The relationship between sex hormones and glycated hemoglobin in a non-diabetic middle-aged and elderly population.

BACKGROUND: Sex hormones are strongly linked to the occurrence and development of diabetes, and influence glycated hemoglobin (HbA1c) levels in diabetic population; but, the relationship between sex hormones and HbA1c in non-diabetic population remains unknown. This study aimed to explore the extent of influence of sex hormones on HbA1c levels in non-diabetic population. METHODS: A total of 1409 non-diabetic subjects, including 601 men and 808 postmenopausal women were recruited from Shanghai community. HbA1c was detected using high performance liquid chromatography, and hemoglobin level was determined by sodium lauryl sulfate colorimetry. Serum estradiol (E2), total testosterone (TT), and sex hormone binding globulin (SHBG) were measured by chemiluminescent microparticle immunoassays. RESULTS: The level of HbA1c was 5.6 (5.4-5.9) % in all subjects, with 5.6 (5.4-5.8) % in men and 5.7 (5.5-5.9) % in postmenopausal women. After adjusting for age, body mass index (BMI), and hemoglobin, E2 was positively correlated with HbA1c in men (r = 0.122, P = .003), and SHBG was inversely correlated with HbA1c (r = - 0.125, P < .001) in women. Other hormones were not correlated with HbA1c (all P > .05). Multivariate linear regression analysis showed that, except for traditional factors, such as age, hemoglobin, and BMI, E2 was another determinant of HbA1c (standardized ß = 0.137, P = .003) in men; besides, in women, SHBG was another determinant of HbA1c (standardized ß = - 0.178, P < .001), except for age and systolic blood pressure. CONCLUSION: After controlling for confounding factors, two sex hormones, as E2 and SHBG could influence HbA1c levels in non-diabetic population.

Targeting the lncRNA DUXAP8/miR-29a/PIK3CA Network Restores Doxorubicin Chemosensitivity via PI3K-AKT-mTOR Signaling and Synergizes With Inotuzumab Ozogamicin in Chemotherapy-Resistant B-Cell Acute Lymphoblastic Leukemia.

Purpose: This study aimed to determine the expression profiles of long non-coding RNA (lncRNA), microRNA (miRNA), and mRNA in chemotherapy-resistant B-cell acute lymphoblastic leukemia (B-ALL). Methods: LncRNA, miRNA, and mRNA profiles were assessed by RNA-seq in diagnostic bone marrow samples from 6 chemotherapy-resistant and 6 chemotherapy-sensitive B-ALL patients. The lncRNA DUXAP8/miR-29a/PIK3CA signaling network was identified as the most dysregulated in chemoresistant patient samples, and its effect on cellular phenotypes, PI3K-AKT-mTOR signaling, and chemosensitivity of doxorubicin (Dox)-resistant Nalm-6 (N6/ADR), and Dox-resistant 697 (697/ADR) cells were assessed. Furthermore, its synergy with inotuzumab ozogamicin treatment was investigated. Results: 1,338 lncRNAs, 75 miRNAs, and 1620 mRNAs were found to be dysregulated in chemotherapy-resistant B-ALL in comparison to chemotherapy-sensitive B-ALL patient samples. Through bioinformatics analyses and RT-qPCR validation, the lncRNA DUXAP8/miR-29a/PIK3CA network and PI3K-AKT-mTOR signaling were identified as significantly associated with B-ALL chemotherapy resistance. In N6/ADR and 697/ADR cells, LncRNA DUXAP8 overexpression and PIK3CA overexpression induced proliferation and inhibited apoptosis, and their respective knockdowns inhibited proliferation, facilitated apoptosis, and restored Dox chemosensitivity. MiR-29a was shown to affect the lncRNA DUXAP8/PIK3CA network, and luciferase reporter gene assay showed direct binding between lncRNA DUXAP8 and miR-29a, as well as between miR-29a and PIK3CA. Targeting lncRNA DUXAP8/miR-29a/PIK3CA network synergized with inotuzumab ozogamicin's effect on N6/ADR and 697/ADR cells. Conclusion: Targeting the lncRNA DUXAP8/miR-29a/PIK3CA network not only induced an apoptotic effect on Dox-resistant B-ALL and restored Dox chemosensitivity via PI3K-AKT-mTOR signaling but also showed synergism with inotuzumab ozogamicin treatment.

