The protective effect of intranasal immunization with influenza virus recombinant adenovirus vaccine on mucosal and systemic immune response.

Influenza virus is a kind of virus that poses several hazards of animal and human health. Therefore, it is important to develop an effective vaccine to prevent influenza. To this end we successfully packaged recombinant adenovirus rAd-NP-M2e-GFP expressing multiple copies of influenza virus conserved antigens NP and M2e and packaged empty vector adenovirus rAd-GFP. The effect of rAd-NP-M2e-GFP on the activation of dendritic cell (DC) in vitro and in vivo was detected by intranasal immunization. The results showed that rAd-NP-M2e-GFP promoted the activation of DC in vitro and in vivo. After the primary immunization and booster immunization of mice through the nasal immune way, the results showed that rAd-NP-M2e-GFP induced enhanced local mucosal-specific T cell responses, increased the content of SIgA in broncho alveolar lavage fluids (BALF) and triggered the differentiation of B cells in the germinal center. It is proved that rAd-NP-M2e-GFP can significantly elicit mucosal immunity and systemic immune response. In addition, rAd-NP-M2e-GFP could effectively protect mice after H1N1 influenza virus challenge. To lay the foundation and provide reference for further development of influenza virus mucosal vaccine in the future.

MYCT1 attenuates renal fibrosis and tubular injury in diabetic kidney disease.

Tubulointerstitial abnormalities contribute to the progression of diabetic kidney disease (DKD). However, the underlying mechanism of the pathobiology of tubulointerstitial disease is largely unknown. Here, we showed that MYCT1 expression was downregulated in in vitro and in vivo DKD models. Adeno-associated virus (AAV)-Myct1 significantly attenuated renal dysfunction and tubulointerstitial fibrosis in diabetic db/db mice and downregulated Sp1 transcription and TGF-ß1/SMAD3 pathway activation. In human proximal tubular epithelial cells, high glucose-induced high expression of SP1 and TGF-ß1/SMAD3 pathway activation as well as overaccumulation of extracellular matrix (ECM) were abrogated by MYCT1 overexpression. Mechanistically, the binding of VDR to the MYCT1 promoter was predicted and confirmed using dual-luciferase reporter and ChIP analysis. VDR transcriptionally upregulates MYCT1. Our data reveal MYCT1 as a new and potential therapeutic target in treating DKD.

Treatment of a patient with severe lactic acidosis and multiple organ failure due to mitochondrial myopathy: A case report.

BACKGROUND: Mitochondrial myopathy is a rare genetic disease with maternal inheritance that may involve multiple organ systems. Due to the lack of typical characteristics, its clinical diagnosis is difficult, and it is often misdiagnosed or even missed. CASE SUMMARY: The patient was a young college student. When he presented at the hospital, he had severe lactic acidosis, respiratory failure, and shock with multiple organ dysfunction syndrome (MODS). He was treated by mechanical ventilation, veno-arterial extracorporeal membrane oxygenation, and other organ support. However, his condition continued to worsen. After a thorough and detailed medical and family history was taken, a mitochondrial crisis was suspected. A muscle biopsy was taken. Further genetic testing confirmed a mitochondrial gene mutation (TRNL1 3243A>G). The final diagnosis of mitochondrial myopathy was made. Although there is no known specific treatment, intravenous methylprednisone and intravenous immunoglobulin were started. The patient's shock eventually improved. The further course was complicated by severe infection in multiple sites, severe muscle weakness, and recurrent MODS. After 2 mo of multidisciplinary management and intensive rehabilitation, the patient could walk with assistance 4 mo after admission and walk independently 6 mo after admission. CONCLUSION: More attention should be paid to mitochondrial myopathy to avoid missed diagnosis and misdiagnosis.

Prolonged fasting induces significant germ cell loss in chickens after hatching.

