目的:按照教育部和学校"停课不停教、停课不停学"的工作部署,为保障学生在疫情期间的学习病理生理学进行线上教学.方法:线上教学应用网络平台与自建课程相结合的方式,在课程设计上,增加小视频,科技前沿、新冠疫情、思政等元素,在实施过程注重知识传授及价值的引领,在教学完成后对教学效果进行及时检验,最后,通过教学反馈进行教学效果反思.结果:教学改革将现代化信息技术与教学相结合,激发学生的学习兴趣,提高了学生的学习深度,促进了教师教学能力的提升.结论:线上教学是教学改革的一个新尝试,为今后的线上线下混合教学奠定了基础.

目的近年来我国的病原生物安全防护工作越来越被重视,所以在医学病原微生物肠道杆菌实验教学中,根据卫生部公布的《人间传染的病原微生物名录》,将肠道致病微生物列为第二类病原微生物,属高致病性病原微生物。方法所以在教学中肠道致病伤寒杆菌减毒株的研究迫在眉睫,特此设计了肠道伤寒杆菌的减毒株实验。结果通过肠道伤寒杆菌减毒株实验,使得肠道伤寒实验无毒性,保持生物学性状。结论避免了学生病原微生物学实验中的感染,提高了病原微生物实验教学的安全性,保证医学高校学生学习到真正的肠道致病菌的生化特性,同时保证了生物安全。

目的:探讨PI3K和BRD4双重抑制剂SF2523对人源脑胶质瘤干细胞TS576增殖的抑制作用,并初步阐明其作用机制.方法:培养人源脑胶质瘤干细胞TS576,将其分为对照组和0.25、0.50、1.00、2.00 μmol·L-1 SF2523组.采用CCK-8法检测各组TS576细胞存活率;将TS576细胞分为对照组和2 μmol·L-1 SF2523组,利用细胞生长计数法检测各组TS576细胞数;将TS576细胞分为对照组和1及2 μmol·L-1 SF2523组,采用流式细胞术检测各组不同细胞周期TS576细胞百分比;将TS576细胞分为对照组和1及2 μmol·L-1 SF2523组,利用Annexin Ⅴ/PI染色法检测各组TS576细胞凋亡率;将TS576细胞分为对照组和1及2 μmol·L-1 SF2523组,采用Western blotting法检测各组TS576细胞中B淋巴细胞瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)和cyclinD1蛋白表达水平.结果:CCK-8检测,作用24、48和72 h时,与对照组比较,0.25、0.50、1.00和2.00 μmol· L-1 SF2523组TS576细胞存活率均明显降低(P＜0.01).细胞生长计数法检测,与对照组比较,2 μmol·L-1 SF2523组细胞数明显减少(P＜0.05或P＜0.01).流式细胞术检测,作用72 h时,与对照组比较,1和2μmol·L-1 SF2523组G1期TS576细胞百分比升高(P＜0.05),S期TS576细胞百分比降低(P＜0.05).Annexin Ⅴ/PI染色检测,作用72 h时,与对照组比较,1和2μmol·L-1 SF2523组细胞凋亡率均明显升高(P＜0.01).Western blotting法检测,作用72 h时,与对照组比较,1和2 μmol·L-1SF2523组TS576细胞中Bax蛋白表达水平升高(P＜0.01),Bcl-2和cyclinD1蛋白表达水平降低(P＜0.05或P＜0.01),Bax/Bcl-2比值升高(P＜0.01).结论:SF2523通过下调cyclinD1表达引起TS576细胞G1期阻滞,通过上调Bax表达、下调Bcl-2表达促进TS576细胞凋亡,从而抑制TS576细胞增殖.

目的：尿素通道蛋白B（ urea transporter B， UT－B）是介导尿素快速跨膜转运的膜蛋白，UT－B基因敲除后可导致老龄小鼠发生心肌肥大，本研究拟探讨UT－B基因敲除后对小鼠线粒体功能的影响。方法：应用试剂盒检测线粒体呼吸链复合体的活性， PCR检测mtDNA 基因D－loop区域多态性／突变情况，流式细胞术检测线粒体膜电位和ROS水平。结果：与野生型小鼠相比， UT－B基因敲除小鼠线粒体复合体I和IV活性明显下降，通过复合体Ⅴ产生的ATP量明显降低；线粒体mtDNA发生多处突变，线粒体损伤标志线粒体DNA／核DNA的比率显著下降。 UT－B基因敲除小鼠线粒体膜电位水平较野生型小鼠明显下降，而心肌ROS水平增加约31．42％。结论：UT－B基因敲除后可导致小鼠线粒体功能障碍，这可能是其老龄后发生心肌肥大的机制之一。

