Re-understanding and focusing on normoalbuminuric diabetic kidney disease.

Diabetes mellitus (DM) has grown up to be an important issue of global public health because of its high incidence rate. About 25% of DM patients can develop diabetic foot/ulcers (DF/DFU). Diabetic kidney disease (DKD) is the main cause of end-stage kidney disease (ESKD). DF/DFU and DKD are serious complications of DM. Therefore, early diagnosis and timely prevention and treatment of DF/DFU and DKD are essential for the progress of DM. The clinical diagnosis and staging of DKD are mostly based on the urinary albumin excretion rate (UAER) and EGFR. However, clinically, DKD patients show normoalbuminuric diabetic kidney disease (NADKD) instead of clinical proteinuria. The old NADKD concept is no longer suitable and should be updated accordingly with the redefinition of normal proteinuria by NKF/FDA. Based on the relevant guidelines of DM and CKD and combined with the current situation of clinical research, the review described NADKD from the aspects of epidemiology, pathological mechanism, clinical characteristics, biomarkers, disease diagnosis, and the relationship with DF/DFU to arouse the new understanding of NADKD in the medical profession and pay attention to it.

Analysis of Vitamin D Components in Serum of Minors by UHPLC-MS/MS.

Serum 25-hydroxyvitamin D [25(OH)D] concentration represents the body's reserves of vitamin D, which is mostly used by clinicians to evaluate the storage status of vitamin D in the body. The present study aimed to investigate the serum vitamin D components in different health status of minors to correctly evaluate the vitamin D storage in vivo. A total of 2,270 minors were included in the study, which was divided into healthy group (1,204 cases) and disease group (1,066 cases, including 270 short stature, 433 respiratory infections, 175 malnutrition and 188 tic disorder subjects). The levels of 25-hydroxyvitamin D2 [25(OH)D2] and 25-hydroxyvitamin D3 [25(OH)D3] were measured by UHPLC-MS/MS in all subjects, and the 25(OH)D3 activity equivalents [25(OH)D3-AE] and 25(OH)D were calculated. In addition, the 3-epi-25-hydroxyvitamin D3 [3-epi-25(OH)D3] concentrations of 278 subjects (including 147 healthy and 131 disease subjects) were measured by random sampling. 25(OH)D2, 25(OH)D3, 25(OH)D and 25(OH)D3-AE levels in disease group were significantly lower than those in healthy group (p<0.001). According to the level of 25(OH)D, the sufficiency of vitamin D [25(OH)D≥30 ng/mL] was 65.4% in healthy group and 50.5% in disease group. When the 25(OH)D2 activity was converted into 25(OH)D3-AE, 53.2% of the patients in the healthy group had sufficiency vitamin D, and 39.1% in the disease group. The 3-epi-25(OH)D3 level in the disease group was significantly lower than that in the healthy group (p<0.001). Not only the 25(OH)D, but also the both of 25(OH)D2 and 25(OH)D3 levels may overestimate the vitamin D status in subjects. For accurate evaluation, at least the serum levels of 25(OH)D2, 25(OH)D3 and 3-epi-25(OH)D3 should be determined simultaneously.

Evaluation of vitamin D storage in patients with chronic kidney disease: Detection of serum vitamin D metabolites using high performance liquid chromatography-tandem mass spectrometry.

BACKGROUND: Vitamin D (VitD) deficiency is extremely common in chronic kidney disease (CKD). However, the current clinical testing of vitamin D is based on the recommended serum 25-hydroxyvitamin D [25(OH)D]. The levels of VitD components in CKD patients are rarely reported. In this study, we tested various VitD components, and used different methods to evaluate the VitD status of CKD patients in vivo. METHODS: Totally 173 CKD patients and 111 control individuals were enrolled. Serum levels of 25(OH)D2, 25(OH)D3, C3-epimers (C3-epi) and free 25(OH)D [f-25(OH)D] were measured. The 25(OH)D2/25(OH)D3 ratio, C3-epi/25(OH)D3 ratio, total 25(OH)D [t-25(OH)D], and bioavailable vitamin D (BAVD) were calculated, respectively. RESULTS: The ratios of 25(OH)D2/25(OH)D3, C3-epi/25(OH)D3, and the level of C3-epi in CKD patients were significantly higher than those in the control group (all P < 0.05). The levels of t-25(OH)D, 25(OH)D3, C3-epi, f-25(OH)D and BAVD in patients with CKD stage 5 were significantly lower than those in stages 2, 3, and 4 (all P < 0.05). The calculated VitD storage according to Method 3 [25(OH)D2/3 + 25(OH)D3] was only 32.95 %, which was lower than the results of 53.76 % by Method 1 [25(OH)D2+ 25(OH)D3+C3-epi] and 48.56 % by Method 2 [25(OH)D2/3 + 25(OH)D3+C3-epi]. In addition, the VitD results calculated by three methods were positively correlated with f-25(OH)D and BAVD, while C3-epi levels were also positively correlated with f-25(OH)D and BAVD. CONCLUSION: Serum levels of t-25(OH)D, 25(OH)D3, C3-epi, f-25(OH)D and BAVD in CKD patients gradually decrease with the progression of CKD stages. Though the results of VitD storage in CKD patients evaluated by different methods are different, simultaneous detection of 25(OH)D2, 25(OH)D3, C3-epi and f-25(OH)D levels and fully estimation of their respective biological activities could accurately evaluate the VitD storage in vivo.

IL-12 could induce monocytic tumor cells directional differentiation.

