Imeglimin profoundly affects the circadian clock in mouse embryonic fibroblasts.

OBJECTIVE: Imeglimin is a novel antidiabetic drug structurally related to metformin. Metformin has been shown to modulate the circadian clock in rat fibroblasts. Accordingly, in the present study, we aimed to determine whether imeglimin can impact the circadian oscillator in mouse embryonic fibroblasts (MEFs). METHODS: MEFs carrying a Bmal1-Emerald luciferase (Bmal1-ELuc) reporter were exposed to imeglimin (0.1 or 1 mM), metformin (0.1 or 1 mM), a nicotinamide phosphoribosyltransferase inhibitor FK866, and/or vehicle. Subsequently, Bmal1-ELuc expression and clock gene mRNA expression levels were measured at 10-min intervals for 55 h and 4-h intervals for 32 h, respectively. RESULTS: Imeglimin significantly prolonged the period (from 26.3 to 30.0 h at 0.1 mM) and dose-dependently increased the amplitude (9.6-fold at 1 mM) of the Bmal1-ELuc expression rhythm; however, metformin exhibited minimal effects on these parameters. Moreover, imeglimin notably impacted the rhythmic mRNA expression of clock genes (Bmal1, Per1, and Cry1). The concurrent addition of FK866 partly inhibited the effects of imeglimin on both Bmal1-ELuc expression and clock gene mRNA expression. CONCLUSION: Collectively, these results reveal that imeglimin profoundly affects the circadian clock in MEFs. Further studies are needed to evaluate whether imeglimin treatment could exert similar effects in vivo.

Molecular complexation properties of Cd2+ by algal organic matter from Scenedesmus obliquus.

A detailed understanding the metals binding with algal organic matter (AOM) is essential to gain a deeper insight into the toxicity and migration of metals in algae cell. However, the molecular complexation mechanism of the metals binding with AOM remains unclear. In this study, cadmium ion (Cd2+) binding properties of AOMs from Scenedesmus obliquus, which included extracellular organic matter (EOM) and intracellular organic matter (IOM), were screened. When Cd2+ < 0.5 mg/L, the accumulation of Cd2+ could reach 40%, while Cd2+ > 0.5 mg/L, the accumulation of Cd2+ was only about 10%. EOM decreased gradually (from 8.51 to 3.98 mg/L), while IOM increased gradually (from 9.62 to 21.00 mg/L). The spectral characteristics revealed that IOM was richer in peptides/proteins and had more hydrophilic than EOM. Both EOM and IOM contained three protein-like components (containing tryptophan and tyrosine) and one humic-like component, and their contents in IOM were higher than that in EOM. The tryptophan protein-like substances changed greatly during Cd2+ binding, and that the tryptophan protein-like substances complexed to Cd2+ before tyrosine protein-like substances in IOM was identified. Moreover, the functional groups of N-H, O-H, and CO in AOM played an important role, and the N-H group was priority to interacts with Cd2+ in the complexing process. More functional groups (such as C-O and C-N) were involved in the metals complexing in EOM than in IOM. It could be concluded that Cd2+ stress promoted the secretion of AOM in Scenedesmus obliquus, and proteins in AOM could complex Cd2+ and alleviate its toxicity to algal cell. These findings provided deep insights into the interaction mechanism of AOM with Cd2+ in aquatic environments.

Prevalence of mental disorders among middle school students in Shaoxing, China.

BACKGROUND: In China, adolescents account for about a quarter of those treated for mental disorders each year, and adolescent mental health issues have become a social hotspot. Although several epidemiological surveys of mental disorders have been conducted in China, no study has yet focused on the prevalence of mental disorders among adolescents in a certain region of Zhejiang. METHODS: In the first stage, 8219 middle school students aged 12-18 years in a city of Zhejiang Province (Shaoxing) were screened with the mental health screening checklist. In the second stage, participants who screened positive were tested with the Mini International Neuropsychiatric Interview for Children and Adolescents (MINI-KID) and the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV). Then, the prevalence of mental disorders were calculated. RESULTS: The overall prevalence in this population was 12.4%, with prevalence rates exceeding 20% in both the 17- and 18-year-old age groups. The most common mental disorders were obsessive-compulsive disorder (OCD) (9.1%) and major depressive disorder (MDD) (8.9%). CONCLUSIONS: Mental disorders are common among middle school students, and girls are at higher risk than boys. As the most prevalent mental disorders, OCD and MDD should receive timely attention, especially for upper grade students.

