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.     

大剂量环磷酰胺冲击治疗三种自身免疫病的临床研究报告

目的　观察大剂量环磷酰胺冲击治疗自身免疫病的长期疗效。方法　选择难治性或复发性原发性血小板减少性紫癜 4 0例 ,难治性或复发性自身免疫性溶血性贫血 10例 ,狼疮性肾炎或重症系统性红斑狼疮 2 0例 ,用CTX 0 5～ 1 5 g/m2 体表面积 ,快速静滴 ,2h滴完 ,同时嘱大量饮水 ,每 4周 1次 ,最多为 4次。治疗前后自身对照。结果　ITP、AIHA、SLE长期疗效上差异均有非常显著意义 (P <0 0 1)。总有效率87%。结论　研究结果表明 ,大剂量CTX冲击治疗自身免疫症长期疗效显著、安全、操作简便、经济适用。     

Therapeutic effects of total coumarins from Urtica dentata Hand on collagen-induced arthritis in Balb/c mice

Ethnopharmacological relevance

Urtica dentata Hand (UDH), the root of Laportea bulbifera (Sieb. et. Zucc.) Wedd, has long been utilized in traditional Chinese medicine for the treatment of rheumatoid arthritis and some other autoimmune diseases. Coumarins are the main active principles contributing to UDH's efficacy, but the mechanisms have not been fully clarified.

Aim of study

To explore effects of total coumarins (TC) isolated from UDH on the development of type II collagen (CII)-induced arthritis (CIA) in Balb/c mice.

Materials and methods

Arthritis was induced in Balb/c mice by immunization with an emulsion of 200 mg CII and complete Freund's adjuvant (CFA). The CIA mice were then given with a suspension of TC or saline by intragastric (i.g.) administration every other day. The incidence and severity of disease and histopathology of inflammation were assessed. Inflammatory response was determined by measuring the levels of different inflammation mediators in serum. The effect of TC on differentiation of CD4⁺CD25⁺ Foxp3⁺Treg cells was examined by flow cytometry. The phenotype of bone marrow-derived dendritic cells (DCs), T-bet mRNA level and IL-12p70 secretion by DCs were also detected.

Results

Pharmacologically, treatment with TC for type II collagen induced arthritis in mice through oral administration displayed significant and dose-dependent drop of clinical arthritis score and paw swelling, compared with the untreated CIA mice. Pathologic changes showed that TC protected tissues against bone destruction, whereas an almost complete destruction occurred in the CIA model group. The protective status was associated with a substantial decrease in the production of IFN-γ and IL-2, an increase of IL-10 and TGF-β and suppressive expression of T-bet in DCs. TC also induced the generation of CD4⁺CD25⁺ Treg cells with a Treg phenotype Foxp3. TC-treated DCs were characterized as low expression of MHC class II and CD86 molecules, as well as a reduction of IL-12p70.

Conclusions

Our data suggest that TC provides substantial therapeutic protection against CIA by eliciting immune tolerance and it would be a valuable candidate for further investigation as a new anti-arthritic agent.     

烟曲霉醇联合环磷酰胺对小鼠LA795肺腺癌转移的抑制作用

背景与目的:血管生成抑制剂联合化疗药物治疗肿瘤成为目前研究热点之一。本研究旨在观察烟曲霉醇(fumagillol,TNP-470)联合环磷酰胺(cyclophosphamide,CTX)对肺腺癌小鼠异体移植转移的协同抑制作用,并初步探讨TNP-470抑制肿瘤转移的相关机制。方法:将40只接种高转移性LA795肺腺癌细胞的T739裸小鼠随机分成5组:对照组、溶剂组、TNP-470组(30mg/kg)、CTX组(40mg/kg)、联合组(TNP-47030mg/kg CTX40mg/kg)。实验3周后,处死全部小鼠。剥离皮下肿瘤称瘤重并计算抑瘤率;取出双肺观察表面肿瘤转移情况,计算肿瘤肺转移发生率,计数各组小鼠肺表面转移结节数及计算出肺表面结节转移抑制率。免疫组化和图像分析系统检测皮下移植瘤中微血管密度(microvesseldensity,MVD)、P-选择素表达并定量分析。结果:联合组抑瘤率(81.5%)明显高于其他各组(P<0.01),Q值等于1.21,说明两药合用具有协同作用。与对照组(12.13±4.02)相比,联合组(1.75±1.71)、TNP-470组(4.75±3.34)、CTX组(8.50±2.67)肺表面转移结节数明显下降;同时TNP-470组和联合用药组皮下肿瘤内MVD、P-选择素表达与对照组相比均下降,差异有显著性(P<0.01),而CTX组对此则无明显影响。结论:TNP-470与CTX对LA795肺腺癌的肺结节转移具有协同抑制作用;TNP-470抑制LA795肺腺癌转移与其抑制肿瘤内P-选择素表达有关。     

