bluuebunny/crossref_metadata_2025 huggingface dataset:
MMP‐9 gene rs3918242 polymorphism increases risk of stroke: A meta‐analysis
Authors: Wang Baohua , Wang Yinhua , Zhao Lei
Published: August 2018

The association between matrix metalloproteinase‐9 (MMP‐9) gene rs3918242 polymorphism and the risk of stroke has been investigated, but the findings have been found to be contradictory. Thus, this association was assessed by meta‐analysis. Potential studies were searched throughout PubMed, Embase, and CNKI. Pooled odds ratios and 95% confidence intervals were calculated. Sixteen original studies analyzing this association were included, involving 3647 stroke patients and 3685 unrelated controls. It was confirmed that MMP‐9 gene rs3918242 polymorphism increased the risk of stroke. Stratification analysis of ethnicity revealed an increased risk of stroke among Asians, but not among Caucasians. Subgroup analysis by the type of stroke showed that MMP‐9 gene rs3918242 polymorphism increased the risk of ischemic stroke. Sensitivity analysis did not draw different findings. In conclusion, MMP‐9 gene rs3918242 polymorphism increases the risk of stroke.
Intracellular protons accelerate aging and switch on aging hallmarks in mice
Authors: Osanai Tomohiro , Tanaka Makoto , Izumiyama Kei , Mikami Kasumi , Kitajima Maiko , Tomisawa Toshiko , Magota Koji , Tomita Hirofumi , Okumura Ken
Published: August 2018

Diet‐induced metabolic acidosis is associated with the impairment of bone metabolism and an increased risk of a number of chronic noncommunicable diseases, such as type 2 diabetes mellitus and hypertension. The serum bicarbonate level is an independent predictor of chronic kidney disease progression. We investigated whether proton accelerates aging by analyzing both coupling factor 6‐overexpressing transgenic (TG) and high salt‐fed mice which display sustained intracellular acidosis, due to enhanced proton import through ecto‐F1Fo complex and/or reduced proton export through Na+‐K+ ATPase inhibition. Both types of mice displayed shortened lifespan and early senescence‐associated phenotypes such as signs of hair greying and alopecia, weight loss, and/or reduced organ mass. In chronic intracellular acidosis mice, autophagy was impaired by regression of Atg7, an increase in nuclear acetylated LC3 II, and acetylation of Atg7. The increase in histone 3 trimethylation at lysine 4 (H3K4me3) and H4K20me3 and the decrease in H3K9me3 and H3K27me3 were observed in the heart and kidney obtained from both TG and high salt‐fed mice. The decrease in lamin A/C, emerin, and heterochromatin protein 1α without changes in barrier‐to‐autointegration factor and high‐mobility group box 1 was confirmed in TG and high salt‐fed mice. Suppression of nuclear histone deacetylase 3‐emerin system is attributable to epigenetic regression of Atg7 and H4K5 acetylation. These findings will shed light on novel aging and impaired autophagy mechanism, and provide implications in a target for antiaging therapy.
Gene therapy in cardiovascular diseases: A review of recent updates
Authors: Gorabi Armita M. , Hajighasemi Saeideh , Tafti Hossein A. , Soleimani Masoud , Panahi Yunes , Ganjali Shiva , Sahebkar Amirhossein
Published: August 2018

Gene therapy is considered as a promising approach for treating cardiac dysfunction. In this review, we evaluated the clinical trials assessing gene therapy in cardiovascular diseases (CVD) from 2000 to 2017. PubMed and ClinicalTrials.gov (only English language) were searched for clinical trials published between January 2000 and May 2017, using the search terms “gene transfer” OR “gene therapy” and “cardiovascular diseases” and related terms. The trials with sample size lower than 10 patients were excluded. Twenty‐six clinical trials on human and animals, including 1543 patients were listed and evaluated. The sample size in 14 trials was lower than 100 patients and in seven trials lower than 20 patients. Eleven trials used plasmid DNA and eight trials used adenovirus, one study used plasmid DNA, adenovirus, and liposome. We detected that gene therapy was a safe approach and improved the symptoms of CVD; however, the effect of gene therapy on the cardiac dysfunction is controversial.
Establishment and characterization of human theca stem cells and their differentiation into theca progenitor cells
Authors: Dalman Azam , Totonchi Mehdi , Valojerdi Mojtaba Rezazadeh
Published: August 2018

In this study, we have characterized the human theca stem cells (hTSCs) and their differentiation into human theca progenitor cells (hTPCs). hTSCs were isolated from the theca layer of small antral follicles (3‐5 mm in size). Alkaline phosphatase activity, cell cycle status, and cell surface markers were evaluated in hTSCs. The differentiation potential of these cells was investigated via differentiation of hTSCs into adipocyte‐, osteocyte‐, and chondrocyte‐like cells. The cells also differentiated into hTPCs. The hTSCs were morphologically similar to human fibroblast cells (hFCs). Some of the cells were positive for alkaline phosphatase activity. The expression of OCT4 in hTSCs was significantly higher than that of human bone marrow mesenchymal stem cells (hBMSCs) and hFCs. To determine the type of OCT4 (isoform A or B), RT PCR was performed. The data showed that OCT‐4A was expressed in hBMSCs and hTSCs but immunofluorescence analyses using the OCT‐4A‐specific and OCT4 antibodies did not show OCT‐4A protein. In addition, cell cycle status showed that the number of hTSCs in the S phase was significantly higher than that of hFCs. CD29, CD44, CD73, CD90, and CD105 were present in hTSCs. Osteogenic, adipogenic, and chondrogenic differentiation was confirmed by cytochemical staining and lineage‐specific transcripts. Our results showed that specific Dulbecco modified Eagle medium F12 culture medium results in the presence of hTPC markers. hTPCs displayed lipid droplets, appropriate gene expression, and secreted dehydroepiandrosterone and estradiol. hTSCs have the ability to differentiate into mesenchymal lineages and hTPCs. This study may provide a novel in vitro model for further investigation of theca cell maturation and differentiation.
Inhibition of lncRNA X inactivate‐specific transcript ameliorates inflammatory pain by suppressing satellite glial cell activation and inflammation by acting as a sponge of miR‐146a to inhibit Nav1.7
Authors: Sun Wenbo , Ma Meina , Yu Hongmei , Yu Hong
Published: August 2018

