[Hepatic Insulin Resistance and Type 2 Diabetes Mellitus].

Insulin resistance (IR) is a pathological reaction of hyperinsulinemia and impaired glucose tolerance caused by decreased sensitivity of target tissues such as liver to insulin.The pathogenesis of IR as a typical pathological feature of type 2 diabetes is the focus of anti-diabetes research.In this paper,we reviewed the molecular mechanisms of glucose and lipid metabolism,oxidative stress,mitochondrial dysfunction,endoplasmic reticulum stress,inflammation,and hepatic IR in the case of type 2 diabetes mellitus,which might provide new ideas and theoretical guidance for the treatment of diabetes mellitus.

Predictive value of plasma neutrophil gelatinase-associated lipocalin for acute kidney injury in intensive care unit patients after major non-cardiac surgery.

AIM: The performance of plasma neutrophil gelatinase-associated lipocalin (pNGAL) for prediction of acute kidney injury (AKI) in non-cardiac surgical patients has not been well described. This study investigates the use of pNGAL for early detection of AKI in patients admitted to an intensive care unit (ICU) after major or ultra-major non-cardiac surgery. METHODS: A total of 151 patients were recruited. Blood samples at 0 h and 6 h post-ICU admission were collected. Primary outcome was occurrence of AKI within 48 h of ICU admission defined using Acute Kidney Injury Network (AKIN) classification. RESULTS: Forty-five (29.8%) patients developed AKI within 48 h of ICU admission. Among them, 22, 14, and nine were classified as AKIN Stage 1, 2, and 3 respectively. pNGAL levels at 0 h and 6 h were significantly related to AKI severity. The AUROC for pNGAL at 0 h and 6 h increased with AKI severity (AKIN stage ≥1 0.671 ± 0.048 and 0.691 ± 0.047; stage ≥2 0.737 ± 0.055 and 0.796 ± 0.048; stage 3 0.829 ± 0.072 and 0.860 ± 0.065, respectively) and requirement of renal replacement therapy (0.880 ± 0.059 & 0.837 ± 0.088). Change of pNGAL from 0 h to 6 h showed no advantage in predictive power compared with pNGAL level at 0 h or 6 h alone. The addition of pNGAL into clinical AKI prediction model could only provide marginal benefit. CONCLUSION: pNGAL correlated with severity of AKI and requirement of renal replacement therapy in ICU patients who received major or ultra-major non-cardiac surgery. However, the benefit of adding pNGAL into clinical AKI prediction model is marginal.

A Brassica napus PHT1 phosphate transporter, BnPht1;4, promotes phosphate uptake and affects roots architecture of transgenic Arabidopsis.

Phosphorus (P) is one of the essential nutrient elements for plant development. In this work, BnPht1;4 gene, encoding a phosphate transporter of PHT1 family, was isolated from Brassica napus. BnPht1;4 possesses the major characteristic of PHT1 high-affinity Pi transporters in plants, such as plasma-membrane localization and 12 transmembrane-spanning domains. Quantitative reverse-transcription PCR analysis and promoter activity assay showed BnPht1;4 was inert in plants under Pi sufficient conditions. However, expression of this gene was remarkably enhanced in roots under Pi deficient conditions. Interestingly, under low Pi conditions, its promoter activity is impaired in tips of elongated roots, suggesting that the high-affinity Pi transporter may be not involved in low Pi response at root tip area. The experimental results also indicated that BnPht1;4 induction by Pi deficiency is dependent on the existence of sugar. In 35S:BnPht1;4 transgenic Arabidopsis, the increase of Pi availability resulted in the change of root architecture under Pi deficient conditions, showing longer primary roots and lower lateral root density than that of wild type. By cis-element analysis, two P1BS and two W-box elements were found in BnPht1;4 promoter. Yeast one-hybrid assay indicated that PHR1 could bind to the BnPht1;4 promoter. P1BS elements in BnPht1;4 promoter are essential for BnPht1;4 induction in Pi starvation response. Furthermore, WRKY75 could bind to the BnPht1;4 promoter, in which W-box elements are important for this binding. These results indicated BnPht1;4 may be dually controlled by two family regulators under low Pi responses. Thus, our data on the regulative mechanism of high-affinity Pi transporter in Pi starvation response will be valuable for B. napus molecular agriculture.

The potential function of IKKα in gastric precancerous lesion via mediating Maspin.