Lymphoma cell-derived extracellular vesicles inhibit autophagy and apoptosis to promote lymphoma cell growth via the microRNA-106a/Beclin1 axis.

Lymphoma is a common malignant tumor globally. Tumor-derived extracellular vesicles (Evs) participate in genetic information exchange between tumor cells. We investigated the role and mechanism of human Burkitt lymphoma cells Raji-derived Evs (Raji-Evs) in lymphoma cells. Effects of Evs on lymphoma cell proliferation, invasion, autophagy, and apoptosis were assessed using Cell Counting Kit-8 method, Transwell assay, laser confocal microscopy, Western blotting, and flow cytometry. microRNA (miR)-106a expression in lymphoma cells was determined using reverse transcription-quantitative polymerase chain reaction and then downregulated in Raji cells and then Evs were isolated (Evs-in-miR-106a) to evaluate its role in lymphoma cell growth. The binding relationship between miR-106a and Beclin1 was verified using RNA pull-down and dual-luciferase assays. Beclin1 was overexpressed in SU-DHL-4 and Farage cells and SU-DHL-4 cell autophagy and apoptosis were detected. The levels of miR-106a and Beclin1 in SU-DHL-4 cells were detected after adding autophagy inhibitors. The tumorigenicity assay in nude mice was performed to validate the effects of Raji-Evs in vivo. Raji-Evs promoted lymphoma cell proliferation and invasion and increased miR-106a. miR-106a knockdown reversed Evs-promoted lymphoma cell proliferation and invasion. miR-106a carried by Raji-Evs targeted Beclin1 expression. Beclin1 overexpression or miR-106a inhibitor reversed the effects of Evs on lymphoma cell autophagy and apoptosis. Autophagy inhibitors elevated miR-106a expression and lowered Beclin1 expression. Raji-Evs-carried miR-106a inhibited Beclin1-dependent autophagy and apoptosis in lymphoma cells, which were further verified in vivo, together with promoted tumor growth. We proved that Raji-Evs inhibited lymphoma cell autophagy and apoptosis and promoted cell growth via the miR-106a/Beclin1 axis.

Corrigendum to "Identification and characterization of the cellular subclones that contribute to the pathogenesis of mantle cell lymphoma" [Genes Dis 6(4) 2019 407-418].

[This corrects the article DOI: 10.1016/j.gendis.2018.12.002.].

Potential of the tumor­derived extracellular vesicles carrying the miR­125b­5p target TNFAIP3 in reducing the sensitivity of diffuse large B cell lymphoma to rituximab.

Diffuse large B­cell lymphoma (DLBCL) is the most common and aggressive form of non­Hodgkin's lymphoma. Extracellular vesicles (EVs) derived from cancer cells are known to modify the tumor microenvironment. The aim of the present study was to investigate the role of miR­125b­3p carried by EVs in DLBCL in vitro and in vivo. TNFAIP3 expression in patient lesions was measured and the upstream miR that regulates TNFAIP3 was predicted using the starBase database. EVs were isolated from DLBCL cells and identified. DLBCL cells were transfected with pcDNA to overexpress TNFAIP3 or inhibit miR­125b­5p expression, incubated with EVs, and treated with rituximab to compare cell growth and TNFAIP3/CD20 expression. DLBCL model mice were administered EVs, conditioned medium, and rituximab to observe changes in tumor size, volume, and weight. TNFAIP3 was downregulated in patients with DLBCL and its levels further decreased in patients with drug­resistant DLBCL. Overexpression of TNFAIP3 in DLBCL cells enhanced the inhibitory effect of rituximab and increased CD20 expression. miR­125b­5p targeted TNFAIP3. Inhibition of miR­125b­5p enhanced the inhibitory effect of rituximab in DLBCL cells. The EV­carried miR­125b­5p reduced the sensitivity of DLBCL cells to rituximab, which was averted by overexpression of TNFAIP3. EVs reduced the sensitivity of DLBCL model mice to rituximab via the miR­125b­5p/TNFAIP3 axis. The study findings indicate that the tumor­derived EVs carrying miR­125b­5p can enter DLBCL cells and target TNFAIP3, thus reducing the sensitivity of DLBCL to rituximab, which may provide a novel therapeutic approach for DLBCL.