Long noncoding RNAs (lncRNA) has been validated to participate in nociception in inflammatory pain, presenting as a potential target against anesthesia. Previous research work confirmed the correlation between lncRNA X inactivate–specific transcript (XIST) and inflammation. However, its role in inflammatory pain is undefined. In animal pain models, voltage‐gated sodium channels (VGSCs) reportedly participate in neural excitation. In this study, we observed the high expression of XIST and VGSC 1.7 (Nav1.7) in the dorsal root ganglion (DRG) of the complete Freund’s adjuvant (CFA)‐induced rat inflammatory pain model. Furthermore, XIST inhibition alleviated pain behavior and the activation of DRG satellite glial cells by suppressing glial fibrillary acidic protein (GFAP) expression, as well as inflammatory cytokine levels of interleukin‐6 and tumor necrosis factor‐α. XIST downregulation increased the mechanical pain threshold in an inflammatory pain model. Moreover, the expression of miR‐146a was decreased in CFA rats. In vitro, XIST acted as a sponge of miR‐146a, which targeted Nav1.7 via bioinformatic prediction, luciferase reporter, and pull‐down assay. More importantly, activation of the Nav1.7 pathway or miR‐146 depression both reversed XIST knockdown–inhibited satellite glial cell activation and inflammatory pain in CFA rats. These results suggest that cessation of XIST may ameliorate inflammatory pain by acting as a sponge of miR‐146a to inhibit Nav1.7, implying a promising strategy against inflammatory pain.
Retracted: Inositol polyphosphate‐4‐phosphatase type II and rucaparib treatment inhibit the growth of osteosarcoma cells dependent on phosphoinositide 3‐kinase/protein kinase B pathway
Authors: Li Dong , Yang Jingyan , Ma Huanzhi , Sun Chengliang , Feng Rongjie
Published: August 2018

Osteosarcoma (OS) is an aggressive malignant tumor of bone, which often occurs in children and adolescents. Currently, the effective method for the treatment of OS is still limited. The study aimed to investigate the synergistic antitumor effect of inositol polyphosphate‐4‐phosphatase, type‐II (INPP4B) and rucaparib on OS cells. The expression levels of INPP4B in OS tissues and OS cell lines were examined by quantitative real‐time polymerase chain reaction and Western blot analysis. SaOS2 and U2OS cells were then transfected with overexpression vector of INPP4B or were treated with different concentrations of rucaparib, and cell viability, cell cycle, and apoptosis were determined by 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyltetrazolium bromide and flow cytometry. Western blot assay uncovered the combined effects of INPP4B and rucaparib on cell cycle, apoptosis and phosphoinositide 3‐kinase/protein kinase B (PI3K/AKT) signal pathway. Further, the tumor formation was examined in vivo. Results showed that INPP4B was low expressed in OS tissues and in OS cell lines. INPP4B overexpression significantly decreased cell viability and induced apoptosis in SaOS2 and U2OS cells. Additionally, rucaparib remarkably reduced cell viability in a dose‐dependent and time‐dependent manner. Meanwhile, rucaparib suppressed cell cycle progression in the S phase and promoted apoptosis in a dose‐dependent manner. Further, combination of INPP4B overexpression and rucaparib declined Myc, cyclin E1 and cyclin D1 expressions, enhanced Bad, Bax, and cleaved‐caspase‐3 expressions, and blocked PI3K/AKT signal pathway in SaOS2 and U2OS cells. Finally, combination of INPP4B overexpression and rucaparib inhibited tumor formation in vivo. The study demonstrated that INPP4B and rucaparib exhibited synergistic antitumor effect by regulating PI3K/AKT pathway in OS cells.
Hsa‐miR‐5582‐3P regulatory effect on TGFβ signaling through targeting of TGFβ‐R1, TGFβ‐R2, SMAD3, and SMAD4 transcripts
Authors: Abedini Bakhshmand Elham , Mohammad Soltani Bahram , Fasihi Ali , Mowla Seyed Javad
Published: August 2018

Transforming growth factor β (TGFβ) signaling pathway which is regulated by factors such as microRNAs (miRNAs) has pivotal roles in various cellular processes. Here, we intended to verify bioinformatics predicted regulatory effect of hsa‐miR‐5582‐3P against TGFβ/SMAD signaling pathway components. Quantitative reverse‐transcription polymerase chain reaction (RT‐qPCR) analysis indicated a negative correlation of expression between hsa‐miR‐5582‐3P against TGFβ‐R1, TGFβ‐R2, SMAD3, and SMAD4 putative target genes in all of tested cell lines. Also, hsa‐miR‐5582‐3P was significantly downregulated in glioma, breast, and ovarian tumor tissues compared with their normal pairs, detected by RT‐qPCR. Then dual luciferase assay supported direct interaction between this miRNA and TGFβ‐R1, TGFβ‐R2, SMAD3, and SMAD4, 3′ untranslated region sequences. Western blot analysis confirmed negative effect of hsa‐miR‐5582‐3P overexpression on at least TGFβ‐R1 expression. Consistently, hsa‐miR‐5582‐3P overexpression brought about downregulation of TGFβ‐R1, TGFβ‐R2, SMAD3, and SMAD4 expression in HCT‐116 cell line, followed by cell cycle arrest in sub‐G1 phase, detected by flow cytometry. Altogether, our data suggest that hsa‐miR‐5582‐3P reduces the TGFβ/SMAD signaling pathway through downregulation of TGFβ‐R1, TGFβ‐R2, SMAD3, and SMAD4 transcripts. These data introduce hsa‐miR‐5582‐3P as a potential tumor suppressors‐miR and a therapy candidate to be tested in cancers in which TGFβ/SMAD is deregulated.
Z‐band and M‐band titin splicing and regulation by RNA binding motif 20 in striated muscles
Authors: Chen Zhilong , Maimaiti Rexiati , Zhu Chaoqun , Cai Hanfang , Stern Allysa , Mozdziak Paul , Ge Ying , Ford Stephen P. , Nathanielsz Peter W. , Guo Wei
Published: August 2018

