发生了不同程度的紊乱。结论本实验通过观测无水乙醇损伤子宫后小鼠动情周期发生紊乱的现象,为临床无水乙醇的使用提供一定的实验基础。     

小鼠生物净化中的动情周期判定及超数排卵

目的 准确判定小鼠动情周期的变化,并在动情周期的不同阶段诱导超数排卵,探索最佳的超排时间,以提高生物净化的成功率.方法 C57BL/6J小鼠每日08：00至13：00,取阴道脱落细胞进行涂片,观察脱落细胞中白细胞、有核细胞和角质化细胞的比例及变化的规律,确定小鼠的动情周期.据判定的动情周期的各阶段,腹腔注射外源激素分别诱导超数排卵后,于输卵管中取卵母细胞计数,并统计分析.结果 根据阴道涂片确定小鼠动情前期（包括过渡期）、动情期（Ⅰ、Ⅱ期）、动情后期和间情期.4～6周龄C57BL/6J小鼠诱导超数排卵的最佳时期是动情前期,平均可获得23.20枚卵母细胞,显著高于其他实验组（P＜0.001）.结论 据阴道涂片观察,C57BL/6J小鼠单个动情周期持续4～10d,动情周期内不同时间段超排效果也不同.     

电针对血管性痴呆小鼠海马神经元c-fos表达的影响

目的观察电针对血管性痴呆(VD)小鼠海马神经元凋亡及相关基因cfos表达的影响。方法以反复脑缺血再灌注法复制VD小鼠模型,电针肾俞、膈俞和百会穴,对照组灌胃喜得镇,分别于治疗1d和7d后,以FCM法检测小鼠海马细胞凋亡率、cfos表达。结果术后1d,模型小鼠海马细胞凋亡率(6.86±1.10)较假手术组(3.59±0.70)明显增高(P<0.01),术后7d(13.49±1.72)较术后1d还高(P<0.01)。术后1d,模型小鼠海马细胞cfos表达(1.08±0.33)较假手术组(1.00±0.25)有升高趋势(P>0.05),7d时(2.65±0.29)则显著升高(与术后1d和假手术组比较,均P<0.01)。电针在两个时点均可上调小鼠海马细胞cfos的表达(2.26±0.31,3.76±0.41),降低细胞凋亡率(3.68±1.48,6.36±1.52),与模型组相比均差异有显著性(P<0.01)。结论电针具有抑制细胞凋亡、保护神经元的作用,这可能是电针改善VD小鼠智能的作用机理之一。     

乙二醇、丙二醇对胎儿卵巢冻存效果的影响

目的比较1.5mol/L的乙二醇和丙二醇两种低温保护剂对胎儿卵巢组织的保存效果。方法采用“一步降温法”对24～28周水囊引产的胎儿卵巢进行冻存,将解冻后的胎儿卵巢组织移植于去势裸鼠的双肾被膜下,分别于移植后2、4及8周取材,通过电镜观察及移植后的组织学、受体鼠血清雌激素水平测定和阴道脱落细胞涂片判断组织细胞活性。结果乙二醇对卵泡形态结构的保存优于丙二醇,后者在卵巢的部分区域可见不可逆性的冷冻损伤;乙二醇组动情周期恢复率为33.3%,移植后动情周期开始天数为(15.00±5.0)d,而丙二醇组动情周期恢复率为16.7%,移植后动情周期开始天数(23.50±6.26)d。移植2周后乙二醇组与丙二醇组的移植物存活率分别为83.3%、66.7%,两组受体鼠血清雌激素水平之间的差异具有统计学意义(P<0.05),异种移植8周后两组移植物的存活率分别为58.3%、33.3%。结论以1.5mol/L的乙二醇作低温保护剂行快速冷冻效果优于1.5mol/L的丙二醇。     