[Effect of Autophagy to Drug Resistance of B-cell Lymphoma Cells].

OBJECTIVE: To investigate the effect of autophagy on the drug resistance of different human lymphoma cells. METHODS: Human Burkitt's lymphoma cell Daudi, human B lymphoma cell SUDHL-4, and human mantle cell lymphoma cell JeKo-1 were taken as the research subjects. The expression of Atg5 was inhibited by the treatments of autophagy inhibitors or stable interference via lentivirus infection. The autophagy activity of B lymphoma cell was changed, and the changes of lymphoma cells to the drug resistance of ADR and VCR was observed. RESULTS: JeKo-1 cells showed the strongest resistance to ADR and VCR, followed by SUDHL-4, and Daudi cells showed the weakest resistance to ADR and VCR. At the same time, JeKo-1 cells showed the strongest autophagy activity, followed by SUDHL-4, and Daudi cells showed the weakest autophagy activity. After the treatments of autophagy inhibitors or stable Atg5 interference, the resistance of lymphoma cells to ADR and VCR was significantly weakened, and there was the positive correlation at the drug resistance and the autophagy activity of B lymphoma cell. CONCLUSION: The higher autophagy activity in lymphoma cells, the lower chemotherapy resistance of the lymphoma cells after autophagy was inhibited.

S100A8 promotes chemoresistance via augmenting autophagy in B­cell lymphoma cells.

B­cell lymphomas (BCLs) are malignant lymphoid tumours originating from the malignant proliferation and transformation of mature lymphocytes at various stages of differentiation and clonal expansion of the lymphatic and circulatory systems. Efforts to control or even eradicate BCLs are frequently hampered by the development of drug resistance. Autophagy is an evolutionarily conserved biological process of the energy metabolism. By degrading intracellular organelles and proteins, autophagy provides cells with biochemical reaction substrates for the maintenance of homeostasis under nutrient deprivation or other stressful conditions. Accumulating evidence indicates that autophagy plays an important role in chemotherapy resistance. S100A8 is an important member of the calcium­binding protein family that plays an important role in regulating tumour resistance to chemotherapy, while the specific molecular regulatory mechanisms remain unclear. In the present study, by employing three BCL cell lines (Daudi, SUDHL­4 and JeKo­1), it was demonstrated that BCL cells with a strong drug resistance also exhibited active autophagy. In addition, S100A8 was found to be crucial for regulating drug resistance and promoting autophagy in BCL cells. Interference of S100A8 significantly downregulated Bcl­2/adenovirus E1B 19­kDa protein­interacting protein 3 located in the mitochondria and endoplasmic reticulum to further inhibit autophagy. In addition, S100A8 interference markedly inhibited the formation of the BECN1­PI3KC3 complex and promoted B­cell lymphoma 2 expression, which collectively inhibited autophagy.

[Biological Characteristics of Immunophenotype, Stem Cell Gene and Colony Forming Ability in Side-Population Cells of Jeko-1 Cell Line].