Titin (TTN) has multifunctional roles in sarcomere assembly, mechanosignaling transduction, and muscle stiffness. TTN splicing generates variable protein sizes with different functions. Therefore, understanding TTN splicing is important to develop a novel treatment for TTN‐based diseases. The I‐band TTN splicing regulated by RNA binding motif 20 (RBM20) has been extensively studied. However, the Z‐ and M‐band splicing and regulation remain poorly understood. Herein, we aimed to define the Z‐ and M‐band splicing in striated muscles and determined whether RBM20 regulates the Z‐ and M‐band splicing. We discovered four new Z‐band TTN splicing variants, and one of them dominates in mouse, rat, sheep, and human hearts. But only one form can be detected in frog and chicken hearts. In skeletal muscles, three new Z repeats (Zr) were detected, and Zr4 to 6 exclusion dominates in the fast muscles, whereas Zr4 skipping dominates in the slow muscle. No developmental changes were detected in the Z‐band. In the M‐band, two new variants were discovered with alternative 3′ splice site in exon363 (Mex5) and alternative 5′ splice site in intron 362. However, only the sheep heart expresses two new variants rather than other species. Skeletal muscles express three M‐band variants with altered ratios of Mex5 inclusion to Mex5 exclusion. Finally, we revealed that RBM20 does not regulate the Z‐ and M‐band splicing in the heart, but does in skeletal muscles. Taken together, we characterized the Z‐ and M‐band splicing and provided the first evidence of the role of RBM20 in the Z‐ and M‐band TTN splicing.
Apoptotic effect of resveratrol on human T‐ALL cell line CCRF‐CEM is unlikely exerted through alteration of BAX and BCL2 promoter methylation
Authors: Zadi Heydarabad Milad , Vatanmakanian Mousa , Abdolalizadeh Jalal , Mohammadi Hamed , Azimi Ako , Mousavi Ardehaie Reza , Movasaghpour Aliakbar , Farshdousti Hagh Majid
Published: August 2018

One of the fundamental barriers leading to failure of leukemia therapy is the resistance against conventional chemotherapies, common modality used to cure leukemia. Having the potential to trigger apoptosis in various human leukemia cell lines, resveratrol is regarded as a robust agent in chemotherapy regimens. The current study was aimed to assess whether the apoptotic effect of resveratrol on T‐cell acute lymphoblastic leukemia cell line, CCRF‐CEM, is exerted through DNA methylation of BAX and BCL2 gene promoters. For this purpose, the CCRF‐CEM cells were treated by resveratrol under standard cell culture. To analyze the promoter DNA methylation changes, we used methylation‐specific polymerase chain reaction technique following the resveratrol treatment at different dosages and time intervals. Based on our previous study, the resveratrol treatment can trigger apoptosis in CCRF‐CEM cell line via upregulation of apoptotic BAX gene and downregulation of antiapoptotic BCL2 gene. Despite these alterations in gene expression, the current study reveals no changes in DNA methylation patterns of subjected genes following the resveratrol treatment. Unchanged status of DNA methylation of BAX and BCL2 genes may suggest that resveratrol causes the gene expression changes through a distinct mechanism which requires further studies to be understood.
Retracted: Knockdown of long noncoding RNA urothelial carcinoma–associated 1 inhibits cell viability, migration, and invasion by regulating microRNA‐182 in gastric carcinoma
Authors: Qin Lei , Jia Zhihua , Xie Dawei , Liu Zhongyuan
Published: August 2018

Background

Long noncoding RNA urothelial carcinoma–associated 1 (UCA1) has been reported to be a vital mediator in various cancers. But, in terms of gastric cancer (GC), the effects of UCA1 on GC cell proliferation, migration, invasion, and apoptosis remain unclear. This study aimed to uncover the potential regulatory mechanism of UCA1 in GC cells.

Methods

The expression level of UCA1 was first examined in the five different cell lines of HEK293, CCL‐153, HUVEC, SUN‐216, and SGC‐7901 using a reverse‐transcriptase quantitative polymerase chain reaction. Then, the vectors of short hairpin UCA1, the microRNA‐182 (miR‐182) mimic/inhibitor, and the pEX‐tissue inhibitor of metalloproteinases 2 (TIMP2)/small interfering TIMP2 were transfected into SUN‐216 and SGC‐7901 cells to alter UCA1, miR‐182, and TIMP2 expression. To investigate the biological functions, cell viability, migration, invasion, and apoptosis were examined by Cell Counting Kit‐8, Transwell, and flow cytometry. The key factors of apoptosis and phosphoinositide 3‐kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase 3β (GSK3β) and nuclear factor κB (NF‐κB) signal pathways were determined by Western blot analysis.

Results

UCA1 was upregulated in SUN‐216 and SGC‐7901 cells than in the other three cell lines of HEK293, CCL‐153, and HUVEC. Knockdown of UCA1 significantly suppressed cell viability, migration, and invasion, and promoted apoptosis by regulating B‐cell lymphoma 2, Bax, and cleaved‐caspase‐3/9 expressions. However, miR‐182 overexpression markedly reversed the regulatory effect of UCA1 knockdown on SUN‐216 and SGC‐7901 cells. TIMP2 was a direct target gene of miR‐182, and TIMP2 overexpression exhibited the same effect of UCA1 knockdown on cell viability, migration, invasion, and apoptosis. Besides, miR‐182 activated PI3K/AKT/GSK3β and NF‐κB signal pathways by regulation of TIMP2.

