Golgi-protein 73 facilitates vimentin polymerization in hepatocellular carcinoma.

Golgi-protein 73 (GP73) is highly expressed in hepatocellular carcinoma (HCC) and, as a secretory protein, it has been proposed as a serum biomarker indicating progression of HCC. The underlying mechanism by which GP73 may promote HCC metastasis is still poorly understood. In this study, we discovered that GP73 interacted with vimentin to facilitate Serine/Threonine-protein phosphatase PP1-alpha (PP1A)-mediated dephosphorylation of vimentin at S56 and facilitated vimentin polymerization, which blocked vimentin degradation via TRIM56-mediated ubiquitin/proteasome-dependent pathway. Strikingly, Clomipramine, a 5-hydroxytryptamine receptor (5-HTR) agonist approved for the treatment of depression, impaired GP73-mediated vimentin polymerization to effectively inhibit metastasis of HCC with high GP73 expression, which provided a new strategy against HCC metastasis. Lastly, it was found that serum GP73 (sGP73) correlated positively with vimentin in primary tissues of HCC, suggesting that sGP73 might serve as a potential serum biomarker for companion diagnosis of HCC with highly expressed vimentin. In summary, this study reveals the process of GP73-mediated vimentin polymerization and proves that Clomipramine serves as a potential drug targeting vimentin for metastatic HCC patients with high sGP73 level.

RNautophagic regulation of DNMT3a-dependent DNA methylation by Linc00942 enhances chemoresistance in gastric cancer.

BACKGROUND: Energy balance has long been known to extend lifespans and inhibit carcinogenesis in multiple species by slowing age-related epigenetic changes while the underlying mechanisms remain largely unknown. Herein, we found that starvation activated autophagy to remodel the DNA methylation profile by inhibiting DNMT3a expression. METHODS: Illumina Infinium MethylationEPIC BeadChip and dot blot assay were performed to quantify the global DNA methylation level. Protein-RNA interactions were validated through RNA immunoprecipitation and RNA pull-down assay. In vitro and in vivo experiments were carried out to testify the effect of DNMT3a on chemoresistance. RESULTS: Autophagy is impaired in chemoresistance which was associated with differential DNA methylation and could be reversed by DNMT3a inhibition. Autophagy activation decreases the expression of DNMT3a mRNA, accompanied with the downregulation of chemoresistance-related Linc00942. Knockdown of Linc00942 reduces DNMT3a expression and genome-wide DNA methylation while Linc00942 overexpression increased DNMT3a expression and correlated hypermethylation in cancer cells and primary tumour tissues. Mechanistically, Linc00942 recruits RNA methyltransferase METTL3 to stimulate N6-methyladenosine (m6A) deposit on DNMT3a transcripts, triggering IGF2BP3/HuR to recognize modified mRNA for reinforced stability. SQSTM1/p62 recruits Linc00942 for autophagic degradation which can be abrogated after autophagy inhibition by p62 knockdown or chloroquine treatment. CONCLUSIONS: Inhibition of autophagy increases Linc00942 expression to promote chemoresistance and autophagy activation or hypomethylating agent decitabine restores chemosensitivity by reducing global DNA methylation. Overall, this study identifies a novel methylation cascade linking impaired RNautophagy to global hypermethylation in chemoresistance, and provides a rationale for repurposing decitabine to overcome chemoresistance in cancer treatment.

Acute necrotising pancreatitis: measurements of necrosis volume and mean CT attenuation help early prediction of organ failure and need for intervention.

