[Effect of Zuogui Jiangtang Qinggan Formula on lipid metabolism in db/db mouse model of type 2 diabetes mellitus complicated with non-alcoholic fatty liver disease].

This study aims to explore the effect of Zuogui Jiangtang Qinggan Formula(ZGJTQGF) on the lipid metabolism in the db/db mouse model of type 2 diabetes mellitus(T2DM) complicated with non-alcoholic fatty liver disease(NAFLD) via the insulin receptor(INSR)/adenosine 5'-monophosphate(AMP)-activated protein kinase(AMPK)/sterol-regulatory element-binding protein 2(SREBP-2) signaling pathway. Twenty-four db/db mice were randomized into positive drug(metformin, 0.067 g·kg~(-1)) and low-(7.5 g·kg~(-1)) and high-dose(15 g·kg~(-1)) ZGJTQGF groups. Six C57 mice were used as the blank group and administrated with an equal volume of distilled water. The mice in other groups except the blank group were administrated with corresponding drugs by gavage for 6 consecutive weeks. At the end of drug administration, fasting blood glucose(FBG) and blood lipid levels were measured, and oral glucose tolerance test was performed. Compared with the blank group, the mice treated with ZGJTQGF showed decreased body mass and liver weight coefficient, lowered levels of FBG, total cholesterol(TC), triglyceride(TG), and low-density lipoprotein(LDL), and weakened liver function. The pathological changes and lipid accumulation in the liver tissue were examined. Western blot was employed to measure the protein levels of INSR, AMPK, p-AMPK, and SREBP-2. Compared with the blank group, the model group showed down-regulated protein levels of INSR and p-AMPK/AMPK and up-regulated protein level of SREBP-2. Compared with the model group, high-dose ZGJTQGF up-regulated the protein levels of INSR and p-AMPK/AMPK and down-regulated the protein level of SREBP-2. Low-dose ZGJTQGF slightly up-regulated the protein levels of INSR and p-AMPK/AMPK and down-regulated the protein level of SREBP-2, without significant differences. The results suggested that ZGJTQGF may alleviate insulin resistance and improve lipid metabolism in db/db mice by activating the INSR/AMPK/SREBP-2 signaling pathway.

Mitochondrial dynamics modulate the allergic inflammation in a murine model of allergic rhinitis.

OBJECTIVE: Allergic rhinitis (AR) is a common allergic disorder, afflicting thousands of human beings. Aberrant mitochondrial dynamics are important pathological elements for various immune cell dysfunctions and allergic diseases. However, the connection between mitochondrial dynamics and AR remains poorly understood. This study aimed to determine whether mitochondrial dynamics influence the inflammatory response in AR. METHODS: In the present study, we established a murine model of AR by sensitization with ovalbumin (OVA). Then, we investigated the mitochondrial morphology in mice with AR by transmission electron microscopy and confocal fluorescence microscopy, and evaluated the role of Mdivi-1 (an inhibitor of mitochondrial fission) on allergic symptoms, inflammatory responses, allergic-related signals, and reactive oxygen species formation. RESULTS: There was a notable enhancement in mitochondrial fragmentation in the nasal mucosa of mice following OVA stimulation, whereas Mdivi-1 prevented aberrant mitochondrial morphology. Indeed, Mdivi-1 alleviated the rubbing and sneezing responses in OVA-sensitized mice. Compared with vehicle-treated ones, mice treated with Mdivi-1 exhibited a reduction in interleukin (IL)-4, IL-5, and specific IgE levels in both serum and nasal lavage fluid, and shown an amelioration in inflammatory response of nasal mucosa. Meanwhile, Mdivi-1 treatment was associated with a suppression in JAK2 and STAT6 activation and reactive oxygen species generation, which act as important signaling for allergic response. CONCLUSION: Our findings reveal mitochondrial dynamics modulate the allergic responses in AR. Mitochondrial dynamics may represent a promising target for the treatment of AR.

[Zuogui Jiangtang Qinggan Prescription promotes recovery of intestinal mucosal barrier in mice with type 2 diabetes mellitus and nonalcoholic fatty liver disease by improving intestinal flora homeostasis].