Germ cell loss is a crucial biological event during germ cell development. The number of female germ cells determines the reproductive performance and egg production of hens. Various intrinsic and extrinsic factors affect germ cell loss, such as germ cell nest breakdown in early life and nutritional deficiencies during daily husbandry. Here, we examined the effect of fasting on the germ cell number of chicks. The results showed that 72 h fasting resulted in a higher germ cell loss than that by 24 h fasting in chicks. The RNA-seq analysis revealed that the genes of ribosome pathway were down-regulated and the biological processes of protein processing in endoplasmic reticulum were inhibited in starved chicks. Furthermore, in female chicks treated with 72 h fasting, the qPCR of ovaries showed down-regulation of ribosome-related genes, and transmission electron microscopy imaging of ovaries showed fewer ribosomes. The blood biochemical indices indicated that 72 h fasting reduced the liver functions and affected the glucose metabolism, lipid metabolites and ion metabolites. In summary, the present results concluded negative impacts on the germ cell pool by prolonged fasting in the early life of chicks and manifested that adequate management should be cared for fasted time for breeding.

YY1-induced upregulation of LncRNA-ARAP1-AS2 and ARAP1 promotes diabetic kidney fibrosis via aberrant glycolysis associated with EGFR/PKM2/HIF-1α pathway.

Objectives: Dimeric pyruvate kinase (PK) M2 (PKM2) plays an important role in promoting the accumulation of hypoxia-inducible factor (HIF)-1α, mediating aberrant glycolysis and inducing fibrosis in diabetic kidney disease (DKD). The aim of this work was to dissect a novel regulatory mechanism of Yin and Yang 1 (YY1) on lncRNA-ARAP1-AS2/ARAP1 to regulate EGFR/PKM2/HIF-1α pathway and glycolysis in DKD. Materials and methods: We used adeno-associated virus (AAV)-ARAP1 shRNA to knocked down ARAP1 in diabetic mice and overexpressed or knocked down YY1, ARAP1-AS2 and ARAP1 expression in human glomerular mesangial cells. Gene levels were assessed by Western blotting, RT-qPCR, immunofluorescence staining and immunohistochemistry. Molecular interactions were determined by RNA pull-down, co-immunoprecipitation, ubiquitination assay and dual-luciferase reporter analysis. Results: YY1, ARAP1-AS2, ARAP1, HIF-1α, glycolysis and fibrosis genes expressions were upregulated and ARAP1 knockdown could inhibit dimeric PKM2 expression and partly restore tetrameric PKM2 formation, while downregulate HIF-1α accumulation and aberrant glycolysis and fibrosis in in-vivo and in-vitro DKD models. ARAP1 knockdown attenuates renal injury and renal dysfunction in diabetic mice. ARAP1 maintains EGFR overactivation in-vivo and in-vitro DKD models. Mechanistically, YY1 transcriptionally upregulates ARAP1-AS2 and indirectly regulates ARAP1 and subsequently promotes EGFR activation, HIF-1α accumulation and aberrant glycolysis and fibrosis. Conclusion: Our results first highlight the role of the novel regulatory mechanism of YY1 on ARAP1-AS2 and ARAP1 in promoting aberrant glycolysis and fibrosis by EGFR/PKM2/HIF-1α pathway in DKD and provide potential therapeutic strategies for DKD treatments.

Tracking the dynamics of female germ cell development during peri-hatch periods using a gene-edited chicken model.

