Inhibition of cathepsin-K and matrix metalloproteinase by photodynamic therapy

ObjectivesThe objective of this study was to determine the effects of antimicrobial photodynamic therapy (aPDT) with indocyanine green (ICG) and toluidine blue (TB) on protease activity (matrix-bound cathepsin K and matrix metalloproteinase (MMP) and dentin bond strength.MethodsCaries-free human third molars were assigned to five groups: 1—control group, 2—application of ICG with activation using an 810 nm diode (aPDT), 3—application of ICG, 4—application of TB with activation using a 660 nm diode (aPDT), and 5—application of TB. For the enzymatic investigation, dentin beams were incubated for either 3 days or 3 weeks. Aliquots of the incubation media were analyzed by ELISA for CTX (C-terminal cross-linked telopeptide of type I Collagen) and ICTP (cross-linked carboxy-terminal telopeptide of type I collagen). For microtensile bond strength testing (μTBS), composite resins were layered onto the tooth surface; the samples were then subjected to μTBS. Kruskall–Wallis and Mann–Whitney U tests were applied for statistical analysis of CTX and ICTP, one way-ANOVA and Tukey’s test were applied for statistical analysis of μTBS.ResultsPretreating the dentin matrices with aPDT decreased the endogenous protease activity. ICG with laser activation resulted in the highest μTBS. Therefore, aPDT should be considered as a treatment method because it can reduce MMP-mediated dentin degradation and increase the μTBS.SignificanceInhibiting endogenous protease activity improves the stability of the dentin–adhesive bond and the durability of the bond strength.     

Characteristic Distribution of Ciguatoxins in the Edible Parts of a Grouper,Variola louti

Ciguatera fish poisoning (CFP) is one of the most frequently encountered seafood poisoning syndromes; it is caused by the consumption of marine finfish contaminated with ciguatoxins (CTXs). The majority of CFP cases result from eating fish flesh, but a traditional belief exists among people that the head and viscera are more toxic and should be avoided. Unlike the viscera, scientific data to support the legendary high toxicity of the head is scarce. We prepared tissue samples from the fillet, head, and eyes taken from five yellow-edged lyretail (Variola louti) individuals sourced from Okinawa, Japan, and analyzed the CTXs by LC-MS/MS. Three CTXs, namely, CTX1B, 52-epi-54-deoxyCTX1B, and 54-deoxyCTX1B, were confirmed in similar proportions. The toxins were distributed nearly evenly in the flesh, prepared separately from the fillet and head. Within the same individual specimen, the flesh in the fillet and the flesh from the head, tested separately, had the same level and composition of toxins. We, therefore, conclude that flesh samples for LC-MS/MS analysis can be taken from any part of the body. However, the tissue surrounding the eyeball displayed CTX levels two to four times higher than those of the flesh. The present study is the first to provide scientific data demonstrating the high toxicity of the eyes.     

Impact of radiotherapy on bone health in women with rectal cancer – A prospective cohort study

IntroductionPelvic radiotherapy (RT) increases the risk of pelvic insufficiency fractures. The aim was to investigate if RT is associated with changes in serum bone biomarkers in women with rectal cancer, and to examine the incidence of radiation-induced bone injuries and the association with bone biomarkers.Material and methodsWomen diagnosed with rectal cancer stage I-III, planned for abdominal surgery ± preoperative (chemo) RT, were prospectively included and followed one year. Serum bone biomarkers comprised sclerostin (regulatory of bone formation), CTX (resorption), BALP and PINP (formation). A subgroup was investigated with annual pelvic magnetic resonance imaging (MRI). The association between RT and bone biomarkers was explored in regression models.ResultsOf 134 included women, 104 had surgery with preoperative RT. The formation markers BALP and PINP increased from baseline to one year in the RT-exposed group (p < 0.001, longitudinal comparison). In the adjusted regression analysis, the mean increase in PINP was higher in the RT-exposed than the unexposed group (17.6 (3.6–31.5) μg/L, p = 0.013). Sclerostin and CTX did not change within groups nor differed between groups. Radiation-induced injuries were detected in 16 (42%) of 38 women with available MRI. At one year, BALP was higher among women with than without bone injuries (p = 0.018, cross-sectional comparison).ConclusionsPreoperative RT was associated with an increase in the formation marker PINP, which could represent bone recovery following RT-induced injuries, commonly observed in participants evaluated with MRI. These findings should be further explored in larger prospective studies on bone health in rectal cancer patients.     