This study aimed to investigate the recovery effect of Zuogui Jiangtang Qinggan Prescription on intestinal flora homeostasis control and intestinal mucosal barrier in type 2 diabetes mellitus(T2DM) with nonalcoholic fatty liver disease(NAFLD) induced by a high-fat diet. NAFLD was established in MKR transgenic mice(T2DM mice) by a high-fat diet(HFD), and subsequently treated for 8 weeks with Zuogui Jiangtang Qinggan Prescription(7.5, 15 g·kg~(-1)) and metformin(0.067 g·kg~(-1)). Triglyceride and liver function were assessed using serum. The hematoxylin-eosin(HE) staining and Masson staining were used to stain the liver tissue, while HE staining and AB-PAS staining were used to stain the intestine tissue. 16S rRNA sequencing was utilized to track the changes in the intestinal flora of the mice in each group. Polymerase chain reaction(PCR) and immunofluorescence were used to determine the protein and mRNA expression levels of ZO-1, Occludin, and Claudin-1. The results demonstrated that Zuogui Jiangtang Qinggan Prescription increased the body mass of T2DM mice with NAFLD and decreased the hepatic index. It down-regulated the serum biomarkers of liver function and dyslipidemia such as alanine aminotransferase(ALT), aspartate transaminase(AST), and triglycerides(TG), increased insulin sensitivity, and improved glucose tolerance. According to the results of 16S rRNA sequencing, the Zuogui Jiangtang Qinggan Prescription altered the composition and abundance of the intestinal flora, increasing the relative abundances of Muribaculaceae, Lactobacillaceae, Lactobacillus, Akkermansia, and Bacteroidota and decreasing the relative abundances of Lachnospiraceae, Firmicutes, Deslfobacteria, Proteobacteria, and Desulfovibrionaceae. According to the pathological examination of the intestinal mucosa, Zuogui Jiangtang Qinggan Prescritpion increased the expression levels of the tight junction proteins ZO-1, Occludin, and Claudin-1, promoted intestinal mucosa repair, protected intestinal villi, and increased the height of intestinal mucosa villi and the number of goblet cells. By enhancing intestinal mucosal barrier repair and controlling intestinal microbiota homeostasis, Zuogui Jiangtang Qinggan Prescription reduces intestinal mucosal damage induced by T2DM and NAFLD.

Acremoniumcapsici and A.guizhouense, two new members of Acremonium (Hypocreales, Sordariomycetes) isolated from the rhizosphere soil of Capsicumannuum.

Two new species, Acremoniumcapsici and A.guizhouense, isolated from the rhizosphere soil of Capsicumannuum, are described and illustrated. Two-locus DNA sequences based on phylogeny, in combination with the morphology of the asexual morph, were used to characterize these species. In the phylogenetic tree, both new species clustered into a monophyletic clade with strong support, distinct from other previously known species of Acremonium. The new species differed from their allied species in their morphology.

Obesity indicators as mediators of the association between age at menopause and blood pressure values.

Having a later age at menopause is associated with having a higher blood pressure (BP) value, but the mediation pathways remain unclear. We quantitatively examined the mediation effects of various obesity indicators using baseline data from phase 4 of the Guangzhou Biobank Cohort Study. The product of coefficients approach and bootstrapping procedures were used to assess the mediation effects of body mass index (BMI), waist circumference (WC), waist-to-hip ratio (WHR) and waist-to-height ratio (WHtR) on the association between age at menopause and BP values. Age, education, occupation, family income, smoking, drinking, diet, physical activity, age at menarche, number of births, fasting glucose, triglycerides, and high-density lipoprotein cholesterol were adjusted as covariates. Of 5429 women with natural menopause, the mean age and menopausal age were 60.0 (standard deviation = 5.8) and 50.3 (3.1) years, respectively. The prevalence of hypertension was 29.6%. In women with a menopausal age of ≥50 years, BMI, WC, WHR and WHtR showed significant mediation effects on the positive association between menopausal age and BP. The adjusted proportion (95% confidence interval) of the mediation effects for those variables were 26.04% (10.40-116.82%), 25.92% (10.19-108.57%), 14.11% (3.59-62.78%), and 23.17% (8.70-95.81%), respectively, for systolic BP values and 22.59% (10.72-53.60%), 20.67% (9.83-49.31%), 9.21% (2.73-23.92%), and 17.19% (7.56-41.31%) for diastolic BP values. In women with a menopausal age of <50 years, no significant association between age at menopause and systolic/diastolic BP values was found. In conclusion, obesity indicators showed significant mediating effects on the association between having a later age at menopause and having a higher BP value. Further longitudinal studies with detailed and accurate measurements of metabolic changes after menopause and sufficient follow-up are warranted to confirm these results. We demonstrated obesity indicators showed significant mediating effects on the association between later age at menopause (≥50 years) and higher BP.