In hens, egg production depends on the development of germ cells in the ovary. Germ cells are established before birth, and their number gradually decreases during their lifespan. Therefore, it is essential to determine the time points of massive germ cell loss and the underlying mechanism. In this study, a gene-edited chicken with mCherry fluorescence specifically expressed in the germline was generated by the integration of the mCherry gene into the 3'-end of the DAZL locus, which facilitated the isolation of germ cells from the gonads of DAZL-mCherry embryos or chicks and quantification using flow cytometry based on the observation of red fluorescence. The results demonstrated the dynamics of germ cell development from embryos at 17 d of hatching (dh) to chickens at 7 d post-hatch (dph) and revealed a substantial loss of germ cells in the late embryonic stage (18 -19 dh) and post-hatch period (2 -3 dph). Additionally, the number of germ cells in DAZL × Guangxi Ma chicken was significantly higher than that in DAZL × Lohmann Pink chicken at 19 dh and 3 dph (P < 0.05). Furthermore, the numbers of germ cells positively correlated with the body weight in DAZL × Lohmann Pink chicken. In conclusion, our results showed the dynamics of germ cell development in chicken ovaries during peri-hatch periods and indicated the time point of substantial germ cell loss. The results provide evidence for further exploration of the underlying mechanism and serve as a reference for chicken breeding and management.

KCNE4 expression is correlated with the pathological characteristics of colorectal cancer patients and associated with the radioresistance of cancer cells.

BACKGROUND: Colorectal cancer (CRC) is a common malignancy, and radioresistance limits the effectiveness of radiotherapy for rectal cancer. This study is performed to investigate the role and regulatory mechanism of Potassium Voltage-Gated Channel Subfamily E Regulatory Subunit 4 (KCNE4) in the radioresistance of CRC cells. METHODS: Immunohistochemical staining results of KCNE4 in normal tissues and CRC tissues were obtained from the Human Protein Atlas (HPA) database. The UALCAN database was used for analyzing KCNE4 mRNA expression in normal tissue samples and CRC tissue samples and its relationship with tumor stage. The relationship of KCNE4 expression with prognosis was analyzed utilizing the data of GEPIA database. LinkedOmics database was searched to analyze the co-expressed gene sets of KCNE4 in CRC, and to analyze the signaling pathways related with KCNE4 in CRC. GO and KEGG enrichment analyses were carried out on the co-expressed genes of KCNE4 with DAVID database. Ionizing radiation (IR)-resistant cell lines (HCT116/IR and HT29/IR) were established; cell viability was assessed via cell counting kit-8 (CCK-8) and EdU assays, and terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay was performed for detecting cell apoptosis. Western blotting was carried out to detect the expressions of p-p85 and p-AKT. RESULTS: KCNE4 was highly expressed in CRC tissues and linked to advanced tumor stage, lymph node metastasis and poor prognosis of CRC patients. KCNE4 overexpression promoted HCT116/IR cell proliferation and inhibited the apoptosis, while KCNE4 knockdown suppressed HT29/IR cell proliferation and facilitated the apoptosis. Furthermore, high KCNE4 expression was associated with the activation of the PI3K/AKT signal pathway. CONCLUSION: KCNE4 is associated with the clinicopathological characteristics of CRC patients, and its high expression level contributes to the radioresistance of cancer cells via activating the PI3K/AKT signal pathway.

Genetically predicted body fat mass and distribution with diabetic kidney disease: A two-sample Mendelian randomization study.

The aim of this study is to apply a Mendelian randomization (MR) design to investigate the potential causal associations between the body mass index (BMI), body fat mass such as trunk fat mass and waist circumference (WC), and diabetic kidney disease (DKD). A two-sample MR study was conducted to obtain exposure and outcome data from previously published studies. The instrumental variables for BMI, trunk fat mass, and WC were selected from genome-wide association study datasets based on summary-level statistics. The random-effects inverse-variance weighted (IVW) method was used for the main analyses, and the weighted median and MR-Egger approaches were complementary. In total, three MR methods suggested that genetically predicted BMI, trunk fat mass, and WC were positively associated with DKD. Using IVW, we found evidence of causal relationships between BMI [odds ratio (OR) = 1.99; 95% confidence interval (CI), 1.47-2.69; p = 7.89 × 10-6], trunk fat mass (OR = 1.80; 95% CI, 1.28-2.53; p = 6.84 × 10-4), WC (OR = 2.48; 95% CI, 1.40-4.42; p = 1.93 × 10-3), and DKD. MR-Egger and weighted median regression also showed directionally similar estimates. Both funnel plots and MR-Egger intercepts showed no directional pleiotropic effects involving the aforementioned variables and DKD. Our MR analysis supported the causal effect of BMI, trunk fat mass, and WC on DKD. Individuals can substantially reduce DKD risk by reducing body fat mass and modifying their body fat distribution.

