CMaf-Inducing Protein Promotes LUAD Proliferation and Metastasis by Activating the MAPK/ERK Pathway.

Objective: Previous studies have shown that cMaf-inducing protein (CMIP) promotes tumorigenesis and progression, however, the role of CMIP in lung adenocarcinoma (LUAD) and its molecular mechanism remain unclear. Methods: In this study, the Human Protein Atlas and Kaplan-Meier Plotter database were used to analyze the expression and prognostic value of CMIP in LUAD. Then, the expression levels of CMIP in LUAD tissues and cells were detected by qRT-PCR and western blot. The lentiviral vector was used to establish a stable transfected cell line, and the transfection efficiency was detected by qRT-PCR. MTT assay, colony formation assay, transwell assay, and wound healing assay were used to evaluate the function of CMIP in LUAD. In addition, the effect of CMIP on the MAPK/ERK pathway in LUAD cells was analyzed by western blot. Results: The expression level of CMIP was significantly increased in LUAD cell and tissue samples, and the high expression of CMIP was associated with overall survival (OS) and progression-free survival (PFS) in LUAD patients. In vitro experiments showed that CMIP overexpression significantly promoted the proliferation, migration, and invasion of A549 cells. CMIP knockout significantly inhibited the proliferation, migration, and invasion of H1299 cells. In addition, it was observed that the expression levels of the MAPK/ERK pathway-related proteins were significantly increased in CMIP-overexpressed A549 cells, and promoted cell proliferation, migration, and invasion, while U0126 could significantly reverse the activation of the MAPK/ERK pathway by CMIP overexpression, and inhibit the proliferation, migration, and invasion of A549 cells. Conclusion: Our study shows that CMIP, as an oncogene, is associated with poor patient prognosis, and may promote the proliferation and metastasis of LUAD by activating the MAPK/ERK pathway. Therefore, CMIP may be a new potential therapeutic target for LUAD.

Inhibitory effect of tanshinone IIA, resveratrol and silibinin on enterovirus 68 production through inhibiting ATM and DNA-PK pathway.

BACKGROUND: Human enterovirus 68 (EV68) is a primary etiological agent for respiratory illnesses, while no effective drug has yet used in clinics largely because the pathogenesis of EV68 is not clear. DNA damage response (DDR) responds to cellular DNA breaks and is also involved in viral replication. Three DDR pathways includes ataxia telangiectasia mutated (ATM), ATM and Rad3-related (ATR), and DNA-dependent protein kinase (DNA-PK). Natural products proved to be an excellent source for the discovery and isolation of novel antivirals. Among them, tanshinone IIA, resveratrol, silibinin, rutin and quercetin are reported to target DDR, therefore their roles in anti-EV68 are investigated in this study. PURPOSE: This study investigated the anti-EV68 ability of various natural compounds related to DDR. STUDY DESIGN AND METHODS: The methods include cell counting, flow cytometry, western blot, Immunofluorescence staining, comet assays, quantitative real-time RT PCR and short interfering RNAs (siRNAs) for analysis of cell number, cell cycle, protein expression, protein location, DNA damage, mRNA level and knock down target gene, respectively. RESULTS: EV68 infection induced DDR. Down-regulation or inhibition of ATM or DNA-PK lowered DDR in EV68-infected cells and mitigated viral protein expression, however, down-regulation or inhibition of ATR unexpectedly up-regulated DDR, and promoted viral protein expression. Meanwhile tanshinone IIA, resveratrol, and silibinin inhibited ATM and/or DNA-PK activation and decreased viral proliferation, while rutin and quercetin inhibited ATR activation and promoted viral production. The role of them in ATM, DNA-PK and ATR activation was consistent with previous reports. CONCLUSION: Tanshinone IIA, resveratrol and silibinin inhibited EV68 proliferation through inhibiting ATM and/or DNA-PK activation, and they were effective anti-EV68 candidates.

The experiences of nurses and midwives who provide surgical abortion care: A qualitative systematic review.