OBJECTIVES: This study explored the early predictive value of volume and mean CT density of necrosis for adverse outcomes in patients with acute necrotising pancreatitis (ANP). METHODS: A total of 155 patients with ANP who underwent CECT within 7 days of symptom onset were included. The necrosis volume, mean CT density, and modified CT severity index (mCTSI) were calculated. C-reactive protein (CRP) and blood urea nitrogen (BUN) levels both 48 h after symptom onset were reviewed. Adverse outcomes were recorded. The predictive value of each indicator was assessed using ROC curve analysis. RESULTS: There were significant associations between necrosis volume and mean CT density and organ failure (OF), persistent OF (POF), and need for intervention (p < 0.001 for all). For predicting OF, the area under the curve (AUC) was significantly higher for necrosis volume than for mCTSI and BUN (AUC: 0.84 vs 0.67, p = 0.0011; 0.84 vs 0.71, p = 0.0193, respectively). For predicting POF and need for intervention, the AUCs for necrosis volume were significantly higher than those for mCTSI (AUC: 0.79 vs 0.66, p = 0.0045; 0.77 vs 0.61, p = 0.0019, respectively), but did not significantly differ from those for CRP and BUN. For predicting OF, a significantly better predictive value was achieved with mean CT density than with mCTSI (AUC: 0.79 vs 0.67, p = 0.0163). There were no significant differences in predictive value between mean CT density, CRP, and BUN. CONCLUSIONS: The volume and mean CT density of necrosis based on CECT can provide early prediction of OF, POF, and need for intervention. KEY POINTS: • Compared to mCTSI, necrosis volume might be used to more accurately diagnose organ failure and persistent organ failure and might be better associated with the need for intervention. • Necrosis volume and mean CT density based on CECT are reliable quantitative predictors for organ failure, persistent organ failure, and intervention in acute pancreatitis.

Magnetic resonance colonography with intestine-absorbable nanoparticle contrast agents in evaluation of colorectal inflammation.

OBJECTIVES: To develop a nanoparticle-based MRI protocol based on transrectal administration of intestine-absorbable nanoparticle contrast agents to evaluate ulcerative colitis (UC). METHODS: Solid lipid nanoparticles (SLNs) were synthesized by loading gadolinium diethylenetriaminepentaacetic acid (Gd-DTPA) and octadecylamine-fluorescein isothiocyanate to produce Gd-FITC-SLNs as T1 contrast agents. Twenty mice with acute UC were divided into four groups: enema with Gd-FITC-SLNs, intravenous injection of Gd-FITC-SLNs, enema with Gd-DTPA, and intravenous injection of Gd-DTPA. Five mice with chronic UC and five mice without UC underwent enema with Gd-FITC-SLNs. Axial T1- and T2-weighted MR images were obtained before and 20, 40, 60, 80,100, and 120 min after enema or intravenous injection of the contrast agent. The signal-to-noise ratios (SNRs) of the colorectal wall were measured in both groups. The MRI findings were correlated with subsequent histological confirmation. RESULTS: At 20 min after enema with Gd-FITC-SLNs, MRI showed the following contrast enhancement pattern: acute UC > normal intestinal wall > chronic UC. A continuous enhancement effect was observed in mice with acute UC, whereas a slight continuous enhancement of the colorectal wall was observed in mice with chronic UC. The normal intestinal wall rapidly metabolized the contrast agent, and the enhancement decreased on sequential scans. There was no significant difference between the SNRs of the intestinal wall at 20 min after intravenous Gd-DTPA and transrectal Gd-FITC-SLN administration. CONCLUSIONS: Enema with Gd-FITC-SLNs may be helpful for the diagnosis and differential diagnosis of acute and chronic UC and can confer the same or better results than with intravenous Gd-DTPA. KEY POINTS: • Enema with Gd-FITC-SLNs may be helpful for the diagnosis and differential diagnosis of acute and chronic UC. • Enema with Gd-FITC-SLNs can achieve the same or better result than that with intravenous Gd-DTPA. • SLN-based MR colonography enhances the colorectal wall inflammation, based on the colonic absorption of the nanoparticle contrast agents.

Identification of tumorigenicity-associated genes in osteosarcoma cell lines based on bioinformatic analysis and experimental validation.

