目的观察使用川芎嗪对SH-SY5Y细胞自噬的影响,探讨其保护神经细胞的可能机制。方法采用Hanks液替代培养基进行饥饿诱导SH-SY5Y细胞72 h发生自噬。分为3组,正常组为完全培养基,模型组采用Hanks液替代培养基进行饥饿诱导,治疗组在诱导自噬前加入川芎嗪与Hanks液共孵育。兔抗人微管相关蛋白1轻链3Ⅱ(microtubule-associ-ated protein 1 light chain 3,MAP1-LC3-Ⅱ)免疫细胞化学染色,激光共聚焦检测各组细胞发生自噬的情况,流式细胞仪检测细胞周期分布和细胞质内钙离子浓度。结果激光共聚焦显微镜下模型组出现典型自噬细胞形态学改变,与模型组相比,治疗组LC3荧光强度明显降低,治疗组细胞质内钙离子浓度组明显降低,治疗组细胞由G0/G1期所占比例减少,进入S期和G2/M期所占比例明显增加。结论川芎嗪可以抑制SH-SY5Y细胞自体吞噬,这一作用可能与川芎嗪抑制胞内钙超载和刺激细胞由G0/G1期进入S期和G2/M期有关。

目的探讨脑缺血再灌注损伤(cerebral ischaemia reperfusion,CIR)海马神经元Bax mRNA表达的变化及川芎嗪对其影响。方法参考Bannisters方法复制大鼠脑缺血再灌注动物模型,然后利用川芎嗪治疗;利用核酸原位杂交技术观察海马神经元BaxmRNA表达的变化。结果治疗组与模型组相比,阳性细胞减少,组间密度和灰度有显著差异(P<0.05)。结论川芎嗪能够下调CIR海马神经元BaxmRNA表达,从而抑制神经元凋亡作用。

目的 探讨金属硫蛋白(MT)在膀胱移行细胞癌和宫颈鳞状细胞癌中表达状况,及其与肿瘤细胞生物学特性的关系.方法 利用SP免疫组织化学技术检测癌组织中MT表达率.结果 MT在膀胱癌和宫颈癌中的阳性率分别为64.86%(48/74)和58.57%(41/70),两者无差异(P>0.05).在宫颈鳞状细胞癌组织,分化程度低和浸润深者,MT阳性率高于中、高分化组和浸润浅组(P<0.01).结论 MT过表达与宫颈癌组织分化程度和浸润深度有关;检测MT表达状况.对临床选择化疗药物有指导意义.

Objective:To study the clinical significance of CD44smRNA expression in cervical intraepithelial neoplasis(CIN) and cervical squamous cell carcinoma(CSC).Methods:The expression of CD44smRNA in 108 cases of CIN and CSC were examined by in situ hybridization(ISH).Results:The positive rate of CD44smRNA in CIN and CSC were 58.06%(36/62) and 21.74%(10/46),and among two group there were no signification different(P0.05).The positive rate of CD44smRNA in grade Ⅲ of CIN was higher than that in grade Ⅰ and grade Ⅱ,but the different were not significantly(P0.05).The positive rate of CD44smRNA in CSC Ⅱ stage and Ⅲ,Ⅳ stage were higher than that in 0-I stage(P0.05).The positive rate of lymph node metastasis group was significantly higher than that in negative lymph node metastasis group(P0.05).Conclusion: The expression level of CD44smRNA was related to CIN and CSC in grade stage,and lymph node metastasis.

目的 探讨尿激酶型纤溶酶原激活剂(UPA)和生存素(Survivin)在乳腺良、恶性肿瘤组织中的表达及临床意义.方法 采用PV免疫组化法,检测77例乳腺癌和22例乳腺腺瘤患者病灶切除标本中的Survivin、UPA表达;分析两者表达与临床病理参数的关系.结果 在腺瘤和腺癌中,Survivin阳性率分别为72.72%、70.13%,两者比较无统计学差异(P＞0.05);UPA阳性率分别为31.28%、58.44%,腺癌阳性率明显高于腺瘤(P＜0.01).腺癌临床分期高、浸润性癌和有淋巴结转移者Survivin、UPA阳性率高(P均＜0.05).结论 乳腺癌患者Survivin、UPA表达率较高,两者呈低度正相关,其表达与临床病理参数有关;检测两者对乳腺癌的靶向治疗和预后判断有指导意义.

冠心病(CHD)是中老年人最常见的心血管疾病,常伴有急性心血缺血、心肌梗死或猝死等急性冠脉综合征(ACS),因此CHD是老年人生命和健康危害极大的常见病.目前研究证明[1,2],CHD的发生、发展与炎症反应密切相关.血清中许多炎性因子含量与CHD有关,其中血清C反应蛋白(CRP)水平,是评价CHD危害程度的重要指标之一,它对ACS的预测价值已日益引起人们的重视,相应地也为动脉粥样硬化的干预治疗提供了潜在的目标.CRP和CHD的相关性日益受到人们的关注.本文就血清CRP生物学特性及其与CHD发生、发展和治疗的相关性加以概述.

