目的 观察芳香烃受体(AhR)及Th17相关细胞因子在类风湿关节炎中的表达水平及其对疾病的预测价值。方法 选择2014年1月—2015年12月于我院就诊的RA患者60例作为观察组,选取同期于我院进行健康体检的正常人60例作为对照组,采用酶联免疫吸附法检测血清中 IL-17、IL-23及AhR的表达水平,并分析其与疾病活动度的关系。结果 RA患者血清IL-17、IL-23及AhR水平高于对照组(P＜0.05)。而根据病情严重程度,RA 非早期组IL-17、IL-23及AhR水平较早期组增高(P＜0.05)。血清 IL-17 水平与除CRP以外的病情活动度指标均呈正相关关系(P＜0.05)。IL-23 水平与SJC、TJC、HAQ 、DAS28 评分呈正相关关系(P＜0.05),但其他指标无明显相关性(P>0.05)。RA患者PBMC中AhR的表达水平与各项临床指标无相关性(P>0.05)。IL-17和IL-23水平与Sharp评分呈正相关关系(r=0.895,P＜0.01;r=0.708,P＜0.01)。AhR的表达与血清IL-17和IL-23水平呈正相关(r=0.415,P＜0.01)。经荧光定量PCR检测结果显示,RA患者AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平高于对照组(P＜0.05)。且RA 非早期组AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平较早期组增高(P＜0.05)。经相关关系检测,RA患者AhR及其下游因子CYP1A1、IL-17、FOXP3 mRNA的表达水平与Sharp评分呈正相关关系(r=0.715,P＜0.01;r=0.734,P＜0.01;r=0.812,P＜0.01;r=0.755,P＜0.01)。结论 Th17相关细胞因子IL-17和IL-23在RA病理生理过程中发挥重要作用,其表达增高,提示关节炎症处于活跃状态,骨质破坏较重,可作为评估RA病情的重要指标。RA患者体内AhR蛋白及其相关下游信号通路均呈高表达状态,AhR通路在RA患者的发病过程中可能发挥关键作用。

Objective To observe the expression level of aroma receptor (AhR) and Th17 related cytokines in rheumatoid arthritis and its predictive value to disease. Methods Sixty patients with RA who were treated in our hospital from January 2014 to December 2015 were selected as the observation group. Sixty healthy subjects were randomly selected as the control group. IL-17, IL-23 and AhR levels in serum were detected by enzyme-linked immunosorbent and the relationship between IL-17, IL-23 and disease activity were analyzed. Results IL-17, IL-23 and AhR levels in serum of RA patients were significantly higher than those in controls (P<0.05). However, the levels of IL-17, IL-23 and AhR levels were significantly higher in the non- early RA group than in the early group (P<0.05), depending on the severity of the disease. There was a positive correlation between serum IL-17 level and disease activity index except CRP (P<0.05). IL-23 level was positively correlated with SJC, TJC, HAQ and DAS28 scores (P<0.05), but no significant correlation was found between other indexes (P>0.05). The expression level of AhR in PBMC of RA patients was not correlated with clinical indexes (P>0.05). IL-17 and IL-23 levels were positively correlated with Sharp score (r=0.895, P<0.01; r=0.708, P<0.01). The expression of AhR was positively correlated with serum IL-17 and IL-23 levels (r=0.415, P<0.01). The expression of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA patients were significantly higher than those in the control group (P<0.05) by fluorescence quantitative PCR. The expression of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA group were significantly higher than those in early group (P<0.05). The expression level of AhR and its downstream factors CYP1A1, IL-17 and FOXP3 mRNA in RA patients were positively correlated with Sharp scores (r=0.715, P<0.01; r=0.734, P<0.01; r=0.812, P<0.01; r=0.755, P<0.01). Conclusion The Th17-related cytokines IL-17 and IL-23 play an important role in the pathophysiology of RA, and the expression of Th17-associated cytokines is increased, suggesting that arthritis is active and bone destruction is serious. The AhR protein and its associated downstream signaling pathways are highly expressed in RA patients, and the AhR pathway may play a key role in the pathogenesis of RA patients.

目的 探讨妇科门诊宫颈癌高危人群采取阴道镜检查后配合宫颈细胞学检查对宫颈癌筛查价值。方法 选取2016年2月—2017年4月我院妇科门诊就诊合并宫颈癌高危患者387例为研究对象,所有患者均行阴道镜检查和宫颈细胞学检查,以最终病理检查为诊断“金标准”,探究阴道镜、宫颈细胞学检查在宫颈癌中筛查灵敏度、特异度及阳性符合率。结果 387例纳入研究宫颈癌高危患者,经病理诊断后明确诊断为宫颈癌51例,占13.18%。细胞学检查在宫颈癌中筛查灵敏度为72.55%,特异度为90.48%,阳性符合率为88.11%。阴道镜检查在宫颈癌中筛查灵敏度为50.98%,特异度为94.64%,阳性符合率为88.89%。宫颈癌筛查中,细胞学检查灵敏度高于阴道镜检查,特异度低于阴道镜检查(P＜0.05),两者检查阳性符合率比较差异无统计学意义(P＞0.05)。结论 妇科门诊宫颈癌高危患者宫颈筛查中,阴道镜筛查具有较高特异度,细胞学检查具有较高灵敏度,联合检查能提高筛查价值。