目的：探讨新型氧化石墨烯纳米粒子运载Stat3－siRNA质粒靶向治疗肝癌的作用。方法：Western blot和半定量RT－PCR检测H22细胞中Stat3的蛋白以及下游基因表达水平。 HE染色观察GO－PEI－PEG－FA的安全性；TUNEL法检测原位肝癌的凋亡情况。结果：经叶酸修饰石墨烯基因载体成功将Stat3－siRNA质粒转染进细胞内；实验组Stat3的mRNA和蛋白表达明显下调；PCNA阳性表达明显下调；原位肝癌的凋亡上调。结论：经叶酸修饰氧化石墨烯纳米粒子是安全并且有效的靶向基因运载工具，可携带Stat3特异性siRNA质粒靶向进入肝癌细胞内，从而发挥抑制肝癌生长的作用。

Objective To construct the recombinant vectors that coexpress wild type p53(wt-p53) and siRNA-mdm2 for gene therapy in androgen independent prostate cancer.Methods The subcloning,T-Acloning,and PCR technique were used to construct the recombinant vectors,named pcDNA3.1-p53,siRNA-mdm2,p53/siRNA-mdm2,and siRNA-scramble.The recombinant expression vectors mentioned above were transfected into PC-3 cells,the expression levels of the mdm2 siRNA and p53 after transfection were detected by the semi-quantitative RT-PCR analysis and Western blotting analysis.MTT assay was used to detect the proliferation inhibition of PC-3 cells after transfection.Results The recombinant plasmids containing both wt-p53 and siRNA-mdm2 were successfully constructed,and confirmed by DNA sequence analysis.After transfected for 48 h,the expression of GFP was observed.The expression level of wt-p53 was increased,and the expression of mdm2 was decreased detected by RT-PCR and Western blotting.MTT assay showed that the inhibitory rate of proliferation of PC-3 cells was 40.4% after transfection.Conclusion The recombinant plasmids containing both wt-p53 and siRNA-mdm2 are successfully constructed,they can remarkablely inhibit the proliferation of PC-3 cells.

Objective To study the mechanism of bladder carcinoma cells apoptosis induced by platelet-derivered endothelial cell growth factor(PDECGF)-small interference RNA(siRNA).MethodssiRNA aiming human PDECGF gene synthesized in vitro was labeled by fluorescence and transfected T24 cells.The PDECGF gene and related genes expressions in T24 cells were detected by RT-PCR and Western blot.DNA ladder assay and acridine orange staining were used to observe the apoptosis in T24 cells.ResultsRT-PCR and Western blot demonstrated that psiPDECGF-2 significantly inhibited the expression of PDECGF in T24 cells;with the inhibition of PDECGF,expression of caspase-3 and p53 in the cells elevated obviously.FCM and acridine orange staining showed that psiPDECGF-2 induced apoptosis of T24 cells in vitro.ConclusionspsiPDECGF-2 could significantly induce T24 apoptosis.The mechanism of the apoptosis resulted from the upregulation of caspase-3 and p53 expression.

我国长期以来被认为是前列腺癌"极低发"国家.2002年国际癌症综合网络中心(IARC)资料显示,中国前列腺癌发病率是1.6/10<'5>PY,死亡率为1.0/10<'5>PY,病死率为0.63.事实上,近20年的临床报告已证明我国临床诊断的前列腺癌在逐年增长,而且,以中晚期病例为主.为改变我国前列腺癌的发病现状,与日本专家合作发现,长春50岁以上人群中有1%的前列腺癌发现率(5/501),经申请得到中日政府官员的理解与支持.

Objective To investigate the effects of zinc on the mRNA expression of MMP-2 and MMP-9 of human ptostate carcinoma cell PC-3M.Methods The effects of zinc with different concentractions on proliferation of PC-3M cell were determined by MTT assay.Transwell chamber was used to investigate the effects of zinc on invasion abilities of PC-3M cell.The mRNA expression of MMP-2 and MMP-9 were detected by RT-PCR synthesis after 24h in culture with zinc.The activation of MMP-2 and MMP-9 was detected by gelatin zynography.Results MTT assay showed that zinc with a certain concertration could inhibit proliferation of PC-3M cell.Under the effect of zinc,the number of PC-3M cell which moved through the matrigel membrane decrease.The result of RT-PCR and gelatin zynography respectively showed the synthesis and activation of MMP-2 and MMP-9 decreased after treated with zinc.d.Conclusion Zinc with a certain concentraction can inhibit the proliferation and invasion abilities of PC-3M cell,and the mechanism is possibly relevant to the inhibitation of the synthesis and activation of MMP-2 and MMP-9.