Replenishment of vitamin A for 7 days partially restored hepatic gene expressions altered by its deficiency in rats.

We investigated the effects of vitamin A (VA) status on metabolism of Zucker rats with different genders and genotypes, and of short-term refeeding of a VA sufficient (VAS) diet on VA deficient (VAD) animals. First, male and female Zucker lean (ZL) and fatty (ZF) rats at weaning were fed a VAD or VAS diet for 8 weeks. Second, male VAD ZL rats were fed a VAS diet for 3 (VAD-VAS3d) or 7 (VAD-VAS7d) days. The body weight (BW), blood parameters, and hepatic expressions of genes for metabolism were determined. VA deficiency reduced BW gain in ZL and ZF rats of either gender. VAD ZL rats had lower plasma glucose, insulin, and leptin levels than VAS ZL rats. VAD-VAS3d and VAD-VAS7d rats had higher plasma glucose, insulin, and leptin levels than that in the VAD rats. The hepatic mRNA levels of Gck, Cyp26a1, Srebp-1c, Igf1, Rarb, Rxra, Rxrg, Pparg, and Ppard were lowered by VA deficiency. Refeeding of the VAS diet for 3 days restored the Gck and Cyp26a1 expressions, and for 7 days restored the Gck, Cyp26a1, Igf1, and Rxrb expressions significantly. The 7-day VA replenishment partially restored the hepatic gene expressions and metabolic changes in VAD ZL rats.

Dietary Lasia spinosa Thw. improves reproductive performance of aged roosters.

The application of artificial insemination is particularly, owing to which breeder animals are considered an important resource in breeding farms. However, the reproductive performance of roosters typically declines with age, and the economic loss experienced by breeders is attributable to this shortened reproductive lifespan. Lasia spinosa Thw. (LST) reportedly improved reproductive capacity in male rodents. The objective of this study was to investigate the effects of LST on the reproductive performance of aged roosters. Male Guangxi Partridge chicken (mean weight, 3032.41 ± 34.48 g; age, 500 days; n = 72) randomly received the following three dietary treatments: LST0 group (a basal diet), LST2 group (a basal diet with 2% LST powder), and LST4 group (a basal diet with 4% LST powder). Computer-aided sperm analysis revealed that dietary LST supplementation significantly improved semen volume, sperm motility, and concentration. Furthermore, the most potent effects were observed in the treatment group with the administration of 2% LST, which significantly improved the weight of the testes. Hematoxylin-eosin staining revealed the increase in diameter of the seminiferous tubule and height of the seminiferous tubule epithelium possibly caused as a result of LST treatment. A significant increase in fructose and glucose concentrations were observed in the testis and seminal plasma; in addition, a significant increase was observed in the α-glycosidase levels in the testis and spermatozoa. However, the monoaldehyde levels in the spermatozoa appeared to decline significantly. Additionally, the fertility rate increased significantly following 2% LST supplementation. RNA-seq analysis revealed that 34 and 16 unigenes were upregulated and downregulated, respectively, in testicular tissues from roosters that received dietary supplementation of 2% LST. The assigned functions of the unigenes revealed that LST primarily influenced the mechanisms underlying catalytic activity and cellular processes. Kyoto Encyclopedia of Genes and Genomes enrichment analysis suggested that spermatogenesis-related pathways were significantly enriched, including ABC transporters, ribosome biogenesis in eukaryotes, and VEGF, cAMP, and ErbB signaling pathways.

Detection of CEA and ProGRP Levels in BALF of Patients with Peripheral Lung Cancer and Their Relationship with CT Signs.