不同剂量顺铂诱导小鼠卵巢功能早衰程度的差异

目的比较不同剂量顺铂诱导小鼠卵巢功能早衰(POF)的差异,提供适宜的POF研究模型。方法 6～8周雌性ICR小鼠腹腔注射不同剂量顺铂,观察一般情况、动情周期和卵巢组织病理学变化,计算小鼠死亡率。结果顺铂按首日给予3mg/kg,再连续6d腹腔注射2mg/kg,小鼠除体重减轻外,一般状况尚可,卵巢损伤相对较轻,停药后14d,50%小鼠动情周期恢复正常,显微镜下可见较多不同发育时期的卵泡及成熟卵泡,小鼠死亡率10%;连续7d给予顺铂3.0mg/kg,小鼠出现精神萎靡、食欲下降、动情周期紊乱,停药后14d动情周期未恢复,且小鼠死亡率达70%,卵巢组织中可见较多闭锁卵泡,基本见不到发育成熟的卵泡。3d给予顺铂3mg/kg,再给予顺铂2.0mg/kg 4d,小鼠的一般状况、动情周期与组织病理学变化介于前两个方案之间,20%的小鼠死亡。结论小鼠前3d和后4d分别给予3mg/kg和2mg/kg顺铂腹腔注射,在高存活率基础上能更好地形成POF病变,是最为可行的诱导POF方案。     

小鼠阴道涂片技术及其染色方法的研究

目的比较研究棉签法、生理盐水冲洗法进行阴道涂片,HE染色法、瑞氏染色法、巴氏染色法染色对C57BL/6小鼠阴道涂片细胞组织染色的效果,为小鼠动情周期的确定寻找便捷、有效的检验方法。方法对10只C57BL/6雌性小鼠分两笼饲养,每日10∶00用棉签法、生理盐水法进行阴道涂片,连续15 d,并使用HE染色法、瑞氏染色法、巴氏染色法,观察记录。结果连续实验生理盐水法优于棉签法,HE染色染料易于获得;瑞氏染色分型清楚,染色佳;巴氏染色形态清晰,分色明显,透明度好。结论大样本量旨在判断动情周期的发生与改变实验应采用瑞氏染色法,可辅以巴氏染色以增强实验动物数据资料的可靠性。     

豫医无毛小鼠寿命观察

目的 :通过对豫医无毛小鼠寿命的观察 ,探讨突变基因对无毛小鼠寿命的影响。方法 :在正常生产繁殖状态下 ,随机选取无毛小鼠雌雄各 50只 ,杂合子有毛小鼠雌雄各 2 5只 ,饲养条件、营养状况、实验方法等均相同 ,观察每只小鼠自然衰老的过程 ,并计算出各自的寿命。结果 :纯合子无毛小鼠平均寿命雌性为 (30 3 .3± 35 .0 )d、雄性为 (2 61 .6± 41 .2 )d ,杂合子有毛小鼠平均寿命雌性为 (382 .7± 35 .7)d、雄性为 (353 .0± 37.2 )d。纯合子无毛小鼠与杂合子同性别有毛小鼠寿命相比差异有统计学意义 (P <0 .0 1 )。结论 :突变基因可能对无毛小鼠的寿命有影响 ,豫医无毛小鼠可作为衰老模型供科研应用     

PGRN对胚胎植入及胎盘血管形成的影响

目的：探讨颗粒前体蛋白（progranulin,PGRN）对小鼠胎盘血管形成的影响。方法：收集2017年1至4月在重庆医科大学附属第一医院行剖宫产手术的子痫前期（pre-eclampsia,PE）组（10例）以及正常对照（normal control,NC）组（12例）的胎盘组织;通过PCR及Western blot比较子痫前期组和正常对照组胎盘组织中PGRN蛋白的表达情况;通过免疫荧光以及免疫组化等方式来对比在孕6.5 d胚胎及孕17.5 d胎盘在PGRN-/-小鼠和正常小鼠血管生成情况;苏木素-伊红染色比较孕6.5 d子宫动脉管径大小及管壁厚度。结果：PGRN在子痫前期组胎盘组织中的表达明显高于正常对照组,差异有统计学意义（1.420±0.255,0.726±0.192,t=4.118,P=0.001）;孕6.5 d时,PGRN-/-小鼠的胚胎个数较正常对照明显减少（5.333±1.154,9.333±1.527,t=3.618,P=0.022）;在孕早期血管标志物SMA蛋白在PGRN-/-小鼠表达较对照组减弱（25 396.600±2 413.606,17 484.980±1 606.019,t=4.727,P=0.009）;PGRN-/-小鼠子宫动脉较正常小鼠子宫动脉直径稍减小,血管肌层厚度减少较明显（675.844±147.635,1 022.234±133.902,t=3.010,P=0.039;10.136±1.634,18.214±2.899,t=4.204,P=0.013）;孕17.5 d,血管标志物SMA、COX2及Tie2蛋白在PGRN-/-小鼠胎盘迷路层表达量比正常对照组减弱（626.467±114.744,5 288.375±871.319,t=9.188,P=0.001;828.298±29.192,1169.734±43.490,t=11.290,P=0.000;335.766±125.570,2 415.752±579.038,t=6.080,P=0.003）。结论：PGRN可能通过参与胚胎和胎盘的血管形成,在妊娠相关并发症的发生中起作用。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