AbstractObjective: To investigate the in vitro biological characteristics of side-population(SP) cells in mantle cell lymphoma(MCL). METHODS: The SP cells in JeKo-1 cells were enriched by continuous culture and sorted by flow cytometry（FCM）, and the expression of CD19 and IgM were analyzed. The differences of gene expression and their self-renewal ability between SP cells and non-SP cells were estimated by qRT-PCR and colony-forming assay(CFA) respectively. RESULTS: SP cells in JeKo-1 cells could be enriched by continuous sorting and in vitro cultureing, and it was found that CD19-/IgM- accounted for (2.79±0.82)%,CD19-/IgM+accounted for (70.99±13.61)%,CD19+/IgM-accounted for (0.55±0.25)%, and CD19+/IgM+accounted for (25.67±14)%. Bmi-1,CD133, C-MYC and Nanog genes were highly expressed in SP cells, but were low expressed in non-SP cells (P<0.05). Compared with non-SP cells, SP cells had stronger colony forming ability (P<0.05). CONCLUSION: JeKo-1 cell line of mantle cell lymphoma contains SP cells, which can be enriched by continuous sorting and culture for research. The SP cells in JeKo-1 contain four different cell populations with different immunophenotypes： CD19-/IgM-、CD19-/IgM+、CD19+/IgM-、CD19+/IgM+, and show stronger gene expression of stem cells and self-renewal ability in vitro.

Three-dimensional printed tissue engineered bone for canine mandibular defects.

BACKGROUND: Three-dimensional (3D) printed tissue engineered bone was used to repair the bone tissue defects in the oral and maxillofacial (OMF) region of experimental dogs. MATERIAL AND METHODS: Canine bone marrow stromal cells (BMSCs) were obtained from 9 male Beagle dogs and in vitro cultured for osteogenic differentiation. The OMF region was scanned for 3D printed surgical guide plate and mold by ProJet1200 high-precision printer using implant materials followed sintering at 1250 °C. The tissue engineered bones was co-cultured with BASCs for 2 or 8 d. The cell scaffold composite was placed in the defects and fixed in 9 dogs in 3 groups. Postoperative CT and/or micro-CT scans were performed to observe the osteogenesis and material degradation. RESULTS: BMSCs were cultured with osteogenic differentiation in the second generation (P2). The nanoporous hydroxyapatite implant was made using the 3D printing mold with the white porous structure and the hard texture. BMSCs with osteogenic induction were densely covered with the surface of the material after co-culture and ECM was secreted to form calcium-like crystal nodules. The effect of the tissue engineered bone on the in vivo osteogenesis ability was no significant difference between 2 d and 8 d of the compositing time. CONCLUSIONS: The tissue-engineered bone was constructed by 3D printing mold and high-temperature sintering to produce nanoporous hydroxyapatite scaffolds, which repair in situ bone defects in experimental dogs. The time of compositing for tissue engineered bone was reduced from 8 d to 2 d without the in vivo effect.

Identification and characterization of the cellular subclones that contribute to the pathogenesis of mantle cell lymphoma.

Mantle cell lymphoma (MCL) is a B-cell malignancy with poor clinical outcome and undefined pathogenesis. Development of clinically relevant cellular models for MCL research is an urgent need. Our preliminary observations lead the development of two novel hypotheses that we tested in this study: 1. multicellular spheroid might be a unique growth mode of early-stage cells in MCL; 2. MCL might be a polyclonal tumor. We made the following original observations that have not been reported: First, we have provided a new experiment method for enriching MCL early-stage cells and characterized the spheroid mode of growth as a unique feature of early-stage MCL cells in cell line as well as in clinical samples. Second, we have established a clinically relevant cellular model of MCL, the JeKo-1-spheroid cell line, that was highly enriched in early-stage sub-clones. JeKo-1-spheroid cells and the spheroid growing cells enriched from MCL patients exhibited comparably enhanced tumorigenic abilities and similar biological features. Third, Immunophenotypic analysis has revealed that MCL may be derived from precursor-B(pre-B), immature-B and mature-B cells, not only the mature-B cells as WHO classified in 2016. Fourth, MCL may be a polyclonal disease composed of CD19-/IgM-, CD19-/IgM+, CD19+/IgM+ three sub-clones, of which the CD19-/IgM+ sub-clone might be the dominant sub-clone with the strongest tumorigenic ability. Fifth, CD19+/IgM- that differentiates MCL and normal B cells may represent a new marker for MCL early detection, minor residual disease monitoring after therapies and prognosis.