目的 探讨ERCC1、RRM1、TS蛋白表达对晚期非小细胞肺癌(NSCLC)个体化治疗的指导意义。方法 收集经病理确诊的晚期NSCLC患者87例,其中67例愿意接受药敏免疫组化检测的患者作为研究组,采用SP法检测肿瘤组织ERCC1、RRM1、TS蛋白表达,并根据蛋白表达情况选择化疗方案;另外20例患者不进行药敏免疫组化检测,以常规吉西他滨联合顺铂方案化疗,以此作为对照组。比较两组患者化疗的有效率,疾病控制率(DCR),并以无进展生存期(PFS)为指标比较患者预后。结果 研究组67例患者中,PR 33例(49.25%),SD 13例(19.4%),PD 21例(31.35%);对照组20例患者中,PR 4例(20%),SD 4例(20%),PD 12例(60%),两组疗效之间有差异( χ²=6.437,P=0.04),研究组DCR为68.6%,高于对照组DCR 40%,差异有统计学意义(χ²=5.372,P=0.034)。研究组患者的中位PFS高于对照组,研究组的PFS为5月,对照组为3月,差异有统计学意义(P＜0.05)。结论 对晚期NSCLC患者进行ERCC1、RRM1、TS药敏蛋白免疫组化检测,指导个体化治疗方案,能提高患者化疗的疾病控制率及延长患者的疾病进展时间。

目的 对比观察单孔、单操作孔及三孔胸腔镜治疗早期非小细胞肺癌(NSCLC)的临床疗效。方法 选择125 例早期NSCLC患者,分为单孔组(38例)单操作孔组(42例)和三孔胸腔镜组(45例),观察3组手术结果和并发症发生率。结果 3组患者均顺利完成手术,无中转开胸。单孔组手术时间长于单操作孔及三孔组,差异有统计学意义(P＜0. 05)。对比所有3组手术患者的术中出血量及淋巴结清扫数目、术后总引流量及引流管留置时间、术后并发症发生率,差异无统计学意义(P＞0. 05)。单孔组及单操作孔组术后疼痛程度评分优于三孔组,差异有统计学意义(P＜0. 05)。结论 单孔及操作孔胸腔镜治疗早期NSCLC已可取代三孔胸腔镜技术,其术后恢复快,疗效确切,其中单孔手术对设备及胸腔镜医师操作技术熟练程度等要求更高,故在设备仍未有突破性的进展时,单操作孔胸腔镜手术可作为治疗早期NSCLC的优先选择。

Objective To compare the clinical effects of uniportal video-assisted thoracic surgery (VATS), single utility port VATS and 3-portal VATS lobectomy for patients with early stage non-small cell lung cancer. Methods Patients were divided into uniportal VATS lobectomy group(n=38), single utility port VATS lobectomy group(n=42) and 3-portal VATS lobectomy group (n=45). The surgical results and complication rates were observed. Results All patients completed the operation successfully, no one was changed to open operation. Operation time in uniportal VATS lobectomy group were longer than single utility port VATS lobectomy group and 3-portal VATS lobectomy group(P＜0. 05). There were no significant differences in intraoperative blood loss, number of lymph node dissection, the amount and time of postoperative extubation, and the incidence of postoperative complications(P＞0. 05). Post-operative pain score were higher in 3-portal VATS lobectomy group than in uniportal VATS lobectomy group and single utility port VATS lobectomy group (P＜0. 05). Conclusion Uniportal VATS lobectomy and single utility port VATS lobectomy can replace the 3-portal VATS lobectomy in treatment of early NSCLC, because of the faster postoperative recovery and curative effect. Uniportal VATS lobectomy requires special equipment and more operation skills, as there is no breakthrough in the equipment, single utility port VATS lobectomy may still be used as the first choice for treatment of early NSCLC.

目的 了解异鼠李素对LPS刺激下THP-1细胞炎症因子IL-6、MCP-1、TNF-α释放的影响及其抗炎特征。方法 用PI单染色法检测其细胞存活率;用ELISA法检测THP-1细胞炎症因子IL-6、MCP-1、TNF-α释放。结果 不同浓度的LPS均能使炎症因子IL-6释放增高,且24 h及48 h间水平无差异,但随着LPS刺激浓度的增高,细胞的坏死数量也会随之升高;不同浓度的异鼠李素能在不同时间点不同程度地抑制炎症因子MCP-1、TNF-α、IL-6的释放。结论 异鼠李素能抑制LPS刺激下THP-1细胞炎症因子IL-6、TNF-α、MCP-1的释放,并呈浓度依赖性。

Objectives Our study is aim to investigate the influence of the isorhamnetin on the releasing of inflammatory factor cytokines of the THP-1 cells. Methods PI single staining method was used to detect the cell survival rate. ELISA was used to detecte the concentrations of inflammatory cytokines(MCP-1、IL-6、TNF-α). Results The concentration of IL-6 in THP-1 cells were increased after stimulated with LPS and there no difference between the 24 h group and 48 h group. The survival rate of THP-1 cells decreased as the concentration of LPS increased.The isorhamnetin in different concentration could inhibit the secretion of inflammatory cytokines in different time. Conclusion It is found in our study that isorhamnetin may inhibit the secretion of MCP-1, IL-6, TNF-α in THP-1 cells stimulated by LPS in a concentration dependent way.

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.