Background: Lung cancer is a common clinical thoracic malignant tumor, which had a serious impact on the safety of patients, currently ranking first in all malignant tumors in morbidity and mortality, with generally less than 5% survival rate in 5 years. Objective: To investigate the relationship and significance between carcinoembryonic antigen (CEA) and precursor gastrin-releasing peptide (ProGRP) changing levels in bronchoalveolar lavage fluid (BALF) and CT imaging features in patients with peripheral lung cancer. Methods: A total of 90 patients diagnosed with peripheral lung cancer as the lung cancer group and 60 patients with benign lung diseases as the control group in our hospital from May 2019 to October 2021 were selected to compare the differences of CEA and ProGRP in BALF by the classification of CT features. Results: The levels of CEA and ProGRP in the BALF of the lung cancer group were significantly higher than those of the control group; the proportion of patients with lobulation sign, burr sign, ground glass sign, pleural effusion, and lesion diameter ≥ 3.0 cm in the lung cancer group was higher than that in the control group; the CEA level in BALF of lung cancer patients with spicule sign, pleural effusion, and lesion diameter ≥ 3.0 cm was significantly higher than that without these symptoms, while ProGRP level in the BALF of lung cancer patients with lobulation sign, burr sign, ground glass sign, pleural effusion, and lesion diameter ≥ 3.0 cm was significantly higher than that of lung cancer patients without these symptoms. Conclusion: The check of CEA and ProGRP in BALF in combination with CT features has a certain clinical value for the diagnosis of lung cancer. At the same time, the increased levels of CEA and ProGRP in BALF have a certain correlation with the changes of malignant signs of lung cancer in CT examination.

Helicobacter pylori promotes invasion and metastasis of gastric cancer by enhancing heparanase expression.

Correction to "Liu LP, Sheng XP, Shuai TK, Zhao YX, Li B, Li YM. Helicobacter pylori promotes invasion and metastasis of gastric cancer by enhancing heparanase expression. World J Gastroenterol 2018; 24: 4565-4577 [PMID: 30386106 DOI: 10.3748/wjg.v24.i40.4565]." In this article, we have identified some of the images in Figure 2A, C, E, G, and I are identical to the images in Figures 1B, 2A, 3B, 3E, and 3G of another paper entitled "Liu L, Zhao Y, Fan G, Shuai T, Li B, Li Y. Helicobacter pylori infection enhances heparanase leading to cell proliferation via mitogenactivated protein kinase signalling in human gastric cancer cells.", which was published by us in the Molecular Medicine Reports in December, 2018 [PMID: 30320396 DOI: 10.3892/mmr.2018.9558]. The reason why we asked to replace the pictures was that when we were simultaneously preparing to submit our two different articles to the World Journal of Gastroenterology (WJG) and Molecular Medicine Reports, we uploaded the wrong pictures to the WJG, which were same as those submitted to the Molecular Medicine Reports. We apologize for this negligence and any inconvenience that this may cause. We would be grateful if you could replace the wrong pictures with the correct ones attached.

Nickel-Catalyzed Asymmetric Hydrogenation of Cyclic Alkenyl Sulfones, Benzo[b]thiophene 1,1-Dioxides, with Mechanistic Studies.

A highly efficient catalytic system based on the cheap transition metal nickel for the asymmetric hydrogenation of challenging cyclic alkenyl sulfones, 3-substituted benzo[b]thiophene 1,1-dioxides, was first successfully developed. A series of hydrogenation products, chiral 2,3-dihydrobenzo[b]thiophene 1,1-dioxides, were obtained in high yields (95-99%) with excellent enantioselectivities (90-99% ee). According to the results of nonlinear effect studies, deuterium-labeling experiments, and DFT calculation investigations, a reasonable catalytic mechanism for this nickel-catalyzed asymmetric hydrogenation was provided, which displayed that the two added hydrogen atoms of the hydrogenation products could be from H2 through the insertion of Ni-H and subsequent hydrogenolysis.

Dietary Lasia spinosa Thw. Improves Growth Performance in Broilers.