AIM: To synthesize qualitative evidence on nurses' and midwives' experiences in the provision of surgical abortion care. We address three specific questions: (a) what are the experiences of nurses and midwives in surgical abortion care? (b) what are their responses and coping strategies? (c) what are the deficiencies in surgical abortion care? DESIGN: Qualitative studies were synthesized using Thomas and Harden's qualitative thematic synthesis method. DATA SOURCES: Electronic databases, including PubMed, Embase, CINAHL, PsycINFO, Scopus and Web of Science were searched. Grey literature using ProQuest was searched. The databases were searched from inception to 5 August 2020. REVIEW METHODS: The SPIDER (Sample, Phenomenon of Interest, Design, Evaluation and Research type) search tool was used in the literature search. Data synthesis was conducted using the three-stage thematic synthesis method described by Thomas and Harden. RESULTS: 966 studies were identified in the initial search and 18 studies were included. Four analytical themes were generated: 'Providing abortion care requires high emotional labour'; 'Professionalism of abortion care providers'; 'Initiatives in professional development' and 'Improving directions for high-quality abortion care'. CONCLUSION: Nurses and midwives indicated that they require support to enhance psychological health and improve professional skills. Hospital managers should organize regular debriefing or structured group workshops for exchange of practical experiences and strengthening emotional support. More research is required to establish comprehensive training related to abortion care for nurses and midwives. The findings demonstrate that optimization of abortion services should start from hospital management models, pain management and bereavement care. IMPACT: Understanding the experiences of nurses and midwives in abortion provision will inform future clinical practice in surgical abortion care, which would be helpful in improving the professionalism and confidence of abortion providers. Our findings have implications for the training, development of policies and standards for surgical abortion care for nurses and midwives.

MiR-92b-3p is Induced by Advanced Glycation End Products and Involved in the Pathogenesis of Diabetic Nephropathy.

PURPOSE: The current study aims to examine the effects of advanced glycation end products (AGEs) on the microRNA (miRNA) expression profile in the kidney tissues of rats. METHODS: Wistar rats were randomly divided into three equal experiment groups: the AGE group, the RSA group, and the control group. The rats in the AGE group and the RSA group were administered with advanced glycation end products (AGEs) and rat serum albumin (RSA) via the tail vein, respectively, whereas the control group received PBS. Total RNA was prepared from the rat kidney tissues, and the miRNA expression profiles in different experiment groups were compared by microarray analysis. The expression levels of selected differential miRNAs were verified by RT-qPCR. Target gene prediction was conducted using algorithms such as TargetScan, miRanda, and PICTar. Functional analysis was performed to determine the putative biological roles of the validated miRNAs. RESULTS: The microarray study revealed 451 upregulated and 320 downregulated miRNAs in the AGE group compared with the RSA group (p < 0.05). Seven miRNAs, including miR-21-5p, miR-92b-3p, miR-140-3p, miR-196a-5p, miR-181b-5p, miR-186-5p, and miR-192-5p, were screened and verified using RT-qPCR, of which, the change of miR-92b-3p was the most obvious according to the miRNA expression different multiple and p < 0.05). Seven miRNAs, including miR-21-5p, miR-92b-3p, miR-140-3p, miR-196a-5p, miR-181b-5p, miR-186-5p, and miR-192-5p, were screened and verified using RT-qPCR, of which, the change of miR-92b-3p was the most obvious according to the miRNA expression different multiple and. CONCLUSION: The results of the current study suggested that miR-92b-3p could mediate AGE-induced development of renal abnormalities through targeting Smad7 in rats with DN.

Effects of keto acid supplements on Chinese patients receiving maintenance hemodialysis: a prospective, randomized, controlled, single-center clinical study.

BACKGROUND: The effects of keto acid (KA) supplements on Chinese patients receiving maintenance hemodialysis (MHD) are unclear. This study aimed to evaluate the effects of KA supplementation on nutritional status, inflammatory markers, and bioelectric impedance analysis (BIA) parameters in a cohort of Chinese patients with MHD without malnutrition. METHODS: This was a prospective, randomized, controlled, single-center clinical study conducted in 2011 till 2014. Twenty-nine patients with MHD were randomly assigned to a control (n = 14) or a KA (n = 15) group. The control group maintained a dietary protein intake of 0.9 g/kg/day. The KA group received additional KA supplement (0.1 g/kg/day). BIA was used to determine the lean tissue mass, adipose tissue mass, and body cell mass. The patients' nutritional status, dialysis adequacy, and biochemical parameters were assessed at the ends of the third and sixth months with t test or Wilcoxon rank-sum test. RESULTS: The daily total energy intake for both groups was about 28 kcal/kg/day. After 6 months, the Kt/V (where K is the dialyzer clearance of urea, t is the dialysis time, and V is the volume of the distribution of urea) was 1.33 ±â€Š0.25 in KA group, and 1.34 ±â€Š0.25 in the control group. The median triceps skin-fold thickness in KA group was 12.00 and 9.00 mm in the control group. In addition, the median hand-grip strength in KA group was 21.10 and 25.65 kg in the control group. There were no significant differences between the groups with respect to the anthropometry parameters, dialysis adequacy, serum calcium and phosphorus levels, inflammatory markers, and amino-acid profiles, or in relation to the parameters determined by BIA. Both groups achieved dialysis adequacy and maintained nutritional status during the study. CONCLUSIONS: In this cohort of Chinese patients with MHD, the patients in the control group whose dietary protein intake was 0.9 g/kg/day and total energy intake was 28 kcal/kg/day, maintained well nutritional status during study period. The KA supplement (0.1 g/kg/day) did not improve the essential amino acid/non-essential amino acid ratio, nor did it change the patients' mineral metabolism, inflammatory parameters, or body compositions.