Conclusion

Knockdown of UCA1 exerts an anticancer effect on GC cells by regulating miR‐182.
Effect of resveratrol on Sertoli cell proliferation
Authors: Gorga A , Rindone GM , Regueira M , Pellizzari EH , Camberos MC , Cigorraga SB , Riera MF , Galardo MN , Meroni SB
Published: August 2018

Resveratrol (RSV), a polyphenolic compound largely found in red grape skin, has been used as a nutritional supplement as it exhibits beneficial health effects, such as anticancer, cardioprotective, antiaging, and anti‐inflammatory. Particularly, it has been shown that it participates in the mechanisms involved in cell proliferation. Sirtuin 1 (SIRT1) is considered a well‐known RSV effector. Noteworthy, Sirt1‐knockout animals are infertile. The aim of this study was, first, to determine whether RSV has any effect on Sertoli cell proliferation and, second, whether SIRT1, a putative target of RSV, is present in immature Sertoli cells. Sertoli cell cultures obtained from 8‐day‐old rats, which actively proliferate, were treated with RSV (10 and 50 µM) under basal and follicle‐stimulating hormone (FSH)‐stimulated conditions. Bromodeoxyuridine (BrdU) incorporation and the expression of cyclins D1, D2, D3, E1, and E2 and the Cip/Kip cell cycle inhibitors p21Cip and p27Kip were analyzed. RSV decreased BrdU incorporation and cyclins D1, D2, E1, and E2 expression and increased p21Cip and p27Kip messenger RNA (mRNA) levels. RSV also decreased FSH‐stimulated BrdU incorporation and cyclins D1 and D2 mRNA levels. The effect of RSV on cMYC was also analyzed. RSV treatment did not modify basal and FSH‐stimulated cMyc expression; however, it inhibited basal and FSH‐stimulated cMYC transcriptional activity, suggesting a role of cMYC in RSV effects. Additionally, Sirt1 was detected in immature Sertoli cells. Altogether, these results suggest that RSV possibly, by activating SIRT1 and regulating cMYC transcriptional activity, participates in the regulation of immature Sertoli cell proliferation.
CASTOR1 suppresses the progression of lung adenocarcinoma and predicts poor prognosis
Authors: Zhou Xuefeng , Cheng Zhenshun , Chen Hao , Shi Shenqi , Wang Xianguo , Orang Matthew , Zhao Jinping
Published: August 2018

As a naturally occurring inhibitor of mammalian target of rapamycin (mTOR), accumulated evidence has confirmed that CASTOR1 plays a pivotal role in regulating the progression of human malignancies. However, the function of CASTOR1 in the development of lung adenocarcinoma is still unclear. Here we report that the expression of CASTOR1 is significantly reduced in lung adenocarcinoma cell lines as compared with that in normal lung epithelial cells. Cellular studies further revealed that ectopic CASTOR1 expression led to a significant suppression of the cellular proliferation, migration, and invasion of PC‐9 cells. To further test the role of CASTOR1 in human patients with lung cancers, tumor tissues derived from lung cancer patients and paired adjacent normal lung tissues were collected for immunohistochemical​​​​​​, biochemical, and molecular biology analysis. Immunoblotting and real‐time quantitative reverse transcription polymerase chain reaction analysis is revealed that CASTOR1 was significantly reduced in tumor tissues as compared with that in paired normal lung tissues. With immunostaining and retrospective analysis of pathological samples from lung cancer patients further revealed that expression of CASTOR1 was negatively correlated with smoking history, TNM stage, lymphnode metastasis, and distant metastasis. Furthermore, Kaplan‐Meier survival analysis indicated that patients with low CASTOR1 expression levels had a significantly worse 5‐year survival rate than those with high CASTOR1 expression. To further explore the molecular actions of CASTOR1 in lung cancer development, biochemical assays were performed and the results suggested that ectopic CASTOR1 expression resulted significant suppression of mTOR and downstream S6K phosphorylation, indicating that CASTOR1 might regulate the progression of lung adenocarcinoma by controlling mTOR activation. Thus, these findings demonstrated that CASTOR1 inhibits the tumorigenesis of lung adenocarcinoma cells and might serve as a potential therapeutic target or prognostic marker for human patients with lung adenocarcinoma.
Targeting axonal degeneration and demyelination using combination administration of 17β‐estradiol and Schwann cells in the rat model of spinal cord injury
Authors: Namjoo Zeinab , Mortezaee Keywan , Joghataei Mohammad T. , Moradi Fateme , Piryaei Abbas , Abbasi Yusef , Hosseini Amir , Majidpoor Jamal
Published: August 2018

Schwann cells (SCs) are known to be responsible for axonal ensheathing and myelination, and their transplantation is used commonly for treatment of spinal cord injury (SCI). 17β‐estradiol (E2) has also reported for its protective roles in neurons in the transplanted SCs to the SCI model. In the current study, we evaluated the roles of E2 administration before SCs transplantation in targeting SCI‐induced axonal degeneration and demyelination. E2 (25 µg/kg, IP) was administered to the male Wistar rats underwent contusive SCI at T10 segment. At 7 days after injury, 1 × 106 SCs were transplanted to the injury epicenter of the spinal cord. The groups were laminectomy, SCI, SCI+E2, and SCI+E2+SCs. Functional recovery was evaluated using the Basso‐Bresnahan‐Beattie locomotor test. Sections from spinal cord were also assessed for histoloical staining, including Luxol fast blue, Bielschowsky's silver and immunofluorescence evaluation of myelin basic protein (MBP). The SCI group showed impaired locomotion in the hind limb, increased number of cavities within spinal cord, low observable numbers of regenerating fibers, and a significant decrease in the rate of expression for MBP. These changes were counteracted in the treatment groups (
P < 0.05 vs SCI) with no significant changes among them. From the results, it may be concluded that application of E2 and SCs could be effective when axons undergo demyelination and degenerative processes, and their combination could partly provide cumulative outcomes.
Chrysophanol induces cell death and inhibits invasiveness via mitochondrial calcium overload in ovarian cancer cells
Authors: Lim Whasun , An Yikyung , Yang Changwon , Bazer Fuller W. , Song Gwonhwa
Published: August 2018

