DNA-Thioguanine Nucleotides as a Marker for Thiopurine Induced Late Leukopenia after Dose Optimizing by NUDT15 C415T in Chinese Patients with IBD.

Thiopurine dose optimization by thiopurine-S-methyltransferase (TPMT) or nudix hydrolase-15 (NUDT15) significantly reduced early leucopenia in Asia. However, it fails to avoid the late incidence (> 2 months). Although laboratory monitoring of 6-thioguanine nucleotides (6TGN) to optimize thiopurine dose was suggested in White patients the exact association between leucopenia and 6TGN was controversial in Asian patients. In the present study, we aimed to explore whether DNA-thioguanine nucleotides (DNA-TGs) in leukocytes, compared with 6TGN in erythrocytes, can be a better biomarker for late leucopenia. This was a prospective, observational study. Patients with inflammatory bowel disease (IBD) prescribed thiopurine from February 2019 to December 2019 were recruited. Thiopurine dose was optimized by NUDT15 C415T (rs116855232). DNA-TG and 6TGN levels were determined at the time of late leucopenia or 2 months after the stable dose was obtained. A total of 308 patients were included. Thiopurine induced late leucopenia (white blood cells < 3.5 × 109 /L) were observed in 43 patients (14.0%), who had significantly higher DNA-TG concentration than those without leucopenia (P = 4.1 × 10-9 , 423.3 (~ 342.2 to 565.7) vs. 270.5 (~ 188.1 to 394.3) fmol/µg DNA). No difference in 6TGN concentrations between leucopenia and non-leucopenia was found. With a DNA-TG threshold of 340.1 fmol/µg DNA, 83.7% of leucopenia cases could be identified. Multivariate analysis showed that DNA-TG was an independent risk factor for late leucopenia. Quantification of DNA-TG, rather than 6TGN, can be applied to gauge thiopurine therapy after NUDT15 screening in Chinese patients with IBD.

Systematic review and meta-analysis: Association of a pre-illness Western dietary pattern with the risk of developing inflammatory bowel disease.

OBJECTIVE: Previous studies have presented conflicting results on Western diets and the risk of inflammatory bowel disease (IBD). This study aimed to evaluate the role of a pre-illness Western dietary pattern in the development of IBD. METHODS: The Western dietary pattern was defined as that met at least two of the following, either a high intake of refined grains, red and processed meat, animal protein, animal fats or high-fat dairy products, or with a low consumption of fruit and vegetables. Four medical databases (PubMed, EMBASE, the Cochrane Library and the China National Knowledge Infrastructure) were searched to identify all relevant references. Risk estimate and corresponding 95% confidence interval (CI) were pooled using a random-effects model. RESULTS: Nine studies (seven case-control studies and two prospective cohorts) were included, with a total of 1491 IBD cases and 53 089 controls. A Western dietary pattern was associated with a risk of all IBD (relative risk [RR] 1.92, 95% CI 1.37-2.68) and separately with Crohn's disease (CD) (RR 1.72, 95% CI 1.01-2.93) and ulcerative colitis (UC) (RR 2.15, 95% CI 1.38-3.34). Subgroup analysis by region showed that a Western dietary pattern was associated with the risk of CD and UC for studies performed in Europe (RR 2.25, 95% CI 1.44-3.50 for CD; RR 2.65, 95% CI 1.61-4.36 for UC). The pooled RR was 2.26 (95% CI 1.42-3.59) in the pediatric CD subgroup. CONCLUSION: This meta-analysis indicates that a pre-illness Western dietary pattern may increase the risk of developing CD and UC.

MiR-449 overexpression inhibits osteogenic differentiation of bone marrow mesenchymal stem cells via suppressing Sirt1/Fra-1 pathway in high glucose and free fatty acids microenvironment.

