目的 探索不同浓度苯妥英钠(PHT)对大鼠牙周膜干细胞(rat periodontal ligament stem cells, rPDLSCs)粘附于牙根面的影响,为PHT应用于牙周重建提供一定的实验依据,为牙周炎的治疗提供了新思路。方法 提取大鼠rPDLSCs,培养并纯化。通过多项诱导分化、表面标记物鉴定后,使细胞在不同浓度PHT刺激条件下,与牙骨质片共同培养,检测粘附于牙骨质片上的细胞量并作比较。结果 20～80 mg/L 浓度范围内的PHT能够促进rPDLSCs的粘附数量,40 mg/L PHT组促进粘附的效果最强。实验组与对照组有显著统计学差异。结论 适合浓度的PHT可以促进rPDLSCs粘附于牙骨质表面,40 mg/L PHT组促进粘附的效果最强。

Objective To investigate the effects of PHT on attachment of rat periodontal ligament stem cells (rPDLSCs) to cementum chips, in order to provide a certain experimental basis for periodontal regeneration and new ideas for therapy of periodontitis. Methods To isolate rPDLSCs from SD rats, culture and purify them. To identify the cells by their apperance, induced multi-direction differentiation potential and cell surface markers. The rPDLSCs were cultured with cementum chips under the action of different concentrations of PHT. Then testing and comparing the amount of cells attached on the cementum chips in different groups. Results The concentraion among 20～80 mg/L of PHT can increase the number of attached cells. 40 mg/L PHT can promote the cells attachment mostly. Conclusion A proper concentration of PHT may promote rPDLSCs to attach to cementum chips′ surface, 40 mg/L PHT may promote the cell attachment mostly.

目的 观察HB13对SD大鼠的一般生殖毒性。方法 SD大鼠,雌雄各100只,分为低、中、高剂量组(20、40和80 mg/(kg·d))和对照组(0.5%羧甲基纤维素钠溶液),每组25只。雄鼠于交配前28 d给药,雌鼠于交配前14天给药,给药至妊娠第7天。雄鼠交配后处死,孕鼠于妊娠第14天颈椎脱臼处死,观察HB13对雌雄鼠一般情况、生育力和胚胎发育的影响。结果 与对照组相比,HB13高、中剂量组雄鼠给药期体重减轻,睾丸系数及附睾系数增大,异常精子率升高;高剂量组还使雄鼠生育力降低。给予高、中剂量HB13的雌鼠妊娠后体重较对照组减轻,着床后丢失率增高,吸收胎数增加,活胎数减少,连胎子宫重降低;高剂量组HB13还使雌鼠生育率及着床数降低,着床前丢失率升高。结论 HB13低剂量对雌、雄性大鼠无一般生殖毒性;中剂量对孕鼠早期胚胎发育有明显干扰作用;高剂量对雌、雄大鼠生育力有显著降低作用并且对早期胚胎发育有明显干扰作用。

Objective To study the general reproductive toxicity of HB13 in SD rats. Methods SD rats, 100 females and 100 males, were divided into low, medium and high dose groups 20, 40 and 80 mg/(kg·d) and the control group (0.5% CMC-Na), The rats were given HB13 for 28 days in male rats and 14 days in female rats respectively before mating, and then mated. The HB13 treatment continued until the 7th day of pregnancy. When the female rats were confirmed pregnant, the male rats were executed and the female rats were executed on the 14th day of pregnancy. Then we can observe the effect of HB13 on fertility and embryonic development in rats. Results Compared with the control group, the body weight of medium, high dose group reduced significantly, the coefficient of testicular and epididymis, abnormal sperm rate increased significantly. High dose group of HB13 also made the fertilityof male rats decline; The body weight, live births uterus weight (including the fetus) of pregnant rats in high and medium group decreased significantly. At the same time, the rate of lost oosperm and absorbed embryo increased. The high dose group of HB13 can also reduce the fertility of female rats. Conclusion The low dose group of HB13 didn't have general reproductive toxicity in rats, the medium dose group impaired early embryonic development, but the high dose group can significantly reduce the fertility of both male and female rats, and can impair the development of embryonic.