This study aimed to evaluate the effects of dietary Lasia spinosa Thw. (LST) powder supplementation on growth performance, blood metabolites, antioxidant status, intestinal morphology, and cecal microbiome in broiler chickens. A total of 400 1-day-old male Guangxi partridge broilers (initial body weight: 42.52 ± 0.06 g) were randomly allotted to 4 dietary treatments: LST0 group (a basal diet), LST1 group (a basal diet with 1% LST powder), LST2 group (a basal diet with 2% LST powder), LST4 group (a basal diet with 4% LST powder), 10 replicates for each treatment, and 10 broilers in each treatment group. Results indicated that the average daily feed intake of broilers during 22-42 days and the average daily gain of chickens during 1-42 days significantly increased by dietary supplementation of LST powder (p < 0.01), while the feed conversion ratio during the overall periods was decreased by dietary supplementation of LST powder (p < 0.01). Except for the levels of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in liver (p > 0.05), the levels of SOD, catalase (CAT) and GSH-Px in serum, liver, and breast muscle were significantly increased in the LST supplemented groups (p < 0.05), while the levels of reactive oxygen species (ROS) and malondialdehyde (MDA) in serum, liver, and breast muscle were significantly decreased in the LST supplemented groups (p < 0.05). Furthermore, the levels of triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) were significantly decreased by the addition of dietary LST powder (p < 0.01), while the levels of HDL-C, Ca, Fe, Mg, and P were linearly increased by the addition of dietary LST powder (p < 0.01). With respect to the gut morphometric, crypt depth was significantly decreased by LST supplementation (p < 0.05), while villus height and the ratio of villus height to crypt depth were notably increased by LST supplementation (p < 0.05). Sequencing of 16S ribosomal RNA (16S rRNA) from the cecal contents of broilers revealed that the composition of the chicken gut microbiota was altered by LST supplementation. The α-diversity of microbiota in broilers was increased (p < 0.05) in the LST1 group, but was decreased (p < 0.05) in the LST2 and LST4 groups compared with the LST0 group. The differential genera enriched in the LST1 group, such as Bacillus, Odoribacter, Sutterella, Anaerofilum, Peptococcus, were closely related to the increased growth performance, antioxidant status, intestinal morphology, Ca, Mg, and reduced blood lipid in the treated broilers.

Injection of Carbon Nanoparticles for Lymph Node Detection After Laparoscopic Colorectal Cancer Surgery.

Colorectal cancer is the third most common cause of cancer-related death in the world because of its poor prognosis and many related complications. In clinical practice, lymph node metastasis is an important index to evaluate the recurrence and survival rate of patients with rectal cancer, it also the main method to determine the treatment plan of patients with rectal cancer. Nano carbon tracer can guide surgeons to clear regional lymph nodes accurately, improve the number of lymph nodes detected, and then improve the accuracy of pathological staging of rectal cancer. It has an important clinical significance in the removal of lymph nodes of middle and low rectal cancer, and provides an important basis for the development of postoperative adjuvant treatment plan. The purpose of this study is to explore the application value of this method in improving the detection rate of lymph nodes in laparoscopic colorectal cancer. According to the research results, the total number of lymphatic gland, the number of small lymphatic gland, the rate of lymph node metastasis, the number of black stained lymphatic gland and the number of lymphatic gland confirmed by pathology in the two groups were better than those in the traditional surgical treatment, which had better application value in the radical operation of colorectal cancer.

Correction to: Matrix metalloproteinase 9 induces endothelial-mesenchymal transition via Notch activation in human kidney glomerular endothelial cells.

An amendment to this paper has been published and can be accessed via the original article.

LncRNA ARAP1-AS2 promotes high glucose-induced human proximal tubular cell injury via persistent transactivation of the EGFR by interacting with ARAP1.

