BACKGROUND: Temozolomide (TMZ) resistance is a significant challenge in treating glioblastoma (GBM). Collagen remodeling has been shown to be a critical factor for therapy resistance in other cancers. This study aimed to investigate the mechanism of TMZ chemoresistance by GBM cells reprogramming collagens. METHODS: Key extracellular matrix components, including collagens, were examined in paired primary and recurrent GBM samples as well as in TMZ-treated spontaneous and grafted GBM murine models. Human GBM cell lines (U251, TS667) and mouse primary GBM cells were used for in vitro studies. RNA-sequencing analysis, chromatin immunoprecipitation, immunoprecipitation-mass spectrometry, and co-immunoprecipitation assays were conducted to explore the mechanisms involved in collagen accumulation. A series of in vitro and in vivo experiments were designed to assess the role of the collagen regulators prolyl 4-hydroxylase subunit alpha 1 (P4HA1) and yes-associated protein (YAP) in sensitizing GBM cells to TMZ. RESULTS: This study revealed that TMZ exposure significantly elevated collagen type I (COL I) expression in both GBM patients and murine models. Collagen accumulation sustained GBM cell survival under TMZ-induced stress, contributing to enhanced TMZ resistance. Mechanistically, P4HA1 directly binded to and hydroxylated YAP, preventing ubiquitination-mediated YAP degradation. Stabilized YAP robustly drove collagen type I alpha 1 ( COL1A1) transcription, leading to increased collagen deposition. Disruption of the P4HA1-YAP axis effectively reduced COL I deposition, sensitized GBM cells to TMZ, and significantly improved mouse survival. CONCLUSION P4HA1 maintained YAP-mediated COL1A1 transcription, leading to collagen accumulation and promoting chemoresistance in GBM.
Temozolomide ; Humans ; Glioblastoma/drug therapy* ; Animals ; Mice ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics* ; YAP-Signaling Proteins ; Hydroxylation ; Dacarbazine/pharmacology* ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Collagen/biosynthesis* ; Collagen Type I/metabolism* ; Prolyl Hydroxylases/metabolism* ; Antineoplastic Agents, Alkylating/therapeutic use*

Objective To establish a rapid and sensitive liquid chromatograph-mass spectrometry (LC-MS/MS) method for the determination of thiabendazole in honeysuckle flowers. Methods The acetic ether was selected as extraction solvent. The mass spectrometer analysis was conducted in the positive ionization electrospray mode using SIM. The transitions m/z 202→175 was used to quantify thiabendazole. Results The satisfactory linearity was obtained in the range of 0.1×10-5-2×10-5mg for thiabendazole (r=0.999 5), and the limit of detection (LOD) of 10.0 ng/ml and the mean recovery of 93.70% were obtained by this LC-MS/MS method. Conclusions The method of LC-MS/MS is sensitive, simple and accurate, and it proved to be suitable for the determination of thiabendazole in Honeysuckle flowers.

Objective:To study the effect of atractylodes lactideⅠ,Ⅱand Ⅲ on the expression of cytokines by inducing M1 type macrophage model. Methods:Apoptotic macrophages were induced by sodium thioglycolate, and then the rat peritoneal inflammatory macrophages were purified by a differential adherence method. The expression of macrophage marker antigen (ED1) was identified by flow cytometry macrophages. Tumor necrosis factor ( TNF-α) , proinflammatory cytokines ( IL-1β) and interleukin-6 ( IL-6 ) were measured by ELISA in vitro. The levels of expression of arginase 1, inducible nitric oxide synthase (iNOS), inflammatory cytokines (IL-1β) chemokine (CCL22) and transforming growth factor (TGF-β) in inflammatory macrophages were determined by RT-PCR. Results:As for the purification of cultured rat inflammatory macrophage expressing ED1, atractylenolideⅠand Ⅲ could reduce the ex-pression levels of iNOS and IL-1βand increase the expression levels of arginase1 CCL22 and TGF-β. The release of inflammatory fac-tor IL-1β decreased, and the release levels of TGF-β and chemokines CCL22 were promoted(P<0. 05). Atropine lactoneⅠ and Ⅲ had significant inhibitory effects on TNF-α, IL-1β and IL-6 in macrophages(P<0. 05). Conclusion: Atractylodes lactone Ⅰand Ⅲ with anti-inflammatory activity can promote the expression of cytokines in inflammatory macrophages, while the change induced by at-ractylaxanthin Ⅱ is not obvious.