Osteosarcoma is the most common primary malignant tumor of bone. Tumorigenic investigation of osteosarcoma cell lines may facilitate preclinical studies of targeted therapy. Therefore, the aim of this study was to explore the tumorigenicity-associated genes in osteosarcoma cells. We found that 138 genes were highly expressed and 86 genes were lowly expressed in highly tumorigenic osteosarcoma cell lines (143B, MNNG/HOS, and SJSA-1) compared with poorly tumorigenic osteosarcoma cell lines (MG-63, Saos-2, and U-2 OS). Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that highly expressed genes were associated with amino acids and energy metabolism, while lowly expressed genes were associated with cell cycle and DNA replication. Gene Ontology (GO) analysis showed that highly expressed genes were associated with endoplasmic reticulum stress response and aggrephagy, whereas lowly expressed genes were correlated with extracellular matrix assembly and DNA damage response. Further analysis identified six highly expressed genes and six lowly expressed genes. Three of highly expressed genes (DDX10, FOXA2, and HEY1) were correlated with poor prognosis, while three of lowly expressed genes (CYP26B1, GP1BB, and IFI44) showed the opposite trend in patients with osteosarcoma. Knockdown of HEY1 significantly inhibited the tumorigenicity of 143B cells in BALB/c nude mice.

Associations Between Genetic Polymorphisms in the VEGFA, ACE, and SOD2 Genes and Susceptibility to Diabetic Nephropathy in the Han Chinese.

Objective: To investigate the association between polymorphisms in the rs2010963 and rs69947 loci of the vascular endothelial growth factor A (VEGFA) gene; the rs4646994 locus of the angiotensin I converting enzyme (ACE) gene; and the rs4880 locus of the superoxide dismutase 2 (SOD2) gene with genetic susceptibility to type 2 diabetic nephropathy (T2DN) in the Chinese Han population. Methods: A total of 650 Chinese Han patients with T2DN and 580 non-nephropathy patients with type 2 diabetes were enrolled in this study. Sanger sequencing was used to detect the genotypes of the rs2010963 and rs69947 loci within VEGFA, the rs4646994 locus of for the ACE gene, and the rs4880 locus of the SOD2 gene in all subjects. Enzyme-linked immunosorbent assays were used to detect VEGFA, ACE, and SOD2 levels in serum. Results: The risk of T2DN was significantly increased (odds ratio [OR] = 1.15, confidence interval [95% CI]: 1.03-1.30) in patients with the GC/CC genotypes compared to those with the GG genotype at the rs2010963 locus of the VEGFA gene under the dominant model of inheritance. Similarly, the risk of T2DN was significantly increased (OR = 1.17, 95% CI: 1.05-1.31) in patients with the CA/AA genotypes compared to those with the CC genotype at the rs69947 locus of the VEGFA gene under the dominant model of inheritance. In addition, the risk of T2DN was increased (OR = 1.57, 95% CI: 1.37-1.74) in patients with the AA genotype compared to the CC/CA genotypes under the recessive model of inheritance. The risk of T2DN was also significantly increased (OR = 1.54, 95% CI: 1.24-1.97) in patients with the ID/DD genotypes compared to those with the II genotype at the rs4646994 locus of the ACE gene under the dominant model; and (OR = 1.41, 95% CI: 1.26-1.57) for DD genotype compared to the II/ID genotypes under the recessive model. We also found the risk of T2DN was significantly increased (OR = 1.25, 95% CI: 1.11-1.39) under the dominant model in patients with the TC/CC genotypes compared to the TT genotype, and for the CC genotype (compared to the TT/TC genotype) (OR = 1.45, 95% CI: 1.18-1.66) under the recessive model at the rs4880 locus of the SOD2 gene rs4880. The haplotypes GAC (OR = 1.17, 95% CI: 1.04-1.29), CAT (OR = 1.12, 95% CI: 1.03-1.60), and CAC (OR = 1.13, 95% CI: 1.01-1.24) constructed from VEGFA gene rs2010963 and rs69947 loci, and the SOD2 gene rs4880 locus were associated with a higher risk for T2DN. Finally, we found that VEGFA protein levels from subjects with the rs2010963 GG genotype and the rs69947 CC genotype were higher in both case groups and the control group than in subjects with rs2010963 GA/AA genotypes and rs69947 CA/AA genotypes, respectively (p < 0.05). The ACE protein levels for variants at the rs4646994 locus showed that the case group and control group subjects with the DD genotype had the highest levels, followed by the ID genotype and the II genotype (p < 0.05). Conclusion: Genetic variation in the VEGFA gene at the rs2010963 and rs69947 loci, the ACE gene at the rs4646994 locus, and the SOD2 gene at the rs4880 locus may increase the risk of developing T2DN.