Risk assessment of malignancy in solitary pulmonary nodules in lung computed tomography: a multivariable predictive model study.

BACKGROUND: Computed tomography images are easy to misjudge because of their complexity, especially images of solitary pulmonary nodules, of which diagnosis as benign or malignant is extremely important in lung cancer treatment. Therefore, there is an urgent need for a more effective strategy in lung cancer diagnosis. In our study, we aimed to externally validate and revise the Mayo model, and a new model was established. METHODS: A total of 1450 patients from three centers with solitary pulmonary nodules who underwent surgery were included in the study and were divided into training, internal validation, and external validation sets (n = 849, 365, and 236, respectively). External verification and recalibration of the Mayo model and establishment of new logistic regression model were performed on the training set. Overall performance of each model was evaluated using area under receiver operating characteristic curve (AUC). Finally, the model validation was completed on the validation data set. RESULTS: The AUC of the Mayo model on the training set was 0.653 (95% confidence interval [CI]: 0.613-0.694). After re-estimation of the coefficients of all covariates included in the original Mayo model, the revised Mayo model achieved an AUC of 0.671 (95% CI: 0.635-0.706). We then developed a new model that achieved a higher AUC of 0.891 (95% CI: 0.865-0.917). It had an AUC of 0.888 (95% CI: 0.842-0.934) on the internal validation set, which was significantly higher than that of the revised Mayo model (AUC: 0.577, 95% CI: 0.509-0.646) and the Mayo model (AUC: 0.609, 95% CI, 0.544-0.675) (P < 0.001). The AUC of the new model was 0.876 (95% CI: 0.831-0.920) on the external verification set, which was higher than the corresponding value of the Mayo model (AUC: 0.705, 95% CI: 0.639-0.772) and revised Mayo model (AUC: 0.706, 95% CI: 0.640-0.772) (P < 0.001). Then the prediction model was presented as a nomogram, which is easier to generalize. CONCLUSIONS: After external verification and recalibration of the Mayo model, the results show that they are not suitable for the prediction of malignant pulmonary nodules in the Chinese population. Therefore, a new model was established by a backward stepwise process. The new model was constructed to rapidly discriminate benign from malignant pulmonary nodules, which could achieve accurate diagnosis of potential patients with lung cancer.

A novel fluorescent nanoprobe that based on poly(thymine) single strand DNA-templated copper nanocluster for the detection of hydrogen peroxide.

In this paper, a label-free fluorescence nanoprobe is constructed based on poly(thymine) single strand DNA-templated Copper nanocluster (denote as: T-CuNCs) for the detection of hydrogen peroxide. In the assay, the fluorescent T-CuNCs will generate though the reaction of Cu2+, poly(thymine) and sodium ascorbate. However, the hydroxyl radical (.OH) will generated in the presence of H2O2, which is able to induced the oxidative lesions of poly(thymine) single chain DNA and lead to the poly(thymine) being splitted into shorter or single oligonucleotide fragments and lose the ability to template the fluorescent T-CuNCs again. Therefore, H2O2 can be detected by monitoring the fluorescence strength change of T-CuNCs. The experimental results show that the fluorescence intensity change of T-CuNCs has fantastic linearity versus H2O2 concentration in the range of 1-30 µM (R2 = 0.9947) and 30-80 µM (R2 = 0.9972) with the limit of detection (LOD) as low as 0.5 µM (S/N = 3). More important, the fluorescent nanoprobe was also successfully utilized on the detection of H2O2 in serum samples. Therefore, a label-free, costless and effective fluorescence method has been established for the detection of H2O2, the intrinsic properties of the nanoprobe endow its more potential applications in chemical and biological study.