安心颗粒对急诊经皮冠状动脉介入术后生活质量影响的研究

目的：评价使用安心颗粒对急诊经皮冠状动脉介入术（PPCI）术后生活质量的影响.方法：将160例接受PPCI的急性ST段抬高型心肌梗死患者随机分为安心颗粒组（术前顿服安心颗粒8.8g,术后安心颗粒4.4 g/次,每日2次）和对照组（仅接受基础药物治疗）.所有患者均服用阿司匹林、氯吡格雷和阿托伐他汀.分别在入院时、出院前1d、出院后180 d时,应用心肌梗死多维度量表（MIDAS）、中文版SF-36评价量表对患者生活质量评分.并观察术后30 d以内的出血并发症、血小板减少症发生情况.结果：入院时和出院前1d,两组患者的心肌梗死MIDAS、SF-36量表评分比较无差异（P＞0.05）;出院后180 d时,与对照组比较,安心颗粒组MIDAS、SF-36评分明显减低（P＜0.05）;组内与入院时比较,两组出院前1d、出院后180 d时,MIDAS、SF-36评分均降低（P＜0.05）.两组患者在随访期间均无大量出血、少量出血、重度和极重度血小板减少症发生,安心颗粒组有4例、对照组有7例发生不明显出血（P＞0.05）.两组发生轻度血小板减少症的患者数比较无差异（P＞0.05）.结论：PPCI使用安心颗粒,能改善急性ST段抬高型心肌梗死患者的生活质量,且不增加出血风险.     

The comparative studies of the influences of Urapidil and Nicardipine on sino-atrial node function, atrio-ventricular node function and hemodynamics

Objective:To investigate the influences of urapidil and nicardipine on rabbit sinus function,atrio-ventricular node function and hemodynamics.Methods:Thirty-two Angora's rabbits were selected and randomly divided into four groups.U1 group:urapidil 0.25 mg/kg;U2 group:urapidil 0.5 mg/kg;N1 group:nicardipine 10 μg/kg;N2 group:nicardipine 20 μg/kg.All these medicine were administrated within 30 seconds.Measurements were taken before and after the administration of urapidil or nicardipine for the following data:mean blood pressure(MAP),heart rate(HR),sino-atrial conduction time(SACT),maximal sinoatrial recovery time(SNRTmax)corrected sinus node recovery time(CSNRT),index of sinus node recovery time(SNRTI),Wenckebach A-V conduction frequency (WB),and P-R interval.Results:Significant MAP and HR changes were identified in all of the four groups before and after administration of both urapidil and nicardipine.No significant changes could be found in the rest of the parameters.Intergroup analysis showed that SACT and CSNRT of N1 and N2 groups were shorter than those of the U2 group(P＜0.01);the MAP decreased(P＜0.01)and the HR increased drastically(P＜0.01).Conclusions:Neither urapidil(0.25 mg/kg,0.5 mg/kg)nor nicardipine(10μg/kg,20μg/kg)has any significant influence on rabbit sinus function or rabbit atrio-ventricular node function.Nicardipine could be a better choice than urapidil for parafunctional sinus node patients.     