A Retrospective Study from a Single Center to Identify Hematological Factors that Distinguish Between Patients with Colorectal Carcinoma and Colorectal Adenoma.

BACKGROUND This retrospective study from a single center in China was conducted using data from medical records between 2012 and 2020, to identify hematological factors that distinguish between patients with colorectal carcinoma (CRC) and colorectal adenoma. MATERIAL AND METHODS In this case-control study, 856 eligible patients were randomly divided into a training set (n=600) and a testing set (n=256). Routine blood parameters, blood coagulation, and liver and kidney function parameters were collected. Univariate and multivariate Cox regression analyses were used to explore diagnostic indicators. The values of the area under the curve and calibration curves were used to evaluate the model. RESULTS The study included 325 colorectal adenoma and 531 CRC patients. The prediction model for diagnosing CRC using hemoglobin-to-platelet ratio, fibrinogen-albumin ratio (FAR), albumin-globulin ratio (A/G), platelet-lymphocyte ratio, carcinoembryonic antigen (CEA), and thrombin time (TT) was developed on the basis of the patients grouped into the CRC and colorectal adenoma groups. The prediction model for diagnosing CRC stage was developed using prothrombin time (PT), TT, CEA, A/G, FAR, and HPR. The prediction model for diagnosing CRC grade was developed using PT, TT, A/G, plateletcrit, FAR, and HPR. The AUCs of the 3 prediction models were [0.848, 95% CI: (0.800-0.896)], [0.806, 95% CI: (0.775-0.836)], and [0.829, 95% CI: (0.797-0.860)] in the testing set. CONCLUSIONS Three diagnostic prediction models for early screening of CRC, stage of CRC, and grade of CRC were established through a panel of readily available hematological parameters, which could provide auxiliary tools for early screening of CRC.

m6A-dependent glycolysis enhances colorectal cancer progression.

BACKGROUND: Epigenetic alterations are involved in various aspects of colorectal carcinogenesis. N6-methyladenosine (m6A) modifications of RNAs are emerging as a new layer of epigenetic regulation. As the most abundant chemical modification of eukaryotic mRNA, m6A is essential for the regulation of mRNA stability, splicing, and translation. Alterations of m6A regulatory genes play important roles in the pathogenesis of a variety of human diseases. However, whether this mRNA modification participates in the glucose metabolism of colorectal cancer (CRC) remains uncharacterized. METHODS: Transcriptome-sequencing and liquid chromatography-tandem mass spectrometry (LC-MS) were performed to evaluate the correlation between m6A modifications and glucose metabolism in CRC. Mass spectrometric metabolomics analysis, in vitro and in vivo experiments were conducted to investigate the effects of METTL3 on CRC glycolysis and tumorigenesis. RNA MeRIP-sequencing, immunoprecipitation and RNA stability assay were used to explore the molecular mechanism of METTL3 in CRC. RESULTS: A strong correlation between METTL3 and 18F-FDG uptake was observed in CRC patients from Xuzhou Central Hospital. METTL3 induced-CRC tumorigenesis depends on cell glycolysis in multiple CRC models. Mechanistically, METTL3 directly interacted with the 5'/3'UTR regions of HK2, and the 3'UTR region of SLC2A1 (GLUT1), then further stabilized these two genes and activated the glycolysis pathway. M6A-mediated HK2 and SLC2A1 (GLUT1) stabilization relied on the m6A reader IGF2BP2 or IGF2BP2/3, respectively. CONCLUSIONS: METTL3 is a functional and clinical oncogene in CRC. METTL3 stabilizes HK2 and SLC2A1 (GLUT1) expression in CRC through an m6A-IGF2BP2/3- dependent mechanism. Targeting METTL3 and its pathway offer alternative rational therapeutic targets in CRC patients with high glucose metabolism.

In Vivo Flow Cytometric Evaluation of Circulating Metastatic Pancreatic Tumor Cells after High-Intensity Focused Ultrasound Therapy.

