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Mitosis and spindle assembly require the microtubule-associated protein Xenopus kinesin-like protein 2 (TPX2). Although TPX2 is highly expressed in several malignant tumor forms, little is known about its role in cancer. In this study, we performed the gene set enrichment analysis of TPX2 in 33 types of cancers and an extensive pan-cancer bioinformatic analysis using prognosis, tumor mutational burdens, microsatellite instability, tumor microenvironment, and immune cell infiltration data. According to the differential expression study, TPX2 was found to be overexpressed across all studied cancer types. Based on the survival analysis, increased TPX2 expression was associated with a poor prognosis for most cancers. The TPX2 expression level was confirmed to correlate with the clinical stage, microsatellite instability, and tumor mutational burden across all cancer types. Furthermore, TPX2 expression has been linked to tumor microenvironments and immune cell infiltration, particularly in bladder urothelial carcinoma, liver hepatocellular carcinoma, lung adenocarcinoma, stomach adenocarcinoma, and uterine corpus endometrial carcinoma. Finally, the gene set enrichment analysis implicated TPX2 in the regulation of aminoacyl tRNA biosynthesis, which is the most important tumor cell cycle signaling pathway. This comprehensive pan-cancer analysis shows that TPX2 is a prognostic molecular biomarker for most cancers and suggests its potential as an effective therapeutic target for the treatment of these diseases.

Aim: This study aimed to develop a predictive model for patients with duodenal carcinoma. Methods: Duodenal carcinoma patients from the Surveillance, Epidemiology, and End Results database (2010–2015) and the First Affiliated Hospital of Nanchang University (2010–2021) were enrolled. A nomogram was constructed according to least absolute shrinkage and selection operator regression analysis, the Akaike information criterion approach and Cox regression analysis. Results: Five independent prognostic factors were significantly associated with the prognosis of the duodenal carcinoma patients. A nomogram was constructed with a C-index in the training and validation cohorts of 0.671 (95% CI: 0.578–0.716) and 0.662 (95% CI: 0.529–0.773), respectively. Conclusion: The established nomogram model provided visualization of the risk of each prognostic factor.

Gamma synuclein (SNCG) is a neuronal protein that is also aberrantly overexpressed in various types of human cancer. SNCG overexpression promotes cancer invasion and metastasis. However, the mechanisms that drive cancer metastasis upon SNCG expression remain elusive. Elucidation of the mechanisms underlying the promotion of cancer metastasis by SNCG may help discover therapeutic avenues for SNCG-overexpressed cancer. Here, we show that SNCG promotes transforming growth factor-β (TGF-β)-induced p38 mitogen-activated protein kinase (MAPK) phosphorylation. Mechanistically, SNCG promotes p38MAPK phosphorylation by interacting with the MAPK kinase 3/6 (MKK3/6) and prevents their degradation. SNCG knockdown leads to a decrease in TGF-β-induced phosphorylation of MKK3/6; and abrogates the induction of matrix metalloproteinase (MMP)-9 expression by TGF-β and its target gene Twist1. Furthermore, p38MAPK inhibition abrogates the promotion of MMP-9 expression and cancer cell invasion by SNCG. Both p38MAPK and MMP inhibitors can suppress the promotion of cancer cell invasion by SNCG. Finally, overexpression of SNCG in liver cancer cells promotes lung metastasis, which can be suppressed by the p38MAPK inhibitor. Together, our data uncover a previously unknown role of SNCG in promoting TGF-β-MKK3/6-p38MAPK signaling. This study highlights the critical role of p38MAPK in the promotion of cancer metastasis by SNCG, and indicates that p38MAPK inhibitor may serve as a potential therapeutic for SNCG-overexpressed cancer.

Background The study aimed to investigate the relationship between changes in corneal epithelial thickness and the outcome of myopic regression after femtosecond laser-assisted in situ keratomileusis (FS-LASIK) and transepithelial photorefractive keratectomy (TPRK). Methods This study included 45 eyes of 25 patients undergoing FS-LASIK and 44 eyes of 24 patients undergoing TPRK. Myopic regression occurred in these patients postoperatively from 8 to 21 months. The corneal epithelial thickness was measured using a spectral-domain optical coherence tomography at the onset of regression, 3 months after treatment, and 3 months after drug withdrawal. Results Compared with that of preoperation, corneal epithelial thickness increased when regression occurred in both groups (all P < 0.05). The thickness of central corneal epithelium in FS-LASIK and TPRK groups reached 65.02 ± 4.12 µm and 61.63 ± 2.91 µm, respectively. The corneal epithelial thickness decreased when myopic regression subsided after 3 months of steroid treatment compared to the onset (P < 0.05). With a decrease in corneal epithelial thickness, the curvature of the anterior corneal surface, central corneal thickness, and refractive power all decreased (all P < 0.05). The corneal epithelial thickness and refractive error remained relatively stable after 3 months of treatment withdrawal (P > 0.05). Conclusion The corneal epithelial thickness determined the outcome of myopic regression similarly in FS-LASIK and TPRK. When the corneal epithelium thickened, regression occurred. After steroid treatment, epithelial thickness decreased whereas regression subsided.

The C-terminal of G protein-coupled receptors is now recognized as being important for G protein activation and signaling function. To detect the role of C-terminal tail in receptor activation, we used the α1b-AR, which has a long C-terminal of 164 amino acids. We constructed the intramolecular FRET sensors, in which the C-terminal was truncated to 10 (∆C-10), 20 (∆C-20), 30 (∆C-30), 50 (∆C-50), 70 (∆C-70), or 90 (∆C-90). The truncated mutants of ∆C-10, ∆C-20, or ∆C-30 cannot induce FRET signal changes and downstream ERK1/2 phosphorylation. However, the truncated mutants of ∆C-50, ∆C-70, or ∆C-90 induce significant FRET signal changes and downstream ERK1/2 phosphorylation, especially ∆C-90. This is particularly true in the case of the ∆C-90, ∆C-70, or ∆C-50 which retained the potential phosphorylation sites (Ser401, Ser404, Ser408, or Ser410). The ∆C-90 showed an increase in agonist-induced FRET signal changes and ERK1/2 phosphorylation in PKC- or endocytosis-dependent and EGFR-, src-, or β-arrestin2-independent.

Illicit drug use in rural and suburban areas of China has not been studied extensively, as most studies have focused on illicit drug use in urban areas. To compare the differences between urban, suburban, and rural drug use, we collected influent samples from 19 urban, 9 suburban, and 18 rural wastewater treatment plants in Dalian, respectively. A method using solid-phase extraction combined with derivatization for gas chromatography -mass spectrometry analysis was applied to detect biomarker concentrations. The concentrations of methamphetamine and morphine ranged from 3.12 to 605 ng/L and

Virus-induced asthma is prevalent among children, but its underlying mechanisms are unclear. Accumulated evidence indicates that early-life respiratory virus infection increases susceptibility to allergic asthma. Nonetheless, the relationship between systemic virus infections, such as enterovirus infection, and the ensuing effects on allergic asthma development is unknown. Early-life enterovirus infection was correlated with higher risks of allergic diseases in children. Adult mice exhibited exacerbated mite allergen-induced airway inflammation following recovery from EV-A71 infection in the neonatal period. Bone marrow-derived macrophages (BMDMs) from recovered EV-A71-infected mice showed sustained innate immune memory (trained immunity) that could drive naïve T helper cells toward Th2 and Th17 cell differentiation when in contact with mites. Adoptive transfer of EV-A71-trained BMDMs induced augmented allergic inflammation in naïve recipient mice, which was inhibited by 2-deoxy-D-glucose (2-DG) pretreatment, suggesting that trained macrophages following enterovirus infection are crucial in the progression of allergic asthma later in life.

