目的：通过研究肝癌组织内转录因子Sp1的表达及其与肝癌患者临床病理特征及预后的关系，探讨转录因子Sp1作为肝癌预后预测指标的可行性。方法：对98例根治性切除术的肝细胞肝癌肿瘤组织芯片进行免疫组化检测Sp1的表达情况，分析其与肝癌患者临床病理特征及预后之间的关系。结果：免疫组化结果显示Sp1在肝癌组织中表达明显高于对应正常肝脏组织，在有微血管侵犯的患者中升高尤其明显。进一步分析显示Sp1表达与肝癌患者术后总体生存率呈负相关，而与肝癌患者术后复发率呈正相关。结论：转录因子Sp1在肝癌中明显高表达，可作为肝癌患者预后的独立预测指标。

Objective: To explore the clinical significance of transcription factor Sp1 expression in hepatocellular carcinoma (HCC) and association between Sp1 expression and survival in HCC patients. Methods:With the use of immunoreactivity, Sp1 expres-sion and its correlation with other clinicopathological characteristics were examined in a tissue microarray that contains samples from 98 HCC patients. Results:HCC tissues expressed markedly higher levels of Sp1 than did adjacent normal liver tissues. Sp1 expression was closely associated with microvascular invasion, which suggests that HCC with more microvascular invasion is prone to have in-creased Sp1 expression. Overexpression of Sp1 correlates with significantly shorter overall survival and higher recurrence rates in HCC patients after curative resection. Conclusion:Transcription factor Sp1 may be an independent prognostic factor for both overall surviv-al and cumulative recurrence rate.

目的 研究磁刺激下健康人咬肌抑制反射(masseter inhibitory reflex,MIR)及其恢复情况,为临床辅助检测颅面部疾病提供依据.方法 选择30名健康人,单脉冲模式磁刺激颏神经,记录咬肌肌电,统计早期静息期(the early silent period,SP1)和晚期静息期(the late silent period,Sp2)的潜伏期及持续时间、SP2相对波幅值；在双脉冲模式的条件刺激和测试刺激间设置不同间隔时间(100、200、300、400、500和600 ms),分析双脉冲模式磁刺激下MIR的恢复情况.结果 健康人SP1潜伏期为12.1(11.1,14.4) ms,持续时间为(17.3±2.9)ms;SP2潜伏期和持续时间分别为(47.7±6.0)和(39.7±13.3) ms;SP2相对波幅值为100.0％.不同间隔双脉冲模式下测试刺激所得SP1有稳定的波段；而SP2发生变化,在间隔时间较短时测试刺激所得SP2的面积减少,随着间隔时间的延长,测试刺激所得SP2面积逐渐恢复,100 ms时SP2面积恢复17.1％,400 ms时恢复93.4％,600 ms时几乎完全恢复.结论 采用单脉冲和双脉冲模式磁刺激激发MIR并分析其恢复情况,可用来评估边缘系统、脑干、三叉神经感觉和运动纤维、咀嚼肌等整个系统的功能状态.

Objective To analyze the masseter inhibitory reflex (MIR) and the recovery cycle of the MIR reflex after magnetic stimulation in normal subjects.Methods In 30 healthy subjects we studied the MIR evoked by single magnetic stimulation in the mental territory.Masseter electromyographic activity,latency and duration of the early silent periods (SP1) and late silent periods (SP2),and SP2 amplitude percent were recorded.Paired stimuli technique was used,conditioning and test stimuli were delivered at different interstimulus intervals (ISI),ie.100,200,300,400,500,and 600 ms,then the recovery cycle of the MIR was analyzed Results Latency of SP1 was 12.1(11.1,14.4) ms,and duration ofthe SP1 was (17.3±2.9) ms.Latency of SP2 was (47.7 ±6.0) ms,and duration of the SP2 was (39.7 ± 13.3) ms.SP2 amplitude percent was 100.0％.With the paired stimuli technique,SP1 of the inhibitory reflex evoked by the test stimuli was found to be stable at every ISIs,but SP2 of the inhibitory reflex evoke

目的 探讨P物质(SP)对骨髓基质干细胞(BMSCs)增殖、分化的影响,并从Wnt/ β-链蛋白信号转导通路角度探讨其可能机制. 方法 取SD大鼠第3代BMSCs,根据加入的刺激物不同将其分为4组(n=3):SP组、SP +SP受体拮抗剂组、SP+ Wnt/ β-链蛋白信号转导通路抑制剂1(DKKl)组、对照组(加入等量磷酸盐缓冲液).培养1、3、5、7、9 d采用Alarmar Blue法检测各组BMSCs的增殖情况;于培养7、14d应用荧光定量核酸扩增(Q-PCR)法检测各组碱性磷酸酶(ALP)、骨钙素基因的表达;于培养1、3、5、7 d采用Western blot法检测各组Wnt/β-链蛋白信号转导通路相关蛋白β-链蛋白、C-myc的表达;于培养3d应用免疫荧光法检测各组β-链蛋白的核转移情况.结果 于培养l、3、5、7、9d,SP组细胞数量大于SP+ SP受体拮抗剂组、SP+ DKK1组和对照组,差异均有统计学意义(P＜0.05).于培养14d,SP+ SP受体拮抗剂组、SP+ DKK1组和对照组的ALP、骨钙素基因表达量均明显低于SP组,差异有统计学意义(P＜0.05).于培养l、3、5、7 d,SP+ SP受体拮抗剂组、SP+ DKK1组及对照组的C-myc蛋白的表达量均低于SP组,差异均有统计学意义(P＜0.05);培养3、5d,SP+SP受体拮抗剂组、SP+ DKK1组和对照组的β-链蛋白的表达量均低于SP组,差异均有统