We examined our hypothesis that high-intensity focused ultrasound (HIFU) treatment of pancreatic ductal adenocarcinoma (PDAC) in nude mice models may lead to an increased occurrence of hematogenous metastasis. The human PDAC cell line BxPC-3 transfected with mCherry was implanted into nude mice to establish orthotopic and subcutaneous xenograft (OX and SX) tumor models. Mice were exposed to HIFU when tumor sizes reached approximately 200-300 mm3 . The OX and SX tumor models were monitored continuously for tumor growth characteristics and hematogenous metastasis using in vivo flow cytometric (IVFC) detection of circulating tumor cells (CTCs) from the pancreas. We chose an appropriate mouse model to further examine whether or not HIFU increases the potential risk of hematogenous metastasis, using IVFC detection. Our results showed that the CTC number was greater in the OX model than in the SX model. The CTC number in the OX model increased gradually over time, whereas the CTC number in the SX model remained low. Therefore, the OX model was better for studying tumor metastasis by IVFC detection. We found significantly decreased CTC numbers and tumor volume after HIFU ablation. Our results showed the applicability of the PDAC OX tumor model for studying the occurrence of tumor metastasis due to the generation of CTCs. HIFU ablation substantially restricted PDAC hematogenous metastasis and provided effective tumor control locally. © 2020 The Authors. Cytometry Part A published by Wiley Periodicals Inc., on behalf of International Society for Advancement of Cytometry.

Autofluorescence of hydrogels without a fluorophore.

Fluorescent hydrogels have recently attracted great attention for medical diagnostics, bioimaging and environmental monitoring. However, additional phosphors or fluorophores are always required to label the hydrogels, and they suffer from marker bleaching, signal drifts, or information misrepresentation. Here we report autofluorescence that universally exists in carbonyl-containing hydrogels without any traditional fluorophore. The fluorescence is successfully employed to self-monitor the gelation process since the fluorescence signal is closely related to the internal structural change of the gels. The crosslinked structure is beneficial to the fluorescence efficiency. Specifically, the fluorescence intensity is amplified with decreasing water content of the gels. The system realizes aggregation-induced emission in a water-deficient environment. The fluorescence is quenched by the addition of some specific metal ions, which can realize the successfully erasure and rewriting of information under visible light and ultraviolet light respectively. We believe that the spontaneous fluorescence of a gel provides the most reliable basis for the detection of a gel structure and opens new prospects in the application of hydrogels.

Identification and characterization of protein phosphorylation in the soluble protein fraction of scallop (Chlamys farreri) byssus.

Protein phosphorylation is a widespread modification that and plays a significant role in marine bioadhesion. The phosphorylated proteins of the barnacle Amphibalanus amphitrite can form strong ionic bonds with mineral surfaces to adapt to marine environments. The adhesion protein PC-3 in the sandcastle worm Phragmatopoma californica contains multipleserine phosphorylations. Interactions between these phosphate groups and the Mg/Ca2+ ions are less soluble at seawater pH, making the cement less fluid and more gel-like. The scallop byssus is characterized by strong wet adhesion performance and substantial byssus secretions. Thus, the excellent underwater adhesion properties of the byssus make it an ideal candidate for studies related to the development of new and versatile composite materials. However, phosphoproteins have not been identified or studied in the scallop Chlamys farreri. Phosphorylated proteins in the C. farreri byssus protein were identified by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and further confirmed by phosphorylation staining and in-gel digestion coupled with mass spectrometric analysis (GeLC-MS/MS). Finally, sequence analyses and potential functional analyses were performed for these newly identified proteins. We have identified previously unreported phosphorylation sites within the C. farreri byssus protein. The results show phosphorylation modifications in all parts of the byssus structure and four foot-specific phosphorylated proteins were verified by two types of mass spectrometry and staining. The annotation of biological functions, based on sequence alignments shows that the protein 40,215.25 is homologous with TIMP-2. Similar to the previously identified TIMP-2-like protein Sbp8-1 in the scallop byssus, it contains an abundance of phosphorylated Cys, which may promote protein polymerization. We speculate that protein 40,215.25 may play an important role in cross-linking and adhesion of the scallop byssus. The phosphorylated protein we have identified in the C. farreri byssus may be related to the formation of protein cross-linkings and adhesion of the scallop foot. Our study lays the groundwork for a better understanding of the adhesion mechanism of the scallop byssus.