Chrysophanol is a phytochemical typically extracted from rhubarb. Similar to other extracts from rhubarb, chrysophanol possesses anticancer activity against diverse cancerous cells. However, the apoptotic effects of chrysophanol in ovarian cancer remain unknown. In the current study, we examined the antitumorigenic activity of chrysophanol in human epithelial ovarian cancer cells, such as ES2 and OVCAR3. Chrysophanol decreased cell viability and increased cell death in a dose‐dependent manner. In addition, chrysophanol markedly increased the intracellular reactive oxygen species production in ES2 cells, but only increased it slightly in OVCAR3 cells. Consistent with increased reactive oxygen species production, extensive lipid peroxidation was detected in chrysophanol‐treated ES2 cells compared with that in untreated cells, whereas lipid peroxidation was unchanged in OVCAR3 cells in response to chrysophanol. Although there were no significant changes in calcium ions in the of ES2 and OVCAR3 cells, the concentration of calcium in the mitochondria increased dose dependent through disruption of the mitochondrial membrane potential in both ES2 and OVCAR3 cells compared with nontreated control cells. Moreover, chrysophanol activated the MAPK signaling pathways in the ovarian cancer cells. In addition, ovarian cancer cell invasiveness was suppressed, which implied that chrysophanol plays a role in preventing ovarian cancer metastasis. In conclusion, chrysophanol exhibits an anticancer effect via mitochondrial calcium overload and MAPK activation, suggesting its potential as a novel anticancer agent for human epithelial ovarian cancer.
EGFL7 promotes hepatocellular carcinoma cell proliferation and inhibits cell apoptosis through increasing CKS2 expression by activating Wnt/β‐catenin signaling
Authors: Li Zhi , Xue Tong‐Qing , Yang Chao , Wang Yun‐Liang , Zhu Xiao‐Li , Ni Cai‐Fang
Published: August 2018

Epidermal growth factor‐like domain multiple 7 (EGFL7) is an important sport stimulating factor and motility related factors significantly enhanced the tumor cell metastasis and overexpressed in many cancers, including hepatocellular carcinoma (HCC), associated with tumorigenesis. However, the molecular mechanism by which EGFL7 regulates HCC cell proliferation and apoptosis and the correlation between EGFL7 and cyclin‐dependent kinases regulatory subunit 2 (CKS2), which is essential for biological function, have not fully explained. In this study, EGFL7 and CKS2 expression in patients with HCC was measured by real‐time polymerase chain reaction and immunohistochemistry. After HCC cells respectively transfected with pLKO.1‐EGFL7‐shRNA, pLVX‐Puro‐EGFL7 recombined vector or CKS2 small interfering RNA, cell counting kit‐8 and flow cytometry was performed to examine the cell proliferation and apoptosis, respectively, and the expression of β‐catenin, CKS2, CDK2, and cleaved caspase‐3 was measured by Western blot analysis. We found that EGFL7 and CKS2 were overexpressed in HCC tissues and a positive correlation was found between them. EGFL7 knockdown markedly inhibited proliferation and promoted apoptosis of HCC cells, along with decreased expression of CKS2 and CDK2, but increased cleaved caspase‐3 expression, while EGFL7 overexpression showed an opposite effect. EGFL7 silencing in nude mice also showed decreased tumor growth and altered protein expression similar to its effect in HCC cells in vitro. Importantly, CKS2 silencing significantly inhibited EGFL7‐induced HCC cell proliferation and protein expression, and Wnt/β‐catenin signaling pathway inhibitor IWR‐1‐endo significantly inhibited CKS2 expression in HCC cells. Taken together, EGFL7 promotes HCC cell proliferation and inhibits cell apoptosis through increasing CKS2 expression by activating Wnt/β‐catenin signaling.
TGF‐β/SMAD signaling inhibits intermittent cyclic mechanical tension‐induced degeneration of endplate chondrocytes by regulating the miR‐455‐5p/RUNX2 axis
Authors: Xiao Liang , Xu Shujuan , Xu Yongming , Liu Chen , Yang Bijing , Wang Jing , Xu Hongguang
Published: August 2018

A mechanical stimulation plays a pivotal role in maintaining normal cartilage function. Our objective was to reveal the mechanism of action of the tension‐sensitive molecule miR‐455‐5p in the degeneration of endplate chondrocytes and to identify whether the transforming growth factor beta (TGF‐β)/SMAD signaling pathway has a regulatory effect on it. The expression profiles of members of the TGF‐β/SMAD pathway, miR‐455‐5p, and RUNX2 were determined by microRNA microarray analysis, reverse transcription quantitative polymerase chain reaction, luciferase reporter assay, and Western blot analysis. Intermittent cyclic mechanical tension (ICMT) induced the degeneration of endplate chondrocytes without affecting their viability. The tension‐sensitive molecule miR‐455‐5p specifically bound to RUNX2, a gene involved in the degeneration of endplate chondrocytes. Activation of the TGF‐β/SMAD signaling pathway upregulated miR‐455‐5p expression and thus inhibited RUNX2 levels. Therefore, the TGF‐β/SMAD signaling pathway inhibits the ICMT‐induced degeneration of endplate chondrocytes by regulating the miR‐455‐5p/RUNX2 axis.
Ionic liquid/single‐walled carbon nanotubes composite film modified carbon‐ceramic electrode as an electrochemical sensor for the simultaneous determination of epinephrine and uric acid
Authors: Ayazi Zahra , Izadyar Shima , Habibi Biuck
Published: August 2018

