星点设计-效应面法优化柴银口服液提取工艺及其稳定性研究

目的 优化柴银口服液提取工艺,明确其成品稳定性。方法 在单因素试验基础上将加水量、提取时间、提取次数作为考察因素,以黄芩苷转移率与澄清度值的总评归一（OD）值为评价指标,采用星点设计-效应面法优化柴银口服液提取工艺,将经优化后工艺条件生产的柴银口服液成品与原工艺成品进行加速与长期稳定性对比。结果 最佳提取工艺参数为加水量8.3倍、提取时间2.2 h、提取3次,此条件下OD值最高为0.702,此条件下生产的柴银口服液在稳定性考察中pH、澄清度、黄芩苷含量及微生物限度等指标均明显优于原工艺成品。结论 星点设计-效应面法优化柴银口服液提取工艺方法可靠、参数精确,能有效提升其价值、延长保质期。     

Transcriptomic and Immunophenotypic Characterization of Tumor Immune Microenvironment in Squamous Cell Carcinoma of the Oral Tongue

The tumor immune microenvironment of oral tongue squamous cell carcinoma may be accountable for differences in clinical behavior, particularly between different age groups. We performed RNA expression profiling and evaluated tumor infiltrating lymphocytes (TILs) and their T-cell subsets in order to assess the functional status of oral tongue squamous cell carcinoma tumor microenvironment and detect potentially clinically useful associations. Archival surgical pathology material from sixteen oral tongue squamous cell carcinoma patients was microscopically evaluated for TIL densities. RNA was extracted from macrodissected whole tumor sections and normal controls and RNA expression profiling was performed by the NanoString PanCancer IO 360 Gene Expression Panel. Immunostains for CD4, CD8 and FOXP3 were evaluated manually and by digital image analysis. Oral tongue squamous cell carcinomas had increased TIL densities, numerically dominated by CD4 + T cells, followed by CD8 + and FOXP3 + T cells. RNA expression profiling of tumors versus normal controls showed tumor signature upregulation in inhibitory immune signaling (CTLA4, TIGIT and PD-L2), followed by inhibitory tumor mechanisms (IDO1, TGF-β, B7-H3 and PD-L1). Patients older than 44 years showed a tumor microenvironment with increased Tregs and CTLA4 expression. Immunohistochemically assessed CD8% correlated well with molecular signatures related to CD8 + cytotoxic T-cell functions. FOXP3% correlated significantly with CTLA4 upregulation. CTLA4 molecular signature could be predicted by FOXP3% assessed by immunohistochemistry (R² = 0.619, p = 0.026). Oral tongue squamous cell carcinoma hosts a complex inhibitory immune microenvironment, partially reflected in immunohistochemically quantified CD8 + and FOXP3 + T-cell subsets. Immunohistochemistry can be a useful screening tool for detecting tumors with upregulated expression of the targetable molecule CTLA4.Electronic supplementary materialThe online version of this article (10.1007/s12105-020-01229-w) contains supplementary material, which is available to authorized users.     

豆根管食通口服液对食管鳞癌细胞miR-377的影响研究

目的观察豆根管食通口服液对食管鳞癌细胞株KYSE450的细胞增殖、细胞凋亡及对miR-377、CD133和VEGF表达的影响。方法先制备豆根管食通口服液含药血清,然后分为高中低三个浓度,分别作用24 h、48 h、72 h,筛选出最佳浓度和最佳作用时间,然后将KYSE450细胞株分为空白组、豆根管食通中剂量组、氟尿嘧啶组。采用MTT方法检测豆根管食通口服液含药血清对KYSE450细胞增殖的影响,采用流式细胞仪检测豆根管食通口服液含药血清对KYSE450细胞凋亡的影响,采用RT-PCR方法检测豆根管食通口服液含药血清对KYSE450细胞miR-377的影响,采用WB方法检测豆根管食通口服液含药血清对KYSE450细胞CD133和VEGF表达的影响。结果筛选出最佳浓度为豆根管食通中剂量组,最佳作用时间为48 h;MTT结果显示,豆根管食通口服液对KYSE450细胞能够明显抑制细胞增殖,豆根管食通口服液含药血清中剂量组的细胞凋亡率为(18.8±2.66)%,高于空白组(P <0.01),促进miR-377表达,与空白对照组相比(P <0.05),抑制CD133表达(P <0.05),抑制VEGF表达(P <0.01)。结论豆根管食通口服液对食管鳞癌KYSE450细胞株有抗增殖、促凋亡的作用,其机制可能是通过上调miR-377的表达,进而抑制CD133和VEGF表达。     