In vitro dentin tubule occlusion by an arginine-containing dentifrice.

PURPOSE: To evaluate the ability of an arginine-containing dentifrice to occlude dentin tubules. METHODS: Dentin discs were divided equally into premolar and molar groups, which were then utilized in three treatment groups: a blank control group (distilled water treatment), a negative control group (common dentifrice with calcium carbonate) and an experimental group [dentifrice with 8% (w/w) arginine]. Each dentin disk was brushed with the dentifrice twice daily for 7 consecutive days. After this period, each disc was separated into two equal halves. One half was used for scanning electron microscopy (SEM) and energy-dispersive spectrometer (EDS) examinations, while the other half was brushed with distilled water twice daily for another 7 days prior to SEM observation. RESULTS: The plugging rate in the arginine dentifrice group was significantly higher and more sustainable than in the negative control group. The surface deposition of calcium and phosphorus on the dentin discs in the arginine dentifrice group was also significantly higher. CLINICAL SIGNIFICANCE: This study provided evidence that using arginine as an active ingredient in dentifrice can improve its ability to occlude dentin tubules, thus supporting future efforts to improve dentin hypersensitivity.

Cell-free DNA derived from cancer cells facilitates tumor malignancy through Toll-like receptor 9 signaling-triggered interleukin-8 secretion in colorectal cancer.

Circulating cell-free DNA (cfDNA) has become a potential diagnostic and prognostic biomarker for colorectal cancer (CRC). In non-cancerous diseases, it has been confirmed that cfDNA can be recognized by Toll-like receptor 9 (TLR9), leading to a significant biological change. Nevertheless, the biological significance of cfDNA and its relationship with TLR9 in tumor malignancy is still unclear. Therefore, the purpose of this study is to explore the biological role of cfDNA in colorectal cancer (CRC). The expression of TLR9 was measured in different CRC cell lines and cancerous samples by RT-PCR or immunohistochemistry, which showed that high expression of TLR9 was significantly correlated with the tumor metastasis, advanced TNM stage and poor prognosis of patients. Then, cfDNA was obtained from fluorouracil (5FU)-induced apoptotic cancer cells in vitro and transfection techniques were used to transfect siRNA and cDNA plasmid for TLR9. Cancer cells were stimulated using isolated cfDNA fragments, and results showed that cfDNA could promote colorectal cancer cell proliferation via TLR9. Meanwhile, we demonstrated that the cfDNA binding to TLR9 could facilitate cell migration and invasion. Finally, we demonstrated that cfDNA initiated downstream TLR9-MyD88 signaling and induced robust release of chemokine interleukin 8 (IL-8), which helped to elucidate the mechanisms underlying these phenomena. Our data suggest that cancer cell-derived cfDNA contributes to cancer progression through activation of TLR9-MyD88 signaling and IL-8 secretion in CRC. These findings provide a novel perspective for understanding of tumor progression and provoke a potential therapeutic target for CRC treatment.

Healing of root and surrounding periodontium after root damage with miniscrew implants: a histomorphologic study in dogs.

OBJECTIVES: The main purpose of this study was to investigate the detailed healing process of the roots and surrounding periodontium [cementum, periodontal ligament (PDL), and bone] at different time points after intentional root damage with miniscrew implants (MSIs). MATERIALS AND METHODS: After cone-beam computed tomography examination and measurement, a total of 78 premolar and molar roots from five beagle dogs were intentionally damaged by implanting miniscrews in the interradicular region. MSIs were immediately removed, and the histological morphology was observed at days 0 and 3 and at weeks 1, 2, 3, 4, 6, 8, and 12 after root injury using haematoxylin and eosin and fluorescence stainings (fluorescence staining was performed at days 28 and 56). RESULTS: An early new attachment of PDL adhering on to the damaged root surface was found at week 2 after root injury. Tissue differentiation of newly formed bone tissue, PDL, and cementum began at week 3. Moreover, the newly formed cementum and bone were constantly forming and mineralising at weeks 4, 6, 8, and 12, and the width of PDL gradually narrowed until close to the normal width at week 12. CONCLUSIONS: This study demonstrated the complete healing process of the roots and surrounding periodontium after root damage with MSIs in dogs when the damage was limited to the cementum or dentin. CLINICAL RELEVANCE: The findings of this study may help provide a better understanding of the detailed healing process in roots and PDLs damaged by MSIs.