[Application of infrared thermal imaging technology in the design of free anterolateral thigh perforator flap transplantation].

OBJECTIVE: To explore clinical effect of infrared thermal imaging technology for the treatment of free anterolateral thigh perforator flap transplantation. METHODS: From June 2014 to June 2018, 31 patients with skin defect at various degrees treated by free anterolateral thigh perforator flap transplantation, including 21 males and 10 females aged from 16 to 59 years old with an average age of(35.3±1.5) years old, the courses of disease ranged from 2 to 4 weeks with an average of (1.8±0.6) weeks. The number of perforating branch, the position of the perforating branch, the perforating branch vitality detected by Doppler blood stream detector and parameters of thermal imaging image in order to guide design of skin flap, and compared results with the data of perforator arteries observed during the operation. RESULTS: Totally 52 branches of perforating arteries were detected by Doppler blood stream detector during operation, and 38 perforator branches were confirmed in operation, the accuracy rate was 73.1%. Thirty-eight branches of perforating arteries were detected by infrared thermography during operation, and 35 branches of perforating branches were confirmed in operation, the accuracy rate was 92.1%; there were statistical difference. The most dynamic perforating pivot found by Doppler blood stream detector was confirmed by intraoperative diagnosis, with an accuracy rate of 80.6%. The most dynamic perforating pivot found by infrared thermography is confirmed by intraoperative diagnosis, with an accuracy rate of 100%; there were statistical difference. Thirty-one flaps were survived without vascular crisis occurred. All patients were followed up from 6 to 18 months with an average of(10.7±1.2) months. The flaps survived with soft texture and good blood circulation, the defect was not bloated, the color of skin flap was basically the same as that of the normal skin, and the limbs appearance and function recovered well. CONCLUSIONS: Infrared infrared thermal imaging technology could be used as a new technology in localization of perforator artery in free anterolateral thigh perforator flap transplantation.

The effects of melamine on the formation of carbon xerogel derived from resorcinol and formaldehyde and its performance for supercapacitor.

Carbon aerogel/xerogel can be easily tuned to have hierarchical pores ranging from micropores to macropores. Nitrogen doping is considered to enhance the wettability and conductivity of the carbon electrode, hence improve the electrochemical performance. To prepare N-doped carbon xerogel and study the effects on the structure and the electrochemical performance of resorcinol and formaldehyde derived carbon xerogel, a series of Resorcinol-Melamine-Formaldehyde derived N-doped carbon xerogel were prepared by a facile sol-gel process and ambient drying method. With the increasing amount of melamine, the inside channels become larger, which contributes to faster ion transport and smaller charge transfer resistance (Rct). The Nitrogen content in carbon xerogel is also increased, enhancing the capacitance of carbon electrode by pseudocapacity effect, while damaging the rate performance by introducing more defects and larger degree of disorder. As a result, the best electrochemical performance is achieved in the RM-6-4 sample (resorcinol:melamine = 6:4), showing the largest capacitance of 139 F g-1.

Magnetic immunoassay using CdSe/ZnS quantum dots as fluorescent probes to detect the level of DNA methyltransferase 1 in human serum sample.