Diabetic osteoporosis is a chronic complication caused by diabetes mellitus, and However, the exact mechanism of diabetes mellitus-induced osteoporosis is still unknown. In this study, we investigate the effect of miR-449 on osteogenic differentiation and its underlying mechanism in human bone marrow-derived mesenchymal stem cells (hBMSCs) with high glucose (HG) and free fatty acids (FFA) treatment. Results showed that after culturing for 14 days, high glucose (HG) and free fatty acids (FFA) treatment dramatically decreased mineralization of human bone marrow-derived mesenchymal stem cells (hBMSCs) compared with cells treated with osteogenic medium (OM) alone. We also found that miR-449 expression was up-regulated during osteogenic differentiation of hBMSCs with HG and FFA treatment. Moreover, during osteogenic differentiation of hBMSCs with HG and FFA treatment, miR-449 mimics notably decreased the alkaline phosphatase (ALP) activity and the mRNA and protein expression levels of runt-related transcription factor 2 (Runx2), ALP, collagen I, osteocalcin (OCN), and bone sialoprotein (BSP), which was remarkably increased by miR-449 inhibitors. Furthermore, miR-449 directly targets Sirt1 by binding to its 3'-UTR. Sirt1 overexpression reverses the suppressive effect of miR-449 mimics on Fra-1 mRNA and protein expression, which was also alleviated by Fra-1 overexpression. In addition, Fra-1 overexpression alleviates the inhibitory effect of miR-449 mimics on the ALP activity and the mRNA and protein of Runx2, collagen I, OCN and BSP. Taken together, our results indicated that miR-449 overexpression inhibited osteogenic differentiation of HG-FFA-treated hBMSCs through the Sirt1/Fra-1 signal pathway. It is conceivable that modulating miR-449 might provide a new therapy for intervention in diabetic osteoporosis.

Association of Bactericidal Dysfunction of Paneth Cells in Streptozocin-Induced Diabetic Mice with Insulin Deficiency.

BACKGROUND Type 1 diabetes mellitus (T1DM) is associated with increased risks of enteric infection. Paneth cells constitute the first line of the gut defense. Little is known about the impact of T1DM on the bactericidal function of intestinal Paneth cells. MATERIAL AND METHODS A T1DM mouse model was induced by intraperitoneal injection of streptozocin. The analysis of intestinal microbiota and the mucosal bactericidal assay were conducted to evaluate intestinal innate defense. Numbers of Paneth cells and their expression of related antimicrobial peptides were analyzed. Expression of total insulin receptor (IR) mRNA and relative levels of IR-A/IR-B were analyzed. The primary mouse small intestinal crypt culture was used to analyze the effect of insulin and glucose on the expression of related antimicrobial peptides of Paneth cells. RESULTS In T1DM mice, bacterial loads were increased and there was an alteration in the composition of the intestinal microflora. Exogenous bacteria had better survival in the small bowel of the T1DM mice. The expression of Paneth cell-derived antimicrobial peptides was significantly decreased in the T1DM mice, although the number of Paneth cells was increased. Relative levels of IR-A/IR-B in Paneth cells of diabetic mice were elevated, but the total IR mRNA did not change. Insulin treatment restored the expression of antimicrobial peptides and normalized the microbiota in the gut of T1DM mice. Subsequently, in vitro culture assay demonstrated that insulin rather than glucose was essential for the optimal expression of Paneth cell-derived antimicrobial peptides. CONCLUSIONS The bactericidal function of intestinal Paneth cells was impaired in STZ-induced diabetic mice, resulting in the altered intestinal flora, and insulin was essential for the optimal expression of Paneth cell-derived antimicrobial peptides.

Overexpression of miR-429 impairs intestinal barrier function in diabetic mice by down-regulating occludin expression.