OBJECTIVE: To know the potential role of IKKα (an NF-κB noncanonical pathway) in gastric precancerous lesion via mediating Maspin. METHODS: Gastric cancer, precancerous lesion and control tissues (chronic non-atrophic gastritis) were collected for determining the expression of IKKα and Maspin by immunohistochemistry. Thereafter, gastric precancerous models were established and divided into the Control group, Model group and Model + shIKKα group. All rats were subjected to observe the pathological changes and ultramicro structure of the gastric mucosa by HE staining or electron microscope, and to measure the serum levels of inflammatory cytokines by ELISA, the expression of apoptosis-related proteins by immunohistochemistry, as well as the expression of IKKα and Maspin by quantitative real-time PCR and Western blotting. RESULTS: Precancerous lesion and gastric cancer tissues manifested significant upregulation of IKKα positive expression, concomitant with downregulation of the positive expression of Maspin, and these changes were more evident in the gastric cancer tissues. In comparison with the Control group, rats in the Model group had significant increases in serum levels of TNF-α, IL-1ß, IL-6 and COX-2, with up-regulations of Bcl-2, CyclinD1, IKKα and p-IKKα, and down-regulations of Bax, Caspase-3 and Maspin. shIKKα treatment attenuate inflammation and apoptosis in gastric precancerous lesion (GPL) rat, with the downregulation of IKKα and p-IKKα, and upregulation of Maspin. CONCLUSION: Inhibiting IKKα, via upregulating Maspin, can mitigate the inflammation and promote cell apoptosis in precancerous rats, thereby delaying the development of the precancerous lesions.

Maspin suppresses cell invasion and migration in gastric cancer through inhibiting EMT and angiogenesis via ITGB1/FAK pathway.

This study aims to investigate how Maspin affects the EMT and angiogenesis of gastric cancer (GC) cells via ITGB1/FAK pathway. Immunohistochemistry was used to evaluate the expressions of Maspin, ITGB1, FAK, E-cadherin, Vimentin, D2-40, and CD34 in GC and adjacent normal tissues from 160 patients. Then, the human GC cells with different degree of differentiation were transfected with Maspin CRISPR activation plasmid, ITGB1 siRNA and/or Maspin siRNA, followed by the following experiments, including qRT-PCR, western blotting, tube formation assay, Transwell assay and wound healing. GC tumor tissues manifested decreased Maspin with the activated ITGB1/FAK pathway. In tumor tissues, Maspin was negatively correlated with the expressions of ITGB1 and FAK, as well as Lauren's classification, differentiation degree, and TNM stage. Besides, Maspin was negatively related with lymphatic vessel density (LVD) and microvessel density (MVD), Vimentin and VEGF, but was positive correlated with E-cadherin. Maspin expression decreased, but ITGB1 and p-FAK expressions increased gradually in MKN-28 (well differentiated), SGC-7901 (moderate differentiated), and MKN-45 (poorly differentiated). Maspin CRISPR and ITGB1 siRNA increased E-cadherin with the decreased Vimentin, VEGF and bFGF, and the reductions of tube length. In comparison with the ITGB1 siRNA group, cells in the Maspin siRNA + ITGB1 siRNA group presented the more evident EMT and angiogenesis. Furthermore, ITGB1 siRNA reduced the malignancies of GC cells, which could be restored by Maspin siRNA. Maspin was downregulated in GC tissues, which could inhibit the EMT and angiogenesis by blocking the ITGB1/FAK pathway, thereby decreasing cell invasion and migration of GC.

Development of Fluorescence In Situ Hybridization as a Rapid, Accurate Method for Detecting Coliforms in Water Samples.

Coliform bacteria are indicators of water quality; however, most detection methods for coliform bacteria are time-consuming and nonspecific. Here, we developed a fluorescence in situ hybridization (FISH) approach to detect four types of coliform bacteria, including Escherichia coli, Klebsiella pneumoniae, Enterobacter aerogenes, and Citrobacter freundii, simultaneously in water samples using specific probes for 16S rRNA. This FISH method was applied to detect coliform bacteria in simulated water and domestic wastewater samples and compared with traditional detection methods (e.g., plate counting, multiple-tube fermentation (MTF) technique, and membrane filter (MF) technique). Optimal FISH conditions for detecting the four types of coliforms were found to be fixation in 3% paraformaldehyde at 4 °C for 2 h and hybridization at 50 °C for 1.5 h. By comparing FISH with plate counting, MTF, MF, and a commercial detection kit, we found that FISH had the shortest detection time and highest accuracy for the identification of coliform bacteria in simulated water and domestic wastewater samples. Moreover, the developed method could simultaneously detect individual species and concentrations of coliform bacteria. Overall, our findings indicated that FISH could be used as a rapid, accurate biosensor system for simultaneously detecting four types of coliform bacteria to ensure water safety.