Objective To explore the effect of substance P (SP) on the proliferation and differentiation of bone marrow stromal stem cells (BMSCs) and its association with Wnt/β-catenin signaling pathway.Methods The third passage BMSCs from SD rats were divided into 4 groups according to the stimulator added:SP,SP + SP receptor antagonist,SP + DKK1 (dickkopf 1) and phosphate buffered saline (PBS) (control).BMSCs were identified using the flow cytometry.At 1,3,5,7 and 9 days,proliferation of BMSCs was detected using Alarmar Blue method.At 7 and 14 days,alkaline phosphatase (ALP) and osteocalcin mRNA were detected using quantitative PCR.At 1,3,5 and 7 days,the protein expression of Wnt/β-catenin signaling including β-catenin and C-myc protein was detected using western blotting.At 3 days,nuclear transfer of β-catenin was investigated using immunofluorescence staining.Results Flow cytometry authenticated BMSCs.At 1,3,5,7 and 9 days,the cells were significantly more in SP group than in c

目的 探讨转录因子Sp1在肾盂尿路上皮细胞癌(肾盂癌)的表达及其临床意义.方法 采用免疫组织化学PV-9000法检测Sp1蛋白在43例肾盂癌标本和11例肾盂正常黏膜组织的表达水平.结果 Sp1蛋白在肾盂癌和黏膜组织中的阳性表达率分别为74.4％(32/43)和0,组间差异有统计学意义(P＜0.05).Sp1在低分级和高分级肾盂癌组织中的阳性表达率分别为59.1％(13/22)和90.5％(19/21),Sp1在浅表性(Tis～T1)和浸润性(T2～T4)肾盂癌组织中的阳性表达率分别为58.3％(14/24)和94.7％(18/19),组间差异有统计学意义(P＜0.05).结论 转录因子Sp1蛋白在肾盂癌组织中呈高表达,且其阳性表达与肿瘤的组织学分级和T分期呈正相关,提示Sp1参与了肾盂癌的发生和进展过程.

Objective To investigate the expression and clinical significance of transcription factor Sp1 in uroepithelial cell carcinoma (UCC) of renal pelvis.Methods The immunohistochemical PV-9000 method was used to study the expression level of Sp1 protein in 43 cases of UCC and 11 cases of normal mucosa in renal pelvis.Results The positive expression rate of Sp1 protein in UCC and normal mucosa of renal pelvis was 74.4％ and 0 (P ＜ 0.05),and that in low-grade and high-grade UCC of renal pelvis was 59.1％ and 90.5％ (P ＜ 0.05),that in superficial and infiltrated UCC of renal pelvis was 58.3％ and 94.7％ (P ＜ 0.05),respectively.Conclusion The positive expression of Sp1 protein is directly related to the histological grades and T stages of UCC in renal pelvis,suggesting Sp1 may participate in the carcinogenesis and progression of UCC.

从海洋球石藻Emiliania huxleyi病毒EhV99B1的基因组中克隆了丝氨酸蛋白酶(Sp)基因(GenBank登录号:KC161207),对该基因的开放阅读框(ORF)进行系统的生物信息学分析,并在大肠杆菌中融合表达,通过亲和层析法获得了纯化的重组Sp。结果表明:EhV99B1-Sp基因的ORF为1 110bp,编码368个氨基酸,蛋白相对分子质量为39.5kDa;该基因片段与GenBank中EhV86-Sp的同源性很高,核苷酸及其对应的氨基酸序列同源性分别为95%和97%,而与其他物种Sp序列的同源性仅为28%~32%,说明其可能是丝氨酸蛋白酶家族中的一个新成员;预测的二级结构特征显示EhV99B1-Sp的蛋白结构域中具有典型的LTAGHC(组氨酸活性位点区域)和AICNGDSGGPLF(丝氨酸活性位点区域)两个丝氨酸蛋白酶催化活性位点的氨基酸基序,是一个两次跨膜蛋白;将该基因在大肠杆菌中进行低温诱导表达,得到分子量为60kDa的重组蛋白,经鲤鱼肌肉丝氨酸蛋白酶(MBSP)抗体检测证实为Sp,且重组蛋白在大肠杆菌细胞中具有明显的生物学活性。本研究结果为进一步探讨EhV99B1-Sp在病毒与宿主相互作用过程中的调节作用及其功能与应用奠定基础。

A serine protease (Sp)gene (GenBank accession number:KC161207)was cloned from the genome of marine microalgal Emiliania huxleyi virus (Coccolithovirus )EhV99B1 isolate.Bioinformatic analysis was pre-formed on its open reading frame and the recombinant protein was expressed in E.coli and purified by affinity chromatography.Results showed that the length of the ORF of EhV99B1-Sp was 1 110 bp,which encoded a pro-tein of 386 amino acids with a molecular mass of 39.5 kDa and pI of 6.255.EhV99B1-Sp shared a high sequence similarity with EhV86-Sp,between whom the similarities of nucleotide sequences and deduced amino acid sequences were 95% and 97%,respectively.However,the sequence similarity between EhV99B1-Sp and serine protease from other organisms was only 28% to 32%.Sequences analysis suggested that EhV99B1-Sp might be a new member of the serine protease family.Secondary structure prediction showed that EhV99B1-Sp protein contained two motifs of typical serine protease catalytic active s