有机锗（Ge—132）增强环磷酰胺抗瘤作用的实验研究

本文观察了有机锗在试验治疗荷移植肝癌小鼠中增强血清超氧化物歧化酶活性及细胞免疫的作用，增强环磷酰胺抗移植肝癌作用以及它本身抗移植肝癌的效果，结果是：高剂量Ｇｅ－１３２（１２．５ｍｇ／次）治疗组的抑癌率为３１％，移植肝癌ＷＢＣ浸润者为７２．７％，血清ＳＯＤ活性分别是３６．４％，８１Ｎｕ／ｍｌ，两组比较差异显著（Ｐ〈０．０５），ＣＴＸ组的抑癌率是３７％，癌体ＷＢＣ浸润者为２７．３％，血清ＳＯＤ活性为１     

来氟米特和环磷酰胺治疗以肾病综合征为主的IgA肾病的对照研究

①目的 比较来氟米特(LEF)和环磷酰胺(CTX)分别联合糖皮质激素治疗以肾病综合征为主要表现的IgA肾病的疗效和安全性.②方法 将56例患者随机平均分为两组,分别给予激素+LEF、激素+CTX,随访28周.检测血常规、尿常规、24小时尿蛋白定量、血清白蛋白(Alb)、谷丙转氨酶、谷草转氨酶、血肌酐(SCr),观察疗效及其不良反应.③结果 LEF组治疗总有效率71.4%,CTX组64.3%,两组比较差异无统计学意义(P>0.05);来氟米特组不良反应总发生率17.9%,环磷酰胺组32.1%,两组比较差异有统计学意义(P<0.05).④结论 LEF联合糖皮质激素治疗以肾病综合征为主要表现的IgA肾病的疗效与CTX联合激素治疗的早期疗效相似,不良反应发生率低,但长期疗效及不良反应仍需进一步观察.     

蜂胶黄酮对环磷酰胺诱导HL7702细胞损伤的保护作用

目的探讨蜂胶黄酮(EPF)对环磷酰胺(CTX)诱导HL7702细胞凋亡的保护作用。方法将HL7702细胞按一定密度分别接种于96孔板和24孔板,培养24 h后,向各孔中分别加入CTX(4 ng/L)、CTX(4 ng/L)+EPF(20、30、40 ng/L),继续培养24 h,进行WST-1、AO/EB双染、Annexin-V-FITC/PI流式细胞染色。采用小鼠随机分组,试验组每天给予EPF 100、200、400 mg/mL灌胃,0.5 mL/支,连续给药10 d,同时注射CTX0.15 g/kg,共3 d。取血清检测过氧化氢酶(CAT)、乳酸脱氢酶(LDH)、肌酸激酶(CK)、谷丙转氨酶(GPT)、谷草转氨酶(GOT)等活性及尿素氮(BUN)含量。结果 EPF与CTX共处理HL7702细胞生长抑制明显改善,当EPF浓度达到40 ng/mL时,促进生长作用呈现正比例曲线特征。试验组凋亡细胞明显减少,绝大多数细胞形态饱满,几乎没有死亡细胞。CTX单独处理的HL7702细胞组24 h晚期凋亡细胞占30.2%,CTX与EPF共处理细胞组凋亡细胞分别占19.2%、13.1%、7.1%。结论 EPF对CTX诱导的HL7702凋亡作用具有拮抗作用,可缓解CTX引起的毒性。     

大鼠体内环磷酰胺和顺铂联合用药的药代动力学初步研究

 目的　建立双波长高效液相色谱同时测定环磷酰胺和顺铂质量浓度的方法，并研究其在大鼠体内的药代动力学。方法　色谱柱为Hypersil-ODS2，流动相为甲醇-水（80∶20），紫外检测器在254 nm和195 nm检测顺铂和环磷酰胺质量浓度。结果　环磷酰胺保留时间为5.15 min，质量浓度线性范围0.5～10 μg／ml（r＝0.9999），平均回收率为97.4 %[相对标准偏差（RSD）<5 %]，检测线0.1 μg／ml（信噪比＞3）；顺铂保留时间为9.26 min，质量浓度线性范围0.1～2.0 μg／ml（r＝0.9998），平均回收率为98.1 %（RSD＜5 %），检测线0.1 μg／ml（信噪比＞3）。结论　该法能有效测定两药浓度，并提供了研究其药代动力学的方法，联合用药使二者消除速度下降，消除半衰期延长，存在交互作用。     