Immature Colon Carcinoma Transcript-1 (ICT1) Expression Correlates with Unfavorable Prognosis and Survival in Patients with Colorectal Cancer.

BACKGROUND: Colorectal cancer (CRC) is the third leading cause of death from cancer worldwide. Immature colon carcinoma transcript-1 (ICT1) has been reported to be correlated with lung cancer; however, whether ICT1 is a functional gene in CRC, as well as the molecular mechanism underlying ICT1 mediation of colorectal-tumor formation, remains unknown. METHODS: The expression of ICT1 was firstly determined by using immunohistochemistry in 861 CRC specimens. The correlation of ICT1 expression with clinicopathological parameters and the survival rate was analyzed. Furthermore, we investigated the effect of ICT1 silencing on CRC cell proliferation and migration by MTT, colony formation, flow cytometry, and transwell in vitro. RESULTS: ICT1 is highly expressed in a cohort of human CRCs, and that higher ICT1 expression may lead to reduced overall survival rate of CRC. Likewise, ICT1 silencing lowered the cell viability through cell-cycle arrest, inhibited cell migration, and induced apoptosis in CRC. We further revealed a novel mechanism in which ICT1 promoted CRC growth via the intracellular AMPK, SAPK/JNK, and PARP signaling pathways. CONCLUSIONS: Our data showed that ICT1 could be an important target for CRC diagnosis and treatment.

Changes in the Expression of miR-34a and its Target Genes Following Spinal Cord Injury In Rats.

BACKGROUND Results from DNA microarray experiments have shown that the expression of miR-34s undergoes significant changes following spinal cord injury (SCI). The present study was designed to detect changes in the expression of miR-34s and its target genes during the acute and sub-acute stages of SCI. MATERIAL AND METHODS Luxol fast blue (LFB) staining for myelin was used to observe the differences in the general morphology of the spinal cord after SCI in a contusion model in rats. qPCR was carried out to determine the expression variation of miR-34s and its target genes during the acute and sub-acute stages of SCI. The mimic technique was used to further confirm the regulatory effect of miR-34a on the potential target genes. RESULTS The expression level of miR-34a decreased immediately after SCI and persisted for 21 days after SCI. The expression level of miR-34c began decreasing at day 1 after SCI and persisted until day 14. The expression level of miR-34b did not undergo significant change after SCI. The results of double immunofluorescence and in-situ hybridization suggested that miR-34a was highly expressed in spinal cord neurons. Based on our bioinformatics analysis, we postulated that miR-34a might participate in post-SCI cell apoptosis by regulating the target gene Notch1, and likely participated in the inflammatory response and glial scar formation by regulating the candidate genes Csf1r and PDGFRa, respectively. The expression levels of the candidate genes Csf1r and PDGFRa were consistent with Notch1 after SCI. The mimic technique further confirmed the regulatory effect of miR-34a on the aforementioned target genes. CONCLUSIONS We postulate that miR-34a and miR-34c might participate in multiple aspects of cytobiological activities following SCI. MiR-34a in particular may participate in cell apoptosis, inflammatory response, and glial scar formation by regulating the target gene Notch1 and candidate target genes Csf1r and PDGFRa respectively.

Post-translational regulation of mitogen-activated protein kinase phosphatase (MKP)-1 and MKP-2 in macrophages following lipopolysaccharide stimulation: the role of the C termini of the phosphatases in determining their stability.