Expression of OPGmRNA and ODFmRNA in the patients of hip fracture due to osteoporosis

Objective：To investigate the gene expression of osteoprotegerin（OPG） and osteoclast differentiation factor（ODF） in the bone tissue of patients with hip fracture due to osteoporosis. Methods：OPGmRNA and ODFmRNA in the bone tissue in 50 cases of osteoporosis sufferers（over 50 years old） with hip fracture（Observer Group） and 30 cases of hip facture sufferers with no osteoporosis（Control group） were analyzed with the Semi-Quantitative RT-PCR method. Results：The mRNA expressed of ODF, OPG were both high in the patients with hip fracture. In the control group, the expression of OPG mRNA was observed, while the expression of ODF mRNA was very slight. Conclusion：Aged patients contained all signals including OPG, ODF that are essential for inducing osteoclastogenesis and promoting bone resorption.     

Dynamic Changes in Serum Estradiol and Progesterone levels in Patients of Premenstrual Syndrome with Adverse Flow of Liver-qi

Objective:To probe into the influence of changes of ovarian hormones on the pathogenesis of the specific sub-type premenstrual syndrome(PMS)and reveal partial microcosmic mechanisms of adverse flow of liver-qi.Methods:Estradiol(E2)and progesterone(P)levels in serum were determined at different phases of menstrual cycle by radioimmunoassay.Results:In the group of PMS with adverse flow of liver-qi.the secretive peak value Of E2 and P at the follicular phase significantly decreased,and the secretive peak value at the luteal phase did not come into being.Conclusions:Low E2 and P secretive peak at the follicular phase and absence of secretive peak at the luteal phase is one of the microcosmic mechanisms of PMS with adverse flow of liver-qi.One of the pathophysiologic mechanisms of specific sub-type PMS is probably the continuous low level of E2and P.     

Real-time Three-dimensional Echocardiography in Assessment of Left Ventricular and Right Ventricular Volumes

Real-time three-dimensional echocardiography (RT3DE)is a new ultrasound technique that enables dynamic threedimensional visualization and quantification of the heart in real time. Investigation of feasibility and methodology of RT3DE in determining left ventricular (LV) and right ventricular (RV) volumes, RT3DE was performed in 35 normal adults using Philips SONOS 7500 system with a 2-4 MHz matrix array transducer. The 60°×60° "pyramid" volume database was obtained and analyzed on a TomTec echo workstation. Both LV and RV volumes were calculated with four 3DE methods (i.e. apical 2, 4, 8, and 16-plane) through manually tracing ventricular endocardial borders in end diastole and end systole. Stroke volumes were then calculated. LV volume was also measured by 2DE Simpson's rule using GE VIVID 7 ultrasound machine.     

SCREENING FOR A 21-CHROMOSOME ABNORMALITY IN PREIMPLANTED EMBRYOS OF ELDERLY WOMEN

Increasing maternal age is the only etiological factor unequivocally linked to Down's syndrome in humans. The occurrence rate of newborns with Down's syndrome is about 1/220 in women over 35 years old. However, the occurrence rate in embryos fertilized in vitro, of the elder woman is unclear. Using FISH we screened the number of chromosome 21 in preimplanted embryos of 5 elderly women (average age, 38.4 years) to study the feasibility and necessity of screening trisomy 21 in embryos in patients over 35 years old at the in vitro fertilization (IVF) center.     

Scientific principles and rigorous processes should be followed in developing clinical guidelines for therapeutic interventions of integrative medicine

A clinical guideline for the therapeutic interventions of integrative medicine may be defined as a written document which states a series of recommendations on therapeutic interventions of integrative medicine for a special disease or condition. The guideline may provide assistance to medical professionals in making clinical decisions aimed at improving the clinical outcome of patients and reducing the costs of medical care（~＇4~. Recommendations issued by a guideline should be based on the best available evidence in both Western and Chinese medicine. For fulfilling this purpose, the development of clinical guidelines for therapeutic interventions in the field of integrative medicine should follow scientific principles and undergo a rigorous processes.