To compare the clinical outcomes, mainly including contrast sensitivity and high-order aberrations (HOAs), between wavefront-optimized (WFO) and corneal wavefront-guided (CWFG) transepithelial photorefractive keratectomy (transPRK) for preoperative HOAs0.35 μm.Qingdao Eye Hospital of Shandong First Medical University, Qingdao, Shandong, China.Prospective randomized controlled study.71 patients with preoperative total ocular and corneal aberrations0.35 μm who underwent transPRK for the treatment of myopia and myopic astigmatism were randomly divided into the aberration optimization mode group (WFO group; 36 eyes) and the corneal wavefront-guided mode group (CWFG group; 35 eyes). Preoperative and postoperative visual outcome, refraction, contrast sensitivity, and HOAs were compared.71 patients (71 eyes) who underwent transPRK were selected. The CWFG group had significantly lower total HOAs and coma values in the corneal aberration compared with the WFO group at 3 ( P = .009; P.001) and 6 months postoperatively ( P = .006; P.001). In addition, the CWFG group had significantly lower total HOAs and coma values in the whole-eye aberration compared with the WFO group at 3 ( P = .044; P = .004) and 6 months postoperatively ( P = .026; P = .001). The CWFG group had significantly better improvement in contrast sensitivity than the WFO group at spatial frequencies of 3 cycles per degree (cpd), 6 cpd, 12 cpd, and 18 cpd ( P = .005, P = .007, P = .001, and P.001, respectively).CWFG transPRK is associated with better visual and refractive outcomes and less HOAs than WFO transPRK in eyes with preoperative aberrations0.35 μm.

Purpose To compare higher-order aberrations (HOAs) after small incision lenticule extraction (SMILE) in patients with and without intraoperative angle kappa adjustments. Methods This is a retrospective case series. One hundred six eyes of 106 patients who underwent SMILE at Tianjin Eye Hospital (Tianjin Medical University, Tianjin, China) for correction of myopia and myopic astigmatism were divided into 2 groups. The first group consisted of eyes with intraoperative angle kappa adjustment and the second group consisted of eyes without adjustment. Preoperative and postoperative visual outcome, refraction, and HOA measurements at 1 and 3 months were compared. Results At the pupil size of 6 mm, vertical coma at 1 and 3 months after SMILE for the angle kappa-adjusted group was 0.153 ± 0.107 and 0.157 ± 0.094 μm, which were significantly lower than those of the nonadjusted group (0.204 ± 0.117 and 0.203 ± 0.113 μm, respectively) (P = 0.026 at 1 mo, P = 0.047 at 3 mo). The change in vertical coma between preoperative and postoperative measurements was 0.011 ± 0.136 and 0.023 ± 0.129 μm at 1 and 3 months postoperatively for the angle kappa-adjusted group, which were lower than those of the nonadjusted group (0.082 ± 0.165 and 0.085 ± 0.150 μm, respectively) (P = 0.023 at 1 mo, P = 0.045 at 3 mo). Subgroup analysis for eyes with large angle kappa demonstrated that the vertical coma was significantly less in the angle kappa-adjusted group at both 1 and 3 months (P = 0.009, P = 0.043, respectively). No significant correlation was observed between angle kappa and HOAs in the angle kappa-adjusted group. Conclusions Adjustment of angle kappa during SMILE resulted in less HOAs. It would provide more insight on how to optimize treatment centration in SMILE.

With rapid economic development, an increasing number of people suffer from mental health diseases, which are gradually receiving the attention of society. However, basic data from surveys of mental disorders are limited. Composite influent samples were collected from 26 wastewater treatment plants in 23 major cities in China. The concentrations of the psychoactive drugs diphenhydramine, fluoxetine, doxepin, imipramine, sulpiride, zolpidem, carbamazepine, and flunitrazepam in the wastewater were determined. The detection frequency of diphenhydramine, sulpiride, and carbamazepine was close to 100 %, whereas that of the compounds was lower than 35 %. Carbamazepine had the highest mean consumption (31.1 mg/d/1000 people), followed by diphenhydramine (10.4 mg/d/1000 people) and sulpiride (11.3 mg/d/1000 people). Wastewater-based epidemiology (WBE) estimates of the average use of the three drugs were lower than those from the drug statistics data. Consumption of diphenhydramine in northern China was higher than that in southern China. A correlation analysis of psychotropic and illicit drugs revealed a correlation between sulpiride and heroin use, which may be related to the adverse effects of sulpiride treatment after heroin withdrawal. Psychotropic drug use is associated with both economic and social factors. We found associations between the use of the three drugs and age, occupation, and obesity, which are risk factors for mental disorders. The results showed that the monitoring of psychotropic drug using WBE has a certain reference value for public health care and for improving the understanding of mental disorders.

Methylisothiazolinone (MIT) is frequently used as antimicrobial in household and industrial products, and poses ecological and health risks to aquatic organisms and humans. In this study, vacuum ultraviolet (VUV)/ultraviolet (UV) irradiation was found highly efficient for removal of MIT. The rate constant of MIT degradation (k

Sildenafil (SIL) is widely used to treat erectile dysfunction. Information on its consumption and the factors influencing its use is limited in China. In this study, we sampled composite influent wastewater samples from 33 Chinese cities and analyzed SIL using liquid chromatography-tandem mass spectrometry. SIL consumption was estimated using wastewater-based epidemiology (WBE) and ranged from 10.6 mg/d/1000 people to 132 mg/d/1000 people, with a mean of 53 mg/d/1000 people. Prescription sales (3570 kg) accounted for 13.3% of the estimated SIL use (26842 kg) in 2018, thereby implying that SIL illicit use was greater than prescription use in China. Some regional differences were observed in SIL use, which was significantly higher in North China than South China (p 0.05), thereby reflecting that the prevalence of SIL was affected by differences in lifestyle and socioeconomic factors. We found significant positive correlations between SIL use and consumption of allopurinol, hydrochlorothiazide, nicotine, and alcohol, thereby suggesting that the prevalence of SIL was associated with the prevalence of gout, hypertension, smoking, and drinking. Moreover, age structures, internet use, and marriage rates were positively correlated with SIL use, whereas the unemployment rate was negatively correlated with SIL use. Our study demonstrates that WBE is valuable for medical research to investigate licit and illicit drug use and to assess the underlying associations of different chemical uses.

Epstein-Barr virus (EBV)-associated gastric carcinoma (EBVaGC) is a distinct entity with a conspicuous tumor microenvironment compared with EBV-negative gastric carcinoma. However, the exact role of EBV in gastric carcinogenesis remains elusive. In the present study, we found that EBV upregulated CXCL8 expression, and CXCL8 significantly promoted vasculogenic mimicry (VM) formation of gastric carcinoma (GC) cells. In accordance with these observations, overexpression of CXCL8 increased cell proliferation and migration of AGS and BGC823 cells, while knockdown of CXCL8 with siRNA inhibited cell proliferation and migration of AGS-EBV cells. In addition, activation of NF-κB signaling was involved in VM formation induced by CXCL8, which was blocked by NF-κB inhibitors BAY 11-7082 and BMS345541. Furthermore, EBV-encoded lncRNA RPMS1 activated the NF-κB signaling cascade, which is responsible for EBV-induced VM formation. Both xenografts and clinical samples of EBVaGC exhibit VM histologically, which are correlated with CXCL8 overexpression. Finally, CXCL8 is positively correlated with overall survival in GC patients. In conclusion, EBV-upregulated CXCL8 expression promotes VM formation in GC

Metformin has been widely used as an oral drug for the treatment of diabetes mellitus. However, its consumption can be influenced by many economic and social factors. In this study, we investigated the spatial consumption pattern of metformin in China through wastewater-based epidemiology (WBE) approach. Influent wastewater samples were collected from 21 wastewater treatment plants (WWTPs) in 19 cities of the northeast China. A method using solid-phase extraction combined with N-Methyl-bis (trifluoroacetamide) derivatization for GC-MS detection was applied for metformin analysis. In 21 days, metformin showed high stability in wastewater at 24 °C and −20 °C. The mean concentrations of metformin in all WWTPs ranged from 2.42 μg L−1 to 53.6 μg L−1. The consumption of metformin was 0.66–15.6 mg d−1 capita−1 with the mean value of 5.54 ± 4.28 mg d−1 capita−1. The prevalence of metformin ranged from 0.09% to 2.10% with an average of 0.74%. Both the consumption and prevalence of metformin displays significant spatial variations in northeast China. A statistical correlation analysis indicated that the consumption of metformin increases with the decrease of per capita disposable income of urban residents. To further predict the use of metformin in China, we developed a regress model and depicted a consumption map. The annual consumption of urban residents in Chinese provinces range from 1085-63,828 kg yr−1 with mean value of 25,347 kg yr−1, which would provide a certain reference value for public health care and diabetes control.