MAPK phosphatases (MKPs) are critical modulators of the innate immune response, and yet the mechanisms regulating their accumulation remain poorly understood. In the present studies, we investigated the role of post-translational modification in the accumulation of MKP-1 and MKP-2 in macrophages following LPS stimulation. We found that upon LPS stimulation, MKP-1 and MKP-2 accumulated with different kinetics: MKP-1 level peaked at ∼1 h, while MKP-2 levels continued to rise for at least 6 h. Accumulation of both MKP-1 and MKP-2 were attenuated by inhibition of the ERK cascade. Interestingly, p38 inhibition prior to LPS stimulation had little effect on MKP-1 and MKP-2 protein levels, but hindered their detection by an M-18 MKP-1 antibody. Studies of the epitope sequence recognized by the M-18 MKP-1 antibody revealed extensive phosphorylation of two serine residues in the C terminus of both MKP-1 and MKP-2 by the ERK pathway. Remarkably, the stability of both MKP-1 and MKP-2 was markedly decreased in macrophages in the presence of an ERK pathway inhibitor. Mutation of the two C-terminal serine residues in MKP-1 and MKP-2 to alanine decreased their half-lives, while mutating these residues to aspartate dramatically increased their half-lives. Deletion of the C terminus from MKP-1 and MKP-2 also considerably increased their stabilities. Surprisingly, enhanced stabilities of the MKP-1 and MKP-2 mutants were not associated with decreased ubiquitination. Degradation of both MKP-1 and MKP-2 was attenuated by proteasomal inhibitors. Our studies suggest that MKP-1 and MKP-2 stability is regulated by ERK-mediated phosphorylation through a degradation pathway independent of polyubiquitination.

Comparison of minimally invasive and open colorectal resections for patients undergoing simultaneous R0 resection for liver metastases: a propensity score analysis.

PURPOSE: The role of minimally invasive colorectal resection for patients undergoing a simultaneous resection for synchronous liver metastases had not been established. This study compared the short- and long-term outcomes between minimally invasive and open colorectal resection for patients undergoing simultaneous resection for liver metastases. METHODS: This study reviewed 101 consecutive patients undergoing simultaneous colorectal resection and R0 resection of synchronous liver metastases between January 2008 and December 2012. In the study, 36 consecutive patients who underwent minimally invasive colorectal resection were matched with 36 patients who had an open approach by propensity scoring. The analyzed variables included patient and tumor characteristics and short-term and long-term outcomes. RESULTS: After propensity score matching, the two groups had similar clinicopathologic variables. No patient undergoing the minimally invasive procedure experienced conversion to the open technique. No postoperative mortality occurred in either group. In the minimally invasive group, the estimated blood loss (P < 0.007), bowel function return time (P < 0.016), and postoperative hospital stay (P < 0.011) were significantly lower than those in the open group, although the operating time was significantly longer (P < 0.001). No significant differences in postoperative complications were observed between the groups. The two groups did not differ significantly in terms of the 5-year overall survival rate (51 vs. 55 %; P = 0.794) and disease-free survival rate (38 vs. 27 %; P = 0.860). CONCLUSION: Minimally invasive colorectal resection with simultaneous resection of liver metastases has an outcome similar to open approach but some short-term advantages.

Aberrant expression of sphingosine-1-phosphate receptor 1 correlates with metachronous liver metastasis and poor prognosis in colorectal cancer.

There are currently no accurate predictive markers of metachronous liver metastasis from colorectal cancer. Recent studies demonstrated that the expression patterns of sphingosine-1-phosphate receptor 1 (S1PR1) are altered in several tumors, but in colorectal cancer, the patterns remain unknown. Our study was designed to evaluate the expression and prognostic significance of S1PR1 protein in patients with colorectal cancer. The expression of S1PR1 was detected using the tissue microarray technique and immunohistochemical method and compared with clinicopathological parameters in 153 colorectal cancer patients. The prognostic value of S1PR1 expression was evaluated by Kaplan-Meier and Cox regression analysis. A molecular prognostic stratification scheme incorporating S1PR1 expression was determined by using receiver operating characteristic (ROC) analysis. S1PR1 was significantly highly expressed in 70.6 % (108/153) of the colorectal cancer lesions compared to their high expressions in only 5.9 % (9/153) of the adjacent non-cancerous tissues. Upregulated expression of S1PR1 was significantly associated with depth invasion and metachronous liver metastasis. Increased S1PR1 expression in colorectal cancer was positively correlated with poor overall survival. Multivariate survival analysis suggested that S1PR1 expression was an independent prognostic indicator for the disease. Applying the prognostic value of S1PR1 density to TNM stage system showed a better prognostic value in patients with colorectal cancer. Aberrant S1PR1 expression in colorectal cancer was associated with metachronous liver metastasis and worse survival outcome, and also, it was an independent prognostic factor. According to our analysis, combined TNM stage and intratumoral expression of S1PR1 demonstrated a better prognostic value than any of these two parameters alone. Conclusively, we suggest that detection and analysis of S1PR1 expression in colorectal cancer tissue might be used for predicting prognosis of colorectal cancer.