Establishment and Characterization of Two Novel Cholangiocarcinoma Cell Lines.

BACKGROUND: Cholangiocarcinoma (CCA) is a rare, aggressive and highly lethal tumor. The disease is very difficult to diagnose and multi-modality treatments are ineffective. To improve our understanding of the biological characteristics of CCA, and facilitate the identification of valid treatments, an in-depth characterization of two novel Chinese patient-derived primary CCA cell lines was performed. METHODS: Two CCA cell lines were developed and labelled ZJU-0826 and -1125. The two cell lines were characterized with respect to phenotypic, molecular, biomarker, functional and histological properties. RESULTS: Two novel cell lines were cultured for 2 years, and maintained for more than 100 passages. They retained their typical biliary epithelial morphology and ultrastructure. The population doubling times of ZJU-0826, and -1125 were 63.84 h and 44.73 h, respectively. The cells exhibited near-triploid karyotypes with complex structural aberrations. ZJU-1125 cells had mutations in TP53 exons. Short tandem repeats genotyping confirmed the human origin and difference between lines. An immunophenotype analysis showed that ZJU-0826 is positive for CD44, CD29, Pdx1, CD236, FoxA1, FoxA2, and Nanog, and ZJU-1125 positive for CD44, CD29, CD133, Pdx1, FoxA1, FoxA2, and Nanog. ZJU-1125 had greater invasion ability in vitro and tumorigenicity than those of ZJU-0826. CONCLUSIONS: Our results confirm the validity of the ZJU-0826 and -1125 as representative models for the elucidation of the molecular pathogenesis of perihilar CCA, and intrahepatic CCA in both in vitro and in vivo studies, respectively.

Combined treatment of cholangiocarcinoma with interventional radiofrequency hyperthermia and heat shock protein promoter-mediated HSV-TK gene therapy.

Cholangiocarcinoma is a most lethal malignancy frequently resistant to chemotherapy. Herpes simplex virus thymidine kinase/Ganciclovir (HSV-TK/GCV) suicide gene therapy is a promising approach to treat different cancers, including cholangiocarcinoma. However drawbacks including low therapeutic gene expression and lack of precise targeted gene delivery limit the wide clinical utilization of the suicide gene therapy. We attempted to overcome these obstacles. We established the "proof-of-principle" of this concept via serial in-vitro experiments using human cholangiocarcinoma cells and then validated the new interventional oncology technique in vivo using mice harboring the same patient derived cholangiocarcinomas. Curative effects were evaluated by magnetic resonance imaging and confirmed by pathology and laboratory examinations. Intratumoral radiofrequency hyperthermia (RFH) significantly elevated the targeted expression of HSV-TK gene and further enhanced the therapeutic effects of direct intratumoral HSV-TK/GCV gene therapy, evident as the least number of survival tumor cells, smallest tumor size, and the highest apoptosis index in the combination treatment of HSV-TK plus RFH, compared to other control treatments. The novel combination of image-guided interventional oncology, RFH technology, and direct gene therapy may be valuable for the effective treatment of cholangiocarcinoma.

Establishment and characterization of three stable Basal/HER2-positive breast cancer cell lines derived from Chinese breast carcinoma with identical missense mutations in the DNA-binding domain of TP53.