Diabetes mellitus (DM) is a group of metabolic diseases characterised by insulin deficiency/resistance and hyperglycaemia. We previously reported the presence of an impaired tight junction and decreased expression of occludin (Ocln) and zonula occludens-1 (ZO-1) in the intestinal epithelial cells (IECs) of type 1 DM mice, but the exact mechanism remains unclear. In this study, we investigated the role of microRNAs (miRNAs) in impairing the tight junction in IECs of DM mice. Using an integrated comparative miRNA microarray, miR-429 was found to be up-regulated in IECs of type 1 DM mice. Then, miR-429 was confirmed to directly target the 3'-UTR of Ocln, although it did not target ZO-1. Moreover, miR-429 down-regulated the Ocln expression in IEC-6 cells in vitro. Finally, exogenous agomiRNA-429 was shown to down-regulate Ocln and induce intestinal barrier dysfunction in normal mice, while exogenous antagomiRNA-429 up-regulated Ocln in vivo and improved intestinal barrier function in DM mice. In conclusion, increased miR-429 could down-regulate the expression of Ocln by targeting the Ocln 3'-UTR, which impaired intestinal barrier function in DM mice.

MEK/ERK pathway activation by insulin receptor isoform alteration is associated with the abnormal proliferation and differentiation of intestinal epithelial cells in diabetic mice.

In previous studies, we have reported the abnormal proliferation and differentiation of intestinal epithelial cells (IECs) in diabetes mellitus (DM) mice. The insulin receptor (IR) and its downstream mitogen-activated protein kinase kinase (MAPKK also known as MEK)/extracellular-regulated protein kinase (ERK) pathway is a classic pathway associated with cell proliferation and differentiation. The purpose of the present study is to investigate the role of the MEK/ERK pathway in abnormal proliferation and differentiation of IECs in DM mice. DM mouse models were induced by intraperitoneal injection of streptozotocin. The expression levels of the IR and its isoforms in IECs of DM mice and in IEC-6 cells were investigated. To ensure that the downstream pathways were monitored, QPCR and Western blotting were performed to detect the expression levels of MEK1/2, ERK1/2, PI3K, and Akt. Moreover, siRNA for IR-A and U0126, a specific inhibitor of MEK, were used to further investigate the relationship between the IR/MEK/ERK pathway and abnormal proliferation and differentiation of IECs in DM mice. In DM mice, excessive proliferation, disturbed differentiation, and a high ratio of IR-A/IR-B were detected in IECs. The expression levels of MEK1, MEK2, and ERK1/2 and their phosphorylated proteins in DM mice were significantly higher than those in the control group (P < 0.05), which could be offset by using siRNA for IR-A. The abnormal proliferation and differentiation of IECs in DM mice were normalized after the in vivo administration of U0126. The abnormal proliferation and differentiation of IECs in DM mice are associated with high IR-A/IR-B ratio and increased IR/MEK/ERK pathway activity.

miRNA-30e regulates abnormal differentiation of small intestinal epithelial cells in diabetic mice by downregulating Dll4 expression.

OBJECTIVES: Depression of the Notch/Hes1 pathway has been reported to play a role in abnormal differentiation of intestinal epithelial cells (IECs) in diabetes mellitus (DM). However, the mechanism by which this pathway influences IEC differentiation has remained unclear. In this study, we have investigated the role of microRNAs (miRNAs) in regulating the Notch/Hes1 pathway in IECs of DM mice. MATERIALS AND METHODS: Integrated comparative miRNA microarray technology was used to determine the expression profile of miRNAs in IECs of DM mice. After bioinformatic analysis, an miRNA with altered expression levels, miRNA-30e, was identified as a candidate for regulating the Notch pathway in DM. A luciferase reporter assay confirmed that miRNA-30e targeted 3'-UTR of the Notch gene. The role of miRNA-30e in regulating Notch signalling was then explored by up- and downregulating its expression in vitro and in vivo. RESULTS: Abnormal differentiation of IECs in DM mice was associated with reduced activity of the Dll4/NICD/Hes1 signal pathway. Based on bioinformatic analyses, increased expression of miRNA-30e was identified as a potential candidate for regulating Notch signalling. miRNA-30e targeted the 3'-UTR of Dll4 and downregulated Dll4 expression in primary IECs and IEC-6 cells. Exogenous miRNA-30e reduced activity of the Dll4/NICD/Hes1 pathway, and induced abnormal differentiation of IECs in normal mice. Conversely, treatment with miRNA-30e antagonist upregu-lated activity of the Dll4/NICD/Hes1 pathway in vivo, and normalized IEC differentiation in DM mice. CONCLUSIONS: Increased levels of miRNA-30e downregulated activity of the Dll4/NICD/Hes1 signalling pathway by targeting the 3'-UTR of Dll4, which contributed to abnormal differentiation in small intestinal epithelia of DM mice.