[Expression and Significance of LncRNA RP11-513G11.1 in Peripheral Blood of Patients with Diffuse Large B-Cell Lymphoma].

OBJECTIVE: To investigate the expression and significance of LncRNA RP11-513G11.1 in peripheral blood of patients with diffuse large B-cell lymphoma (DLBCL), and to analyze its correlation with clinicopathological features and prognosis of patients. METHODS: The serum samples of 93 patients with DLBCL(DLBCL group) and 62 normal persons (control group) were collected from the Department of Hematology, Southwest Medical University. The expression of RP11-513G11.1 in serum samples was detected by real-time fluorescence quantitative PCR, the relationship between the RP11-513G11.1 expression with clinicopathological characteristics and prognosis was analyzed. RESULTS: Compared with normal control group, the expression of RP11-513G11.1 significantly increased in DLBCL patients (P<0.001). The expression of RP11-513G11.1 not related with the age, sex, course of treatment and germinal center B-cell-like lymphoma(GCB) subtypes of the patients, but it related with the diameter of tumor,Ann Arbor stage,B symptoms,chemosensitivity and the international prognostic index(IPI) (P<0.05). The progression-free survival time and overall survival time of patients, whom with high expression of RP11-513G11.1 were significantly shorter than those of RP11-513G11.1 low expression（P<0.001）. The median progression-free survival time and overall survival time of chemotherapy-sensitive patients were significantly longer than those of chemotherapy-resistant patients (P<0.001). Univariate analysis and multivariate Cox regression analysis showed that Ann Arbor stage, RP11-513G11.1 expression, IPI and chemosensitivity were also the independent factors affecting the prognosis of DLBCL patients（P<0.05）. CONCLUSION: RP11-513G11.1 is highly expressed in patients with DLBCL, which is related with the prognosis of DLBCL patients.

Role of hyperglycaemia in the relationship between serum osteocalcin levels and relative skeletal muscle index.

BACKGROUND & AIMS: Studies have shown that osteocalcin is involved in energy metabolism and is sufficient to prevent age-related muscle loss. The present study investigated the association of serum osteocalcin levels with muscle mass and the influence of metabolic factors on this association in humans. METHODS: A total of 1742 middle-aged and elderly subjects (median age: 61.2 years; interquartile range: 56.9-65.3 years) were enrolled from Shanghai communities, including 775 men and 967 postmenopausal women. Serum osteocalcin levels were measured by an electrochemical immunoluminescence assay. An automatic bioelectric impedance analyser (BIA) was used to measure body compositions. Relative skeletal muscle index (SMI) was calculated using the BIA equation from Janssen et al. RESULTS: SMI was significantly higher in men than in postmenopausal women (37.30% [35.14%-39.63%] versus 27.72% [25.99%-29.66%], p < 0.001). Increasing SMI was associated with decreases in the frequency of overweight/obesity, central obesity, dyslipidaemia, elevated blood pressure, and hyperglycaemia (all p < 0.001). Serum osteocalcin levels were positively correlated with SMI in both men and women, regardless of treatment as a categorical or continuous variable (all p < 0.001). However, after accounting for confounding variables, the relationship remained only in men with hyperglycaemia (standardized ß = 0.068, p = 0.024). Among men with isolated impaired glucose tolerance, the odds ratio of increased SMI was 2.861 in the fourth osteocalcin quartile compared with the lowest (p = 0.046). Multiple stepwise regression revealed that each standard deviation (SD) increase of serum osteocalcin levels resulted in an increase of 0.131 SD in SMI (p = 0.024). CONCLUSION: Serum osteocalcin levels were positively related to SMI in men with hyperglycaemia, especially in those with isolated impaired glucose tolerance. No association was detected between serum osteocalcin levels and SMI in postmenopausal women.