BACKGROUND: DNA methyltransferase 1 (DNMT1), a dominant enzyme responsible for the transfer of a methyl group from the universal methyl donor to the 5-position of cytosine residues in DNA, is essential for mammalian development and closely related to cancer and a variety of age-related chronic diseases. DNMT1 has become a useful biomarker in early disease diagnosis and a potential therapeutic target in cancer therapy and drug development. However, till now, most of the studies on DNA methyltransferase (MTase) detection have focused on the prokaryote MTase and its activity. METHODS: A magnetic fluorescence-linked immunosorbent assay (FLISA) using CdSe/ZnS quantum dots as fluorescent probes was proposed for the rapid and sensitive detection of the DNMT1 level in this study. Key factors that affect the precision and accuracy of the determination of DNMT1 were optimized. RESULTS: Under the optimal conditions, the limit of detection was 0.1 ng/mL, the linear range was 0.1-1,500 ng/mL, the recovery was 91.67%-106.50%, and the relative standard deviations of intra- and inter-assays were respectively 5.45%-11.29% and 7.03%-11.25%. The cross-reactivity rates with DNA methyltransferases 3a and 3b were only 4.0% and 9.4%, respectively. Furthermore, FLISA was successfully used to detect the levels of DNMT1 in human serum samples, and compared with commercial enzyme-linked immunosorbent assay (ELISA) kits. The results revealed that there was a good correlation between FLISA and commercial ELISA kits (correlation coefficient r=0.866, p=0.001). The linear scope of FLISA was broader than ELISA, and the measurement time was much shorter than ELISA kits. CONCLUSION: These indicated that the proposed FLISA method was sensitive and high throughput and can quickly screen the level of DNMT1 in serum samples.

MiR-155 up-regulated by TGF-ß promotes epithelial-mesenchymal transition, invasion and metastasis of human hepatocellular carcinoma cells in vitro.

It has previously been reported that microRNA (miR)-155 is linked to the recurrence and prognosis of hepatocellular carcinoma (HCC) following liver transplantation. However, the role of miR-155 in the invasion and metastasis of HCC cells remains largely unclear. The aim of this study was to investigate the expression of miR-155 in HCC cells and its role in the invasion and migration of HCC cells in vitro. We found that the level of expression of miR-155 in HCC tissues and cells was significantly increased compared with non-tumorous adjacent tissues. Further study revealed that recombinant human transforming growth factor-ß (TGF-ß1) up-regulated the expression of miR-155 in HCC cells in vitro. Further, the overexpression of miR-155 in HCC cell line Huh-7 led to increased levels of cell invasion and migration compared with untreated control Huh-7 cells. MiR-155-overexpressed Huh-7 cells also exhibited altered levels of expression of certain cellular adhesion molecules related to epithelial-mesenchymal transition (EMT), including low levels of CDH1 and higher levels of FN1, SNAI1 and ZEB1, compared with control Huh-7 cells. Moreover, it was found that the overexpression of miR-155 and of TGF-ß1 protein decreased the expression of E-Cadherin and increased the expression of Vimentin in Huh-7 cells. These results indicate that an increased level of miR-155 in HCC cells, possibly due to stimulation by TGF-ß1, accelerates the process of EMT, promotes cellular invasion and migration in vitro, and thereby further promotes the progression of HCC.

[Research on the Combined Detection of Urine UmAlb and Urinary Nephrin, Podocalyxin in Podocyte of MKR Mice with Diabetic Nephropathy].

OBJECTIVE: To explore the combined detection of urine UmAlb and urinary nephrin (Unephrin), podocalyxin (UPCX) in podocyte of MKR mice with diabetic nephropathy. METHODS: Thirty 8 weeks old MKR mice were randomly divided into two groups as follows: negative control group, DN model group, and another 15 wild C57 mice were used as normal control. Mice in DN model group were received unilateral nephrectomy and high-fat diet feed for 2 months. The morphological structure changes of the podocytes were observed by transmission electron microscopes. The levels of FBG were detected by electrochemical detection method, The nephrin and PCX protein expression were measured by western blotting. The levels of UmAlb, Unephrin and UPCX were detected by ELISA. RESULTS: The podocyte damage in the mice of DN model group increased significantly when compared with normal control. As compared with normal control, FBG in the model group increased significantly (P<0. 01), the expression level of nephrin and PCX in Renal Tissue and Unephrin, UPCX, and urine UmAlb were also increased significantly (P<0. 01). CONCLUSION: The level of Unephrin and UPCX were positive correlated with the level of urine UmAlb, the loss of podocyte strcture protein might be one of the mechanism in leading proteinuria in diabetic nephropathy.