BACKGROUND: Basal/human epidermal growth factor receptor (HER)2-positive (HER2+) breast cancer is resistant to monoclonal antibody (herceptin) treatment. There are currently only three basal/HER2+ breast cancer cell lines available, but they are not from Chinese populations. METHODS: Three immortalized cell lines (ZJU-0327, ZJU-0725, and ZJU-1127) were established from invasive ductal breast carcinoma tissue of two patients treated by surgical resection at our center. The cell lines were characterized in terms of histology, therapeutic response, and biomarker expression. Their tumorigenic potential was evaluated in an athymic nude (BALB/C nu) mouse xenograft model. Cell authentication testing by the techniques of short tandem repeat. RESULTS: ZJU-0327, ZJU-0725, and ZJU-1127 cell lines were maintained for more than 110 passages in vitro. The cells grew as monolayers; showed typical epithelial morphology and ultrastructure; were polyploid; had doubling times of 18, 57.5, and 18 h, respectively; had a near-tetraploid (ZJU-0327 and ZJU-1127) or aneuploid (ZJU-0725) karyotype with structural aberrations and tumor protein 53 mutation; insensitive to chemotherapeutic drugs and/or radiation; show high invasiveness and tumorigenicity in mice; and had no mycoplasma contamination. The cell lines were basal/HER2+, expressed cluster of differentiation, and were associated with poor prognosis. Cell authentication testing by the American Type Culture Collection confirmed the human origin of the cell lines, which did not match those in existing databases. CONCLUSIONS: The three novel basal/HER2+ breast cancer cell lines recapitulating the malignant characteristics of the parent tumor's, and can be useful for clarifying the molecular pathogenesis of basal/HER2+ breast cancer.

Fe3+@polyDOPA-b-polysarcosine, a T1-Weighted MRI Contrast Agent via Controlled NTA Polymerization.

α-Amino acid N-thiocarboxyanhydrides (NTAs) are promising cyclic monomers to synthesize polypeptides and polypeptoids via controlled ring-opening polymerizations. Superior to N-carboxyanhydrides requiring protection on hydroxyl groups, NTAs are able to tolerate such nucleophiles. In this work, we report the synthesis of NTA monomers containing unprotected phenolic hydroxyl groups of 3,4-dihydroxy-l-phenylalanine (DOPA) and l-tyrosine (Tyr). Their controlled ROPs and sequential copolymerizations with polysarcosine (PSar) yield PDOPA, PTyr, and PDOPA-b-polysarcosine (PDOPA-b-PSar) products quantitatively with designable degrees of polymerization. Micellar nanoparticles of Fe3+@PDOPA-b-PSar have been prepared thanks to the strong chelation of iron(III) cation by catechol ligands that act as T1-weighted magnetic resonance imaging (MRI) contrast agents. For instance, Fe3+@PDOPA10-b-PSar50 exhibits higher longitudinal relaxivity (r1 = 5.6 mM-1 s-1) than commercial Gd3+-based compounds. Effective MRI contrast enhancement in vivo of nude mice with a moderate duration (150 min) and 3D magnetic resonance angiography in rabbit illustrated by using volume rendering and maximal intensity projection techniques ignite the clinical application of Fe3+-based polypept(o)ides in diagnostic radiology as Gd-free MRI contrast agents.

Kidins220/ARMS Expression Confers Proliferation But Independent of Self-Renewal in Mouse Embryonic Stem Cells.

Embryonic stem cells (ESCs) are characterized by their ability to self-renew and their potential to differentiate into any cell type. Therefore, identification of novel molecular markers to verify the pluripotent status of mouse ESCs (mESCs) is of great significance. Kinase D interacting substrate of 220 kDa (Kidins220)/ankyrin repeat-rich membrane spanning (ARMS) plays a crucial role in the integration of growth factor receptor pathways during embryonic development. However, the role of Kidins220/ARMS in ESCs is still unknown. To elucidate the effects of Kidins220/ARMS on ESCs, we performed a knockdown of the Kidins220/ARMS gene by RNA interference. To our surprise, downregulation of Kidins220/ARMS did not alter the pluripotent state of mESCs. In contrast, it was essential for the proliferation and survival of ESCs. Furthermore, downregulation of the ARMS gene limited the migration of embryoid body cells derived from mESCs. This study indicates novel roles of Kidins220/ARMS in ESCs, which may represent valuable targets for future clinical applications of ESCs.