目的 探索内源性神经干细胞在大鼠海马可溶性因子中的体外发育归宿及分化鉴定。方法 显微镜下分离Wistar大鼠海马组织放置于低温DMEM/12培养基,低温振荡2小时后高速离心(15000 g),获取实验所用海马组织可溶性因子。取材出生1天的Wistar乳鼠海马中的内源性神经干细胞(endogenous neural stem cells, ENSCs),将ENSCs分别于含海马可溶性因子终浓度为0(对照组)、50、100、200、400 μl/mL的无血清DMEM/F12培养基中培养6天并每日观察,使用免疫细胞化学、Western Blot印记技术比较各组ENSCs中Nestin、CD133的表达量;同时计量并比较各组ENSCs成球个数,以探索在模拟颅内微环境情况下,ENSCs发育、归宿及分化。进一步于最适宜的海马可溶性因子终浓度中分化神经球,对分化的细胞行神经元特异性蛋白入(如:β-tubullin III、MAP2)及胶质细胞特异性蛋白(如:GFAP、S100及p75 NGFR)免疫细胞化学检测。结果 大鼠ENSCs在培养基中呈单细胞漂浮生长,球形; ENSCs于海马可溶性因子各实验分组中培养第2天呈细胞球状态,对照组中无细胞球形成(与100 μl/mL组比较,P₁=0.00),100 μl/mL组与对照组比较有统计学意义(P₁=0.00<0.05);至第6天,在100 μl/mL组中的细胞球数量明显多于其余各组(P₁'=P₂'=P₃'=P₄'=0.00)。在免疫细胞化学检测中,100 μl/mL组中细胞球表达干细胞高亲和蛋白Nestin、CD133阳性,Western Blot免疫印迹检测其中Nestin、CD133蛋白高于对照组。进一步分化试验中,细胞球呈贴壁生长的单细胞状态、有突起伸出、长梭形,免疫细胞化学检测分化的细胞表达胶质细胞特异性蛋白GFAP、S100、p75NGFR阳性,但不表达神经元特异性蛋白β-tubullin III与MAP2。结论 大鼠ENSCs在终浓度为100 μl/mL的HSF作用下,可促进 ENSCs的增殖分裂;ENSCs在同样浓度下的HSF中可进一步分化为表达GFAP、S100、p75NGFR阳性的胶质样细胞;100 μl/mL的HSFS是ENSCs的一种生理性诱导剂或参与促进ENSCs增殖、分化及通过细胞替代或因子分泌等机制修复神经损伤。

Objective The aim of this study was to explore induction and differentiation of endogenous neural stem cells(ENSCs) in the hippocampus soluble factors(HSF) from the hippocampus of adult Wistar rats by mimicking an intracranial microenvironment. Methods After Wistar rats sacrificed, the hippocampus tissue was obtained in cold DMEM/F12. After centrigued and filtered, the HSF was stored at -20℃. The ENSCs was obtained from the hippocampus tissue of a neonate Wistar rat. Collected the tissue, digested and obtained the ENSCs. After we observed the morphology, the ENSCs were cultured in different concentration (0、50、100、200、400 μl/mL) of HSF for 6 days, and compared the expression of Nestin and CD133 by immunocytochemistry. Meanwhile,we compared the Nestin and CD133 protein by western blot. And then we explored the optimal concentration of HSF by the numbers of all groups on the second and sixth day. Furthermore, we did the differentiated experiment using the same concentration of HSF. Results The number of neurospheres in the 100 μg/mL group was significantly higher than those in the other groups on the 6th day. Immunofluorescence revealed that the neurospheres from ENSCs in the 100 μg/mL group more highly expressed nestin and CD133 than control. This result was confirmed by western blot analysis. The neurospheres can differentiate into glia-like cells in 100 μg/mL HSF and 1% FBS expressing GFAP, S100 and P75 NGFR by immunofluorescence. Conclusion These data indicated that HSF alone, mimicking a destination of ENSCs in vitro, could induce and differentiate neurospheres from ENSCs, as a new method to get NSCs and glia-like cells differentiated from ENCs to repair the diseases of center nervous system.