Macrophages promote coal tar pitch extract-induced tumorigenesis of BEAS-2B cells and tumor metastasis in nude mice mediated by AP-1.

BACKGROUND: We sought to evaluate the role of tumor associated macrophages (TAMs) on the promotion of coal tar pitch extract (CTPE)-induced tumorigenesis of human bronchial epithelial cells (BEAS-2B) and tumor metastasis in nude mice, and related mechanisms. MATERIALS AND METHODS: BEAS-2B cells were first treated with 2.4 mg/mL CTPE for 72 hours. After removal of CTPE, the cells were continuously cultured and passaged using trypsin-EDTA. THP-1 cells were used as macrophage-like cells. BEAS-2B cells under different conditions (n=6/ group) were injected into the back necks of nude mice, and alterations of tumor xenograft growth, indicative of tumorigenicity, and tumor metastasis were determined. Pathological changes (tumor nests and microvascular lesions) of HE-stained tumor tissues were also evaluated. The expression of AP-1(c-Jun) in xenografts and metastatic tumors was determined using immunohistochemistry. RESULTS: Tumor size and weight in nude mice transplanted with the mixture of CTPE-induced passage 30 BEAS-2B and THP-1 cells (2:1) were increased compared to those from the CTPE-treated BEAS-2B cells at passage 30 alone at different observation time points. Tumor metastasis to lymph nodes and liver was only detected after transplantation of a mixture the two kinds of cells. The numbers of tumor nests and microvascular lesions, and the expression levels of AP-1 (c-Jun) in tumors from the mixture of two kinds of cells were increased apparently in contrast to those in tumor from the CTPE-treated BEAS-2B cells of passage 30 alone. In addition, there was positive correlation between AP-1 (c-Jun) expression level and the number of microvascular lesions, or between AP-1 (c-Jun) expression level and tumor metastasis in these two groups. CONCLUSIONS: TAMs not only facilitate tumorigenesis transformation of CTPE-induced BEAS-2B cells, but also promote tumor growth, angiogenesis and metastasis in nude mice in vivo, which may be mediated by AP-1.

[Effect of Jiangang Yishen Recipe on high insulin induced cell proliferation of human glomerular mesangial cells and the expression of insulin receptor substrate 1 and phosphatidylinositol-3-kinase].

OBJECTIVE: To investigate the effect of Jiangtang Yishen Recipe (JTYSR) on high insulin induced cell proliferation of human glomerular mesangial cells (HMCs) and the expression of insulin receptor substrate 1 (IRS-1) and phosphatidylinositol-3-kinase (PI-3K). METHODS: HMCs were divided into 4 groups, i.e., the negative control group, the high insulin model group, the JTYSR group, and the LY294002 group. The concentration of insulin, JTYSR, and LY294002 was respectively confirmed by pre-experiment. Different culture solution was respectively added for different groups. RPMI1640 culture solution was added for HMCs in the negative control group, while HMCs in the rest 3 groups were cultured by 100 nmol/L insulin for 24 h. Meanwhile, HMCs from the JTYSR group and the LY294002 group were exposed to 125 mg/L JTYSR and 80 micromol/L LY294002 respectively for further 48 h. The proliferation of HMCs was detected by MTT and flow cytometry. The protein expression of IRS-1 and PI-3K in HMC was detected by immunohistochemical assay and Western blot. Results The proliferation of HMCs induced by high insulin could be significantly lowered, and the protein expression of IRS-1 and PI-3K could be down-regulated in the JTYSR group and the LY294002 group (P <0.01). Compared with the LY294002 group, the protein expression of IRS-1 and PI-3K could be slightly down-regulated in the JTYSR group (P <0.05). CONCLUSION: JTYSR could lower high insulin induced proliferation of HMCs, and its mechanism might be related to insulin signaling pathway.

Identification of differentially expressed genes that potentially confer pest resistance in transgenic ChIFN-γ tobacco.