Carthamus tinctorius L. Extract ameliorates cerebral ischemia-reperfusion injury in rats by regulating matrix metalloproteinases and apoptosis.

We investigate the protective effect of Carthamus tinctorius L. (CTL, also known as Honghua in China or Safflower) on cerebral ischemia-reperfusion and explored the possible mechanisms on regulating apoptosis and matrix metalloproteinases (MMPs). High-performance liquid chromatography method with diode array detection analysis was established to analyze the components of CTL. Middle cerebral artery occlusion rats model was established to evaluate Neurological Function Score and hematoxylin-eosin staining, as well as triphenyltetrazolium was used to examine the infarction area ratio. Transferase-mediated dUTP nick-end labeling was performed for the apoptosis. Apoptosis-related factors, including B-cell lymphoma-2 (Bcl-2), Bax and Caspase3, and MMPs-related MMP2, MMP9, tissue inhibitor of metalloproteinases 1 (TIMP1) in ischemic brain, were assayed by Western blot, reverse transcription polymerase chain reaction, and immunohistochemistry. The data showed that CTL (2, 4 g crude drug/kg/d) treatment could significantly reduce the ischemic damage in brain tissue and improve a significant neurological function score. In addition, CTL could also attenuate apoptosis degree of brain tissues and regulate Bcl-2, Bax, and Caspase 3 and also have a significant decrease on MMP-9 expression, followed by a significant increase of TIMP1 protein expression. These findings indicated that regulation of CTL on apoptosis and MMPs contributed to its protective effect on ischemia/reperfusion injury.

Single-nucleotide polymorphisms of APE1 associated with risk and prognosis of vitiligo in a Han Chinese population.

BACKGROUND: The single-nucleotide polymorphisms (SNPs) of apurinic/apyrimidinicendonuclease 1 (APE1), which has been implicated in cancers and the DNA base excision repair (BER) process, have not been thoroughly investigated in association with the risks of oxidative stress-related vitiligo. OBJECTIVES: The aim of this study is to investigate associations between APE1 single-nucleotide polymorphisms 141T >G and 1349T >G and risk and prognosis of vitiligo. MATERIAL AND METHODS: From June 2013 to June 2015, a total of 460 vitiligo patients were randomly recruited as a case group; 200 of these patients received narrow bound ultraviolet B (NB-UVB) treatment. Meanwhile, 460 healthy controls were included as a control group. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was performed to explore the distribution frequencies of genotypes. RESULTS: Significant differences were detected between the case group and the control group in the frequencies of the 141T >G and 1349T >G genotypes. At 141T >G, compared with patients carrying the TG + GG genotype, male patients carrying the TT genotype aged more than 20 years with active non-segmental vitiligo, without a family history of vitiligo or other autoimmune diseases, exhibited an increased risk of vitiligo. Binary logistic regression analysis demonstrated that the TT genotype at 141T >G and the non-TT genotype at 1349T >G were independent risk factors for vitiligo development. At 1349T >G, compared with patients carrying the TT genotype, male patients carrying the TG + GG genotype aged more than 20 years with active non-segmental vitiligo, without a family history of vitiligo or other autoimmune diseases, exhibited an increased risk of vitiligo. Moreover, patients carrying 141TG + GG or 1349 TT genotypes had better photochromic effects, lower cumulative radiation doses, shorter treatment times, and earlier first photochromic times.

[GITR/GITRL expression in peripheral blood of rheumatoid arthritis].