Cerebrotendinous Xanthomatosis: Molecular Pathogenesis,Clinical Spectrum,Diagnosis, and Disease-Modifying Treatments

Cerebrotendinous xanthomatosis (CTX) is an autosomal recessive lipid storage disorder caused by mutations in the CYP27A1 gene, which encodes the mitochondrial enzyme sterol 27-hydroxylase. Decreased sterol 27-hydroxylase activity results in impaired bile acid synthesis, leading to reduced production of bile acids, especially chenodeoxycholic acid (CDCA), as well as elevated serum cholestanol and urine bile alcohols. The accumulation of cholestanol and cholesterol mainly in the brain, lenses, and tendons results in the characteristic clinical manifestations of CTX. Clinical presentation is characterized by systemic symptoms including neonatal jaundice or cholestasis, refractory diarrhea, juvenile cataracts, tendon xanthomas, osteoporosis, coronary heart disease, and a broad range of neuropsychiatric manifestations. The combinations of symptoms vary from patient to patient and the presenting symptoms, especially in the early disease phase, may be nonspecific, which leads to a substantial diagnostic delay or underdiagnosis. Replacement of CDCA has been approved as a first-line treatment for CTX, and can lead to biochemical and clinical improvements. However, the effect of CDCA treatment is limited once significant neuropsychiatric manifestations are established. The age at diagnosis and initiation of CDCA treatment correlate with the prognosis of patients with CTX. Therefore, early diagnosis and subsequent treatment initiation are essential.     

Associations between Postprandial Gut Hormones and Markers of Bone Remodeling

Gut-derived hormones have been suggested to play a role in bone homeostasis following food intake, although the associations are highly complex and not fully understood. In a randomized, two-day cross-over study on 14 healthy individuals, we performed postprandial time-course studies to examine the associations of the bone remodeling markers carboxyl-terminal collagen type I crosslinks (CTX) and procollagen type 1 N-terminal propeptide (P1NP) with the gut hormones glucose-dependent insulinotropic polypeptide (GIP), glucagon-like peptide 1 (GLP-1), and peptide YY (PYY) using two different meal types—a standardized mixed meal (498 kcal) or a granola bar (260 kcal). Plasma concentrations of total GIP, total GLP-1, total PYY, CTX, and P1NP were measured up to 240 min after meal intake, and the incremental area under the curve (iAUC) for each marker was calculated. The iAUC of CTX and P1NP were used to assess associations with the iAUC of GIP, GLP-1, and PYY in linear mixed effect models adjusted for meal type. CTX was positively associated with GIP and GLP-1, and it was inversely associated with PYY (all p < 0.001). No associations of P1NP with GIP or GLP-1 and PYY were found. In conclusion, the postprandial responses of the gut hormones GIP, GLP-1, and PYY are associated with the bone resorption marker CTX, supporting a link between gut hormones and bone homeostasis following food intake.     

黄蘑多糖对荷瘤小鼠化疗的减毒增效作用

目的研究黄蘑Hohenbuehelia serotina多糖对环磷酰胺(CTX)化疗H22荷瘤小鼠的减毒和增效作用。方法建立小鼠体内移植性肝癌模型;小、中、大剂量治疗组分别ip黄蘑多糖提取物20、40、80mg/kg,连续给药10d,测定荷瘤小鼠的生命延长率和化疗药物CTX的毒副作用以及黄蘑多糖对CTX的减毒和增效作用。同时检测黄蘑多糖对淋巴细胞转化率、IL-2水平等免疫系统功能的影响。结果中、大剂量黄蘑多糖具有抑制肿瘤生长,延长荷瘤小鼠的生存率的作用。与CTX伍用可发挥协同作用,提高抑瘤率(P<0.05、0.01);提高荷瘤小鼠生存质量,体重、白细胞计数及免疫器官指数,与CTX阳性对照组比较均显示显著差异(P<0.05、0.01);实验结果显示黄蘑多糖与CTX伍用具有良好的减毒和增效作用,可增强荷瘤小鼠机体免疫力,提高淋转和IL-2的水平(P<0.05、0.01)。结论黄蘑多糖作为生物反应调节剂可提高机体的免疫力,增强CTX的抗肿瘤作用,同时减轻CTX的毒性。     

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.     

Release of ICTP and CTX telopeptides from demineralized dentin matrices: Effect of time,mass and surface area

Objective

The present study evaluated the influence of time, mass and surface area of demineralized dentin collagen matrices on telopeptides release. The hypotheses tested were that the rates of ICTP and CTX release by matrix bound endogenous proteases are 1) not time-dependent, 2) unrelated to specimen mass, 3) unrelated to specimen surface area.