甲醇沉淀-高效液相色谱法同时测定奶茶中5种添加剂

目的建立甲醇沉淀-高效液相色谱同时测定奶茶中安赛蜜、苯甲酸、糖精钠、山梨酸和咖啡因的方法。方法样品加甲醇沉淀旋涡混合提取后,离心分离,取上清液,过0.22μm滤膜,高效液相色谱检测。色谱条件:使用Agilent Poroshell 120 EC-C₁₈(3 mm×150 mm,2.7μm)色谱柱,以甲醇和0.02 mol/L乙酸铵溶液为流动相,梯度洗脱,流速为1.0 ml/min,柱温为35℃,检测波长为240 nm。结果 5种添加剂在1.0μg/ml~50.0μg/ml线性良好,r>0.999 9,方法检出限为0.002 g/kg~0.005 g/kg,测定限为0.006 g/kg~0.015 g/kg。对饮料进行高、中、低3个浓度水平测定,回收率为93.36%~104.5%,相对标准偏差(RSD)为1.0%~3.4%。结论本方法样品预处理简单、分离效果好、准确度高,适用于奶茶中安赛蜜、苯甲酸、糖精钠、山梨酸和咖啡因的同时检测。     

A comparison of the pro-inflammatory, NF-κB-dependent cytokines: TNF-alpha, IL-1-alpha, IL-6, and IL-8 in different oral fluids from oral lichen planus patients

To explore the feasibility of detection of the level of NF-κB-dependent cytokines in oral fluids from patient with oral lichen planus (OLP) for clinical application, 13 OLP subjects were enrolled in the study as were 13 age–sex-matched controls. In each subject, the whole unstimulated saliva (WUS), mixture of saliva and isotonic saline oral rinse (Saliva-NaCl), and lesion tissue transudates (TT) were collected by standard techniques. The level of cytokines, TNF-alpha, IL-1-alpha, IL-6, and IL-8 in three types of oral fluids was determined by ELISA. In the three types of oral fluids, a significantly higher level of these cytokines was detected in OLP patients than in normal controls. These results indicate that NF-κB-dependent inflammatory cytokines may be detected at increased levels in certain oral fluids which may have diagnostic and prognostic potential for monitoring disease activity and making therapeutic decisions in patients with OLP.     

正交偏光镜下人体胆汁液晶光学性质的研究

本文报导了在正交偏光镜下，对人体胆汁液晶的光学性质的研究，结果表明，胆汁液晶是一种非均质体，在正交偏光镜下，呈现出特殊的干涉图样和四次消光现象，光性符号是单轴正性的，干涉色级是一级灰白，双折射率Δｎ＝０．０１１－０．０１７。     

Copper-inhibition of the growth of oral streptococci and actinomyces

Copper ions were found to inhibit the rate of growth in broth culture of Streptococcus mitis, Streptococcus mutans, Streptococcus salivarious, Streptococcus sanguis, Actinomyces viscosus and Actinomyces naeslundii. In all cases, 10^?3 M copper inhibited the rate of growth, whereas 10^?4 M and lower concentrations had little or no effect. At the concentrations used in mouthwashes one mode of action of copper ions is to reduce the rate of growth of oral bacteria in vitro.     