Currently, immunotherapy is considered as the fourth major modality of cancer treatment except surgery, chemotherapy and radiotherapy. The new therapeutic approach based on immune checkpoint inhibitors is a landmark innovation. Strategies considering checkpoint inhibitors have shown good anti-tumor effect by targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) and programmed cell death protein 1 (PD-1) or programmed cell death ligand 1 (PD-L1). Moreover, DNA mismatch repair-deficient tumors appear to be potential candidates for these therapies. This review summarizes the discussion and oral presentations in the annual meeting of American Society of Clinical Oncology (ASCO) and ASCO-gastrointestinal cancer (GI) in 2016 and provides an update on immunotherapy in gastrointestinal cancers.

Objective:To study the content determination method for the effective components in WudangⅡFlos lonicerae Caulis to lay foundation for the quality evaluation. Methods: An ultrasonic method was used. The effects of extraction solvent, ultrasonic time, ultrasonic power and ratio of solid to liquid on the contents of rutin and mignonette nucleoside were studied, and the extraction conditions were optimized by a 4-factor and 3-level orthogonal experiment. The chromatographic conditions were as follows:a Phenome-nex Luna-C18(250 mm ×4.60 mm, 5 μm) column was adopted for chlorogenic acid, and a Fortis Xi Phenyl column (250 mm × 4. 6 mm, 5 μm) was used for rutin, loganin and luteoloside;the mobile phase was acetonitrile (B)-0. 4% phosphoric acid (C) solu-tion (15 ∶85) for chlorogenic acid and loganin, and acetonitrile (B) -0. 5% glacial acetic acid aqueous solutjion (D) with gradient e-lution for rutin and luteoloside;the column temperature was 30℃, and the detection wavelength was 327,237,354 and 348 nm, re-spectively. Results:The optimum extraction conditions for rutin and luteoloside from WudangⅡFlos lonicerae Caulis were as follows:the extraction solvent was 60% ethanol, the solid-liquid ratio was 1 ∶30, the ultrasonic power and the ultrasonic time were 350 W and 50 min for rutin, and 250W and 60min for luteoloside. The content of chlorogenic acid, loganin, rutin and luteoloside was 10. 27, 6. 33, 0. 401 and 0. 450 mg·g-1 in the samples, respectively. Conclusion:The method is simple and convenient, accurate and re-producible, which can be used to control the quality of WudangⅡFlos lonicerae Caulis and provide reference for the further develop-ment.

As a new type of antitumor therapy, the use of immune checkpoint inhibitors is a landmark innovation in the treatment of malignant tumors. Previous studies suggest that the immune checkpoint inhibition might be also effective in patients with gastrointestinal cancer. In order to improve the efficacy of immunotherapy, several different strategies are currently under evaluation. In this review, the related discussions in European Organization for Research and Treatment of Cancer (EORTC) Gastrointestinal Tract Cancer Translational Research Meeting in 2014 and American Society of Clinical Oncology (ASCO) annual meeting in the resent two years are summarized. Moreover, the paper reviews the latest clinical research progress about the immunotherapy of gastrointestinal cancers from five aspects: The infiltration of immune cells and new molecular targets, immune checkpoint inhibitors, genetic marker for immunotherapy, combination of checkpoint inhibitors and oncolytic virus, and individualized immunotherapy.