Diabetes mellitus increases the risk of colorectal neoplasia: An updated meta-analysis.

OBJECTIVE: Recent studies proved that patients with diabetes were at significantly higher risk of developing colorectal cancer. However, the association between diabetes mellitus and the risk of colorectal adenoma remains undefined. Thus we conducted an updated meta-analysis to identify the association between diabetes mellitus and the risk of colorectal neoplasia including adenoma and cancer. METHODS: We conducted a search in databases including Pubmed, Web of Science, EMBASE Databases, Cochrane CENTRAL, Wanfang Data, and CNKI database. Case-control and cohort studies were included. All articles were published before January 2015 and the quality of each study was evaluated by the Newcastle-Ottawa Scale. Odds ratios (ORs) or relative risks (RRs) and its corresponding 95% confidence intervals (CIs) for each study were calculated and summary relative risk estimates with corresponding 95% CIs were generated using the random-effects model. Heterogeneity and publication bias were assessed. RESULTS: Twenty-nine articles including ten case-control studies and nineteen cohort studies were included in this meta-analysis. In a pooled analysis of all studies, diabetes mellitus was associated with increased risk of colorectal neoplasia (RR=1.35, 95% CI=1.28-1.42). The risk increased significantly for both colorectal cancer (RR=1.37, 95% CI=1.30-1.45) and adenoma (RR=1.26, 95% CI=1.11-1.44). Subgroup analyses on study design, gender, geographical region, and type of diabetes mellitus further evidenced these findings. CONCLUSIONS: Diabetes mellitus was associated with an increased risk of colorectal neoplasia. Not only the increased risk of colorectal cancer but also the higher risk of adenoma was identified in patients with diabetes mellitus.

Rebamipide Promotes the Regeneration of Aspirin-Induced Small-Intestine Mucosal Injury through Accumulation of ß-Catenin.

BACKGROUND: The effect of rebamipide on repairing intestinal mucosal damage induced by nonsteroidal anti-inflammatory drugs and its mechanism remain unclear. In this study, we sought to explore the mechanism whereby rebamipide could promote the regeneration of aspirin-induced intestinal mucosal damage. METHODS: BALB/c mice were administered aspirin (200 mg/kg/d) for 5 days to induce acute small intestinal injury (SII). Subsequently, SII mice were treated with rebamipide (320 mg/kg/d) for 5 days. The structure of intestinal barrier was observed with transmission electron microscope, and Zo-1 and occludin expressions were detected. The proliferative index was indicated by the percentage of proliferating cell nuclear antigen positive cells. The prostaglandin E2 (PGE2) levels in the small intestine tissues were measured by an enzyme immunoassay. The mRNA and protein expression levels of cyclooxygenase (COX) and ß-catenin signal were detected in the small intestine using quantitative PCR and Western blot, respectively. RESULTS: COX expression was significantly down-regulated in aspirin induced SII (P < 0.05). In SII mice treated with rebamipide, histopathological findings of aspirin-induced intestinal inflammation were significantly milder and tight junctions between intestinal epithelial cells were improved significantly. The proliferative index increased after rebamipide treatment when compared with that in the control mice. The expressions of COX-2, ß-catenin, and c-myc and the PGE2 concentrations in small intestinal tissues were significantly increased in mice with rebamipide treatments (P < 0.05). CONCLUSION: Rebamipide administration in aspirin-induced SII mice could improve the intestinal barrier structure and promote the regeneration of small intestinal epithelial injury through up-regulating COX-2 expression and the accumulation of ß-catenin.

[Ecological characteristics of Zostera japonica population in Swan Lake of Rongcheng, Shandong Province of China].