MicroRNAs in the Progress of Diabetic Nephropathy: A Systematic Review and Meta-Analysis.

BACKGROUND: We conducted a systematic review and meta-analysis of existing literature to evaluate the different outcomes of microRNAs (miRNAs) in diabetic nephropathy (DN), including urinary albumin excretion rates, urinary albumin creatinine rates, glomerular filtration rate, HbAc1, and creatinine. METHODS: Electronic databases including PUBMED, MEDLINE, and EMBASE were searched for eligible publications to July 2018. The following comparisons between treatment groups were included: normal group versus DN group; control group versus micro/macroalbuminuria group. RESULTS: Twelve eligible studies that included 2500 participants were finally recruited in this meta-analysis. Fifteen miRNAs (miRNA-21, miRNA-181b, miRNA-194, miRNA-30, miRNA-215, and others) were upregulated whereas seven miRNAs (miRNA-26a, miRNA-126, miRNA-424, miRNA-574-3p, miR-223, miR-155, and miR-192) were downregulated in the DN group compared with control groups. The miR-133b, miR-342, miR-30, miR-192, miR-194, and miR-215 were significantly correlated in urinary albumin excretion rates (r=0.33, 95% CI= 0.26-0.39). miR-192, miR-217, miR-15b, miR-34a, and miR-636 were correlated with urinary albumin creatinine rates (r=0.69; 95% CI=0.12-0.92), while miR-133b, miR-345, miR-33, miR-326, miR-574-3p, miR-126, miR-217, miR-15b, miR-34a, and miR-636 were significantly correlated with HbAc1 (r =0.23, 95% CI = 0.15-0.31). There were twelve miRNAs that were closely related to the glomerular filtration rate (r=0.28, 95% CI =0.21-0.34). Creatinine (r=0.33, 95% CI = 0.22-0.40) was significantly different between normal and DN groups. CONCLUSIONS: The meta-analysis acquired the correlations between miRNAs and outcomes including UAER, UACR, eGFR, HbAc1, and creatinine in DN. It suggested that miRNAs may participate in the pathogenesis of DN process.

Pathogenesis of steroid-resistant asthma and the influence of vitamin D / 中国当代儿科杂志

Glucocorticoid (GC) is currently the most effective drug for controlling persistent asthma; however, there is a significant difference in the response to GC among patients with asthma. Steroid-resistant asthma is one of the subtypes of asthma and has poor response to high-dose GC treatment. It may affect the quality of life of patients and even threaten their lives. Therefore, it is of great significance to explore the pathogenesis of steroid-resistant asthma and related targeted treatment strategy. In recent years, a variety of pathogeneses have been found to participate in the development and progression of steroid-resistant asthma, including the reduction in the binding between GC receptor and GC, the increase in the expression of GC receptor β, over-activation of nuclear transcription factor activating protein 1 and nuclear factor-κB, abnormality in histone acetylation, and immune-mediated cytokine dysregulation. In addition, many studies have shown that vitamin D can improve the sensitivity to GC among patients with steroid-resistant asthma. This article reviews the pathogenesis of steroid-resistant asthma and the influence of vitamin D.

Activation of phenylpropanoid pathway and PR of potato tuber against Fusarium sulphureum by fungal elicitor from Trichothecium roseum.