目的:探讨尿激酶型纤溶酶原激活剂(urokinase type plasmivnogen activator,UPA)和细胞粘附因子标准型(Cell coheresive factor,CD44s)在宫颈癌中表达的临床意义.方法:收集宫颈癌标本86例、瘤样变36例,利用免疫组织化学技术检测UPA和CD44s阳性表达率,并分析它们与癌瘤生物学特性之间的相关性.结果:CD44s在瘤样变中阳性率为19.44%(7/36),在宫颈癌阳性率为45.35%(39/86);UPA在宫颈癌和CIN中阳性率分别为56.98%(49/86)和27.78%(10/36);CD44s和UPA在宫颈癌中的阳性率明显高于瘤样变中的阳性率(P<0.05),在Ⅲ、Ⅳ期组明显高于Ⅰ、Ⅱ期组(P<0.05),有淋巴结转移组阳性率明显高于无转移组(P<0.05).结论:UPA和CD44s高表达与癌发生及转移有相关性,检测癌UPA和CD44s对提示患者预后和开展靶向化学治疗有指导作用.

Objective:To investigate the expression and biological significance of Ang-2 and VEGF of breast cancer.Methods:The expression of Ang-2 and VEGF in 76 specimens of breast carcinoma and 36 cases brest adenoma were examined by SABC immunohistochemical method,and the relationship between Ang-2 as well as VEGF and the clinical pathology was analyzed.Results:The positive rate of Ang-2 were 67.11%,51/76)in breast carcinoma,that the positive rate was significantly higher than that(33.33%,7/36)in breast adenoma(P<0.05),and The positive rate of VEGF in breast carcinoma was 60.52%(51/76),that the positive rate was significantly higher than that(19.44%,7/36)in breast adenona(P<0.01)the expression of Ang-2 and VEGF were not associated with patients'age(P>0.05).The positive rates of Ang-2 and VEGF in stageⅢ and Ⅳgroup were significantly higher than those in stageⅠandⅡgroup(P<0.05).The positive rate of Ang-2 and VEGF in lymph node metastasis group were significantly higher than that in non lymph node metastasis group(P<0.01).Conclusion:There is an abnormal expression of Ang-2 and VEGF in breast carcinoma,and which were associated with the prognosis of patients.The Ang-2 and VEGF were examined guidance for target chemotherapy.

乳腺癌是女性最常见的恶性肿瘤之一,其发病率居女性恶性肿瘤之首.乳腺癌的发生发展受多基因的调控,但其生物学特性至今尚未清楚.目前已发现癌基因的激活与抑癌基因的失活等因素与乳腺癌有关,进一步研究这些因素在乳腺癌的表达及相互作用,对揭示乳腺癌发生防治的机制有着重要的意义.

目的 探讨血管生成素(Ang-2)、血管内皮生长因子(VEGF)和尿激酶纤维酶原激活剂(UPA)在乳腺癌中表达的意义.方法 利用SP免疫组织化学方法, 检测89例乳腺癌和36例乳腺腺瘤中Ang-2、VEGF和UPA的表达.结果 Ang-2、VEGF在Ⅲ、Ⅳ期乳腺癌阳性率明显高于Ⅰ、Ⅱ期(P<0.05). 在浸润性乳腺癌组三者的阳性率明显高于非浸润癌组(P<0.01).Ang-2及UPA在淋巴结转移组阳性率明显高于无转移组(P<0.05).Ang-2与VEGF及UPA、VEGF与UPA在乳腺癌中表达呈相关,相关系数分别为0.952、0.480和0.725.结论 Ang-2,VEGF和UPA在乳腺癌高表达与患者预后不良有关,在乳腺癌浸润和转移过程中三者又协同作用.

BACKGROUND:Currently,there are plenty of the investigations regarding the induced differentiation of neural stem cells(NSCs)in vitro,but the process of differentiation is difficult to control,many methods are complicated to operate,and the proportions of differentiation are very low.OBJECTIVE:To discuss in vitro cultivation of rat embryonic forebrain NSCs and to observe the differentiation rule of NSCs.METHODS:The forebrains were isolated from fetal rats under aseptic conditions,to prepare monocell suspension,and cultured with DMEM/F12 medium containing N2 at the density of 1×1011/L.In the process of cultivating,BrdU was added to mark NSC spheres.The induced differentiation experiment was divided into three groups:polylysine culture plate,gelatin plate and no plate culture groups.20% volume fraction of embryonic bovine serum was used to stimulate the differentiation.Immunohistochemistry method was applied to detect the level of nestin and BrdU,as well as NSCs differentiation into nerve cells in the serum induction.RESULTS AND CONCLUSION:The cells presented the NSC-like growth and continuous proliferative capacity,they can subculture.Nestin and BrdU were both positive in passaged neurospheres.The inducing differentiation effect of the polylysine culture plate group and the gelatin plate group was greater than no plate culture group(P < 0.01),and the polylysine culture plate group had a little higher percentage of neurons differentiating than the gelatin plate group(P > 0.05).The result of immunohistochemistry was glial fibrillary acidic protein-positive and microtubule-associated protein-2-positive.Rat embryonic forebrain is full of NSCs.The polylysine and the gelatin as cells holder in inducing NSCs differentiation can improve NSCs differentiation and mostly differentiate into the astrocytes through differentiation observation.