Interleukin-12 (IL-12), a member of interleukin family, plays a critical role in immune responses and anti-tumor activity. In this study, the effects of IL-12 on monocytic tumor cell lines differentiation to macrophagocyte and its likely mechanism was investigated. We examined the differentiation markers, morphological and functional changes, and possible mechanism in IL-12-treated THP-1 and U937 cells. It was found that IL-12 could up-regulated macrophage surface marker CD68 and CD11b expression in a time-dependent manner. Morphologically, after IL-12 treatment, THP-1 and U937 cells became round or irregular shape, even stretched many cell membrane protuberances; some cell nuclei became fuzzy or completely disappeared, and the chromatin appeared dense and cordlike. Furthermore, IL-12-induced monocytic tumor cell differentiation was accompanied by the growth arrest with G1-phase accumulation and S-phase reduction; apoptosis increased with anti-apoptosis protein Bcl-2 down-expression and pro-apoptosis protein Fas up-regulation, and enhanced phagocytosis function. The IL-12-induced macrophage differentiation of THP-1 and U937 cells was associated with the up-regulation of c-fms expression and the CSF-1R Tyr 809 site phosphorylation. These findings have revealed that IL-12 could induce monocytic tumor cells directional differentiation into macrophage-like cells, and its mechanism is possible connected with the up-regulation of c-fms expression and the phosphorylation of CSF-1R Tyr-809 site.

IL-12 regulates B7-H1 expression in ovarian cancer-associated macrophages by effects on NF-κB signalling.

BACKGROUND AND AIM: B7-H1, a co-inhibitory molecule of the B7 family, is found aberrantly expressed in ovarian cancer cells and infiltrating macrophage/dendritic-like cells, and plays a critical role in immune evasion by ovarian cancer. IL-12, an inducer of Th1 cell development, exerts immunomodulatory effects on ovarian cancer. However, whether IL-12 regulates B7-H1 expression in human ovarian cancer associated-macrophages has not been clarified. Therefore, we investigated the effects of IL-12 on the expression of B7-H1 in ovarian cancer-associated macrophages and possible mechanisms. METHODS: PMA induced THP-1-derived macrophages or human monocyte-derived macrophages were treated with recombinant IL-12 (rIL-12) or infected with adenovirus carrying human IL-12 gene (Ad-IL-12-GFP) for 24 h, then cocultured with the SKOV3 ovarian cancer cell line for another 24 h. Macrophages were collected for real-time PCR and Western blot to detect the expression of B7-H1, and activation of the NF-κB signaling pathway. Moreover, supernatants were collected to assay for IL-12, IFN-γ and IL-10 by ELISA. In addition, monocyte-derived macrophages treated with IFN-γ were cocultured with SKOV3 and determined for the expression of B7-H1. Furthermore, the expression of B7-H1 in monocyte-derived macrophages was also evaluated after blocking NF-κB signaling. RESULTS: The expression of B7-H1 was significantly upregulated in monocyte-derived macrophages treated with rIL-12 or Ad-IL-12-GFP compared with the control groups (p<0.05), accompanied by a remarkable upregulation of IFN-γ (p<0.05), a marked downregulation of IL-10 (p<0.05) and activation of NF-κB signaling. However, the upregulation of B7- H1 was inhibited by blocking the NF-κB signaling pathway (p<0.05). Expression of B7-H1 was also increased (p<0.05) in monocyte-derived macrophages treated with IFN-γ and cocultured with SKOV3. By contrast, the expression of B7-H1 in THP-1-derived macrophages was significantly decreased when treated in the same way as monocyte-derived macrophages (p<0.05), and IL-10 was also significantly decreased but IFN-γ was almost absent. CONCLUSIONS: IL-12 upregulates the expression of B7-H1 in monocyte-derived macrophages, which is possible though inducing the secretion of IFN-γ and further activating the NF-κB signal pathway. However, IL-12 downregulates the expression of B7-H1 in THP-1-derived macrophages, associated with a lack of IFN-γ and inhibition of expression of IL-10.

Integrated analysis of long non-coding RNAs and mRNA expression profiles reveals the potential role of lncRNAs in gastric cancer pathogenesis.

Long non-coding RNAs (lncRNAs) have been shown to play a critical role in cancer biology and are frequently aberrantly expressed. Despite their important role in pathology, little is known mechanistically regarding their role in gastric cancer (GC) pathogenesis. To characterize the role of lncRNAs in GC pathogenesis, 8 paired human GC tissue samples and matched adjacent normal tissue were examined. Large scale expression profiling of lncRNA and mRNA was performed utilizing microarray technology and validated by qPCR. Differentially expressed lncRNAs were subjected to bioinformatic analysis to predict target genes, followed by the integration of differentially expressed mRNA data and GO and network analysis to further characterize potential interactions. In our study, 2,621 lncRNAs and 3,121 mRNAs were identified to be differentially expressed (≥2.0-fold change) in GC samples relative to their matched counterparts. lncRNA target prediction revealed the presence of 221 potential lncRNA-mRNA target pairs for the 75 differentially expressed lncRNAs and 60 differentially expressed genes. KEGG pathway analysis showed that these target genes were significantly enriched in 7 different pathways, of which the p53 signaling pathway was the most significant and has been previously implicated in GC pathogenesis. Construction of a lncRNA-mRNA correlation network revealed 10 differentially expressed lncRNAs potentially regulating the p53 signaling pathway. Overall, this is the first study perform global expression profiling of lncRNAs and mRNAs relating to GC. These results may provide important information for further insights into the pathogenesis of GC and provide potential targets for future therapeutics.