Chicken interferon-γ (ChIFN-γ) is both an inhibitor of viral replication and a regulator of numerous immunological functions. However, since little is known about the mechanisms underlying the insect-resistance of transgenic ChIFN-γ, a transgenic ChIFN-γ tobacco line was employed in the present study to explore this mechanism. A cDNA microarray (with 43,760 unigenes) was used to analyze the gene expression profiles of transgenic and wild-type (WT) tobacco leaves at two different growth stages. Compared with the WT, 1529 and 405 expressed sequence tags were significantly up- or downregulated on days 119 and 147, respectively. The differentially expressed genes (DEGs) are involved in metabolic regulation, cell division and differentiation, material synthesis and transport, signal transduction, and protein synthesis and degradation. Candidate genes that may increase cell density, thicken cell walls, promote secondary metabolite synthesis, and mediate plant hormone-induced resistance responses were used to identify the ChIFN-γ-mediated insect-resistance mechanisms. The insect-resistance of transgenic ChIFN-γ tobacco possibly involves unknown signaling pathways, which may directly or indirectly affect DEG expression-mediating genes. The degree of pest resistance increased as the plants grew. Three genes likely to be related to jasmonic acid- or salicylic acid-dependent plant defense responses, including CAF 1, Cop 8/CSN, and HD, are implicated in the insect-resistance of the transgenic plants. The mechanism of transgenic ChIFN-γ tobacco resistance also involves RPS20 and other genes that induce microRNA-based gene regulation. The ChIFN-γ-mediated DGEs contribute to insect-resistance in transgenic ChIFN-γ tobacco, which provides new insight into the role of ChIFN-γ.

Transcriptional and posttranscriptional regulation and endocytosis were involved in zinc oxide nanoparticle-induced interleukin-8 overexpression in human bronchial epithelial cells.

Inhaled zinc oxide nanoparticles (ZnO-NPs) can induce lung inflammation through released inflammatory mediators, such as interleukin 8 (IL-8), from airways. However, the mechanisms underlying ZnO-NP-induced IL-8 gene expression have not been fully characterized. The transcription inhibitor actinomycin D (Act D) and the BEAS-2B cells stably overexpressing wild-type or mutated IL-8 promoter at the NFκB or C/EBPß binding site were used to determine the involvement of transcriptional mechanisms. The effect of ZnO-NPs on IL-8 mRNA stability was examined using mRNA decay assay. The phagocytosis inhibitor cytochalasin B (CB) was utilized to define the role of endocytosis in ZnO-NP-induced IL-8 expression. In addition, the solubility of ZnO-NPs in culture medium was assessed using atomic absorption spectroscopy. Exposure to ZnO-NPs significantly increased the expression of IL-8 mRNA and protein in a dose-dependent manner. Pretreatment with Act D blocked ZnO-NP-induced IL-8 expression. Both NFκB and C/EBPß transcription factors were required for ZnO-NP-induced IL-8 transcription. mRNA decay assay showed that ZnO-NP stimulation delayed IL-8 mRNA degradation in BEAS-2B cells. Pretreatment of BEAS-2B cells with CB blocked ZnO-NP-induced IL-8 expression by 30 %. Exposure to ZnO-NPs induced IL-8 gene expression through transcriptional activation and mRNA stabilization. Internalization of nanoparticles was partially involved in ZnO-NP-induced IL-8 expression.

Expression and significance of Rac1, Pak1 and Rock1 in gastric carcinoma.

AIMS: Rac1, Pak1 and Rock1 are indicators related to gastric cancer invasion and metastasis, but few reports discuss all three kinds of protein in research on gastric cancer invasion and metastasis. The aim of this study was to investigate the expression and clinical significance of Rac1, Pak1 and Rock1 in gastric carcinoma. METHODS: Rac1, Pak1 and Rock1 expression in 158 cases of gastric carcinoma were investigated via immunohistochemical staining and clinical analysis. RESULTS: The positive expression rates of Rac1, Pak1 and Rock1 in normal tissue, intraepithelial neoplastic tissues and gastric carcinoma showed an increasing trend (P < 0.05). Their expression in lymph node metastasis was significantly higher than in patients with lymph-node metastasis than in those without lymph nodes metastasis (P < 0.05). Their expression in tumor (TNM stages III and IV) were significantly higher than that in stages I and II (P < 0.05). Rac1, Pak1 and Rock1 expression did not differ significantly with patients' sex (P > 0.05). CONCLUSION: Positive rates of Rac1, Pak1 and Rock1 expression in normal tissue, dysplasia and gastric carcinoma show an increasing trend and are correlated with tumor lymph node metastasis and TNM stage. Rac1, Pak1 and Rock1 may be important biomarkers of gastric carcinoma invasion and metastasis.