目的 利用RNA干扰技术在裸鼠膀胱癌动物模型中通过抑制靶向沉默血小板源性内皮生长因子基因(PDECGF)的表达,探讨其在活体动物肿瘤组织中的作用.方法 将psiPDECGF-1及-2稳定转染的阳性T24细胞株及其他对照组直接接种于裸鼠皮下,观察肿瘤细胞生长及成瘤情况、测定肿瘤体积的变化;免疫组化检查PDECGF蛋白表达;TUNEL法检测肿瘤细胞凋亡.结果 裸鼠体内成瘤实验发现siRNA2组第21天时成瘤率只有50%,而其他各组成瘤率为100%.siRNA2组生长速度明显慢于其他3组,siRNA2组不仅成瘤延迟,且生长速度明显慢于其他3组(P＜0.01).免疫组化证实siRNA2组肿瘤组织PDECGF 蛋白表达最低; TUNEL检测siRNA2组细胞凋亡数明显多于其他3组 (P＜0.05).结论 PDECGF siRNA载体稳定转染细胞株体内实验可见膀胱癌细胞成瘤率降低,成瘤延迟及肿瘤生长速度减慢,癌细胞凋亡增多.PDECGF siRNA在裸鼠体内依然对膀胱癌细胞有明显抑制作用.

目的 验证信号传导与转录激活因子(STAT)3、血管内皮生长因子(VEGF)、存活素(Survivin)在膀胱肿瘤细胞的表达并揭示其与膀胱肿瘤病理分级及预后的关系.方法 通过免疫组织化学的方法验证3种物质在膀胱肿瘤的表达并进行分析.结果 膀胱肿瘤STAT3表达阳性组的病理分级较高(大部分位于Ⅱ级以上)、5年生存率较低(31.1%),同时均有VEGF、Survivin的异常表达.结论 STAT3在膀胱肿瘤异常表达时会导致肿瘤的恶性程度增高、预后更差,且有可能是通过其下游的VEGF、Survivin来起作用.

目的沉默人膀胱移行细胞癌细胞T24的STAT3基因,并探讨其对T24细胞生长抑制及诱导凋亡的作用机制。方法利用含有STAT3 mRNA的siRNA的pSilencer3.1-siRNA-STAT3重组质粒转染T24细胞;采用Western印迹、RT-PCR等技术观察重组质粒对STAT3基因表达的影响;用MTT法观察重组质粒对T24细胞的体外生长抑制作用;用流式细胞术(FCM)及吖啶橙染色检测重组质粒的诱导凋亡作用。结果重组质粒成功转染T24细胞;Western印迹、RT-PCR证实重组质粒在蛋白及mRNA水平都显著抑制STAT3基因的表达水平,抑制率为59%~72%;MTT及FCM证实重组质粒可显著抑制T24细胞的体外生长并诱导细胞凋亡,抑制率及凋亡率分别为53%和17.3%。结论pSilencer3.1-siRNA-STAT3重组质粒可抑制STAT3在人膀胱肿瘤细胞的表达,抑制肿瘤细胞的生长,促进其凋亡。

目的探讨pSilencerl.0-U6-siRNA-STAT3重组质粒对人前列腺癌细胞PC3裸鼠移植瘤生长的抑制作用。方法采用裸鼠前列腺癌模型模拟siRNA-STAT3基因治疗实验,观察siRNA-STAT3重组质粒的抗瘤疗效,Western blot法检测重组质粒对STAT3基因表达的影响,HE及Tunel染色方法观察肿瘤组织形态及细胞凋亡情况。结果肿瘤接种后第49天重组质粒组肿瘤体积为（391±56）mm 3 ,盐水对照组为（1111±182）mm 3 ,空质粒对照组为（981±169）mm 3 ,与对照组相比,差异有统计学意义（P<0.01）;重组质粒抑制瘤体内STAT3及pTyr-STAT3基因表达率分别为77%及68%,与对照组比较,差异有统计学意义（P<0.01）。HE染色证实重组质粒组出现大片细胞核固缩、破碎等凋亡征象,Tunel染色显示癌组织出现大量细胞凋亡。结论pSilencerl.0-U6- siRNA-STAT 3重组质粒能明显抑制人前列腺癌细胞PC3裸鼠移植瘤的生长,具有良好的应用前景。

目的探讨血清铜、锌含量与前列腺癌之间的关系，并将其与血清前列腺特异性抗原（PSA）比较，探讨其在前列腺癌诊断中价值比较。方法采用原子吸收分光光度法测定76例前列腺癌患者、125例前列腺增生患者和191例健康男性的血清铜、锌含量。结果前列腺癌组血清锌明显低于健康对照组，血清铜和铜/锌比值明显高于健康对照组（P〈0.05）。前列腺癌组血清铜及铜/锌比值与前列腺增生组比较未见显著性差异;前列腺癌组血清锌低于前列腺增生组（P〈0.05）。前列腺增生组血清锌与健康对照组比较未见显著性差异;前列腺增生组血清铜和铜/锌比值高于健康对照组（P〈0.05）。PSA在4.0-10.0ng/ml灰色区域的前列腺癌组，血清锌ROC-AUC（receiver operator characteristic curve-area under curve）为66%，高于PSA的ROC-AUC。结论血清铜、铜/锌比值升高及血清锌降低可能是前列腺癌发生的危险因素，血清锌在PSA（4.0～10.0）ng/ml灰色区域的前列腺癌诊断效率高于PSA，其可能为前列腺癌的诊断提供有价值的指标。