Heterogeneity of insulin resistance and beta cell dysfunction in gestational diabetes mellitus: a prospective cohort study of perinatal outcomes.

BACKGROUND: Insulin resistance and beta cell dysfunction were reported to be responsible for gestational diabetes mellitus (GDM). However, little is known about the heterogeneity of these factors and its influences on perinatal outcomes. We investigated whether subtypes of insulin resistance and beta cell dysfunction in gestational diabetes mellitus have different impacts on perinatal outcomes. METHODS: In this prospective cohort study, we followed 554 pregnant women and glucose challenge test was performed at 24-28th weeks of their gestation. Women with plasma glucose ≥ 7.8 mmol/L would be included and advised to undergo the diagnostic 75-g, 3-h oral glucose tolerance test. According to indices of measuring insulin resistance or beta cell function were below the 25th percentile of women with normal glucose tolerance (NGT), women with GDM were defined as three subtypes: GDM with the beta cell dysfunction, GDM with the insulin resistance defect or GDM with both traits mentioned above (GDM-mixed). Perinatal outcomes were documented. RESULTS: The levels of prepregnancy and maternal BMI in the GDM-mix group were higher compared to women in the NGT group (23.2 ± 4.0 vs 20.8 ± 3.7 kg/m2, P < 0.001; 24.5 ± 4.3 vs 21.8 ± 3.4 kg/m2, P < 0.001, respectively). Furthermore, women in GDM-mix group more likely to be subjected to LGA (P = 0.008) adverse perinatal outcomes (P = 0.005), although these differences were normalized after adjusting age, prepregnancy and maternal BMI (GDM-mix vs. NGT: P = 0.141 for LGA and P = 0.186 for adverse outcomes). On the other hand, all perinatal outcomes were similar between other two GDM subgroups and NGT group. CONCLUSIONS: Women with GDM display respective characteristics on metabolism disorders and confer discriminating risks of adverse perinatal outcomes because of this heterogeneity.

Sec23a mediates miR-200c augmented oligometastatic to polymetastatic progression.

BACKGROUND: Cancer treatment is based on tumor staging. Curative intent is only applied to localized tumors. Recent studies show that oligometastatic patients who have limited number of metastases may benefit from metastasis-directed local treatments to achieve long-term survival. However, mechanisms underlying oligometastatic to polymetastatic progression remains elusive. METHODS: The effects of miR-200c and Sec23a on tumor metastasis were verified both in vitro and in vivo. The secretome changes were detected by mass spectrometry. FINDINGS: We established a pair of homologous lung-metastasis derived oligometastatic and polymetastatic cell lines from human melanoma cancer cell line M14. Using the two cell lines, we have identified Sec23a, a gene target of miR-200c, suppresses miR-200c augmented oligometastatic to polymetastatic progression via its secretome. Firstly, miR-200c over-expression and Sec23a interference accelerated oligometastatic to polymetatic progression. Secondly, Sec23a functions downstream of miR-200c. Thirdly, mass spectrometric analysis of the secretory protein profile suggests that Sec23a-dependent secretome may impact metastatic colonization by modifying tumor microenvironment. Fourthly, the survival analysis using The Cancer Genome Atlas database shows Sec23a as a favorable prognostic marker for skin cutaneous melanoma, supporting the clinical relevance of our findings. INTERPRETATION: The finding that Sec23a is a suppressor of oligometastatic to polymetastatic progression has clinical implications. First, it provides a new theoretical framework for the development of treatments that prevent oligometastasis to polymetastasis. Second, Sec23a may be used as a favorable prognostic marker for the selection of patients with stable oligometastatic disease for oligometastasis-based local therapies of curative intent. FUND: National Natural Science Foundations of China.