目的 探讨小脑延髓池注射纳洛酮对心肺复苏大鼠脑神经保护的作用机制。方法 将30只雄性SD大鼠随机分为假手术组、常规复苏组和纳洛酮复苏组。采用窒息法建立大鼠心脏骤停模型,复苏的同时给予药物治疗。恢复自主循环(ROSC)后24 h取脑组织,荧光定量PCR法检测脑组织c-Fos mRNA表达水平,免疫组化法检测脑组织c-Fos蛋白的表达。结果 与常规复苏组比较,纳洛酮可显著降低大鼠脑组织c-Fos mRNA及蛋白表达量(P<0.01)。结论 小脑延髓池注射纳洛酮可及时有效的作用于c-Fos基因,发挥脑神经保护作用。

Objective To investigate the neuroprotective mechanism of naloxone injected into cisterna magna on cerebral ischemia-reperfusion. Methods Thirty adult male SD rats were randomly divided into sham group, conventional cardiopulmonary resuscitation (CPR) group and naloxone CPR group. Asphyxiation was used to set up rat cardiac arrest model, and corresponding drugs were given when the resuscitation was carried out. The Brain tissues were taken at 24 h after restoration of spontaneous circulation(ROSC). Fluorescence quantitative polymerase chain reaction (PCR) and immunohistochemical was used to detect the expression of c-Fos proteins was used to detect the expression of c-Fos mRNA level. Results Compared with the conventional CPR group, Naloxone could significantly decrease the expression of c-Fos protein and c-Fos mRNA in rat brain. Conclusion Naloxone injected into cisterna magna can promptly and effectively act on c-Fos gene, playing a neuroprotective role.

目的 探讨糖尿病胃轻瘫大鼠不同血糖水平对Cajal间质细胞(ICC)的影响及其机制。方法 选择雌性Wista大鼠60只进行随机分组,实验组40只,对照组20只。实验组糖尿病Wista大鼠模型以单次腹腔注射链脲佐菌素法诱导。免疫组织化学荧光染色检测不同血糖浓度大鼠胃ICC数量及网络结构。结果 实验组大鼠血糖浓度高于对照组,ICC数量,低于对照组,且比较差异有统计学意义(P<0.05)。实验组大鼠中血糖浓度越高,ICC数量越低,说明血糖浓度升高可能与平滑肌及神经末梢之间缝隙连接的减少及其ICC网络的超微结构损伤及异常有关。结论 DM小鼠胃组织中血糖水平的升高,可能是DM胃中ICC数量减少的原因;外源性降低血糖能改善DM相关的胃肠道ICC病变。

Objective To observe the effects of glucose fluctuations on Cajal interstitial cells (ICC) of rats with diabetic gastroparesis(DGP) and its mechanistic. Methods 60 Wistar rats were selected and randomly divided into two groups. 20 rats in experimental group and 40 rats in control group. Used immunofluorescence staining to detect the amount of gastric ICC and network structure in DGP rats with different glucose levels. Results The blood glucose concentration in the experimental group was significantly higher than that in the control group, the amount of ICC in the experimental group was significantly lower than that in the control group(P<0.05). The amount of ICC decreased with the increase of glucose levels. In the experimental group, The gap junctions between smooth muscle and nerve endings, ultrastructural damage and abnormalities of the ICC network were probably related to glucose level. Conclusion The increase of glucose level was probably the cause of the decrease of the amount in ICC. Exogenousy decrease glucose levels probably can help to improve the lesion of ICC with DGP.