The persistent transactivation of epidermal growth factor receptor (EGFR) causes subsequent activation of the TGF-ß/Smad3 pathway, which is closely associated with fibrosis and cell proliferation in diabetic nephropathy (DN), but the exact mechanism of persistent EGFR transactivation in DN remains unclear. ARAP1, a susceptibility gene for type 2 diabetes, can regulate the endocytosis and ubiquitination of membrane receptors, but the effect of ARAP1 and its natural antisense long non-coding RNA (lncRNA), ARAP1-AS2, on the ubiquitination of EGFR in DN is not clear. In this study, we verified that the expression of ARAP1 and ARAP1-AS2 was significantly up-regulated in high glucose-induced human proximal tubular epithelial cells (HK-2 cells). Moreover, we found that overexpression or knockdown of ARAP1-AS2 could regulate fibrosis and HK-2 cell proliferation through EGFR/TGF-ß/Smad3 signalling. RNA pulldown assays revealed that ARAP1-AS2 directly interacts with ARAP1. Coimmunoprecipitation, dual-immunofluorescence and ubiquitination assays showed that ARAP1 may maintain persistent EGFR activation by reducing EGFR ubiquitination through competing with Cbl for CIN85 binding. Taken together, our results suggest that the lncRNA ARAP1-AS2 may promote high glucose-induced proximal tubular cell injury via persistent EGFR/TGF-ß/Smad3 pathway activation by interacting with ARAP1.

Ursolic Acid Treatment Alleviates Diabetic Kidney Injury By Regulating The ARAP1/AT1R Signaling Pathway.

PURPOSE: This study aimed to investigate whether ursolic acid (UA) mitigates renal inflammation, oxidative stress and fibrosis by regulating the angiotensin II type 1 receptor-associated protein (ARAP1)/angiotensin II type 1 receptor (AT1R) signaling pathway and subsequently alleviating renal damage. METHODS: db/db mice were divided randomly into a diabetic nephropathy (DN) group and a UA treatment group. Light microscopy and electron microscopy were used to observe pathological changes in renal tissues. Immunohistochemistry (IHC) was employed to examine changes in the expression of ARAP1, AT1R, 8-hydroxydeoxyguanosine (8-OHdG), NADPH oxidase 2 (NOX2), the extracellular matrix protein fibronectin (FN), IL-1ß and IL-18 in renal tissues. Western blotting and RT-qPCR were used to detect the respective changes in the protein and mRNA levels of ARAP1, AT1R, NOX4, NOX2, transforming growth factor-ß1 (TGF-ß1), FN, collagen IV, IL-1ß and IL-18 in renal tissues and mesangial cells. In addition, immunofluorescence staining was employed to examine changes in FN and NOX2 expression in mesangial cells. RESULTS: UA treatment effectively reduced the body weights and blood glucose levels of db/db mice (p<0.05) as well as the urinary albumin/creatinine ratio (p<0.05). In addition, the renal tissue lesions and glomerulosclerosis index of the db/db mice were significantly improved after treatment (p<0.01). Histochemical analysis results showed significantly lower expression levels of ARAP1, AT1R, FN, NOX2, 8-OHdG, IL-1ß and IL-18 in renal tissues in the UA treatment group than in the DN group. Western blotting and RT-qPCR data also revealed UA-induced decreases in the renal levels of the ARAP1, AT1, NOX4, NOX2, TGF-ß1, FN, collagen IV, IL-1ß and IL-18 proteins in vivo and/or in vitro (p<0.01). ARAP1 knockdown effectively reduced the expression of NOX2 and FN in vitro. CONCLUSION: UA alleviated renal damage in type 2 diabetic db/db mice by downregulating proteins in the ARAP1/AT1R signaling pathway to inhibit extracellular matrix accumulation, renal inflammation, fibrosis and oxidative stress.

Dynamic contrast enhanced­magnetic resonance imaging for the early evaluation of the response to docetaxel in rats with epithelial ovarian cancer.