Traceability to a primary reference measurement procedure (ID-LCMS); A key step in validating the clinical accuracy and safety of hospital blood glucose monitoring systems.

OBJECTIVE: A key step in the evaluation of the accuracy of blood glucose monitoring systems (BGMS) is using a comparator method aligned to a high order definitive reference method. We describe how we achieved traceability to an isotope dilution liquid chromatography mass spectrometry (ID-LCMS) method. We used ID-LCMS to evaluate the accuracy and specificity of two hospital BGMS used in China. METHOD: ID-LCMS was used to verify the calibration alignment of the laboratory plasma hexokinase reference method using NIST standard reference material and clinical samples. The ID-LCMS aligned hexokinase method was used to evaluate the clinical accuracy of two BGMS in hospitalized patients. System accuracy was evaluated using Chinese consensus guidelines. BGMS accuracy was also assessed with interference factors known to be present in critically ill patients' blood. RESULTS: The laboratory plasma hexokinase reference method was shown to calibrate closely with ID-LCMS. Two BGMS demonstrated good correlation with this reference method. Only one BGMS met the Chinese guidelines. The interference factors didn't influence this BGMS but adversely affected the clinical accuracy of the other. CONCLUSIONS: We advocate that our IDMS calibration alignment approach for ensuring the accuracy of the glucose reference method should be adopted in evaluations assessing the accuracy of blood glucose monitoring systems.

Postload Glycated Albumin as an Alternate Measure for Diabetes Screening in a Chinese Population.

In previous epidemiological screening in China, glycated albumin (GA) was mostly detected during the fasting state. This strict restriction causes some problems with diabetes screening. It is unclear if GA could help improve the efficiency of screening for diabetes for subjects who are not in the fasting state. The present study analyzed the differences between fasting and postload (30, 60, 120, and 180 min) GA levels. A total of 691 participants were enrolled in the present study. The Bland-Altman difference plots revealed that 95.4, 94.8, 93.6, and 93.9% of data points were within the limits of agreement for each time point. The receiver operating characteristic curve showed that the areas under the curve (AUC) for baseline GA and postload GA for every time point were 0.822 (95% CI 0.791-0.849), 0.821 (95% CI 0.790-0.848), 0.833 (95% CI 0.803-0.860), 0.840 (95% CI 0.811-0.867), and 0.840 (95% CI 0.810-0.867), with sensitivities of 67.5, 68.1, 69.3, 71.6, and 69.3%, respectively. There was no difference between the baseline and postload GA levels in either AUC or sensitivity (all p > 0.05). In conclusion, postload serum GA levels were in good agreement with those at baseline, and thus, it may be reasonable to employ nonfasting measurements of GA levels for diabetes screening.

Early elevated alkaline phosphatase increases the risk of large-for-gestational-age birth weight in pregnant women with normal glucose tolerance.

AIMS: The aim of this study was to assess the association between levels of alkaline phosphatase (ALP) in early pregnancy and the incidence of large-for-gestational-age (LGA) neonates in pregnant women without gestational diabetes mellitus. METHODS: A prospective cohort was carried out in 544 women and their biochemical parameters including serum ALP and demographic characteristics were collected in 13-16th gestational week. At 24-28th weeks of gestation, 50 g oral glucose challenge test and oral 75 g glucose tolerance test was performed. LGA was defined as birth weight ≥ 90th percentile for completed week of gestational age based on the sex-specific growth curves. Logistic regression and receiver operating characteristic analysis were utilized to identify independent risk factors and odds ratio among ALP quartiles for incidence of LGA. RESULTS: Women diagnosed as LGA held higher level of ALP than women in non-LGA group (P = 0.008). Moreover, ALP (odds ratio (OR) 1.05 [95% confidence interval (CI): 1.00, 1.10]) was the independent risk factors associated with LGA. Compared with ALP quartile 1, women in quartile 4 had more than 2.5-fold increased odds of LGA (OR 3.78, 95% CI: 1.10, 13.02), and the risk reached 4 times after adjusting several covariates (OR 4.15, 95% CI: 1.14,15.13). CONCLUSIONS: A significantly increased risk of LGA was associated with higher serum concentrations of ALP in pregnant women with NGT, even it is in normal reference range.