Epstein-Barr virus (EBV) is a tumor virus that is associated with a variety of neoplasms, including EBV-associated gastric carcinoma (EBVaGC). Recently, EBV was reported to generate various circular RNAs (circRNAs). CircRNAs are important regulators of tumorigenesis by modulating the malignant behaviors of tumor cells. However, to date, the functions of ebv-circRNAs in EBVaGC remain poorly understood. In the present study, we observed high ebv-circRPMS1 expression in EBVaGC and showed that ebv-circRPMS1 promoted the proliferation, migration, and invasion and inhibited the apoptosis of EBVaGC cells. In addition, METTL3 was upregulated in GC cells overexpressing ebv-circRPMS1. Mechanistically, ebv-circRPMS1 bound to Sam68 to facilitate its physical interaction with the METTL3 promotor, resulting in the transactivation of METTL3 and cancer progression. In clinical EBVaGC samples, ebv-circRPMS1 was associated with distant metastasis and a poor prognosis. Based on these findings, ebv-circRPMS1 contributed to EBVaGC progression by recruiting Sam68 to the METTL3 promoter to induce METTL3 expression. ebv-circRPMS1, Sam68, and METTL3 might serve as therapeutic targets for EBVaGC.

Penisarins A (

Purpose The aim of this study was to assess the impact of cataract progression using the Haigis formula-calculated intraocular lens (IOL) power and investigate the accuracy of IOL power measured at different time points. Methods This prospective study was performed on 75 eyes of 75 patients who underwent uneventful cataract surgery. Preoperative ocular parameters including axial length (AL), keratometry (K), anterior chamber depth (ACD), corneal astigmatism, corrected distance visual acuity (CDVA) and uncorrected distance visual acuity (UDVA) examined at the two time points, more than 3 months preoperatively and preoperative 1 day were compared. The ocular parameters measured in the two time points were used to calculate the predicted implanted IOL power and the actual IOL power was chosen on the basis of parameters measured earlier before surgery using the Haigis formula. The mean numerical error (MNE) and mean absolute error (MAE) predicted by the two time points were also compared. Results There were significant differences in the ACD, IOL power, UDVA and CDVA (P0.05) during the average of 5.6 months before surgery. No statistically significant difference was detected in MNE (P>0.05), while the MAE had a significant difference in the two time points (P Conclusion The IOL power measured earlier before surgery might result in a higher accuracy and the postoperative refractive outcome tended towards emmetropia.

Despite aggressive treatment approaches, muscle-invasive bladder urothelial carcinoma (MIBC) patients still have a 50% chance of developing general incurable metastases. Therefore, there is an urgent need for candidate markers to enhance diagnosis and generate effective treatments for this disease. We evaluated four mRNA microarray datasets to find differences between non-MIBC (NMIBC) and MIBC tissues. Through a gene expression profile analysis via the Gene Expression Omnibus database, we identified 56 differentially expressed genes (DEGs). Enrichment analysis of gene ontology, Kyoto Encyclopedia of Genes and Genomes, and Reactome pathways revealed the interactions between these DEGs. Next, we established a protein-protein interaction network to determine the interrelationship between the DEGs and selected 10 hub genes accordingly. Bladder urothelial carcinoma (BLCA) patients with COL1A2, COL5A1, and COL5A2 alterations showed poor disease-free survival rates, while BLCA patients with COL1A1 and LUM alterations showed poor overall survival rates. Oncomine analysis of MIBC versus NMIBC tissues showed that COL1A1, COL5A2, COL1A2, and COL3A1 were consistently among the top 20 overexpressed genes in different studies. Using the TCGAportal, we noted that the high expression of each of the four genes led to shorter BLCA patient overall survival. It was evident that BLCA patients with an elevated high combined gene expression had significantly shorter overall survival and relapse-free survival than those with low combined gene expression using PROGgeneV2. Using Gene Expression Profiling Interactive Analysis, we noted that COL1A1, COL1A2, COL3A1, and COL5A2 were positively correlated with each other in BLCA. These genes are considered as clinically relevant genes, suggesting that they may play an important role in the carcinogenesis, development, invasion, and metastasis of MIBC. However, considering we adopted a bioinformatic approach, more research is crucial to confirm our results. Nonetheless, our findings may have important prospective clinical implementations.

To explore the efficacy difference of electroacupuncture at lowerA total of 63 patients with gastroparesis were randomly divided into a lowerAfter treatment, the total GCSI scores, TElectroacupuncture at lower

c-Met hyperactivity has been observed in numerous neoplasms. Several researchers have shown that the abnormal activation of c-Met is mainly caused by transcriptional activation. However, the molecular mechanism behind this transcriptional regulation is poorly understood. Here, we suggest that Smad3 negatively regulates the expression and activation of c-Met via a transcriptional mechanism. We explore the molecular mechanisms that underlie Smad3-induced c-Met transcription inhibition. We found in contrast to the high expression of c-Met, Smad3 showed low protein and mRNA levels. Smad3 and c-Met expressions were inconsistent between lung cancer tissues and cell lines. We also found that Smad3 overexpression suppresses whereas Smad3 knockdown significantly promotes Epithelial-Mesenchymal Transition and production of the angiogenic factors VEGF, CTGF and COX-2 through the ERK1/2 pathway. In addition, Smad3 overexpression decreases whereas Smad3 knockdown significantly increases protein and mRNA levels of invasion-related β-catenin and FAK through the PI3K/Akt pathway. Furthermore, using the chromatin immunoprecipitation analysis method, we demonstrate that a transcriptional regulatory complex consisting of HDAC1, Smad3 and mSin3A binds to the promoter of the c-Met gene. By either silencing endogenous mSin3A expression with siRNA or by pretreating cells with a specific HDAC1 inhibitor (MS-275), Smad3-induced transcriptional suppression of c-Met could be effectively attenuated. These results demonstrate that Smad3-induced inhibition of c-Met transcription depends on of a functional transcriptional regulatory complex that includes Smad3, mSin3A and HDAC1 at the c-Met promoter. Collectively, our findings reveal a new regulatory mechanism of c-Met signaling, and suggest a potential molecular target for the development of anticancer drugs.

We fabricated nanomaterials comprising amino-functionalized and nitrogen-doped graphene quantum dots (amino-N-GQDs) and investigated their photostability and intrinsic luminescence in the near-infrared spectrum to determine their suitability as contrast agents in two-photon imaging (TPI). We observed that amino-N-GQDs with a higher amount of bonded nitrogen and amino-functionalized groups (6.2%) exhibited superior two-photon properties to those with a lower amount of such nitrogen and groups (4.9%). These materials were conjugated with polymers containing sulfur (polystyrene sulfonate, PSS) and nitrogen atoms (polyethylenimine, PEI), forming amino-N-GQD&ndash, PSS&ndash, PEI specimens (amino-N-GQD-polymers). The polymers exhibited a high quantum yield, remarkable stability, and notable two-photon properties and generated no reactive oxygen species, rendering them excellent two-photon contrast agents for bioimaging. An antiepidermal growth factor receptor (AbEGFR) was used for labeling to increase specificity. Two-photon imaging (TPI) of amino-N-GQD (6.2%)-polymer-AbEGFR-treated A431 cancer cells revealed remarkable brightness, intensity, and signal-to-noise ratios for each observation at a two-photon excitation power of 16.9 nJ pixel&minus, 1 under 30 scans and a three-dimensional (3D) depth of 105 &micro, m, indicating that amino-N-GQD (6.2%)-polymer-AbEGFR-treated cells can achieve two-photon luminescence with 71 times less power required for two-photon autofluorescence (1322.8 nJ pixel&minus, 1 with 500 scans) of similar intensity. This economy can minimize photodamage to cells, rendering amino-N-GQD-polymers suitable for noninvasive 3D bioimaging.