Dynamic contrast enhanced­magnetic resonance imaging (DCE­MRI) contributes to the early detection and prediction of responses to chemotherapy in cancer. The aim of the present study was to investigate the feasibility of quantitative DCE­MRI parameters for noninvasively predicting the early response to DTX in epithelial ovarian cancer (EOC). In the present study, using 7,12­dimethylbenz (A) anthracene, orthotopic EOC was induced in Sprague Dawley rats. Rats with EOC were treated with docetaxel (DTX) on day 0. DCE­MRI was applied on days 0, 3, 7, 14 and 21. On day 21, the treated tumor types were categorized into sensitive and insensitive groups according to their change in size. Quantitative DCE­MRI parameters were used to assess the early response to therapy. The experiment was performed again, the treatment group was divided into sensitive and insensitive groups according to their initially obtained cut-off values, and histopathological analyses were performed. Comparing the sensitive group with the insensitive group, there were significant differences in the percentage change in the volume transfer constant (Ktrans), rate constant (kep) and initial area under the curve (IAUC) from day 3 and tumor size from day 14. During the early stages of treatment (on day 3), the percentage change of Ktrans combined with kep produced an AUC of 1, and a sensitivity and specificity of 100 and 100%, respectively, using a cut-off value of a 17.59% reduction in Ktrans and kep. From day 7, there were significant differences in the quantitative index percentage change in angiogenesis in the sensitive group compared with the insensitive group. The percentage change in Ktrans, kep and IAUC were positively correlated with the percentage of change in tumor size and angiogenesis, and negatively correlated with the percentage of change in necrosis. The results of the present study indicated that quantitative DCE­MRI parameters were superior to imaging tumor size for the early detection and prediction of the response to DTX chemotherapy in EOC.

Diffusion-weighted imaging and diffusion kurtosis imaging for early evaluation of the response to docetaxel in rat epithelial ovarian cancer.

BACKGROUND: To investigate diffusion-weighted magnetic imaging (DWI) and diffusion kurtosis magnetic imaging (DKI) for the early detection of the response to docetaxel (DTX) chemotherapy in rat epithelial ovarian cancer (EOC). METHODS: 7,12-Dimethylbenz[A]anthracene was applied to induce orthotopic EOC in Sprague-Dawley rats. Rats with EOC were treated with DTX on day 0 (treatment group) or were left untreated (control group). DWI and DKI were performed on days 0, 3, 7, 14 and 21 after treatment. On day 21, the tumors were categorized into the sensitive and insensitive groups according to the size change. The cutoff values of the DWI and DKI parameters for the early response were determined. The experiment was repeated, and the treatment group was divided into the sensitive and insensitive groups according to the initially obtained cutoff values. The DWI and DKI parameters were correlated with tumor size, proliferation, apoptosis and tumor necrosis. RESULTS: In the sensitive vs. insensitive or control group, significant differences were found in the Δ% of the DWI and DKI parameters (ADC, D and K) from day 3 and in tumor size from day 14. Early on day 7, the Δ% of K had an AUC of 1 and sensitivity and specificity values of 100% and 100%, respectively, to detect the response to DTX using a cutoff value of 19.03% reduction in K. From day 7, significant differences were found in the Δ% of Ki-67 and CA125 in the sensitive vs. control group and from day 14 in the sensitive vs. insensitive group. From day 14, there were significant differences in the Δ% of Bcl-2, apoptosis and tumor necrosis in the sensitive vs. control or insensitive group. The Δ% values of ADC and D were negatively correlated with the Δ% values of tumor size, Ki-67, CA125 and Bcl-2 and were positively correlated with the Δ% values of apoptosis and tumor necrosis. The Δ% of K was positively correlated with the Δ% values of tumor size, Ki-67, CA125 and Bcl-2 and was negatively correlated with the Δ% values of apoptosis and tumor necrosis. CONCLUSIONS: DWI and DKI parameters, especially K, are superior for imaging tumor size for the early detection of the response to DTX chemotherapy in induced rat EOC.