Anti-inflammatory diterpenoids from the root bark of Acanthopanax gracilistylus.

Five new ent-pimarane (1-3, 7, and 8) and three new ent-kaurane diterpenoids (4-6) and a new oleanane triterpene acid (9), together with 22 known compounds, were isolated from the root bark of the medicinal herb Acanthopanax gracilistylus. The structures of 1-9 were established based on the interpretation of high-resolution MS and 1D- and 2D-NMR data. The absolute configurations of 7 and 11 were determined by single-crystal X-ray diffraction and electronic circular dichroism analysis. Compounds 7 and 8 represent rare naturally occurring structures based on the devinyl ent-pimarane skeleton. Compounds 3, 10, 14, 16, and 17 exhibited potent inhibitory effects on the release of interleukin-1ß (IL-1ß), interleukin-8 (IL-8), and tumor necrosis factor (TNF-α) in lipopolysaccharide-stimulated peripheral blood mononuclear cells.

[Evaluation of the rationality of current T staging of gastric cancer with transverse mesocolon invasion].

OBJECTIVE: To evaluate the rationality of T staging of gastric cancer with transverse mesocolon invasion. METHODS: Data of 808 patients with primary gastric cancer undergoing surgical treatment was screened from the Data base of Gastric Cancer of Sun Yat-sen University, from April 1996 to October 2009. According to the information of transverse mesocolon invasion, all cases were divided into groups NOI (T4a stage, non organ invasion, n = 638), NTMI (T4b stage, non transverse mesolon invasion, with organ invasion, n = 126), and TMI (transverse mesocolon invasion, n = 44). The clinicopathological features, surgical procedure and prognosis were compared among the three groups. RESULTS: No significant difference was found in gender, age, lymph node metastasis, hepatic metastasis, tumor's Borrmann type, histological type, differentiation degree, value of serum CEA among the 3 groups (all P > 0.05). In the groups NOI, NTMI and TMI, the ratio of mean tumor diameter ≥ 5 cm was 39.0% (249/638), 61.1% (77/126) and 54.5% (24/44), respectively; the ratio of distal metastasis was 11.9% (76/638), 30.2% (38/126) and 43.2% (19/44), respectively; the ratio of peritoneal metastasis was 8.2% (52/638), 26.2% (33/126) and 38.6% (17/44), respectively; the ratio of TNM IV stage was 25.4% (162/638), 84.7% (107/126) and 93.7% (41/44), respectively; and the ratio of radical resection was 92.0% (587/638), 69.8% (88/126) and 77.3% (34/44), respectively; all with significant differences (P < 0.01), and the results of pairwise comparisons (Bonferroni correction, significant level α = 0.05/3 = 0.0167) showed that these parameters were significantly different between groups NOI and TMI (P < 0.0167), but non-significant between groups NTMI and TMI (P > 0.0167). The median survival time was 42.0, 16.4 and 19.0 months in the groups NOI, NTMI and TMI, respectively (P < 0.01), and the results of pairwise comparison showed that the prognosis were significant different between the groups NOI and TMI (P < 0.01), but non-significant between the groups NTMI and TMI (P > 0.05). In the cases who received radical resection, the median survival time was 47.9, 23.5 and 21.4 months in the groups NOI, NTMI and TMI, respectively (P < 0.01), and the results of pairwise comparison showed that the prognosis was significantly different between the groups NOI and TMI (P < 0.05), but not significant between groups NTMI and TMI (P > 0.05). CONCLUSIONS: The tumor size, distal meatastasis, peritoneal metastasis, TNM stage, surgical procedure and prognosis of gastric cancer with transverse mesocolon invasion are similar to that of T4b gastric cancer, but are significantly different from that of T4a gastric cancer. Gastric cancer with transverse mesocolon invasion should be reclassified as T4b stage.

[The efficacy and safety of methotrexate in refractory Crohn's disease].