OBJECTIVE:To study the effects of Tibetan medicine Zuotai on the activities of cytochrome oxidase (CYP1A2) and drug metabolism enzyme N-acetyltransferase 2(NAT2)in rats. METHODS:70 SD rats were equally randomized into a normal control (normal saline) group,the groups of single administration of low,middle and high-dose (1.2,3.8 and 12 mg/kg) Zuotai and the groups of multiple administrations thereof(once daily for 12 consecutive days). The rats were given drugs ig. caffeine(25 mg/kg)was given ig to the rats in the normal control group and the groups of single administration on the 2nd day,and to those in the groups of multiple administrations on the 13th day. 5 h later,their urine was collected and added with vitamin C based on 10 mg/ml. High performance liquid chromatography (HPLC) was adopted to determine the cafeine metabolites contents of 5-acetami-do-6-formamido-3-methyl-uric acid(AFMU),1-methylxanthine(1X),1-methyl-uric acid(1U)and 1,7-dimethyl uric acid(17U) in rats’urine,and the activities of CYP1A2 and NAT2 were reflected through (AFMU+1X+1U)/17U and AFMU/(AFMU+1X+1U). RESULTS:Compared with normal control group,the(AFMU+1X+1U)/17U and AFMU/(AFMU+1X+1U)in rats were de-creased,namely the activities of CYP1A2 and NAT2 were lower in the groups of single administration of middle-dose Zuotai and multiple administrations of middle and high-dose Zuotai than in the normal control group. There was statistical difference (P<0.05). CONCLUSIONS:Zuotai can obviously inhibit the activities of CYP1A2 and NAT2 in rats.

To study the effect of Tibetan medicine Zuotai on the activity, protein and mRNA expression of CYP1A2 and NAT2, three different doses (1.2, 3.8 and 12 mg x kg(-1)) of Zuotai were administrated orally to rats once a day or once daily for twelve days, separately. Rats were administrated orally caffeine (CF) on the second day after Zuotai administration, and the urine concentration of CF metabolite 5-acetylamino-6-formylamino-3-methyl-uracil (AFMU), 1-methyluric acid (1U), 1-methylxanthine (1X), 1, 7-dimethylxanthine (17U) at 5 h after study drug administration was determined by RP-HPLC. The activity of CYP1A2 and NAT2 was evaluated by the ratio of metabolites (AFMU+1X+1U)/17U and the ratio of AFMU/(AFMU+1X+1U), respectively. The protein and mRNA expression of CYP1A2 and NAT2 were determined by ELISA and RT-PCR method, respectively. After single administration of Zuotai 3.8 mg x kg(-1) and repeated administration of Zuotai 3.8 and 12 mg x kg(-1), the activity of CYP1A2 and NAT2 decreased significantly compared with control group and there was no significant difference between other dose group and control group. The protein expression of CYP1A2 was significant lower than that in control group after repeated administration of Zuotai 12 mg x kg(-1), and the mRNA expression of CYP1A2 decreased significantly compared with that of control group after single administration of Zuotai 3.8 mg x kg(-1) and repeated admistration of Zuotai 12 mg x kg(-1), separately. The protein expression of NAT2 decreased significantly compared with that of control group after single and repeated administration of Zuotai 3.8 mg x kg(-1), respectively, and the mRNA expression of CYP1A2 decreased significantly compared with control group after single administration of Zuotai 3.8 mg x kg(-1). This study found that Tibetan medicine Zuotai had significant effect on the activity, protein and mRNA expression of CYP1A2 and NAT2.

Objective To evaluate usefulness of high-resolution MRI in quantitative analysis of femoral micro-structure.Methods This study included 110 cases which were divided into 4 groups according to the results of DXA.High-resolution MRI of left hip joint wasperformed on a 1.5T super-conductive MR scanner.3 ROIs were selected on the largest oblique-coronal images of femur to gain thebinary images derived from software processing and 30 parameters were analyzed.Results The parameters to be of statistical significance were 77% and 67%,respectively.Full points of structural parameters with significant difference were 6,4~6 points were 60% and 2~3 points were 40%.Trabecula connective parameters such as trabecula area,mean framework length and mean trabecula perimeter had positive correlation with bone density,r=0.547～0.722,0.58～0.654 and 0.573～0.688,P

0.05).Results No side effect was recorded.100% of images were with excellent quality.Conclusion Patients with previous endovascular stent surgery can receive 1.5T high field MRI examination safely.Usually the examination can be performed in 4~8 weeks after the surgery,but in special case,earlier examination is also acceptable.