In this study, a large area of well preserved Zostera japonica dominated meadow was found in a coastal lagoon, Swan Lake, in Rongcheng of Shandong Province. Due to its unique geographical position and high biomass, this meadow may act as a typical Z. japonica bed in the coastal area of Shandong. From September, 2011 to October, 2012, an annual investigation was conducted on the Z. japonica and its habitats in east coast of Swan Lake, and the distribution of the Z. japonica and its habitats ecological characteristics were preliminarily understood. The major ingredients of sediments particles in the Z. japonica bed was sand (81%) and silt (14%). The C and N contents in the sediments were the highest in winter, and the C/N ratio was the highest in autumn. The shoot density, shoot height, and biomass of the Z. japonica were all significantly correlated with water temperature (P < 0.05). There was an obvious change in the Z. japonica growth among seasons, with the peak biomass obtained in August-September. The C and N contents and C/N in Z. japonica leaves also varied with seasons. The leaf C content was significantly higher in autumn than in spring and summer (P < 0.05), the leaf N content was significantly lower in summer than in spring and autumn (P < 0.01), whereas the leaf C/N ratio was significantly higher in summer than in spring (P < 0.05). The annual carbon sequestration by the Z. japonica in the Swan Lake was estimated to be 111.4 g C x m(-2).

Functional outcome of en bloc resection and osteoarticular allograft reconstruction with locking compression plate for giant cell tumor of the distal radius.

BACKGROUND: Giant cell tumors of the distal radius at Campanacci grade III are particularly challenging to treat. We have treated 15 cases of giant cell tumor of the distal radius by en bloc excision and osteoarticular allograft reconstruction with locking compression plate (LCP). The purpose of this study was to assess the intermediate outcomes of all patients treated with this surgery. METHODS: From July 2002 to January 2009, we followed up 15 patients with giant cell tumors of the distal radius who were treated with en bloc excision and osteoarticular allograft reconstruction with LCPs that were long enough to approach the distal end of the allograft. All of the cases were evaluated based on clinical and radiologic examinations, the passive range of motion of the wrist joint, complications, Mayo wrist score, and short form (SF)-36. RESULTS: The clinical follow-up time after reconstruction averaged 5.2 years. The mean resected length of the radius was 8.1 cm. One patient had tumor recurrence in the soft tissues after 3 years (recurrence rate 6.67 %). No patient had allograft bone fracture, nonunion, or metastases. Subchondral bone alterations and joint narrowing were present in all cases, with 1 patient suffering from the pain, but the pain could be endured without the need for analgesics. The average range of motion of the wrist was 46.7° of dorsiflexion, 33.3° of volar flexion, 61.3° of supination, and 72.3° of pronation. The mean Mayo wrist score was 70 and the mean modified SF-36 score was 71. CONCLUSIONS: En bloc excision and osteoarticular allograft reconstruction with an appropriate LCP for a Campanacci grade III giant cell tumor of the distal radius result in a reasonable functional outcome at intermediate follow-up evaluation. This method can excise the tumor integrally with a low rate of recurrence, good function, and a satisfactory range of motion.

[Comparison of clinicopathological features and prognosis in familial and sporadic gastric cancer].

OBJECTIVE: To analyze the clinicopathological characteristics and prognosis of familial gastric cancer and to improve the treatment outcome. METHODS: Clinical data of 67 patients with familial gastric cancer and 820 patients with sporadic gastric cancer in the Second Affiliated Hospital of Dalian Medical University from 1995 to 2005 were retrospectively analyzed. RESULTS: Compared to sporadic gastric cancer, the percentage of familial gastric cancer patients less than 45 years old was higher (34.3% vs. 14.6%). Early gastric cancer(23.9% vs. 13.8%), diffuse gastric cancer(79.1% vs. 29.0%), and lymph node metastasis (91.0% vs. 70.9%) were more common in patients with familial cancer(P<0.05). The 5-year survival rate of familial gastric cancer patients was lower than that of patients with sporadic gastric cancer(20.5% vs. 45.1%)(P<0.05). CONCLUSIONS: Familial gastric cancer has characteristics of younger onset age, advanced disease staging, higher positive lymph node ratio and poorer prognosis. Therefore, early diagnosis should be emphasized in the management of familial gastric cancer.