Radiofrequency heat-enhanced direct intratumoral chemotherapy for prostate cancer.

A novel, minimally invasive interventional technique, radiofrequency heat (RFH), has been suggested to improve the efficacy of chemotherapy for solid organ tumors. However, the treatment for prostate cancer has not been completely characterized. The aim of the present study was to investigate the in vitro and in vivo efficiency of chemotherapy in combination with RFH for the treatment of prostate cancer. The following four treatment groups were included: i) No treatment (control); ii) RFH-only; iii) chemotherapy (docetaxel)-only; and iv) combination therapy of docetaxel and RFH in human prostate cancer (HPC) cell lines and mice with HPC xenografts. In the in vitro experiments, a heating guidewire was attached under the bottom of the last chamber of the four-chamber cell culture slide, and was then connected to a radiofrequency (RF) generator. In the in vivo experiments, a tumor model was generated by subcutaneously injecting human prostate cancer cells into 24 male nu/nu mice. RFH was conducted by inserting the 0.022-inch heating-guidewire into the tumor. The follow-up magnetic resonance imaging demonstrated a significant reduction in the average tumor size in animals treated with combination therapy compared with those receiving RFH-only and chemotherapy-only. The number of apoptotic cells and the average apoptotic index of the combination therapy group were significantly higher compared with those of the other three treatment groups. In conclusion, the results of the present study suggested that RFH is able to increase the therapeutic efficiency of docetaxel in prostate cancer, and this study serves as a foundation for the future development of an interventional molecular image-guided local treatment strategy for prostate cancer that integrates RF technology, interventional oncology and direct intratumoral chemotherapy, as a replacement for systemic chemotherapy.

Radiofrequency hyperthermia promotes the therapeutic effects on chemotherapeutic-resistant breast cancer when combined with heat shock protein promoter-controlled HSV-TK gene therapy: Toward imaging-guided interventional gene therapy.

OBJECTIVE: Gene therapy is a frontier in modern medicine. In the present study, we explored a new technique for the effective treatment of multidrug-resistant (MDR) breast cancer by combining fully the advantages of multidisciplinary fields, including image-guided minimally invasive interventional oncology, radiofrequency technology, and direct intratumoral gene therapy. RESULTS: Combination treatment with PHSP-TK plus RFH resulted in significantly higher TK gene transfection/expression, as well as a lower cell proliferation rate and a higher cell apoptosis index, than those of control groups. In vivo validation experiments with MRI confirmed that combination therapy resulted in a significant reduction of relative tumor volume compared with those of control animals, which was supported by the results of histologic and apoptosis analyses. MATERIALS AND METHODS: The heat shock protein promoter (PHSP) was used to precisely control the overexpression of thymidine kinase (TK) (PHSP-TK). Serial in vitro experiments were performed to confirm whether radiofrequency hyperthermia (RFH) could enhance PHSP-TK transfection and expression in a MDR breast cancer cell line (MCF7/Adr). Serial in vivo experiments were then carried out to validate the feasibility of the new technique, termed interventional RFH-enhanced direct intratumoral PHSP-TK gene therapy. The therapeutic effect of combination therapy was evaluated by MRI and confirmed by subsequent laboratory correlation. CONCLUSIONS: This study has established "proof-of-principle" of a new technique, interventional RFH-enhanced local gene therapy for MDR breast cancer, which may open new avenues for the effective management of MDR breast cancers via the simultaneous integration of interventional oncology, RF technology, and direct intratumoral gene therapy.

3-aminopyrazolopyrazine derivatives as spleen tyrosine kinase inhibitors.