The induced resistance of potato tuber (Solanum tuberosum cv. Xindaping) tissue against Fusarium sulphureum by a fungal elicitor from the incompatible pathogen Trichothecium roseum and its possible mechanism were studied. The results showed that the lesion development of the wound-inoculated potato tuber was significantly reduced by treatment with the fungal elicitor from T. roseum (P < 0.05). Inoculation with F. sulphureum on the 16th day after treatment with the fungal elicitor80 at 15.0 µg/ml had the best resistant effect in the potato tuber, with the diameter being only reduced by 47 % that of the control. In addition, the results also showed that the potato tuber treated with the fungal elicitor80 could systemically induce lignin deposition, total phenolic content, flavonoid content and defense enzymes, including three keys phenylpropanoid pathway (PAL, 4CL and C4H) and pathogenesis-related (GLU and CHT) enzymes. The fungal elicitor80 also enhanced the up-regulation of the transcription and expression of PAL, C4H, 4CL, GLU and CHT genes. The treatment with the fungal elicitor80 + F. sulphureum caused the marked and/or prompt enhancement of all indexes when compared to treatment with the fungal elicitor80 or inoculation with the pathogen alone. The results suggested that the fungal elicitor of T. roseum could significantly enhance defense responses in potato tuber against dry rot mainly due to the up-regulation of the transcription and expression of resistance-related genes as well as increasing the activity of resistance-related enzymes and antifungal compounds.

Salvianolic acid A improves intestinal motility in diabetic rats through antioxidant capacity and upregulation of nNOS.

OBJECTIVE: This study aimed to detect the effect of a new herbal extract salvianolic acid A (SalA) on gastrointestinal complications in diabetic rats. METHODS: Altogether 80 rats were divided randomly into five groups, including normal control (NC) group, high-fat (HF) diet group, diabetes mellitus (DM) control group, and DM treated with SalA (0.1 mg/kg and 0.3 mg/kg) groups, respectively. DM was induced by feeding the rats with HF diet and the administration of streptozotocin (30 mg/kg). Four weeks after the establishment of the DM model, the rats received SalA or double distilled water for 8 weeks. After the evaluation of intestinal motility, the animals were sacrificed and their intestines were isolated and collected. The levels of advanced glycation end-products (AGE) and malondialdehyde (MDA) were detected. Protein gene product 9.5 (PGP9.5) and neuronal nitric oxide synthase (nNOS) expressions in the intestine were also detected. RESULTS: Compared with the NC and HF rats, the DM control rats showed significantly increased blood glucose level and decreased weight. Compared with the DM control group, SalA did not influence their weight and blood glucose level, but significantly reduced the levels of AGE and MDA. Intestinal transit was promoted by SalA in diabetic rats, and the expressions of PGP9.5 and nNOS in the intestine were both upregulated. CONCLUSION: The effect of SalA on the intestinal motility of diabetic rats might be due to its antioxidant capacity and restoring nNOS expression.

[Optimization of Supercritical Fluid Extraction of Total Flavonoids and Monomer Compositions from Taxus Remainder Extracts Free of Taxoids].

Objective: To optimize the supercritical fluid extraction technology of flavonoids from taxus remainder extracts free of taxoids( TREFT) by response surface methodology( RSM). Methods: By using a central composite design( CCD) in four factors and five levels to optimize the extraction parameters. The effects of extraction time, temperature, pressure and different concentration of ethanol and their interaction on the extraction rate of total flavonoids, amentoflavone, quercetin and ginkgetin which extracted from TREFT by supercritical fluid CO2 were investigated. Results: Under optimal conditions of extraction time for 2. 2 h,and the extraction temperature was 46. 4 ℃,the extraction pressure was 22. 6 MPa, and 80. 7% ethanol, the index experimental of total flavonoids and monomer compositions was 98. 75. Conclusion: The process of flavonoids extraction from TREFT is stable and feasible, which can provide technical support for the resource utilization of TREFT.

[Effects of exogenous carnitine on function of respiratory chain and antioxidant capacity in mitochondria of myocardium after exhaustive running in rats].