In the present work, a carbon–ceramic electrode (CCE) modified by a composite of an ionic liquid and single‐walled carbon nanotubes (IL‐SWCNT/CCE) was prepared for the electrochemical determination of epinephrine (EP) in the presence of uric acid (UA). The morphology and surface characteristics of the prepared modified CCE were studied using scanning electron microscopy (SEM). Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were used for electrochemical studies and determinations. Their results showed that the IL‐SWCNT/CCE has high electrocatalytic activity toward the oxidation of EP. The electrochemical behavior of EP is closely dependent on the pH; in basic media, its oxidation process is irreversible and there is no cathodic peak in reverse scan, while in acidic media the electrochemical behavior of EP becomes reversible. DPV measurements showed that the oxidation peak current was linear in the concentration range 1.0 × 10−7 to 2.0 × 10−4 M, and the detection limit was 2.8 × 10−8 M. Also, it was confirmed that the IL‐SWCNT/CCE can be used for the simultaneous measurement of EP and UA. The obtained results indicate that the IL‐SWCNT/CCE has high stability, reproducibility, and repeatability. Finally, the developed electro‐analytical method was successfully applied for the determination of EP in human serum and urine samples without any interference.
Effect of a functional variant of tumor necrosis factor‐β gene in temporomandibular disorders: A pilot study
Authors: Yerliyurt Kaan , Nursal Ayse Feyda , Tekcan Akin , Karakus Nevin , Tumer Mehmet K. , Yigit Serbulent
Published: August 2018

Background

Temporomandibular disorders (TMD) are a group of conditions that cause chronic orofacial pain. The tumor necrosis factor β (TNF‐β) is a proinflammatory cytokine that is involved in the various aspects of the inflammatory process including organization and maintenance, and in the arrangement of cells at the inflammation site. The purpose of this study was to evaluate the correlation between TNF‐β +252A/G (rs909253) variant and susceptibility to TMD in a Turkish cohort.

Methods

The study included 104 patients (26 males, 78 females) with TMD and 126 healthy controls (44 males, 82 females). The TNF‐β +252A/G variant analysis was based on Polymerase Chain Reaction‐Restriction Fragment Length Polymorphism (PCR‐RFLP).

Results

There was no deviation from HWA for TNF‐β +252A/G variant in patient and control groups. There was significant difference in genotype and allele frequencies between patient group and control group in terms of TNF‐β +252A/G variant, respectively (P = 0.010, 0.015). A significant increase in the TNF‐β +252 AG genotype and G allele frequencies were observed in TMD patients compared to healthy controls. The individuals with GG genotype and G allele had an increased risk of developing TMD. A statistically significant association was observed when the patients were compared with the controls according to AA genotype vs AG+GG genotypes (P = 0.002, OR: 2.23, 95% CI:1.31‐3.82). TNF‐β +252A/G genotype distribution was associated with chewing problems (P = 0.046).

Conclusions

In conclusion, our results provided evidence that TNF‐β +252A/G variant may contribute to TMD development in a Turkish cohort. Further studies are needed to confirm this observation.
Comparative evaluation of in‐house Carba NP test with other phenotypic tests for rapid detection of carbapenem‐resistant Enterobacteriaceae
Authors: Bir Raunak , Mohapatra Sarita , Kumar Amarjeet , Tyagi Sonu , Sood Seema , Das Bimal Ku , Kapil Arti
Published: August 2018

Background

The prevalence of carbapenem‐resistant Enterobacteriaceae (CRE) is alarming worldwide causing serious infections. Rapid and accurate identification of CRE is crucial to reduce the mortality and morbidity. In this study, we tried to develop an in‐house Carba NP test for detection of CRE and evaluate its performance with others.

Methods

A prospective study was conducted with 40 nonrepeating Enterobacteriaceae isolates over a period of 3 months. All the isolates were screened for carbapenem resistance as per CLSI 2016 guidelines followed by PCR for blaNDM‐1, blaOXA‐48, blaKPC, blaVIM, and blaIMP genes. All the isolates were subjected to five phenotypic tests, that is, in‐house Carba NP (iCarba NP), commercial Carba NP (cCarba NP), Blue‐Carba, modified Hodge test (MHT), and CHROMagar.

Results

Among the 40 isolates, 87.5% were identified as Escherichia coli, 7.5% were Klebsiella pneumoniae, 2.5% were Enterobacter cloacae, and 2.5% were Citrobacter freundii. Thirty‐three of 40 (82.5%) isolates were found to harbor one or more resistant genes. Considering PCR to be the gold standard test, sensitivity of the phenotypic methods for CRE detection ranged from 63.6% (MHT) to 96.9% (CHROMagar). Both cCarba NP and iCarba NP observed to have highest specificity. The performance of iCarba NP was found comparable with cCarba NP by kappa score 1 and found approximately 10 times less expensive than cCarba NP.

Conclusion

CHROMagar was observed most sensitive assay for detection of CRE followed by both Carba NP tests. iCarba NP was proved cheaper and equally good as cCarba NP for detection of CRE.
Trust, sense of community, and civic engagement: Lessons from Australia
Authors: Miranti Riyana , Evans Mark
Published: August 2018

This article examines the association between trust, sense of community, and civic engagement, with a particular focus on the socioeconomic characteristics of individuals, using data from the Household, Income, and Labor Dynamics in Australia Survey. Through the use of panel logit and binary panel data models, we draw three core observations. First, there is evidence that trust is associated with civic engagement in Australia, particularly in the case of volunteering. Trust is a prerequisite for women to participate in volunteering but not for men, where a sense of community matters more. Second, a high sense of community matters for both types of participation, political and volunteering. Third, the relationships between trust, sense of community, and civic engagement are present among the Baby Boomer and Generation X generations but not the Generation Y generation, which participates differently. The study makes an important contribution to the literature by unmasking the gender and generation stories and debunking popular myths about the unwillingness of new Australians to engage in associative behavior.
Peptidyl‐prolyl cis–trans isomerase NIMA interacting 1 regulates skeletal muscle fusion through structural modification of Smad3 in the linker region
Authors: Islam Rabia , Yoon Heein , Shin Hye‐Rim , Bae Han‐Sol , Kim Bong‐Soo , Yoon Won‐Joon , Woo Kyung‐Mi , Baek Jeong‐Hwa , Lee Yun‐Sil , Ryoo Hyun‐Mo
Published: August 2018