目的 初步探讨黄芩苷防治支气管哮喘的作用机理。方法 用卵蛋白致敏大鼠制备支气管哮喘动物模型,经黄芩苷干预治疗,运用免疫组化法及Western Blot法检测各组大鼠肺组织匀浆中p38 MAPK磷酸化蛋白表达量。结果 两种检测方法均显示,p38 MAPK磷酸化蛋白水平在模型组中有明显的增加,地塞米松组、黄芩苷高剂量组和低剂量组的p38 MAPK磷酸化蛋白水平均低于模型组(P<0.05)。结论 黄芩苷能有效治疗哮喘的作用与抑制哮喘大鼠p38 MAPK信号通路的表达密切相关。

Objective To explore the mechanism of Baicalin in treatment of bronchia asthma. Methods Animal models of bronchia asthma were made in rats sensitized with egg albumen. After the treatment of Baicalin, immunohistochemistry and western-blot methods were used to test expression quantity of phosphorylated p38 protein of lung tissue in all groups of guinea rats. Results Our data confirmed that the level of phosphorylated p38 protein increased significantly in model group, but it decreased in hexadecadrol group, high dose and low dose Baicalin group (P<0.05). Conclusion The effects of Baicalin in asthma model were associated with inhibition of P38 MAPK signal pathways in a dose-dependent manner.

心肌梗死模型在心肌梗死病理发生机制、新药研发研究中占有重要地位。目前冠状动脉结扎法制作大鼠心肌梗死模型,模型稳定,操作简单,造价便宜,效果可靠,为最主要的心肌梗死造模方法,制作方法及具体操作中存在各自的优势和不足,影响着模型的进一步推广,有待继续进行改进和深入研究。

目的 探讨雷公藤甲素防治大鼠青光眼术后滤过泡纤维化的可行性。方法 选取Wistar大鼠100只分为两组,对照组和观察组各50例。采用房水释放联合激光房角光凝法建立青光眼大鼠模型,然后所有大鼠均进行青光眼手术。手术后,观察组大鼠使用雷公藤甲素预防治疗青光眼术后滤过泡纤维化,对照组大鼠使用5-氟尿嘧啶预防治疗青光眼术后滤过泡纤维化。观察比较防治滤过泡纤维化效果。结果 观察组大鼠的眼压在手术后第1天与对照组相比无差异(P＞0.05),在第6天、14天低于对照组(P＜0.05),观察组大鼠滤过泡面积在手术后第1天、6天、14天均小于对照组大鼠(P＜0.05);观察组大鼠的治疗后滤过泡分型Ⅰ型和Ⅱ型均优于对照组,Ⅲ型和Ⅳ型均低于对照组(P＜0.05);观察组术后不良反应发生率为12.16%,低于对照组22.86%(P＞0.05)。结论 雷公藤甲素防治大鼠青光眼术后滤过泡纤维化效果明显,且安全性较高,值得临床广泛运用推广。

Objective To investigate the feasibility of triptolide in prevention and cure rats glaucoma surgery fibrosis. Methods 100 cases of Wistar rats were divided into two groups, with 50 cases in the control group and the observation group.Glaucoma rat model were built by aqueous release combined with laser photocoagulation, and all rats underwent glaucoma surgery. After surgery, the rats in the observation group were observed their triptolide preventive treatment in glaucoma surgery fibrosis, the control rats were observed their 5-fluorouracil preventive treatment in glaucoma surgery fibrosis.The effects of prevention and treatment of bleb fibrosis were compared. Results The intraocular pressure of rats in observation group in the first day after surgery compared with the control group has no significant difference (P>0.05), on the 6th、 14th day it was lower than the control group rats(P<0.05). Filtration area in the observation group on first, 6th day, 14th days after surgery, was less than the control rats(P<0.05); In the observation group, the type Ⅰ and type Ⅱ of filtering bleb were better than those of the control group, the type Ⅲ and type Ⅳ were lower than those of the control group (P<0.05);The adverse reaction rate was 12.16% in observation group, it was lower than the control group 22.86% (P<0.05). Conclusion Triptolide in prevention and cure of rats glaucoma surgery fibrosis is obvious, and high security. It is worthy of promotion.