Spleen tyrosine kinase is a new promising target for drug discovery to treat human cancer and inflammatory disorders. A series of pyrazolopyrazine-3-amine and pyrazolopyrimidine-3-amine derivatives was designed and synthesized as new spleen tyrosine kinase inhibitors. The efforts yielded compound 6h with promising spleen tyrosine kinase inhibition in both enzymatic and B-lymphoma cell proliferation assays. Additionally, compound 6h dose dependently inhibited the activation of spleen tyrosine kinase signal in human B-cell lymphoma cells. Compound 6h might serve as a lead for further development of new spleen tyrosine kinase inhibitors.

Gadolinium-Loaded Solid Lipid Nanoparticles as a Tumor-Absorbable Contrast Agent for Early Diagnosis of Colorectal Tumors Using Magnetic Resonance Colonography.

Magnetic resonance (MR) contrast agents focusing on special functions are required to improve cancer diagnosis, particularly in the early stages. Here, we designed multifunctional solid lipid nanoparticles (SLNs) with simultaneous loading of gadolinium (Gd) diethylenetriaminepentaacetic acid (Gd-DTPA) and octadecylamine fluorescein isothiocyanate (FITC) to obtain Gd-FITC-SLNs as a tumor-absorbable nanoparticle contrast agent for the histological confirmation of MR imaging (MRI) findings. Colorectal tumors were evaluated in vitro and in vivo via direct uptake of this contrast agent, which displayed reasonable T1 relaxivity and no significant cytotoxicity at the experimental concentrations in human colon carcinoma cells (HT29) and mouse colon carcinoma cells (CT26). In vitro cell uptake experiments demonstrated that contrast agent absorption by the two types of cancer cells was concentration-dependent in the safe concentration range. During in vivo MRI, transrectal infusion of Gd-FITC-SLNs showed more significant enhancement at the tumor site compared with the infusion of Gd-DTPA in female C57/BL mice with azoxymethane/dextran sulfate sodium-induced colorectal highgrade intraepithelial neoplasia. Subsequent confocal fluorescence microscopy demonstrated Gd-FITC-SLNs as highly concentrated green fluorescent spots distributed from the tumor capsule into the tumor. This study establishes the "proof-of-principle" of a new MRI technique wherein colorectal tumors are enhanced via direct absorption or uptake of the nanoparticle contrast agent.

Hybrid pyrimidine alkynyls inhibit the clinically resistance related Bcr-Abl(T315I) mutant.

A series of pyrimidine alkynyl derivatives were designed and synthesized as new Bcr-Abl inhibitors by hybriding the structural moieties from GNF-7, ponatinib and nilotinib. One of the most potent compounds 4e strongly suppresses Bcr-Abl(WT) and Bcr-Abl(T315I) kinase with IC50 values of 5.0 and 9.0 nM, and inhibits the proliferation of K562 and murine Ba/F3 cells ectopically expressing Bcr-Abl(T315I) cells with IC50 values of 2 and 50 nM, respectively. It also displays good pharmacokinetics properties with an oral bioavailability of 35.3% and T(1/2) value of 48.7 h, and demonstrates significantly suppression on tumor growth in xenografted mice of K562 and Ba/F3 cells expressing Bcr-Abl(T315I). These inhibitors may serve as lead compounds for further developing new anticancer drugs overcoming the clinically acquired resistance against current Bcr-Abl inhibitors.

Adeno-associated virus-mediated cancer gene therapy: current status.

Gene therapy is one of the frontiers of modern medicine. Adeno-associated virus (AAV)-mediated gene therapy is becoming a promising approach to treat a variety of diseases and cancers. AAV-mediated cancer gene therapies have rapidly advanced due to their superiority to other gene-carrying vectors, such as the lack of pathogenicity, the ability to transfect both dividing and non-dividing cells, low host immune response, and long-term expression. This article reviews and provides up to date knowledge on AAV-mediated cancer gene therapy.

Radiofrequency heat-enhanced chemotherapy for breast cancer: towards interventional molecular image-guided chemotherapy.