Gene editing is a genetic manipulation technology which utilizes bacterial nucleases to accurately and efficiently modify DNA or RNA. Gene editing has broad applications in basic research, breeding, and drug screening, and it is gaining validity and applicability to the therapy of many diseases especially genetic-based disease. In this review, we summarize the development of gene editing technology, its different strategies and applications in the treatment of disease, and the research of gene editing therapy for genetic diseases (including base editor and epigenetic regulation) in the treatment of disorders and diseases of the blood system, liver, muscle and nervous system. Finally, we discuss the future development prospects of gene editing therapy.

KRAS is a member of the murine sarcoma virus oncogene-RAS gene family. It plays an important role in the prevention, diagnosis and treatment of tumors during tumor cell growth and angiogenesis. KRAS is the most commonly mutated oncogene in human cancers, such as pancreatic cancers, colon cancers, and lung cancers. Detection of KRAS gene mutation is an important indicator for tracking the status of oncogenes, highlighting the developmental prognosis of various cancers, and the efficacy of radiotherapy and chemotherapy. However, the efficacy of different patients in clinical treatment is not the same. Since RNA interference (RNAi) technologies can specifically eliminate the expression of specific genes, these technologies have been widely used in the field of gene therapy for exploring gene function, infectious diseases and malignant tumors. RNAi refers to the phenomenon of highly specific degradation of homologous mRNA induced by double-stranded RNA (dsRNA), which is highly conserved during evolution. There are three classical RNAi technologies, including siRNA, shRNA and CRISPR-Cas9 system, and a novel synthetic lethal interaction that selectively targets KRAS mutant cancers. Therefore, the implementation of individualized targeted drug therapy has become the best choice for doctors and patients. Thus, this review focuses on the current status, future perspective and associated challenges in silencing of KRAS with RNAi technology.

Monitoring the consumption of pharmaceuticals and licit drugs is important for assessing the needs of public health owing to the impact on individuals as well as society. The present work applied wastewater-based epidemiology to profile the spatial patterns of metformin, nicotine, and caffeine use and their correlations. Influent wastewater samples were collected from 27 wastewater treatment plants in 22 typical Chinese cities that covered all geographic regions of the country. The consumption of metformin ranged from 0.02 g/d/1000 inh to 8.92 g/d/1000 inh, whereas caffeine and nicotine consumption ranged from 4.33 g/d/1000 inh to 394 g/d/1000 inh and 0.17 g/d/1000 inh to 1.88 g/d/1000 inh, respectively. There were significant regional differences in the consumption of caffeine, with the highest consumption in East China and the lowest consumption in Northeast China. The consumption and concentration of caffeine were related to the gross domestic product and per capita disposable income of urban residents, respectively. There was a correlation between the concentrations of caffeine and cotinine (a nicotine metabolite), thereby indicating that individuals that use one of these substances are likely to use the other substance. A significant relationship was found between the concentration of metformin and cotinine, thereby implying that the use of tobacco may be correlated with type 2 diabetes. Co-analysis of these substances in wastewater may provide a more accurate picture of substance use situations within different communities and provide more information on human health, human behavior, and the economy. This report describes the newest study related to the consumption of metformin among the general population in China.

Background Wastewater-based epidemiology (WBE) has become a useful tool in long-term or short-term continuous monitoring of illicit drugs consumption over the world. Methods We investigated the trend of methamphetamine (METH) use between 2015 and 2018 through WBE in Dalian, a typical Chinese city. Samples were collected in 11 municipal wastewater treatment plants (WWTPs). An analytical method, solid-phase extraction combined with trifluoroacetic anhydride derivatization prior to gas chromatography–mass spectrometry (GC–MS) analysis was applied to detect METH concentrations. Results During the sampling period, the METH concentrations increased slowly from 315 ± 243 ng/L in 2015 to 523 ± 549 ng/L in 2016, followed by a significant decrease with the concentrations 188 ± 187 ng/L in 2017 and 54.6 ± 42.9 ng/L in 2018. Ammonium nitrogen (NH4-N) was applied to estimate population size. The average coefficient of variation for population in 11 WWTPs was 35.3 ± 8.9%, reflecting the dynamic variations of population effectively. For METH consumption, there was a gradual increase from 2015 (231 mg/day/1000 people) to 2016 (414 mg/day/1000 people) and a significant linear decrease to 2017 (206 mg/day/1000 people) and 2018 (53.9 mg/day/1000 people). The prevalence of METH increased from 2015 (0.78%) to 2016 (1.06%), then decreased to 2017 (0.55%) and 2018 (0.17%), showed similar trends with the consumption. Conclusions The obvious reduction trends of METH consumption via WBE over the period in Dalian provides objective evidence for declined METH consumption in local population. The reduction is probably due to the severe crack-down of illicit drugs by the government.

To classify the dietary patterns in preschoolers and explore the correlation between dietary patterns and executive functions.Between April 2014 and June 2015, 12363 preschoolers were selected from 91 kindergartens of Ma 'anshan City. The classification of dietary patterns and executive functions by Brief-P were evaluated. Unconditional binary logistics regression model was used to analyze the effects of different dietary patterns and executive functions in preschool chidlren.The higher the parents' education level, the lower the executive functions scores, the difference was statistically significant in preschoolers( P0. 05). The less intake of "processed model", "drinks model"and "snacks model", the more "vegetarian model"and "health model"intake, executive functions score was lower, and the difference was statistically significant( P0. 05). Logistic regression analysis showed that dietary patterns of"processed model"( OR = 1. 44, 95% CI 1. 20-1. 72), "drinks model"( OR = 1. 22, 95% CI 1. 03-1. 46) and "snacks model"( OR = 1. 28, 95% CI 1. 08-1. 52) were risk factors of cognitive functions, and the dietary patterns of "vegetarian model"( OR = 0. 80, 95% CI 0. 67-0. 95) and "health model "( OR = 0. 84, 95% CI 0. 71-0. 98) were protective factors.The present study suggests that cognitive functions might be closely related with dietary pattern. The more "vegetarian model"and "health model"intake, the better cognitive functions developed in preschool children.

With the improvement of living standard, gestational diabetes mellitus (GDM) incidence is increasing every year. We observed the effects of abnormal 75 g oral glucose tolerance test (OGTT) at different time points on neonatal complications and neurobehavioral development in GDM. A total of 144 newborns whose mothers were diagnosed with GDM and received prenatal examination and childbirth in our hospital from October 2015 to April 2016, were observed in this study. Pregnant women underwent 75 g OGTT and the blood glucose level was recorded on an empty stomach, as well as postprandial 1 and 2 hours, respectively. Based on the frequency of 75 g OGTT-abnormal time points, the pregnant women were divided into group 1 (OGTT abnormality at 1 time point), group 2 (OGTT abnormality at 2 time points), and group 3 (OGTT abnormality at 3 time points). Neonatal behavioral neurological assessment (NBNA) was performed on the 3 groups, respectively. In the total score of NBNA, there was a significant difference among the 3 groups (F = 17.120, P = .000), and there were significant differences between the 3 groups (all P

Mammalian target of rapamycin (mTOR) is a core component of raptor-mTOR (mTORC1) and rictor-mTOR (mTORC2) complexes that control diverse cellular processes. Both mTORC1 and mTORC2 regulate several elements downstream of type I insulin-like growth factor receptor (IGF-IR) and insulin receptor (InsR). However, it is unknown whether and how mTOR regulates IGF-IR and InsR themselves. Here we show that mTOR possesses unexpected tyrosine kinase activity and activates IGF-IR/InsR. Rapamycin induces the tyrosine phosphorylation and activation of IGF-IR/InsR, which is largely dependent on rictor and mTOR. Moreover, mTORC2 promotes ligand-induced activation of IGF-IR/InsR. IGF- and insulin-induced IGF-IR/InsR phosphorylation is significantly compromised in rictor-null cells. Insulin receptor substrate (IRS) directly interacts with SIN1 thereby recruiting mTORC2 to IGF-IR/InsR and promoting rapamycin- or ligand-induced phosphorylation of IGF-IR/InsR. mTOR exhibits tyrosine kinase activity towards the general tyrosine kinase substrate poly(Glu-Tyr) and IGF-IR/InsR. Both recombinant mTOR and immunoprecipitated mTORC2 phosphorylate IGF-IR and InsR on Tyr1131/1136 and Tyr1146/1151, respectively. These effects are independent of the intrinsic kinase activity of IGF-IR/InsR, as determined by assays on kinase-dead IGF-IR/InsR mutants. While both rictor and mTOR immunoprecitates from rictor(+/+) MCF-10A cells exhibit tyrosine kinase activity towards IGF-IR and InsR, mTOR immunoprecipitates from rictor(-/-) MCF-10A cells do not induce IGF-IR and InsR phosphorylation. Phosphorylation-deficient mutation of residue Tyr1131 in IGF-IR or Tyr1146 in InsR abrogates the activation of IGF-IR/InsR by mTOR. Finally, overexpression of rictor promotes IGF-induced cell proliferation. Our work identifies mTOR as a dual-specificity kinase and clarifies how mTORC2 promotes IGF-IR/InsR activation.