Cobratoxin inhibits pain-evoked discharge of neurons in thalamic parafascicular nucleus in rats: Involvement of cholinergic and serotonergic systems

The present study investigated the inhibitory effect of cobratoxin (CTX) on pain-evoked discharge of neurons in thalamic parafascicular nucleus (Pf) of rats and analyzed some of the mechanisms involved in this effect. Intracerebroventricular injection (icv) of CTX at 0.56, 1.12 and 4.50 μg/kg resulted in a dose-dependent inhibitory effect on the pain-evoked discharges of Pf neurons. The inhibition of pain-evoked discharges of Pf neurons by CTX at high dose (4.50 μg/kg) persisted at least for 2 h, while the inhibitory effect of morphine (40 μg) persisted no longer than 30 min. The inhibitory effect of CTX was reversed by pretreatment with atropine (icv, 5 μg). In contrast, icv injection of naloxone (4 μg) had no effect on CTX-induced inhibition. Furthermore, pretreatment with parachlorophenylalanine, a specific inhibitor of tryptophan hydroxylase, also significantly attenuated the inhibitory effect of CTX. The results suggested that: (a) CTX has a dose-dependent inhibitory effect on pain-evoked discharges of Pf neurons, confirming electrophysiologically the antinociceptive action of CTX; (b) the inhibitory effect of CTX has a longer duration compared to that of morphine; (c) central cholinergic and serotonergic systems, but not opioidergic system, are involved in the inhibitory effect of CTX.     

甲基莲心碱增强环磷酰胺诱导小鼠Lewis肺癌细胞凋亡的作用

目的评价甲基莲心碱(Nef)对环磷酰胺(CTX)诱导小鼠Lewis肺癌细胞凋亡作用的影响。方法Lewis肺癌移植瘤小鼠40只,雄性,体重18～22g,随机分成CTX+Nef、CTX、Nef、NS4个组,每组10只。通过检测瘤重抑制率及瘤细胞凋亡率,观察Nef联合CTX对小鼠Lewis肺癌的疗效和诱导癌细胞凋亡的影响。结果CTX+Nef、CTX组的瘤重抑制率分别为80.08%、58.95%,明显高于Nef组(6.78%),P<0.01;而CTX+Nef组较CTX组也有增高(P<0.05);流式细胞术检测CTX+Nef、CTX组的癌细胞凋亡率分别为53.50%、25.92%,明显高于Nef、NS组(7.09%,5.63%),P<0.01;CTX+Nef组也高于CTX组(P<0.05);TUNEL法检测CTX+Nef、CTX组的癌细胞凋亡指数分别为(10.24%±11.08%),(5.36%±6.24%),较Nef、NS组明显增加(1.45%±1.98%,1.03%±1.65%),P<0.05;CTX+Nef组的凋亡指数较CTX组也有明显增加(P<0.05)。结论甲基莲心碱能增强环磷酰胺诱导的小鼠Lewis肺癌细胞凋亡作用,且两者具有协同效应。     

每周小剂量化疗治疗高龄晚期非小细胞肺癌的临床研究

目的 探讨每周小剂量化疗方案对高龄晚期非小细胞肺癌的疗效和毒性反应。方法 42例高龄晚期非小细胞肺癌患者，随机分成两组。治疗组22例，CTX200mg／m^2／dl，EPI30mg／m^2／dl，CBP100mg／m^2／dl；每周重复一次，三周后休一周，为一周期，三周期后评定疗效和毒性反应。对照组20例，CTX500mg／m^2／dl，EPI60mg／m^2／dl，CBP100mg／m^2／dl-3；每28d重复一次，重复三次后评定疗效和毒性反应。结果 治疗组有效率40．9％，对照组有效率45．0％，二组比较差异无显著性（P〉0．05）。毒性反应：治疗组13．63％，对照组75．00％，二组比较差异有著性（P〈0．05）。结论 每周小剂量化疗方案对高龄晚期非小细胞肺癌的疗效肯定，毒性反应轻，适合高龄晚期患者。