Breast cancer is the most common malignancy in women worldwide. Recent developments in minimally invasive interventional radiology techniques have significantly improved breast cancer treatment. This study aimed to develop a novel technique for the local management of breast cancers using radiofrequency heat (RFH). We performed both in vitro experiments using human breast cancer cells and in vivo validation in xenograft animal models with magnetic resonance imaging (MRI) and pathological correlation to investigate the feasibility of our approach. Four treatment groups, including (1) no treatment (control), (2) RFH-only, (3) chemo (doxorubicin)-only, and (4) combination therapy with both doxorubicin and RFH, were conducted in each experiment. In vitro combination therapy significantly decreased breast cancer cell proliferation while increased their apoptosis index compared to the other three groups. MRI demonstrated a significant tumor size reduction in animals treated with combination therapy compared to those receiving other treatments in vivo. Such result was further confirmed by pathological examination. In conclusion, our findings suggests that RFH can enhance the therapeutic efficiency of doxorubicin on breast cancers, thus establishing the basis for future development of interventional molecular image-guided local chemotherapy for breast malignancies.

Mesenchymal-like progenitors derived from human embryonic stem cells promote recovery from acute kidney injury via paracrine actions.

BACKGROUND AIMS: The engraftment of mesenchymal stem cells (MSCs) is reported to promote recovery of renal function in animal models of acute kidney injury (AKI). However, it is unknown whether mesenchymal-like progenitors (MPs) derived from human embryonic stem cells (hESCs) can mediate similar therapeutic effects. We investigated the responses of recipient renal tissue to engraftment of hESC-MPs and underlying mechanisms of these effects. METHODS: We measured blood urea nitrogen and creatinine levels of AKI mice with hESC-MPs transplantation and control mice. We performed renal morphology analysis by immunohistochemistry and electron microscopy to confirm the renoprotective effects of engrafted hESC-MPs. Proliferation, apoptosis and gene expression of tubular cells were also monitored by immunohistochemistry and real-time quantitative polymerase chain reaction to investigate the mechanisms that occurred. RESULTS: After transplantation of hESC-MPs into mice with cisplatin-induced AKI, improvements in renal function and recovery from tubular epithelial cell injury were observed. Engrafted hESC-MPs were localized to areas of injured kidney 5 days after cisplatin induction, where they promoted tubular cell proliferation and decreased kidney cell apoptosis. The beneficial effect was further confirmed by the capability of the engrafted cells to up-regulate renal gene expression of anti-inflammatory cytokines and pro-survival cytokines. Meanwhile, infusion of these cells reduced renal gene expression of pro-inflammatory cytokines and monocyte chemotactic protein-1, a chemokine that stimulates monocyte and macrophage infiltration. CONCLUSIONS: Our results show that infused hESC-MPs may promote recovery from AKI by regulating related cytokines.

Taurine postponed the replicative senescence of rat bone marrow-derived multipotent stromal cells in vitro.

The aging of many mammalian tissues is associated with replicative decline in somatic stem cells. Postponing this decline is a direct way of anti-aging. Bone marrow-derived multipotent stromal cells (BMSCs) hold promise for an increasing list of therapeutic uses due to their multilineage potential. Clinical application of BMSCs requires abundant cells that can be overcome by ex vivo expansion of cells, but often facing the replicative senescence problem. We demonstrated that taurine exhibited anti-replicative senescence effect on rat BMSCs by promoting colony forming unit-fibroblast formation and cell proliferation, shortening cell population doubling time, enormously inhibiting senescence-associated beta-galactosidase activity and slowing the loss of differentiation potential, while having no significant effect on the maximum passage number and total culture time, and slight influences on the cell surface CD molecules expressions. Taurine is a quite safe antioxidant and nutrient extensively used in food addition and clinical treatment. These suggested that taurine is a promising anti-replicative senescence additive for ex vivo expansion of BMSCs in experimental and clinical cell therapies.