Myoblast fusion is critical for muscle growth, regeneration, and repair. We previously reported that the enzyme peptidyl‐prolyl cis–trans isomerase NIMA interacting 1 (Pin1) is involved in osteoclast fusion. The objective of this study was to investigate the possibility that Pin1 also inhibits myoblast fusion. Here, we show the increased number of nuclei in the Pin1+/− mice muscle fiber compared to that in wild‐type mice. Moreover, we show that low dose of the Pin1 inhibitor dipentamethylene thiuram monosulfide treatment caused enhanced fusion in C2C12 cells. The R‐Smads are well‐known mediators of muscle hypertrophy and hyperplasia as well as being substrates of Pin1. We found that Pin1 is crucial for maintaining the stability of Smad3 (homologues of the Drosophila protein, mothers against decapentaplegic (Mad) and the Caenorhabditis elegans protein Sma). Our results show that serine 204 within Smad3 is the key Pin1‐binding site during inhibition of myoblast fusion and that both the transforming growth factor‐β receptor and extracellular signal‐regulated kinase (ERK)‐mediated phosphorylation are required for the interaction of Pin1 with Smad3. These findings suggest that a precise level of Pin1 activity is essential for regulating myoblast fusion during myogenesis and muscle regeneration.
Cytosolic sodium regulation in mouse cortical astrocytes and its dependence on potassium and bicarbonate
Authors: Noor Zinnia N. , Deitmer Joachim W. , Theparambil Shefeeq M.
Published: August 2018

Sodium plays a major role in different astrocytic functions, including maintenance of ion homeostasis and uptake of neurotransmitters and metabolites, which are mediated by different Na+‐coupled transporters. In the current study, the role of an electrogenic sodium‐bicarbonate cotransporter (NBCe1), a sodium‐potassium‐chloride transporter 1 (NKCC1) and sodium‐potassium ATPase (Na+‐K+‐ATPase) for the maintenance of [Na+]i was investigated in cultured astrocytes of wild‐type (WT) and of NBCe1‐deficient (NBCe1‐KO) mice using the Na+‐sensitive dye, asante sodium green‐2. Our results suggest that cytosolic Na+ was higher in the presence of CO2/HCO3− (15 mM) than CO2/HCO3−‐free, HEPES‐buffered solution in WT, but not in NBCe1‐KO astrocytes (12 mM). Surprisingly, there was a strong dependence of cytosolic [Na+] on the extracellular [HCO3−] attributable to NBCe1 activity. Pharmacological blockage of NKCC1 with bumetanide led to a robust drop in cytosolic Na+ in both WT and NBCe1‐KO astrocytes by up to 6 mM. There was a strong dependence of the cytosolic [Na+] on the extracellular [K+]. Inhibition of the Na+‐K+‐ATPase led to larger increase in cytosolic Na+, both in the absence of K+ as compared with the presence of ouabain and in NBCe1‐KO astrocytes as compared with WT astrocytes. Our results show that cytosolic Na+ in mouse cortical astrocytes can vary considerably and depends greatly on the concentrations of HCO3− and K+, attributable to the activity of the Na+‐K+‐ATPase, of NBCe1 and NKCC1.
Splicing factor poly(rC)‐binding protein 1 is a novel and distinctive tumor suppressor
Authors: Guo Jihua , Jia Rong
Published: August 2018

A lot of evidence has been found on the link between tumorigenesis and the aberrant expression of splicing factors. A number of splicing factors have been reported to be either oncogenic or overexpressed in cancer cells. However, splicing factors can also play negative roles in tumorigenesis. In the current review, we focus on splicing factor poly(rC)‐binding protein 1 (PCBP1), a novel tumor suppressor that is characterized by downregulation in many cancer types and shows inhibition of tumor formation and metastasis. Notably, the messenger RNA levels of PCBP1 are not significantly decreased in most cancer types. In fact, PCBP1 protein is often degraded or shows a loss‐of‐function through phosphorylation in cancer cells. PCBP1 is highly homologous to its family member, PCBP2. Interestingly, PCBP2 appears to be an oncogenic splicing factor. A growing body of evidence has shown that PCBP1 regulates alternative splicing, translation, and RNA stability of many cancer‐related genes. Taking together, PCBP1 has distinctive tumor suppressive functions, and increasing PCBP1 expression may represent a new approach for cancer treatment.
Thrombospondin‐4 critically controls transforming growth factor β1 induced hypertrophic scar formation
Authors: Qian Wei , Li Ning , Cao Qian , Fan Jufeng
Published: August 2018

Transforming growth factor β (TGF‐β) is a growth factor presenting important functions during tissue remodeling and hypertrophic scar (HS) formation. However, the underlying molecular mechanisms are largely unknown. In this study, we identified thrombospondin‐4 (TSP‐4) as a TGF‐β1 target that essentially mediates TGF‐β1‐induced scar formation both in vitro and in vivo. The expression of TSP‐4 was compared on both mRNA and protein levels between hypertrophic scar fibroblasts (HSFs) and normal skin fibroblast (NFs) in response to TGF‐β1 treatment. Two signaling molecules, Smad3 and p38, were assessed for their importance in regulating TGF‐β1‐mediated TSP‐4 expression. The significance of TSP‐4 in controlling TGF‐β1‐induced proliferation, invasion, migration, and fibrosis in HSFs was analyzed by knocking down endogenous TSP‐4 using small hairpin RNA (shRNA) (TSP‐4 shRNA). Finally, a skin HS model was established in rats and the scar formation was compared between rats treated with vehicle (saline), TGF‐β1, and TGF‐β1 + TSP‐4 shRNA. The TSP‐4 level was significantly higher in HSFs than in NFs and TGF‐β1 more potently boosted TSP‐4 expression in the former than in the latter. Both Smad3 and p38 essentially mediated TGF‐β1‐induced TSP‐4 expression. TSP‐4 shRNA significantly suppressed TGF‐β1‐stimulated proliferation, invasion, migration, or fibrosis of HSFs in vitro and drastically improved wound healing in vivo. TGF‐β1, by activating both Smad3 and p38, induces TSP‐4, which in turn not only presents a positive feedback regulation on the activation of Smad3 and p38, but also essentially mediates TGF‐β1‐induced HS formation. Targeting TSP‐4 thus may benefit HS treatment.
2 dyn/cm2 shear force upregulates kruppel‐like factor 4 expression in human chondrocytes to inhibit the interleukin‐1β‐activated nuclear factor‐κB
Authors: Chang Shun‐Fu , Huang Kuo‐Chin , Chang Hsin‐I , Lee Ko‐Chao , Su Yu‐Ping , Chen Cheng‐Nan
Published: August 2018