OBJECTIVE: To explore the expression of GITR/GITRL in rheumatoid arthritis(RA) and its correlation with clinical features of the disease. METHODS: Fifty three RA patients, 35 osteoarthritis (OA) patients and 35 healthy volunteers were included in the study. The real-time-fluorescence quantitative PCR method was used to analyze the expression level of GITR/GITRL mRNA. Using fluorescence-activated cell sorting(FACS), the expression of GITR on CD4+CD25+ T cells in peripheral blood monouuclear cells (PBMC) was detected. The levels of Anti-CCP antibody, C-reactive protein(CRP), erythrocyte sedimentation rate(ESR) and rheumatoid factor (RF) of the RA patients were measured at the same time. RESULTS: It was shown that the level of GITR/GITRL mRNA and the percentage of GITR+ T cells in the CD4+CD25+ T cell subpopulation[12.38+/-5.72/16.41+/-10.16,(28.12+/-16.85)%] were significantly higher than those of the OA group [9.59+/-5.87/12.09+/-7.53,(21.01+/-14.42)%,P<0.05] and control group [8.75+/-3.78/11.99+/-8.42,(19.98+/- 9.40)%,P<0.05]. The expression of GITR/GITRL from RA patients was positively associated with the level of DAS28 and ESR (r=0.361,r=0.427,P<0.01). CONCLUSION: The GITR/GITRL expression of RA patients was higher than that of other groups which may play an important role in the development of RA.

Comparison of serum human epididymis protein 4 and carbohydrate antigen 125 as markers in ovarian cancer: A meta-analysis.

Ovarian cancer (OC) is the third most common type of gynecological cancer. Measurements of human epididymis protein 4 (HE4) levels have been suggested for improving the specificity of the laboratory identification of OC. For this meta-analysis, the Medline, Embase and Cochrane databases were searched to identify relevant studies. All the included studies for diagnostic performance were combined with sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnostic odds ratios (DORs) with 95% confidence intervals (CIs), summary receiver operating characteristic (SROC) curves and areas under the SROC curves (AUC). A total of 25 studies including 4,729 patients were identified as eligible for inclusion in the final analysis. The pooled sensitivities and respective 95% CIs for HE4 and carbohydrate antigen 125 (CA125) were 0.74 (0.72-0.76) and 0.74 (0.72-0.76), respectively. The pooled specificities and respective 95% CIs for HE4 and CA125 were 0.90 (0.89-0.91) and 0.83 (0.81-0.84), respectively. The summary DORs and 95% CIs for HE4 and CA125 were 43.35 (29.13-64.51) and 17.06 (10.97-26.51), respectively and the AUCs for HE4 and CA125 were 0.8915 and 0.8538, respectively. In total, 9 studies investigated the diagnostic accuracy of HE4 combined with CA125 for the diagnosis of OC. The pooled sensitivity and 95% CIs of HE4, CA125 and HE4+CA125 in this subgroup were 0.71 (0.67-0.75), 0.74 (0.69-0.78) and 0.90 (0.87-0.92), respectively; the pooled specificity and 95% CIs of HE4, CA125 and HE4+CA125 were 0.92 (0.90-0.94), 0.73 (0.69-0.76) and 0.85 (0.82-0.87), respectively. The diagnostic accuracy of HE4 in distinguishing OC from other benign gynecological diseases was found to be to be superior to that of CA125 and the combination of HE4 and CA125 may enhance the diagnostic sensitivity.

Brassica napus PHR1 gene encoding a MYB-like protein functions in response to phosphate starvation.

Phosphorus (P) is one of the essential nutrient elements for plant development. In this work, BnPHR1 encoding a MYB transcription activator was isolated from Brassica napus. The characterization of nuclear localization and transcription activation ability suggest BnPHR1 is a transcriptional activator. The tissue expression and histochemical assay showed that BnPHR1 was predominantly expressed in roots and modulated by exogenous Pi in transcriptional level in roots under Pi deficiency conditions. Furthermore, overexpression of BnPHR1 in both Arabidopsis and B. napus remarkably enhanced the expression of the Pi-starvation-induced genes including ATPT2 and BnPT2 encoding the high-affinity Pi transporter. Additionally, BnPHR1 can in vivo bind the promoter sequence of ATPT2 and BnPT2 in both Arabidopsis and B. napus. Possibly, due to the activation of ATPT2 and BnPT2, or even more high-affinity Pi transporters, the excessive Pi was accumulated in transgenic plants, resulting in the crucially Pi toxicity to cells and subsequently retarding plant growth. Given the data together, BnPHR1, as crucial regulator, is regulated by exogenous Pi and directly activates those genes, which promote the uptake and homeostasis of Pi for plant growth.