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 探讨红细胞分布宽度(RDW)与老年重症社区获得性肺炎(CAP)患者病情严重程度及预后的相关性。方法 将103例老年重症CAP患者按照近期预后分为存活组(n=83例)及死亡组(n=20例),按照RDW的水平分为RDW≥14.5%组(n=83例)和RDW＜14.5%组(n=20例);记录患者的一般临床资料及相关实验室检查指标,比较各组之间的差异,并利用Logistic回归模型分析老年重症CAP患者近期死亡的危险因素。结果 死亡组中的患者入院APACHEⅡ评分、CRP、PCT、RDW水平均高于存活组,差异有统计学意义(P＜0.05)。RDW 异常率随着PSI 级别的增高而增高,RDW 异常率分别为PSIⅠ-Ⅱ级7.32%(3/41、 PSI Ⅲ级16.67%(6/36)、PSI Ⅳ级39.13%(9/23)、PSIⅤ级 66.67%(2/3),差异有统计学意义(P＜0.05)。Spearsman相关性分析显示:RDW与APACHEⅡ评分、CRP、PCT、PSI评分呈正相关(r_s分别为=0.353,0.363,0.432,0.362,P均＜0.05)。多因素Logistic回归分析显示: RDW(OR=2.024,P＜0.05)是老年重症CAP患者近期死亡的独立危险因素。结论 RDW水平随着老年重症CAP患者病情严重程度的增加而增加,RDW增高亦是患者近期死亡的高危因素。

目的 构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型。方法 采用低浓度加量持续诱导法,诱导吉西他滨耐药乳腺癌细胞4T1耐药株,命名为4T1/Gem;CCK-8法测定4T1与4T1/Gem细胞的增殖抑制率,计算耐药指数; Western blot法检测细胞P-gp蛋白表达;B超引导下注射4T1/Gem细胞悬液诱导裸鼠肝脏成瘤;HE染色观察肿瘤组织病理情况,免疫组化法检测瘤组织ER、PR、HER2、Ki-67和P-gp蛋白的表达。结果 经过14个月的诱导成功建立4T1/Gem细胞株,可在含40 μg/mL的Gem培养液中稳定生长。4T1/Gem细胞耐药指数为4T1细胞的788.547倍。与亲代相比,4T1/Gem处于G1期和G2期的细胞增加,S期细胞减少;上调P-gp蛋白的表达。4T1/Gem细胞成功建立裸鼠乳腺癌肝转移模型,瘤组织中ER、PR、HER2蛋白阴性表达,Ki-67阳性10％和P-gp蛋白阳性表达。结论 成功构建吉西他滨耐药乳腺癌细胞4T1耐药株并建立裸鼠乳腺癌肝转移模型,为开发治疗乳腺癌肝转移化疗耐药的药物提供实验基础。

Objective To construct a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem) and establish a nude mouse model of breast cancer with hepatic metastatic. Methods A gemcitabine-resistant variant of the breast cancer 4T1 cell line was induced by gradually increasing the concentration of gemcitabine; this variant is referred to in this study as 4T1/Gem. The proliferation suppression rates of 4T1 and 4T1/Gem cells were determined by using the CCK-8 essay to evaluate the drug resistance indices of the cell lines. Western blot analysis was used to detect P-gp protein expression. Under ultrasonography, a 4T1/Gem cell suspension was injected into nude mice to induce liver tumors. H&E staining was used to observe tumor pathology, and immunohistochemistry was used to detect the expression of ER, PR, HER-2, Ki-67, and P-gp. Results After 14 months of induction, a 4T1/Gem cell line is established successfully. The cell line can grow stably in culture liquid containing 40 μg/ml gemcitabine. The drug resistance index of 4T1/Gem is 788.547. Compared with the 4T1 cell line, the 4T1/Gem cell line can upregulate P-gp protein expression and successfully establish a nude mouse model of breast cancer with hepatic metastatic. ER, PR, and HER-2 proteins exhibit negative expression in the tumor tissue. The positive expression of P-gp and 10% of Ki-67 proteins is also observed. Conclusion This study successfully constructs a gemcitabine-resistant variant of the breast cancer cell line (4T1/Gem)and establishes a nude mouse model of breast cancer with hepatic metastatic, thereby providing an experimental basis for developing and treating a drug-resistant variant of breast cancer.

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.

肺动脉高压是一类发病率低,但常引起右心衰竭等最终导致患者死亡的严重肺血管疾病,其形成的主要病理改变是肺血管重构和肺血管收缩,多种细胞因子异常作用参与发病,对该作用机制的研究成为了治疗疾病,改善疾病预后的关键。

Pulmonary hypertension is a kind of low incidence, but it is often caused by right heart failure and other serious pulmonary vascular disease. The main pathological changes of pulmonary vascular include remodeling and pulmonary vascular contraction, and many kinds of cytokines are involved in the pathogenesis of disease, which is the key to improve the prognosis of the disease.