Neuralized1a regulates asymmetric division in mouse Lewis lung carcinoma cells.

Asymmetric division (ASD), the unique characteristic of normal stem cells, is regarded as a stemness marker when applied to the study of cancer stem cells (CSCs). However, the role of ASD in the self-renewal of CSCs and its regulation remain largely unknown. Here, we first established a mouse Lewis lung carcinoma CSC cell line that could undergo asymmetric division (LLC-ASD cells) derived from the parental mouse Lewis lung carcinoma cancer cells (LLC-Parental cells). In vitro assessment of stemness by RT-qPCR and western blot analysis of stem cell markers, clonogenic assay (p < 0.001), single cell spheroid formation assay (p < 0.05) and 96-well-plate single-cell cloning assay (p < 0.01) indicated that the LLC-ASD cells exhibited stronger stemness features in comparison to the LLC-Parental cells. In vivo, tumorigenicity of LLC-ASD cells, transplanted subcutaneously to the nude mice, was increased compared to that of LLC-parental cells (p < 0.05). Further, Neuralized1a, a regulator of ASD in normal stem cells, was highly expressed in the LLC-ASD cells. Silencing Neuralized1a expression in LLC-ASD cells by siRNA weakened the stemness features measured by the in vitro assays listed above (p < 0.05). The tumorigenic ability was also decreased in the nude mice upon Neuralized1a silencing (p < 0.05). Collectively, the present study suggests that Neuralized1a regulates the stemness of LLC-ASD cells which could be the new marker and therapeutic target of CSCs.

Verification of a novel point-of-care HbA1c device in real world clinical practice by comparison to three high performance liquid chromatography instruments.

INTRODUCTION: A real world clinical study was designed and conducted to evaluate the performance of a novel point-of-care device for determination of glycated haemoglobin A1c (HbA1c), A1C EZ 2.0, in daily clinical practice. MATERIALS AND METHODS: Five hundred and fourteen subjects were included in this study, and divided into three groups. HbA1c was measured by A1C EZ 2.0 and three different high performance liquid chromatography (HPLC) devices: Bio-Rad Variant II Turbo, Tosoh HLC-723 G8 and Premier Hb9210 separately. Precision of A1C EZ 2.0 was also evaluated. RESULTS: Results obtained from A1C EZ 2.0 and all HPLC devices are correlated. Passing-Bablok regression analysis shows the equation of A1C EZ 2.0 results against the mean of HPLC devices with corresponding 95% confidence intervals (95% CI) for the intercept and slope is y = 0.10 (- 0.17 to 0.10) + 1.00 (1.00 to 1.04) x. Bland-Altman difference plot shows that the mean relative difference between A1C EZ 2.0 and Variant II Turbo, G8, Hb9210 and all HPLC results is 2.5%, 0.6%, 0.4% and 1.1%, respectively. In addition, 121 pairs of results determined by using both venous and capillary blood prove that the difference of two kinds of blood sample causes no notable variation when measured by A1C EZ 2.0. Precision study gives 2.3% and 1.9% of total coefficient of variation for normal and abnormal HbA1c sample in A1C EZ 2.0. CONCLUSIONS: HbA1c values measured by A1C EZ 2.0 were in good accordance with the results obtained with the reference HPLC devices.