The shear force effect on human chondrocytes is time and magnitude dependent. Recently, kruppel‐like factor (KLF) 4 has been identified as a pleiotropic protein and its activity in cells is dependent on different stimuli and/or cell types. The role of KLF4 in chondrocytes is still unclear and there has been no report determining whether shear force regulates KLF4 levels in chondrocytes. Hence, this study was carried out to investigate the role of KLF4 in human chondrocytes under shear force stimulation and the underlying mechanism. Human primary and SW1353 chondrocytes were used in this study. The shear forces at 2, 5, or 15 dyn/cm2 intensity were applied to both types of human chondrocytes. The specific small interfering RNAs, activators, and inhibitors were used to study the detailed mechanism of shear force. The presented results showed that 2, but not 5 and 15, dyn/cm2 shear force increases KLF4 expression in human primary and SW1353 chondrocytes. Extracellular signal‐regulated kinase 5 induced peroxisome proliferator‐activated receptor γ transcription activity to increase KLF4 transcription. Moreover, the KLF4 induction in human chondrocytes in response to 2 dyn/cm2 shear force could attenuate interleukin (IL)‐1β‐stimulated nuclear factor‐κB activation. These results elucidate the role of KLF4 in antagonizing the effect of IL‐1β in human chondrocytes under 2 dyn/cm2 shear force stimulation and provide a possible mechanism to demonstrate the protection of moderate forces or exercises in cartilage.
Aggregation of human dental pulp cells into 3D spheroids enhances their migration ability after reseeding
Authors: Hsieh Hao‐Ying , Young Tai‐Horng , Yao Chung‐Chen , Chen Yi‐Jane
Published: August 2018

Multicellular three‐dimensional (3D) spheroids allow intimate cell–cell communication and cell–extracellular matrix interaction. Thus, 3D cell spheroids better mimic microenvironment in vivo than two‐dimensional (2D) monolayer cultures. The purpose of this study was to evaluate the behaviors of human dental pulp cells (DPCs) cultured on chitosan and polyvinyl alcohol (PVA) membranes. The protein expression of hypoxia‐inducible factor 1‐α (HIF‐1α) and vascular endothelial growth factor (VEGF), and the migration ability of the DPCs from 2D versus 3D environments were investigated. The results showed that both chitosan and PVA membranes support DPCs aggregation to form multicellular spheroids. In comparison to 2D cultures on tissue culture polystyrene, DPC spheroids exhibited higher protein expression of HIF‐1α and VEGF. The treatment with YC‐1 (inhibitor to HIF‐1α) blocked the upregulation of VEGF, indicating a downstream event to HIF‐1α expression. When DPC spheroids were collected and subjected to the transwell assay, the cells growing outward from 3D spheroids showed greater migration ability than those from 2D cultures. Moreover, DPCs aggregation and spheroid formation on chitosan membrane were abolished by Y‐27632 (inhibitor to Rho‐associated kinases), whereas the inhibitory effect did not exist on PVA membrane. This suggests that the mechanism regulating DPCs aggregation and spheroid formation on chitosan membrane is involved with the Rho‐associated kinase signaling pathway. In summary, the multicellular spheroid structure was beneficial to the protein expression of HIF‐1α and VEGF in DPCs and enhanced the migration ability of the cells climbing from spheroids. This study showed a new perspective in exploring novel strategies for DPC‐based research and application.
Exosome‐encapsulated miR‐6089 regulates inflammatory response via targeting TLR4
Authors: Xu Donghua , Song Meiyan , Chai Chunxiang , Wang Jinghua , Jin Chengwen , Wang Xiaodong , Cheng Min , Yan Shushan
Published: August 2018

Exosome‐encapsulated microRNAs (miRNAs) have been identified as potential biomarkers in autoimmune diseases. However, little is known about the role of exosome‐delivered miRNAs in rheumatoid arthritis (RA). In this study, we investigated the profile of specific exosomal miRNAs by microarray analysis of serum exosomes from three patients with RA and three healthy controls. Quantitative real‐time PCR (qRT‐PCR) was performed to validate the aberrantly expressed exosomal miRNAs. A total of 20 exosome‐encapsulated miRNAs were identified to be differently expressed in the serum of patients with RA compared with controls. Interestingly, we found that exosome‐encapsulated miR‐6089 was significantly decreased after validation by qRT‐PCR in serum exosomes from 76 patients with RA and 20 controls. Besides, miR‐6089 could inhibit lipopolysaccharide (LPS)‐induced cell proliferation and activation of macrophage‐like THP‐1 cells. TLR4 was a direct target for miR‐6089. MiR‐6089 regulated the generation of IL‐6, IL‐29, and TNF‐α by targetedly controlling TLR4 signaling. In conclusion, exosome‐encapsulated miR‐6089 regulates LPS/TLR4‐mediated inflammatory response, which may serve as a novel, promising biomarker in RA.