OBJECTIVE: To investigate the effect of exogenous carnitine on function of respiratory chain and antioxidant capacity in mitochondria of myocardium in rats. METHODS: Forty male Wistar rats were randomized to 4 groups (n = 10): static control (C), supplementation of carnitine (LC), exercise-training (T) and training with supplementation of carnitine (TLC). LC and TLC animals were perfused carnitine by the dose of 300 mg/kg bw x d. T and TLC animals were forced to performed 6-week treadmill training. Heart were prepared immediately after exhaustive running. Myocardium mitochondria was extracted by differential centrifugation. Spectrophotometric analysis was used to evaluate activities of respiratory chain complex (C) I -IV and superoxide dismutase (SOD), malondialdehyde (MDA) level in myocardium mitochondria. RESULTS: To compare with C group, C I and C IV activity in LC, T and TLC group were increased significantly (P < 0.05, P < 0.01), CII and C III activity in T and TLC group were increased significantly (P < 0.05, P < 0.01); to compare with LC group, C I - IV activity in TLC group were increased significantly (P < 0.05, P < 0.01); to compare with T group, CI and C IV activity in TLC group increased significantly (P < 0.05). To compare with C group, SOD activity increased remarkably, MDA was remarkably lower (P < 0.05, P < 0.01) in LC, T and TLC group; To compare with LC and T group, SOD activity increased remarkably, MDA was remarkably lower (P < 0.05) in TLC group. CONCLUSION: Carnitine and training could improve function of respiratory chain and increased antioxidant capacity in myocardium mitochondria, there was better function of cooperation between carnitine and training.

Inhibitory effects of salvianolic acid B on CCl(4)-induced hepatic fibrosis through regulating NF-κB/IκBα signaling.

ETHNOPHARMACOLOGICAL RELEVANCE: Hepatic fibrosis, a precursor of liver cirrhosis, is a consequence of severe liver damage that occurs in many patients with chronic liver diseases. Salvianolic acid B (SA-B) is one of water soluble compounds derived from Salvia miltiorrhiza Bunge (Danshen in Chinese) widely used for chronic liver diseases. In this study we investigated the protective effects of SA-B on CCl(4)-induced hepatic fibrosis. MATERIALS AND METHODS: Hepatic fibrosis in rats was induced by carbon tetrachloride (CCl(4)). Rats were divided into four groups, including normal controls (N group), model (M group), low SA-B of 10mg/kg body weight (L group), or high SA-B of 20mg/kg body weight (H group). After 6 weeks, macroscopic features of the liver and weight ratio of liver to body were measured. Liver fibrosis of the rats was evaluated by HE and Massion staining. Activities of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) and total bilirubin (TBIL) were checked with automated biochemistry analyzer. Serum levels of hyaluronic acid (HA), type IV collagen (IV-C), Laminin (LN) and procollagen III peptide (PIIIP) were detected by radioimmunoassay (RIA). The expression of NF-κB and IκBα was detected by western blotting. RESULTS: SA-B was shown to reduce CCl(4)-induced hepatic fibrosis in rats. The serum levels of ALT, AST, and TBIL were significantly lower in the SA-B treatment groups than in the M group. Compared the M group, the serum levels of HA, LN, IV-C and PIIIP were decreased markedly after treatment with SA-B, especially in the H group. Treatment with SA-B at 10-20mg/kg (L and N groups, respectively) dose-dependently decreased the expression of NF-κB in the nucleolus and increased the expression levels of NF-κB and IκBα protein in the cytoplasm compared to that of the M group. CONCLUSIONS: This study reveals that SA-B could prevent the progression of liver angiogenesis and alleviate liver fibrosis possibly by regulating the expression of NF-κB and IκBα.

Screening for novel lead compounds increasing insulin expression in medullary thymic epithelial cells.

Insulin expression in the thymus has been implicated in regulating the negative selection of autoreactive T cells and in mediating the central immune tolerance to pancreatic beta-cells. Thymic insulin expression modulation might be an important drug target for preventing type 1 diabetes. We performed a high-throughput screening to identify compounds with such activity. A reporter plasmid was constructed with the human insulin promoter sequence including a short allele of the upstream variable number tandem repeat (VNTR) sequence (32 repeats), subcloned into the pGL4.17 vector. The plasmid was stably transfected into an insulin-transcribing medullary thymic epithelial cell (mTEC) line. Primary high-throughput screening assays were carried out by stimulating with candidate compounds for 24h, and the activity of luciferase was measured. Positive compounds were further validated by real-time PCR. Of 19,707 compounds, we identified one compound that could enhance mTEC insulin expression, as confirmed by real-time PCR. We also observed that transfection with the autoimmune regulator (AIRE) increased endogenous AIRE expression in mTECs. Our insulin-VNTRI-promoter reporter system is consistent with the insulin expression regulation in mTECs, and one compound that was identified could increase insulin expression in mTECs. A positive feedback effect of AIRE in mTECS was observed. Whether these efforts in murine thymus cells apply to humans remains to be determined.