[Changes of centrosome and related protein in malignant transformation of BEAS-2B cell induced by coal tar pitch smoke extracts].

OBJECTIVE: To analyze the centrosome abnormalities in the malignant transformation of human bronchial epithelial cells (BEAS-2B) induced by coal tar pitch smoke extracts and to investigate the role and action mechanism of centrosome in the lung cancer induced by coal tar pitch. METHODS: Medium-temperature coal tar pitch smoke extracts were used to treat immortalized human bronchial epithelial cells (BEAS-2B) and establish a malignant transformation model. The treated BEAS-2B cells were used as exposure group, and solvent control group and normal control group were also set for passage culture. The changes of centrosome in BEAS-2B cells seeded on coverslips were evaluated by indirect immunofluorescence assay. The mRNA expression of p53, p21, and cyclin E in BEAS-2B cells was measured by real-time quantitative RT-PCR, and their protein levels in BEAS-2B cells seeded on coverslips were measured by semiquantitative immunohistochemical analysis. RESULTS: The overall rate of centrosome abnormalities in BEAS-2B cells at passage 20 was 6.56±1.01% in the exposure group, significantly higher than those in the normal control group (3.40±0.86%) and solvent control group (3.14±0.59%) (P < 0.05). In addition, the exposure group had a significantly higher overall rate of centrosome abnormalities in BEAS-2B cells at passage 30 compared with the normal control group and solvent control group (22.39±9.5% vs 4.34±1.04%, P < 0.05; 22.39±9.5% vs 4.33±1.20%, P < 0.05). Compared with the normal control group and solvent control group, the exposure group had significantly decreased mRNA and protein expression of p53 and significantly increased mRNA and protein expression of cyclin E in BEAS-2B cells at passages 20 and 30 (P < 0.05). CONCLUSION: Centrosome abnormalities occur before the malignant transformation in BEAS-2B cells treated with coal tar pitch smoke extracts, and they may be mediated by the p53/p21/cyclin E signaling pathway.

[Expression and significance of DNA methyltransferase in sera of patients with lung cancer].

OBJECTIVE: To explore the protein expressions of DNMT1, DNMT3a and DNMT3b in sera of lung cancer patients. METHODS: Enzyme-linked immunosorbent assay (ELISA) method was used to measure the protein expressions of DNMT1, DNMT3a and DNMT3b in 136 lung cancer patients hospitalized at Department of Respiratory Diseases, First Affiliated Hospital, Zhengzhou University during September 2012 to June 2013. And 147 healthy controls were selected from a population of physical examination at Sixth People's Hospital of Zhengzhou. And the relationship was analyzed between protein expressions of DNMT1, DNMT3a and DNMT3b and clinic characteristics of lung cancer. RESULTS: The protein expressions of DNMT1, DNMT3a and DNMT3b in patients with lung cancer (15 ± 10, 997 ± 76 , 302 ± 25) were higher than those of the controls (13 ± 10, 344 ± 93, 108 ± 22). And there were statistical significance (t = 3.28, 62.51, 37.27; P = 0.021, 0.000, 0.000). The results of Logistic regression show that the protein expressions DNMT1, DNMT3a and DNMT3b increased morbidity for lung cancer (χ(2) = 14.811, 26.768, 12.057; P = 0.000, 0.000 0.001), especially so for DNMT1 (OR = 1.545, 95%CI: 1.238-1.928). No correlation existed between the protein expressions of DNMT1, DNMT3a, DNMT3b and histological types or stages (P > 0.05). CONCLUSIONS: The high protein expressions of serum DNMT1, DNMT3a and DNMT3b increase morbidity for lung cancer. And these markers may predict the early occurrence of lung cancer.