目的:研究尿纤溶酶原激活物(urinary plasminogen activator,UPA)在宫颈鳞状细胞癌(cervical squamous cell cancer,CSC)和典型子宫内膜腺样癌(tipical endonetrioid adenocarcinoma,TEA)表达的临床意义.方法:收集宫颈鳞状细胞癌标本86例、典型子宫内膜腺样癌58例.利用免疫组织化学技术检测UPA阳性表达率,并分析它们与癌瘤生物学特性之间的相关性.结果:UPA在CSC和TEA中阳性率分别为56.98%(49/86)和58.62%(34/58),两组差异无显著性(P>0.05); UPA在宫颈鳞癌Ⅲ、Ⅳ期组的阳性率明显高于0期、Ⅰ期(P<0.05);内膜腺样癌组分化低者阳性率明显高于分化高者(P<0.05).UPA在CSC和 TEA中,有淋巴结转移组阳性率明显高于无转移组(P<0.05).结论:UPA高表达与子宫癌分期(或分化程度)及转移有相关性.

目的 探讨孕激素受体(PR)、表皮生长因子受体(EGFR)和血管内皮生长因子(VEGF)在乳腺癌表达的意义和相关性.方法 采用SP免疫组织化学技术,检测40例乳腺腺瘤和88例乳腺癌中PR、EGFR和VEGF阳性率.结果 PR,EGFR和VEGF在乳腺腺瘤中的阳性率分别为5%,20%和37.5%,EGFR在乳腺癌中阳性率为57.95(51/88);VEGF阳性率为60.22%(53/88);PR阳性率为48.86%(43/88).在浸润性乳癌和有转移组EGFR和VEGF阳性率均高于非浸润组和无转移组(P<0.05).PR 与 EGFR相关系数为0.11,PR与VEGF相关系数0.045,EGFR和VEGF相关系数为0.52.结论 EGFR和VEGF过表达与乳腺癌发生和转移呈显著正相关.PR与EGFR和VEGF呈低度正相关,检测PR、EGFR和VEGF将对乳癌靶向治疗有指导作用.

Objective:To investigate the significance of MGMT and EGFR expression in gastric cancer.Methods: The expressions of MGMT and EGFR in gastric cancer of 69 cases were examined by SP immunohistochemical method.Results:The positive rates of MGMT and EGFR were 52.17%(36/69) and 59.12%(41/69) in gastric cancer.Their positive expression of MGMT and EGFR were correlated with lymph node(P<0.05).The positive rates of MGMT and EGFR in cases with lymph node metastasis were significantly higher than without ones(P<0.05).The expression of EGFR was higher in the group of cancer cell invasion depth.The expression of MGMT was higher in the group of lower differration degree.MGMT and EGFR were postively related(r =0.2723).Conclusion:Petecton of The expression of MGMT and EGFR in gastric cancer has guidance role to judge the degree of maliguancy and to make the clinical scheme of therapy.

背景:成年人中枢神经系统再生困难,颅脑损伤后,损伤灶周边区域神经细胞的存活数量直接影响患者的预后。如何有效地使移植入创伤性脑损伤灶周边的神经干细胞存活分化,是目前神经修复再生研究的重点。目的:探讨神经干细胞移植入大鼠创伤性脑损伤灶周边的成活、迁移和分化情况。方法:利用无血清培养技术,加入表皮生长因子、碱性成纤维生长因子诱导刺激大鼠胚胎源性前脑神经干细胞生长增殖,并在体外进行克隆培养,移植前行BrdU标记,采用免疫组化和免疫荧光法检测其增殖特性和多向分化潜能,并观察其移植到Fenney’s落体脑损伤模型鼠脑皮质内的成活和迁移情况。结果与结论:免疫组化及免疫荧光检测结果显示克隆细胞球呈nestin和BrdU阳性,分化后呈NSE,GFAP,MAP-2阳性。免疫组化及荧光双标检测结果显示移植后7,14d损伤灶周边散在BrdU阳性细胞,并且GFAP阳性细胞增多。提示前脑神经干细胞在体外培养中能够增殖,并分化为神经元和神经胶质细胞,移植后能够在创伤性脑损伤灶周边存活和迁移,形态上显示出与脑组织整合的特点。