支气管哮喘是由多种细胞和和细胞因子参与的气道慢性炎症性疾病,白细胞介素、干扰素、集落刺激因子、肿瘤坏死因子、趋化因子等细胞因子在哮喘发生发展过程中发挥着重要作用。

目的 对比紫杉醇脂质体(LEP)与紫杉醇(PTX)联合顺铂(DDP)治疗晚期非小细胞肺癌(NSCLC)的临床疗效及安全性。方法 晚期NSCLC患者48例,随机分为对照组和试验组,对照组采用紫杉醇175 mg/m²,试验组采用紫杉醇脂质体175 mg/m²,均联合顺铂75 mg/m²化疗,21天为1个周期,治疗2个周期后评价疗效,记录近期疗效与治疗期间不良反应。结果 近期疗效:对照组有效率37.50%,疾病控制率为79.17%,试验组有效率为41.67%,疾病控制率为83.33%,两组差异均无统计学意义(P>0.05)。不良反应:白细胞减少、贫血及血小板减少的发生率两组差异无统计学意义(P>0.05),脱发和恶心、呕吐的发生率两组差异亦无统计学意义(P>0.05),但试验组皮疹、呼吸困难、肌肉痛及周围神经炎的发生率明显低于对照组,差异有统计学意义(P<0.05)。结论 紫杉醇脂质体治疗晚期NSCLC与紫杉醇疗效相当,但周围神经炎及过敏反应较紫杉醇为轻。

Objective To compare the efficacy and safety of paclitaxel liposome combined with cisplatin and paclitaxel combined with cisplatin in the treatment of advanced non-small cell lung cancer (NSCLC). Methods 48 patients with advanced NSCLC were randomized into two groups, experimental group were given paclitaxel liposome at 175 mg/m²,and control group were given paclitaxel at 175 mg/m². Both groups combined with DDP at 75 mg/m² per cycle every 21 days.The efficacy and safety were evaluated after two cycles. Results The overall response rate was 37.50% in experimental group and 41.67% in control group, and the disease control rate was 79.17% in experimental group and 83.33% in control group. There was no significant difference between two groups(P>0.05). Though there was no significant difference in incidence of neutropenia,anemia, thrombocytopenia and alopecia, nausea and vomiting, but the occurred rates of rash、muscle pain and peripheral neuritis were significantly lower in experimental group than those in control group. Conclusion In the treatment of advanced NSCLC, both paclitaxel liposome combined with cisplatin and paclitaxel combined with cisplatin have similar efficacy, but paclitaxel liposome can significantly reduce the incidence of peripheral neuritis and serious hypersensitive reactions.

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 探讨儿童肾病综合征(NS)的红细胞分布宽度(RDW)与肾功能损害的关系。方法 收集168例NS患者作为观察组,根据eGFR分期分为三组,按照起病时长4月为界限分为两组;选健康儿童100例为对照组。检测各组血常规、肝肾功能等,采用SPSS 21.0软件进行统计分析。结果 ①与对照组相比,NS患儿的RBC[(4.86±0.69)vs(4.32±0.48)],RDW[(13.39±1.69)vs(12.99±1.04)]升高,差异有统计学意义(P<0.05);②RDW在肾功能3期(14.60±1.36)较2期(12.84±0.79)升高,在起病时长≤4月患儿(13.66±1.78)较健康儿童(12.99±1.04)上升,差异有统计学意义(P<0.05);红细胞平均容积(MCV)在起病时长≤4月较对照组下降,起病时间>4月组较起病时长≤4月上升,差异有统计学意义(P<0.05)。结论 儿童NS患者的RBC、RDW较健康儿童升高,RDW在肾功能3期较2期升高。

Objective To investigate the relationship between red cell distribution width(RDW) and renal function damage in children with nephrotic syndrome(NS). Methods 168 NS patients were chosen as the observation group, divided into three groups by eGFR level, while also divided into two groups by onset duration of 4 months. 100 healthy children were included as the control group. The blood routine, biomedical function of liver and kidney were detected. Data were analyzed by SPSS 21.0. Results ① Compared to control group, NS patients have higher levels of RBC[(4.86±0.69) vs (4.32±0.48)] and RDW[(13.39±1.69) vs (12.99±1.04)], P<0.05; ② RDW is higher in the third phase of renal function than the second [(14.60±1.36) vs(12.84±0.79)], and also higher in the onset duration of less than 4 months group than the control group [(13.66±1.78) vs (12.99±1.04)], P<0.05; Mean corpuscular volume is lower in the onset duration group of less than 4 months than the control group, and higher in the onset duration group more than 4 months than the onset duration group of less than 4 months, P<0.05. Conclusion The RBC, RDW are higher in children NS patients than in the healthy children; RDW is higher in the third renal function than the second renal function.

目的 探讨白癜风初诊患者外周血细胞因子水平与临床分型、皮损面积的相关性。方法 收集我院2009年7月—2013年7月4年间收治的初诊白癜风患者78例及健康志愿者37例,用ELISA检测外周血血清IL-6、IL-2, IFN-γ、TNF-α浓度并判断其与白癜风分型、皮损程度的相关性。结果 与对照组相比,白癜风各分型组的IL-6、IL-2水平均升高,其中,散在性的升高幅度最为明显。IL-6及IL-2与皮损面积呈正相关,其相关系数分别为0.664及0.483,P值分别为0.021及0.014。结论 IL-2及IL-6在白癜风的发病过程中起重要作用,且在不同的分型中的表达存在差异,并与皮损面积呈正相关。

Objective To evaluate the correlation of cytokine level of peripheral blood cells, clinical classification and skin lesion in preliminary diagnosis of vitiligo. Methods A total of 78 patients with vitiligo of preliminary diagnosis and 37 health volunteers were collected from July 2009 to July 2013. The peripheral blood were collected for ELISA measurement of IL-6, IL-2, IFN-γand TNF-α, and evaluated whether the cytokines concentration was correlated to clinical classification and skin lesion. Results Compared to control group, IL-6 and IL-2 in all of classification of vitiligo was higher, and the most increase was observed in scattered group. Both IL-6 and IL-2 showed the positive correlation with skin lesion (CI: 0.664 and 0.483, P=0.021 and 0.014). Conclusion IL-2 and IL-6 maybe play a key role in vitiligo etiology, and showed different level in different classification of vitiligo. Besides, they showed a positive correlation with skin lesion.