Transforming growth factor β (TGF-β) is critical for embryonic development, adult tissue homeostasis, and tumor progression. TGF-β suppresses tumors at early stage, but promotes metastasis at later stage through oncogenes such as Twist1. Gamma-synuclein (SNCG) is overexpressed in a variety of invasive and metastatic cancer. Here, we show that TGF-β induces SNCG expression by Smad-Twist1 axis, thus promoting TGF-β- and Twist1-induced cancer cell migration and invasion. We identify multiple Twist1-binding sites (E-boxes) in SNCG promoter. Chromatin immunoprecipitation and luciferase assays confirm the binding of Twist1 to the E-boxes of SNCG promoter sequence (−129/−1026 bp). Importantly, the Twist1-binding site close to the transcription initiation site is critical for the upregulation of SNCG expression by TGF-β and Twist1. Mutations of Twist1 motif on the SNCG promoter constructs markedly reduces the promoter activity. We further show that TGF-β induces Twist1 expression through Smad thereby enhancing the binding of Twist1 to SNCG promoter, upregulating SNCG promoter activity and increasing SNCG expression. SNCG knockdown abrogates TGF-β- or Twist1-induced cancer cell migration and invasion. Finally, SNCG knockdown inhibits the promotion of cancer metastasis by Twist1. Together, our data demonstrate that SNCG is a novel target of TGF-β-Smad-Twist1 axis and a mediator of Twist1-induced cancer metastasis.

Integrin αV gene expression is often dysregulated in cancers especially in hepatocellular carcinoma (HCC); however, the mechanism of regulation is poorly understood. Here, it is demonstrated that sulfatide activated integrin αV gene transcription, through histone H3K9/14 acetylation at the promoter, and high integrin αV expression are closely associated with poor prognosis. To elucidate the mechanism of regulation of acetylation, sulfatide-bound proteins were screened by mass spectrometry (MS), and bromodomain containing protein 1 (BRD1) was identified as an interacting protein that also colocalized with sulfatide in HCC cells. BRD1 was also formed a complex with Sp1, which was recruited to the integrin αV gene promoter. Sulfatide was also found to induce BRD1, monocytic leukemia zinc finger (MOZ) and histone acetyltransferase binding to ORC1 (HBO1) acetyltransferase multiprotein complex recruitment to the integrin αV promoter, which is responsible for histone H3K9/14 acetylation. Finally, knockdown of BRD1 limited sulfatide-induced H3K9/14 acetylation and occupancy of MOZ or HBO1 on integrin αV gene promoter. Implications: This study demonstrates that sulfatide interaction with BRD1 mediates acetylation and is important for regulation of integrin αV gene expression. Mol Cancer Res; 16(4); 610–22. ©2018 AACR.

The natural agent rhein is an ananthraquinone derivative of rhubarb, which has anticancer effects. To determine the mechanisms underlying the anticancer effects of rhein, we detected the effect of rhein on several oncoproteins. Here, we show that rhein induces β‐catenin degradation in both hepatoma cell HepG2 and cervical cancer cell Hela. Treatment of HepG2 and Hela cells with rhein shortens the half‐life of β‐catenin. The proteasome inhibitor MG132 blunts the downregulation of β‐catenin by rhein. The induction of β‐catenin degradation by rhein is dependent on GSK3 but independent of Akt. Treatment of HepG2 and Hela cells with GSK3 inhibitor or GSK3β knockdown abrogates the effect of rhein on β‐catenin. GSK3β knockdown compromises the inhibition of HepG2 and Hela cell growth by rhein. Furthermore, rhein dose not downregulate β‐catenin mutant that is deficient of phosphorylation at multiple residues including Ser33, Ser37, Thr41 and Ser45. Moreover, rhein induces cell cycle arrest at S phase in both HepG2 and Hela cells. Intraperitoneal administration of rhein suppresses tumour cells proliferation and tumour growth in HepG2 xenografts model. Finally, the levels of β‐catenin are reduced in rhein‐treated tumours. These data demonstrate that rhein can induce β‐catenin degradation and inhibit tumour growth.

c-Jun N-terminal kinases (JNK) are members of the mitogen-activated protein kinase (MAPK) family that have important roles in signal transduction. The small molecule SP600125 is widely used in biochemical studies as a JNK inhibitor. However, recent studies indicate that SP600125 may also act independent of JNK. Here, we report that SP600125 can induce Src, type I insulin-like growth factor receptor (IGF-IR), Akt and Erk1/2 phosphorylation. Notably, these effects are independent of its inhibition of JNK. Inhibition of Src abrogates the stimulation of IGF-IR, Akt and Erk1/2 phosphorylation. IGF-IR knockdown blunts the induction of both Akt and Erk1/2 phosphorylation by SP600125. Moreover, combination of SP600125 and the Src inhibitor saracatinib synergistically inhibits cell proliferation. We conclude that SP600125 can activate Src-IGF-IR-Akt/Erk1/2 signaling pathways independent of JNK.

Background Attention-deficit/hyperactivity disorder (ADHD) among children is an increasing public health concern. The identification of behavioral risk factors, including sleep quality, has important public health implications for prioritizing behavioral intervention strategies for ADHD. Herein, this study aimed to investigate the prevalence of high levels of ADHD symptoms and to explore the association between sleep schedules, sleep-related problems and ADHD symptoms among preschoolers aged 3 to 6 years in mainland China. Methods A cross-sectional study was conducted, comprising a large sample of 15,291 preschoolers in Ma’anshan city of Anhui Province in China. ADHD symptoms were assessed by the 10-item Chinese version of the Conners Abbreviated Symptom Questionnaire (C-ASQ). Sleep-related variables included caregivers’ responses to specific questions addressing children’s daytime and nighttime sleep schedules, as well as sleep-related behaviors. Data on other factors were also collected, such as socio-demographic characteristics, TV viewing duration on weekdays and weekends, and outdoor activities. Logistic regression models were used to analyze the relationships between sleep schedules, sleep-related problems and ADHD symptoms. Results Approximately 8.6% of the total sample of preschoolers had high levels of ADHD symptoms, with boys having higher levels than girls (9.9% vs. 7.2%). In the logistic regression analysis, after adjusting for TV viewing duration, outdoor activities, and socio-demographic characteristics, delayed bedtime was significantly associated with a risk of high levels of ADHD symptoms, with odds ratios (OR) of 2.50 [95% confidence interval (CI): 2.09 ~ 3.00] and 2.04 (95% CI: 1.72 ~ 2.42) for weekdays and weekends, respectively. Longer time falling asleep (≥ 31 min) (OR = 1.76, 95% CI: 1.47 ~ 2.11), no naps (OR = 1.57, 95% CI: 1.34 ~ 1.84) and frequent sleep-related problems (OR = 4.57, 95% CI: 3.86 ~ 5.41) were also significantly associated with an increased risk of high levels of ADHD symptoms, while longer sleep duration (> 8.5 h) was associated with a decreased risk of high levels of ADHD symptoms (OR = 0.76, 95% CI: 0.67~ 0.87). Conclusions ADHD symptoms are prevalent in preschoolers in Ma’anshan region, China. Undesirable sleep schedules and sleep-related problems among preschoolers confer a risk of ADHD symptoms, highlighting the finding that beneficial and regular sleep habits potentially attenuate ADHD symptoms among preschoolers.