[Bioremediation potential of Apostichopus japonicus (Selenka) in coastal bivalve suspension aquaculture system].

Suspension aquaculture of filter-feeding bivalves can produce large amount of faeces and pseudofaeces (biodeposits) that may impact aquaculture environment, while deposit-feeding sea cucumbers may effectively utilize such particulate wastes and act as a scavenger in mariculture system. In this paper, the ingestion, growth, and excretion of deposit-feeder Apostichopus japonicus were investigated in situ seasonally to evaluate its bioremediation potential of a suspension aquaculture system of filter-feeding bivalves. The results showed that A. japonicus could grow well in newly designed culture nets, with its maximum specific growth rate being 0.34% d(-1). The A. japonicus could effectively use the biodeposits generated by co-cultured bivalves, and the ingestion rate at 21.2 degrees C in summer, 19.2 degrees C in autumn, and 7.7 degrees C in winter was 0.1746, 0.0989, and 0.0050 g g(-1) d(-1), respectively. A. japonicus could promote the regeneration of nutriens in biodeposits via the excretion of considerable amount of dissolved N and P, and the excretion also showed obvious seasonal fluctuation. The extrapolation based on the in situ investigation results showed that when co-cultivated with bivalves in lantern nets, A. japonicus would ingest 4.5-159.6 kg hm(-2) d(-1) of dry biodeposits and excrete 1,382.5-3,678.1 mmol hm(-2) d(-1) of NH4(+)-N and 74.6-335.7 mmol hm(-2) d(-1) of PO4(3-)-P, indicating that the deposit-feeding A. japonicus had a great bioremediation capability in suspension aquaculture systems. The integrated model of deposit-feeding A. japonicus and filter-feeding bivalve could not only benefit the economy, but also sustain the environment.

[Photodegradation of bisphenol A in presence of Suwannee River fulvic acid].

Under simulated solar irradiation, the degradation of bisphenol A (BPA) was studied in the presence of Suwannee River Fulvic Acid (SRFA). The results demonstrate that the photodegradation of BPA followed a pseudo-first-order kinetics and the photodegradation rate increased rapidly with increasing initial concentration of SRFA. Hydroxyl radical, singlet oxygen were found in the SRFA solutions of BPA with molecular probes and the technique of electronic spin resonance. The electronic energy transfer of triplet state fulvic acid was also studied with various aerated conditions. The results showed that the photodegradation of BPA was related with triplet state fulvic acid. The photodegradation products of BPA in the presence of SRFA were identified with GC/MS methods. The photodegradation pathways of BPA were also discussed.

Comparison of effectiveness of different ashing auxiliaries for determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method.

An improved method for the determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method is proposed. The recoveries of phosphorus (P) from selected inorganic and organic P-containing compound standards after ignition with different auxiliaries, such as MgSO(4), Mg(NO(3))(2), MgCl(2), Mg(Ac)(2) and CaCl(2), were compared. We found that the phosphorus from most compound standards could not be completely recovered when these compounds were ignited (450-500 degrees C) with the MgSO(4) as auxiliary and the baked residue was extracted with 0.2 mol x l(-1) HCl for 30 min at 80 degrees C or at room temperature. P recoveries, for example, were poor, less than 85%, if pyrophosphate and metaphosphate were ignited with the addition of MgSO(4) prior to the extraction of the baked residue with 0.2 mol x l(-1) HCl at 80 degrees C for 30 min. In contrast, MgCl(2), Mg(Ac)(2) and CaCl(2), as well as Mg(NO(3))(2), could all yield complete P recoveries at routine ashing temperatures (450-500 degrees C). The results demonstrate that MgCl(2) is a more effective auxiliary agent for the determination of phosphorus in natural waters, aquatic organisms and sediments by ignition method than MgSO(4) which is commonly used.