Objective:To study the clinical significance of P-gplycoprotein(P-gp)and lung resistance protein(LRP)expression in breast cancer.Methods:The LRP and P-gp were detected by immunohistochemistry in 86 cases breast cancer.Results:The positive rates of P-gp and LRP were 29.07%(25/86)and 69.77%(60/86)respectively in breast cancer.The positive rates of the infiltrating cancer group and lymph node metastasis were higher(P0.05).Conclusion:The positive rates of LRP were signification higher than P-gp expression(P0.05).The expressions of P-gp and LRP is lower concerned to breast cancer.The detection on expressions of P-gp and LRP in breast cancer play a role for planning chemotherapy regimen.

Objecive:To study the clinincal significance of angiopoietin-2(Ang-2) and epidermal growth factor receptor family,EGFR and C-erbB-2 gene protien in breast carcinoma.Methods: Thirty six cases of adenoma and 86 cases of breast cancer were tested for expression of Ang-2,EGFR and C-erbB-2 by SP immunohistochemistry.Results:The positive rates of Ang-2,EGFR and C-erbB-2 were 63.95%(55/86),56.98%(49/86) and 70.93(61/86) separetly in breast cancer,and the difference had not significance(P0.05).Their possitive rates in breast carcinoma were higher than that in adenoma obviously(P﹤0.001).The expression rates of Ang-2,EGFR and C-erbB-2 in the lymphnode metastasis group and invasive cancer group were higher than nonlymphnode metastasis and noninivasive cancer group(P0.01).EGFR was highly related with C-erbB-2 in the expression of breast carcinoma(r=0.73).Ang-2 was low related with C-erbB-2 in the expression of breast carcinoma.Conclusion:Ang-2,EGFR and C-erbB-2 play roles in occurrence of breast cancer.

谷胱甘肽转移酶(GST)有5个亚型,其中胎盘型GST(GSTπ)占90%左右,在上皮肿瘤内表达较高,而且GSTπ高表达与肿瘤发生和耐药有密切关系~([1～3]),因此GST被称为肿瘤标志物或耐药相关基因.目前关于GSTπ在上皮性肿瘤表达的报道不多,本文采用SP免疫组织化学技术,检测了胃肠腺癌和乳腺癌中GSTπ的表达,分析了GSTπ过表达与癌细胞生物学特性的关系和导致肿瘤耐药机制,旨在为临床治疗提供依据.

目的 观察金属硫蛋白(MT)和DNA修复酶(MGMT)在胃、结直肠癌中的表达临床意义和它们的相关性.方法 采用S-P免疫组织化学方法 ,检测了143例胃、结直肠癌中MT和MGMT的表达状况.结果 MT在胃和结直肠癌中的总阳性率为58.74%(84/143),在胃腺癌中阳性率为48.48%(32/66);在结直肠癌中阳性率为67.53%(52/77).MGMT在胃和结直肠癌中的总阳性率为51.05%(73/143),在胃癌和结直肠癌中阳性率分别为43.94%(29/66)和57.14%(44/77).且MT与MGMT在胃癌和结直肠癌中呈正相关(r=0.35,0.23,P<0.05).结论 MT和MGMT在消化管癌中有较高表达,检测癌瘤中MT和MGMT对制定化疗方案有指导意义.

目的 观察康脑液1号对SD大鼠脑缺血再灌注后病灶和神经胶质纤维酸性蛋白(glial fibrillary acdic protein,GFAP)表达的影响,探讨其对脑缺血再灌注保护作用的可能机制.方法 将180只大鼠随机分为5组,每组36只.分别为假手术组,模型(缺血再灌注)组,盐酸法舒地尔注射液组,康脑液1号A组,康脑液1号B组.采用栓线法复制SD大鼠大脑中动脉梗死模型,分别于缺血2h后再灌注1、7、14 d.观察康脑液1号对大鼠脑缺血再灌注神经功能、梗死面积和病理形态学的影响.采用免疫组织化学法检测缺血半暗带和灶中心区星形胶质细胞GFAP表达的变化.结果 模型组与假手术组相比,GFAP表达阳性细胞数增多;康脑液1号可不同程度地降低实验性脑缺血大鼠的神经功能评分(P<0.01)、减小梗死面积和减轻病理形态改变(P<0.05);康脑液1号各组大鼠脑组织缺血半暗带GFAP表达减少(P<0.05),而梗死灶中心GFAP表达却增多(P<0.05);康脑液1号A组与B组比较差异无统计学意义(P>0.05).结论 康脑液1号可能通过调节脑缺血再灌注后GFAP表达而促进脑损伤修复及重塑,从而发挥脑保护作用.