目的 观察并评估内毒素性急性肺损伤大鼠吸入一氧化氮后外周血中内皮祖细胞和炎症介质的变化情况。方法 90只SPF级健康大鼠分为3组,A组为正常对照组(n=30),B组为急性肺损伤组(ALI)(n=30), C组为一氧化氮(NO)组(n=30)。分别计算各组外周血内皮祖细胞(Endothelial progenitor cells,EPCs) 数量,同时监测肺组织中白细胞介素-10(Interleukin-10,IL-10)水平和髓过氧化物酶(Myeloperoxidase,MPO)活性。结果 我们成功建立了大鼠的ALI肺损伤模型, C组EPCs数量、MPO活性上升幅度均小于B组、而IL-10上升水平均高于B组,差异有统计学意义(P<0.05)。结论 大鼠吸入一氧化氮可减轻内毒素所致急性肺损伤程度,其机制可能与外周血中内皮祖细胞数量及MPO水平下降和IL-10水平上升有关。

Objective To investigate the effect of nitric oxide(NO) inhalation in endotoxin-induced acute lung injury mice. Methods Ninety SPF mice were randomly assigned to the normal group(group A), ALI group(group B)and ALI+NO group(group C). The number of endothelial progenitor cells was counted and the level of Interleukin-10(IL-10) and myeloperoxidase (MPO) were measured. Results Endotoxin administration resulted in pulmonary edema. The pulmonedema was lightened and the level of MPO were decreased by the inhalation of nitric oxide while the level of IL-10 increased. Conclusion NO inhalation can mitigate acute lung injure. The decline of EPCs and MPO and the increase of IL-10 may be one of the mechanism.

目的 评价实时荧光定量PCR分析低氧性肺动脉高压大鼠肺动脉平滑肌基因表达时12个候选内参基因表达的稳定性,获得最适合的内参基因。方法 以低氧性肺动脉高压大鼠肺动脉平滑肌为研究对象,选择文献报道的常用12种内参基因为候选内参基因,利用geNorm和NormFinder程序分析实时荧光定量PCR数据,筛选出最适内参基因。结果 12个候选内参基因在低氧性肺动脉高压大鼠肺动脉平滑肌表达稳定性由强到弱顺序为:TBP>B2M>HPRT1>HMBS>RPL13a>18sRNA>PPIA>ACTB>GUSB>TFRC>GAPDH>PGK1,平均表达稳定度(M值)均<0.5,geNorm和NormFinder评估后推荐使用TBP和B2M一起作为该研究时的内参基因。结论 同时使用TBP和B2M是实时荧光定量PCR分析低氧性肺动脉高压大鼠肺动脉平滑肌基因表达的最适合内参基因,为低氧性肺动脉高压相关基因研究提供最优内参基因。

Objective To compare and select the suitable reference genes in real-time quantitative PCR analysis of rat pulmonary artery smooth muscle cells mRNA expression level of pulmonary hypertension. Methods To choose appropriate reference gene, the expression of twelve commonly use housekeeping genes were examined in rat pulmonary artery smooth muscle cells of hypoxia-induced pulmonary hypertension by using geNorm and NormFinder programs. Results The expression consistency of 12 genes was (from high to low): TBP>B2M>HPRT1>HMBS>RPL13a>18sRNA>PPIA>ACTB>GUSB>TFRC>GAPDH>PGK1. The average expression stability(M) values of them were low than 0.5. TBP and B2M reference genes were recommended to use in the same condition. Conclusion TBP and B2M reference genes were the most suitable combination of the reference genes for real-time quantitative PCR analysis in rat pulmonary artery smooth muscle cells of hypoxia-induced pulmonary hypertension.