口矫器治疗阻塞性睡眠呼吸暂停的机制及疗效预测

目的观察一种下颌前移性口矫器治疗阻塞性睡眠呼吸暂停(OSA)的疗效并探讨其治疗原理,探讨应用颌面结构分析法来预测口矫器的疗效。　方法70例经多导睡眠仪(PSG)检查确诊的OSA患者,男性63例,女性7例,平均年龄(50.5±11.6)岁,呼吸暂停指数(AI)(34.9±21.3)次/h,夜间最低血氧饱和度(66.3±16.5)%。经口腔科会诊后制作口矫器,治疗3个月后复查PSG检查。每例患者治疗前均在带及不带口矫器两种状态下摄头颅侧位片作颌面结构分析。　结果治疗后AI由(34.9±21.3)次/h减至(15.6±19.2)次/h(P<0.000 1),夜间最低血氧饱和度由(66.3±16.6)%升至(74.1±15.5)%(P<0.001)。共有45例患者AI减少≥50%。反应良好组患者缩颌较明显,前上面部长度较长,置入口矫器后下颌显著前突、上气道径增宽、长度缩短、上气道面积增大。　结论口矫器对部分OSA患者具有确切疗效,其治疗作用可能主要与改善睡眠期上气道周围结构关系,保持气道通畅有关。     

沙利霉素对口腔鳞癌细胞增殖和凋亡的影响

目的:探讨沙利霉素(salinomycin)对口腔鳞癌细胞增殖和凋亡的影响,并初步探讨沙利霉素对信号通路的影响。方法:培养口腔鳞状细胞癌细胞系CAL-27,将1、2、4、8、16、32 μmol/L沙利霉素和1.25、2.5、5、10、20、40、80 μmol/L顺铂与CAL-27细胞共同培养,24 h和48 h后用细胞计数试剂盒-8(cell counting kit-8,CCK-8)法检测沙利霉素和顺铂对CAL-27细胞增殖的影响;0、2、4、8 μmol/L沙利霉素和0、5、10、20 μmol/L顺铂与CAL-27细胞共培养48 h后,通过流式细胞术检测沙利霉素和顺铂对CAL-27细胞周期的影响,蛋白免疫印迹法(Western blot)检测CAL-27细胞中天冬氨酸特异性半胱氨酸蛋白酶-3(cysteine-containing aspartate-specific proteases-3,Caspase-3)、天冬氨酸特异性半胱氨酸蛋白酶-9(cysteine-containing aspartate-specific proteases-9,Caspase-9)、脱氧核糖核酸(deoxyribonucleic acid,DNA)修复酶(poly ADP-ribose polymerase,PARP)、蛋白激酶B(protein kinase B, Akt)和磷酸化蛋白激酶B (phosphorylated protein kinase B,p-Akt)的表达。结果:CCK-8实验表明沙利霉素和顺铂均能显著抑制口腔鳞状细胞癌CAL-27细胞增殖,且抑制作用呈时间依赖性和药物浓度依赖性,但是相对于临床一线化疗药物顺铂而言,沙利霉素对CAL-27细胞增殖的抑制效果更加显著(P<0.001)。细胞周期检测表明,与加入二甲基亚砜(dimethyl sulfoxide,DMSO)的对照组相比,8 μmol/L沙利霉素与CAL-27细胞共同培养48 h后,细胞休眠期/DNA合成前期的CAL-27细胞比例明显升高(40.40%±1.99% vs.64.46%±0.90%,P<0.05), DNA合成期和DNA合成后期/有丝分裂期的CAL-27细胞比例出现降低(24.32%±2.30% vs.18.73%±0.61%,P<0.05,35.01%±1.24% vs.16.54%±1.31%,P<0.05);顺铂对CAL-27细胞周期没有特异性改变。蛋白免疫印迹法结果显示,沙利霉素在上调CAL-27细胞中Caspase-3和 Caspase-9蛋白表达(P<0.05)的同时下调PARP、Akt和p-Akt蛋白的表达(P<0.05)。结论:相对于顺铂而言,沙利霉素对CAL-27细胞增殖有更强的抑制作用,并且能将口腔鳞状细胞癌CAL-27细胞周期阻滞在细胞休眠期/DNA合成前期,同时能够诱导CAL-27细胞发生凋亡,这一机制可能和Akt/p-Akt 信号通路相关。