To investigate the related influencing factors of preschool children's emotional and behavioral problems in early life and explore the associations between the symptoms of depression or anxiety during pregnancy and emotional and behavioral problems in preschool children.Based on the Ma'anshan Birth Cohort Study of the China-Anhui Birth Cohort Study (C-ABCS), women were recruited at their first clinical visit between October 2008 and October 2010 in four municipal medical and health institutions of Ma'anshan City, a total of 5 084 pregnant women and 4 669 singletons live births were included in the birth cohort. Women completed measures of depressive (Self-Rating Anxiety scale) and anxious (Center for Epidemiologic Studies Depression) symptoms in pregnancy. By the age of 3-6 follow-up, 3 653 children were followed with completed information between April 2014 and April 2015, strengths and difficulties questionnaires were used to assessed offspring emotional and behavioral problems. Logistics regression was used to investigate the relationship between the symptoms of depression or anxiety during pregnancy and emotional and behavioral problems in preschool children.The detected rates of emotional symptoms, conduct problems, hyperactivity and peer problems in preschool children were 6.3% (229/3 653), 7.5% (274/3 653), 7.6% (278/3 653) and 2.8% (103/3 653), while 7.6% (277/3 653) for total difficulties, 10.9%(398/3 653) for prosocial behavior and 27.4%(981/3 557) for impact respectively. Prevalence of anxiety and depression in the first trimester was 2.7%(100/3 653) and 4.7%(171/3 653) respectively, and in the second trimester was 2.0%(66/3 375) and 3.6%(122/3 375) respectively. After we controlled the confoundings of gestation age, place of residence, family income, maternal education, paternal education, premature birth and folic acid supplement before pregnancy, multinomial logistic regression analysis showed that the risk of children's emotional symptoms in maternal anxiety in both first-trimester and second-trimester group was higher than the group of no depression and anxiety symptoms, and OR(95%CI) was 5.90(2.00-17.48). Compared with whose mother no depression in both first-trimester and second-trimester, the risk of children's emotional symptoms in maternal depression in both first-trimester and second-trimester group was higher, and OR(95% CI) was 3.07 (1.30-7.28). And the risk of children's total difficulties of maternal anxiety in second-trimester was 2.27 (95%CI: 1.10-4.71) times of no anxiety in second-trimester. While the risk of children's total difficulties of maternal depression in second-trimester was 2.20 (95%CI: 1.24-3.93) times of no depression in second-trimester. Maternal emotional symptoms were not significant associations with conduct problems, hyperactivity, peer problems and prosocial behaviors (P0.05).There was a negative impact of maternal anxiety and depression symptoms during pregnancy on emotional and behavioral problems in preschool children. These findings highlight the need for additional clinical and research attention to both maternal depression and anxiety in pregnancy, which may be helpful to reduce the incidence of children's emotional and behavioral problems and act as an important measure in prevention.

To analyze the associations between pregnancy-related anxiety and the prevalence of subthreshold autism trait (SAT) in preschool children.Baseline data came from the Ma'anshan Birth Cohort Study, a part of the China-Anhui Birth Cohort Study (C-ABCS). All the participants were enrolled among pregnant women who received prenatal health care in 4 municipal medical centers during Oct. 2008 to Oct. 2010. A total of 5 084 pregnant women were recruited at the beginning and 4 669 singleton live births were included until childbirth. The situation about pregnancy-specific anxiety during trimester and third trimester of women were evaluated by Pregnancy-specific Anxiety Questionnaire (PAQ). Between April 2014 and April 2015, the cohort was followed up again, and the Clancy Autism Behavior Scale (CABRS) filled out by parents was used for telling the SAT children from the healthy children among 3 663 preschool children. Univariate and binary regression model was used to estimate associations between the pregnancy-related anxiety during trimester and third trimester and the subthreshold autism trait in children.During the pregnancy, the detected rates of women with pregnancy-specific anxiety in trimester and the third trimester were 25.5%(935/3 663), 13.9%(501/3 592) respectively, and the detected rate of maternal pregnancy-specific anxiety in both periods was 7.7%(278/3 592). There were 290 positive children with SAT and the detection rate was 7.9%. After controlling possible confounding factors including children's genders, place of residence, supplement folic acid during pregnancy, preterm birth, exposure to second-hand smoke during pregnancy, the father (mother) cultural levels, the father (mother) nature of work and family income, the results of multinomial logistic regression analysis showed that maternal pregnancy-specific anxiety in trimester was the risk factor for SAT in preschool children (OR=1.51, 95% CI: 1.11-2.04), and there was no association between maternal pregnancy-specific anxiety in the third trimester and SAT in preschool children (OR=1.36, 95% CI: 0.82-2.22). Compared with the single function of maternal pregnancy-specific anxiety in trimester or the third trimester for SAT in preschool children, maternal pregnancy-specific anxiety in both periods presented a joint action that increasing the risk for SAT (OR=2.02, 95% CI: 1.36-2.98).Maternal pregnancy-related anxiety was a risk factor for subthreshold autism trait in preschooler children. Pregnant women should try to keep a good mental state to create a good environment for fetal growth.

To examine the effect of maternal pre-pregnancy body mass index (BMI) and gestational weight gain (GWG) with childhood overweight and adiposity, and to explore possible early life risk factors for obesity in preschool children.Basic information of pregnant women and gestation period came from the Ma'anshan Birth Cohort Study, a part of the China-Anhui Birth Cohort Study (C-ABCS). Pregnant women in routine health care from four municipal medical and health institutions were enrolled voluntarily during October 2008 and October 2010 in Ma'anshan City. A total of 5 084 pregnant women and 4 669 singleton live births were included in this study. Between April 2014 and April 2015, 3 797 children were followed up. Children whose BMI were85th percentiles for age and genders of World Health Organization (WHO) reference were considered as overweight, and95th percentiles for age and genders cut-off values were considered as obesity (pathological and secondary causes of obesity were excluded). Gestational weight gain was defined according to the Institute of Medicine (IOM) guidelines. Univariate and binary regression model analysis was used to examine the effect of pre-pregnancy BMI and GWG with childhood overweight and adiposity.Of the 3 797 pregnant women, the prevalence of underweight, normal weight, overweight and obesity were respectively 22.6% (n=858), 70.3% (n=2 671), 6.2% (n=234) and 0.9% (n=34). There were 3 563 pregnant women who were obtained gestational weight gain data, the prevalence of inadequate GWG, appropriate GWG, excessive GWG were respectively 12.4% (n=443), 25.9% (n=922) and 61.7% (n=2 198). The prevalence of overweight and obesity were 11.5% (n=437) and 10.8% (n= 411) in preschool children, respectively. After adjusting confounding factors including age at delivery, genders of children, children age, birth weight, breastfeeding and household economic status, binary logistic regression analysis showed that pre-pregnancy overweight and obesity(OR=2.01, 95% CI: 1.53-2.65), excessive GWG(OR=1.65, 95% CI: 1.35-2.03) were risk factors for overweight and obesity, and pre-pregnancy underweight was protective factor for childhood overweight and obesity (OR=0.49, 95% CI: 0.39-0.62). Joint associations of pre-pregnancy BMI and inappropriate GWG were also noticed in the study: compared to only pre-pregnancy higher BMI or excessive GWG or indequate GWG, combination of high pre-pregnancy BMI and excessive GWG or high pre-pregnancy BMI and inadequate GWG, adverse effects on childhood overweight and obesity were much higher,OR (95%CI) values were 2.90(1.97-4.28), 3.17(1.44-6.97) respectively.Both high pre-pregnancy BMI and inappropriate GWG are associated with greater offspring BMI. Pregnant women should achieve appropriate weight gain and help prevent obesity in their children.