Objective To study the mechanism of Prostate cancer cells apoptosis induced by STAT3(Signal transducer and activator of transcription 3)-siRNA(small interference RNA).Methods The pair of DNA template coding siRNA against STAT3 was synthesized to reconstruct pSilencer 2.1-U6-STAT3-siRNA,which was transfected into PC3 cells.The STAT3 expression in PC3 cells were detected by Western blot and Northern blot analyses apoptosis related genes Bcl-2,C-Myc and Cyclin D1 expression were detected in the transfected cells.FCM and acridine orange were used to observe the apoptosis in PC3 cells.Results Western blot and Northern blot analyses demonstrated that pSilencer 2.1-U6-siRNA-STAT3 could significantly inhibit the expression of STAT3 in PC3 cells;with the inhibition of Stat3,expression of Bcl-2,C-Myc and Cyclin D1 in the cells reduced obviously.FCM and acridine orange visualized pSilencer 2.1-U6-STAT3-siRNA could induce apoptosis of PC3 cells in vitro.Conclusion pSilencer 2.1-U6-STAT3-siRNA could significantly induce PC3 apoptosis;The mechanism of the apoptosis resulted from the downregulation of Bcl-2,C-Myc and Cyclin D1 expression.

目的研究survivin-siRNA对前列腺癌裸鼠移植瘤生长的作用,探讨survivin与前列腺癌发生、发展的关系。方法人前列腺癌细胞PC-3M荷瘤裸鼠18只随机分为3组:mock组(n=6),pSi-scrambled组(n=6)和pSi-sur2组(n=6)。通过电子穿孔仪将质粒转入裸鼠移植瘤内,20d后处死动物,计算抑瘤率。利用Western blot检测survivin蛋白表达水平,免疫组化SP法检测增殖指数变化;TUNEL法检测肿瘤细胞凋亡的变化。结果体内实验表明,pSi-sur2组与其他两对照组比较裸鼠移植瘤生长速度明显减慢(P<0.01),肿瘤组织survivin蛋白表达及肿瘤增殖指数显著降低(P<0.01),而肿瘤凋亡显著增加(P<0.01)。结论通过RNA干扰技术抑制survivin的表达,在裸鼠体内可显著抑制前列腺癌生长。

目的 探讨血清铜、锌含量与前列腺癌之间的关系,并将其与血清前列腺特异性抗原(PSA)比较,探讨其在前列腺癌诊断中价值比较.方法 采用原子吸收分光光度法测定76例前列腺癌患者、125例前列腺增生患者和191例健康男性的血清铜、锌含量.结果 前列腺癌组血清锌明显低于健康对照组,血清铜和铜/锌比值明显高于健康对照组(P＜0.05).前列腺癌组血清铜及铜/锌比值与前列腺增生组比较未见显著性差异;前列腺癌组血清锌低于前列腺增生组(P＜0.05).前列腺增生组血清锌与健康对照组比较未见显著性差异;前列腺增生组血清铜和铜/锌比值高于健康对照组(P＜0.05).PSA在4.0-10.0ng/ml灰色区域的前列腺癌组,血清锌ROC-AUC(receiver operator characteristic curve-area under curve)为66%,高于PSA的ROC-AUC.结论 血清铜、铜/锌比值升高及血清锌降低可能足前列腺癌发生的危险因素,血清锌在PSA(4.0～10.0)ng/ml灰色区域的前列腺癌诊断效率高于PSA,其可能为前列腺癌的诊断提供有价值的指标.

Objective To establish the prostate cancer orthotopic(o.t.)implantation model in C57BL6 syngenic mice and discuss its value of application.Methods The o.t .implantation model was established by subcapsular implantation of histologically intact tumor tissues harvested f rom a subcutaneous tumor by RM-1 cells into the protate gland of C57BL6 syngenic mice using microsyringe.The subcutaneous(s.b.)implantation model was used as control.The rate of metastasis and the survival time of these two models were compared.Results Metastasis occurred in all mice of the o.t .model with the highest rate of pelvic lymph node(16/16)and the lung(8/16).No metastasis was seen in the control group except local growth and infiltration.The survival time of o.t .model was(17.0±3.5)days,markedly shorter than that of s.b.model(33.0 ±5.6)days.Conclusion The prostate cancer o.t .implantation model is an optimal clinical animal model to observe metastasis that mimics the natural situation of prostate cancer in human body quite well.