OBJECTIVE: To assess the efficacy and safety of methotrexate(MTX) on refractory Crohn's disease(CD). METHODS: A total of 35 consecutive refractory CD patients in the First Affiliated Hospital of Sun Yat-Sen University treated with MTX were retrospectively analyzed. Clinical data from June 2004 to December 2012 were collected from the database of inflammatory bowel disease (IBD) center. Clinical responses and drug side effects were recorded and analyzed. RESULTS: Thirty-five refractory CD patients were identified including 23 cases intolerant to azathioprine (AZA)/6-mercaptopurine(6-MP), six cases ineffective to AZA/6-MP, 19 cases dependent on steroid. After treatment of MTX for 12 weeks [15(5-20) mg/week], a clinical response was obtained in 80% patients (28/35), including 51.4% (18/35) in remission and 28.6% (10/35) in improvement. The median Crohn's disease activity index (CDAI) scores at the onset and 12 weeks after MTX therapy were 99.2 (75.8, 174.7) and 61.5 (36.0, 106.6) respectively. The median single dose and duration of MTX were 15 (5-20) mg/week and 6.0(0.5-53.0) months respectively. The median cumulative dose was 480 (20-2615) mg. Among the 26 patients dependent on steroid, 21 achieved discontinuation of steroid with a median time of 10 (6-20) weeks after treatment of MTX. Side effects were recorded in 12 patients (34.3%), but usually mild and improved after drug with drawal. CONCLUSIONS: MTX is an effective and steroid-sparing agent for refractory CD. Side effects of MTX are mild and tolerable.

[Clinical characteristics and risk factors of refractory Crohn's disease].

OBJECTIVE: To analyze the clinical characteristics and risk factors of refractory Crohn's disease (CD). METHODS: All clinical data of confirmed consecutive CD patients were collected from our hospital between January 2003 and June 2013. The patients' demographic data, clinical features, therapeutic regimens and laboratory examinations were analyzed. A multivariate Logistic regression was performed to identify the risk factors of refractory CD. RESULTS: (1) A total of 402 confirmed CD patients were recruited for analysis. The prevalence of refractory CD was 33.8% (136/402). The rates of steroid-dependency was 37.0% (97/262) in 262 patients with a history of steroid use and the rate of thiopurines ineffectiveness was 26.9% (79/294) in 294 patients with a history of thiopurines-use; (2) Univariate analysis showed that disease location (L3 type), abdominal pain, diarrhea, fever, abdominal tenderness, perianal lesion, steroid use, AZA/6-MP use, leucocyte, hemoglobin (Hb), platelet level and high-sensitivity C-reactive protein (HsCRP) were significantly different between refractory and non-refractory CD patients (all P < 0.05) . Multivariate Logistic regression showed that steroid use (OR = 6.516, 95% CI: 2.884-14.722, P = 0.000) and low Hb (OR = 1.023, 95% CI: 1.008-1.037, P = 0.002) were independent risk factors related to refractory CD; (3) Univariate analysis showed that Hb level, erythrocyte sedimentation rate (ESR) were significantly different between steroid-dependent and non-steroid-dependent groups (all P < 0.05) . Multivariate Logistic regression showed that only low Hb level (OR = 1.021, 95% CI: 1.006-1.036, P = 0.005) was an independent risk factor related to steroid-dependency; (4) Univariate analysis showed that disease location (L3 type), perianal lesion, abdominal pain, diarrhea, fever, abdominal tenderness, platelet level, steroid use, steroid-dependency were significantly different between thiopurines-ineffective and thiopurines-effective groups (all P < 0.05) . Multivariate Logistic regression showed that perianal lesion (OR = 2.085, 95% CI: 1.007-4.039, P = 0.029), abdominal tenderness (OR = 2.943, 95% CI: 1.452-5.964, P = 0.003) and steroid-dependency (OR = 3.599, 95% CI: 1.847-7.013, P = 0.000) were independent risk factors related to thiopurines-ineffectiveness. CONCLUSIONS: Nearly one third CD patients became refractory during the course of disease. Low Hb and steroid use are independent risk factors. Low Hb is an independent risk factor related to steroid-dependency. Perianal disease, abdominal tenderness and steroid-dependency are independent risk factors related to thiopurines- ineffectiveness.