To examine the effect of parents' occupational and life exposure during six months before pregnancy on executive function of preschool children.Pregnant women involved in the study came from the Ma'anshan Birth Cohort Study,a part of the China-Anhui Birth Cohort Study. Between October 2008 and October 2010, pregnant women who accepted pregnancy care in four municipal medical and health institutions in Ma'anshan city were recruited as study objects. A total of 5,084 pregnant women and 4,669 singleton live births entered in this cohort. Between April 2014 and April 2015, a total of 3,803 pre-school children were followed up. Finally, except 32 preschool children did not have EF evaluation result, there were 3,771 children included in this study. By using self-designed " Maternal health handbook", we researched parents' general demographic characteristics, and life and occupational exposure during six months before pregnancy. To research preschool children's executive function, we used the Behavior Rating Inventory of Executive Function-Preschool Version (BRIEF-P). Univariate and multivariate statistical method was used to analyze the association of parents' life and occupational exposure during six months before pregnancy and preschool children's EF.3,771 preschool children's detected rate of inhibitory self-control index (ISCI), flexibility index (FI), emergent metacognition index (EMI) and global executive composite (GEC) dysplasia were 4.8% (182), 2.3% (88), 16.5% (623) and 8.6% (324) respectively. During six months before pregnancy, children whose parents were lived in a noise environment (OR=1.86, 95% CI: 1.36-2.54), whose maternal were exposed to pesticides were the risk of ISCI dysplasia(OR=3.60, 95% CI: 1.45-8.95). During six months before pregnancy, children whose maternal were exposed to pesticides (OR=6.72, 95% CI: 2.50-18.07) and whose father were exposed to occupational lead (OR=2.10, 95% CI: 1.25-3.54) were the risk of FI dysplasia. During six months before pregnancy, children whose parents were lived in a noise environment (OR=1.42, 95%CI: 1.18-1.71) and whose father were exposed to occupational lead (OR=1.30, 95%CI: 1.02-1.65) were the risk of EMI dysplasia. During six months before pregnancy, children whose parents were lived in a noise environment (OR=1.58, 95% CI: 1.24-2.01) and whose maternal were exposed to pesticides (OR=2.39, 95% CI: 1.02-5.58) were the risk of GEC dysplasia.The development of executive function is worse among preschool children whose parents live in noise environment, mother exposed to pesticides, and father exposed to occupational lead during six months before pregnancy.

Human plasma membrane-associated sialidase (Neu3) is one of several sialidases that hydrolyze sialic acids in the terminal position of the carbohydrate groups of glycolipids and glycoproteins. Neu3 is mainly localized in plasma membranes and plays crucial roles in the regulation of cell surface functions. In this study, we investigated the effects and molecular mechanisms of Neu3 on cell invasion and migration in vivo and in vitro. Initially, we found that the levels of Neu3 expression were higher in prostate cancer tissues and cell lines than in normal prostate tissues based on RT-PCR and Western blotting analyses. We then applied a Neu3 siRNA approach to block Neu3 signaling using PC-3M cells as model cells. Transwell invasion assays and wound assays showed significantly decreased invasion and migration potential in the Neu3 siRNA-transfected cells. RT-PCR and Western blotting analyses revealed that Neu3 knockdown decreased the expressions of the matrix metalloproteinases MMP-2 and MMP-9. In vivo, mice injected with PC-3M cell tumors were evaluated by SPECT/CT to determine the presence of bone metastases. Mice treated with attenuated Salmonella carrying the Neu3 siRNA developed fewer bone metastases than mice treated with attenuated Salmonella carrying a control Scramble siRNA, attenuated Salmonella alone or PBS. The results for bone metastasis detection by pathology were consistent with the data obtained by SPECT/CT. Tumor blocks were evaluated by histochemical, RT-PCR and Western blotting analyses. The results revealed decreased expressions of MMP-2 and MMP-9 at the mRNA and protein levels. Taken together, the present findings suggest that Neu3 is a promising molecular target for the prevention of prostate cancer metastasis.

The development of malignant prostate cancer involves multiple genetic alterations. For example, alterations in both survivin and p53 are reported to have crucial roles in prostate cancer progression. However, little is known regarding the interrelationships between p53 and survivin in prostate cancer. Our data demonstrate that the expression of survivin is inversely correlated with that of wtp53 protein (r(s)=0.548) in prostate cancer and in normal prostate tissues. We have developed a therapeutic strategy, in which two antitumor factors, small interfering RNA-survivin and p53 protein, are co-expressed from the same plasmid, and have examined their effects on the growth of PC3, an androgen-independent prostate cancer cell line. When p53 was expressed along with a survivin-specific short hairpin RNA (shRNA), tumor cell proliferation was significantly suppressed and apoptosis occurred. In addition, this combination also abrogated the expression of downstream target molecules such as cyclin-dependent kinase 4 and c-Myc, while enhancing the expression of GRIM19. These changes in gene expression occurred distinctly in the presence of survivin-shRNA/wtp53 compared with control or single treatment groups. Intratumoral injection of the co-expressed construct inhibited the growth and survival of tumor xenografts in a nude mouse model. These studies revealed evidence of an interaction between p53 and survivin proteins plus a complex signaling network operating downstream of the wtp53-survivin pathway that actively controls tumor cell proliferation, survival and apoptosis.

Aberrant activation of NF-kappaB has been proposed as the major cause of chemoresistance in lung cancer. Low-dose chemotherapeutic agents with limited toxicity and achieving profoundly enhanced efficacy by blocking NF-kappaB activation may be a useful strategy in cancer therapy. Thus, this study was performed to explore the effect of parthenolide, a natural NF-kappaB inhibitor, on human lung cancer A549 cells treated with low-dose oxaliplatin, as well as to determine the potential mechanisms involved. We incubated A549 cells with different concentrations of parthenolide in the absence or presence of a low-dose of oxaliplatin for 48 h. Then, cell proliferation was determined by MTT assay, and flow cytometry was used to study apoptosis. PGE(2) production in culture supernatants was detected by competitive ELISA, while expression of NF-kappaB/p65, COX-2, caspase-3 and caspase-9 proteins were analyzed by Western blot. Finally, compared to parthenolide or oxaliplatin alone, significant improvements in cell apoptosis and growth inhibition indexes were observed in the combined treatment. NF-kappaB/p65, COX-2, and PGE(2) expression were suppressed by the co-application; meanwhile, caspase-3 and caspase-9 proteins were obviously activated. These findings indicate that parthenolide could markedly enhance sensitivity of A549 cells to low-dose oxaliplatin by inhibiting NF-kappaB activation and inducing apoptosis. Parthenolide in combination with a low dose of oxaliplatin may be a beneficial chemotherapeutic strategy for patients who cannot tolerate the severe side effects of the drug at therapeutic concentrations.

AflatoxinB1 (AFB1) is well known as a potent carcinogen. Epidemiological studies have shown an association between AFB1 exposure and lung cancer in humans. AFB1 can induce the mutations of genes such as tumor suppressor p53 through its metabolite AFB1-8,9-exo-epoxide, which acts as a mutagen to react with DNA. In addition, recent study demonstrates AFB1 positively regulates type I insulin-like growth factor receptor (IGF-IR) signaling in hepatoma cells. The current study aims to determine the effects of AFB1 on Src kinase and insulin receptor substrate (IRS) in lung cancer cells and the effects of AFB1 on lung cancer cell migration. To this end, the effects of AFB1 on IRS expression, Src, Akt, and ERK phosphorylation were measured by Western blot analysis. The migration of lung cancer cells was detected by wound-healing assay. AFB1 downregulates IRS1 but paradoxically upregulates IRS2 through positive regulation of the stability of IRS2 and the proteasomal degradation of IRS1 in lung cancer cell lines A549 and SPCA-1. In addition, AFB1 induces Src, Akt, and ERK1/2 phosphorylation. Treatment of lung cancer cells with Src inhibitor saracatinib abrogates AFB1-induced IRS2 accumulation. Moreover, AFB1 stimulates lung cancer cell migration, which can be inhibited by saracatinib. We conclude that AFB1 may upregulate IRS2 and stimulate lung cancer cell migration through Src.

To determine the inhibitory effect of the synthetic STAT3 siRNA on the expression of STAT3 gene in human laryngeal cancer cell lines Hep2 and to investigate the effect of STAT3 siRNA on growth and apoptosis in Hep2 cells. A pair of DNA templates coding siRNA against STAT3-mRNA was synthesized to reconstruct plasmid of pSilencer1.0-U6 siRNA-STAT3. Hep2 cells were transfected with RPMI-1640 media (untreated), plasmid (empty), and STAT3 siRNA, respectively. Northern blot and Western blot analysis of STAT3 and pTyr-STAT3 expression in Hep2 cells and Western blot analysis of Bcl-2 expression in the Hep2 cell was performed 72 h after transfection. MTT, flow cytometry, and AO/EB assay were used for determination of cells proliferation and apoptosis in Hep2 cells. pTyr-STAT3 was markedly expressed in untreated Hep2 cells and the vector-treated Hep2 cells, whereas pTyr-STAT3 expression was significantly reduced in STAT3 siRNA-transfected Hep2 cells, indicating that STAT3 siRNA inhibited the activity of STAT3. Transfection of Hep2 cells with STAT3 siRNA significantly inhibited STAT3 expression at both mRNA and protein level in Hep2 cells and the inhibition was characterized by time-dependent transfection. Treatment of Hep2 cells with STAT3 siRNA resulted in dose-dependent growth inhibition of Hep2, this significantly increased apoptotic cell rate, and decreased Bcl-2 expression level in Hep2 cells. STAT3 siRNA had an effect on induction of either early or late stage apoptosis. This study demonstrates that STAT3 siRNA effectively inhibits STAT3 gene expression in Hep2 cells leading to growth suppression and induction of apoptosis in Hep2 cells. The use of siRNA technique may provide a novel therapeutic approach to treat laryngeal cancer and other malignant tumors expressing constitutively activated STAT3.

It has been demonstrated that shape, area and depth of the optic disc are relevant indices of diabetic retinopathy. In this paper, we present a new fundus optic disc localization and segmentation method based on phase congruency (PC). Firstly, in order to highlight the optic disc, channel images with the highest contrast between optic disc and background are selected in LAB, YUV, YIQ and HSV spaces respectively. Secondly, with the use of PC, features of four selected channel images can be extracted. Multiplication operation is then used to enhance PC detection results. Thirdly, window scanning and gray accumulating are utilized to locate the optic disc. Finally, iterative OTSU automatic threshold segmentation and Hough transform are performed on location images, before the final optic disc segmentation result can be obtained. The experimen- tal results showed that the proposed method can effectively and accurately perform optic disc location and segmentation.

Glycogen synthase kinase-3 (GSK3) has either tumor-suppressive roles or pro-tumor roles in different types of human tumors. A number of GSK3 targets in diverse signaling pathways have been uncovered, such as tuberous sclerosis complex subunit 2 and β-catenin. The O subfamily of forkhead/winged helix transcription factors (FOXO) is known as tumor suppressors that induce apoptosis. In this study, we find that FOXO binds to type I insulin-like growth factor receptor (IGF-IR) promoter and stimulates its transcription. GSK3 positively regulates the transactivation activity of FOXO and stimulates IGF-IR expression. Although kinase-dead GSK3β cannot up-regulate IGF-IR, the constitutively active GSK3β induces IGF-IR expression in a FOXO-dependent manner. Serum starvation or Akt inhibition leads to an increase in IGF-IR expression, which could be blunted by GSK3 inhibition. GSK3β knockdown or GSK3 inhibitor suppresses IGF-I-induced IGF-IR, Akt, and ERK1/2 phosphorylation. Moreover, knockdown of GSK3β or FOXO1/3/4 leads to a decrease in cellular proliferation and abrogates IGF-I-induced hepatoma cell proliferation. These results suggest that GSK3 and FOXO may positively regulate IGF-I signaling and hepatoma cell proliferation.

Recently, microRNAs (miRNA), small noncoding RNAs, have taken center stage in the field of human molecular oncology. However, their roles in tumor biology remain largely unknown. According to the assumption that miRNAs implicated in a specific tumor phenotype will show aberrant regulation of their target genes, we introduce an approach based on the miRNA target–dysregulated network (MTDN) to prioritize novel disease miRNAs. Target genes have predicted binding sites for any miRNA. The MTDN is constructed by combining computational target prediction with miRNA and mRNA expression profiles in tumor and nontumor tissues. Application of the proposed method to prostate cancer reveals that known prostate cancer miRNAs are characterized by a greater number of dysregulations and coregulators and the tendency to coregulate with each other and that they share a higher proportion of targets with other prostate cancer miRNAs. Support vector machine classifier, based on these features and changes in miRNA expression, is constructed and gives an average overall prediction accuracy of 0.8872 in cross-validation tests. The classifier is then applied to miRNAs in the MTDN. Functions enriched by dysregulated targets of novel predicted miRNAs are closely associated with oncogenesis. In addition, predicted cancer miRNAs within families or from different families show combinatorial dysregulation of target genes, as revealed by analysis of the MTDN modular organization. Finally, 3 miRNA target regulations are verified to hold in prostate cancer cells by transfection assays. These results show that the network-centric method could prioritize novel disease miRNAs and model how oncogenic lesions are mediated by miRNAs, providing important insights into tumorigenesis. Mol Cancer Ther; 10(10); 1857–66. ©2011 AACR.

Our objective was to evaluate cell growth and death effects by inhibiting Murine Double Minute 2 (MDM2) expression in human prostate cancer cells overexpressing the wild-type (WT) p53 gene. Prostate PC-3 tumor cells were transfected with a plasmid containing either mdm2 small interfering (Si-mdm2) or the WT p53 gene (Pp53) alone, or both (Pmp53), using Lipofectamine in vitro and attenuated Salmonella enterica serovar Typhi vaccine strain Ty21a (Salmonella Typhi Ty21a) in vivo. Cell growth, apoptosis, and the expression of related genes and proteins were examined in vitro and in vivo by flow cytometry and Western blot assays. We demonstrated that human prostate tumors had increased expression of MDM2 and mutant p53 proteins. Transfection of the PC-3 cells with the Pmp53 plasmid in vitro offered significant inhibition of cell growth and an increase in apoptotic cell death compared with that of the Si-mdm2 or Pp53 group. These effects were associated with up-regulation of p21 and down-regulation of hypoxia-inducible factor 1α expression in Pmp53-transfected cells. To validate the in vitro findings, the nude mice implanted with PC-3 cells were treated with attenuated Salmonella Typhi Ty21a carrying the plasmids, which showed that the Pmp53 plasmid significantly inhibited the tumor growth rate in vivo compared with that of the Si-mdm2 or Pp53 plasmid alone. Tumor tissues from mice treated with the Pmp53 plasmid showed increased expression of p21 and decreased expression of hypoxia-inducible factor 1α proteins, with an increased apoptotic effect. These results suggest that knockdown of mdm2 expression by its specific small interfering RNA with overexpression of the WT p53 gene offers synergistic inhibition of prostate cancer cell growth in vitro and in vivo.

Synergistic regulations among multiple microRNAs (miRNAs) are important to understand the mechanisms of complex post-transcriptional regulations in humans. Complex diseases are affected by several miRNAs rather than a single miRNA. So, it is a challenge to identify miRNA synergism and thereby further determine miRNA functions at a system-wide level and investigate disease miRNA features in the miRNA–miRNA synergistic network from a new view. Here, we constructed a miRNA–miRNA functional synergistic network (MFSN) via co-regulating functional modules that have three features: common targets of corresponding miRNA pairs, enriched in the same gene ontology category and close proximity in the protein interaction network. Predicted miRNA synergism is validated by significantly high co-expression of functional modules and significantly negative regulation to functional modules. We found that the MFSN exhibits a scale free, small world and modular architecture. Furthermore, the topological features of disease miRNAs in the MFSN are distinct from non-disease miRNAs. They have more synergism, indicating their higher complexity of functions and are the global central cores of the MFSN. In addition, miRNAs associated with the same disease are close to each other. The structure of the MFSN and the features of disease miRNAs are validated to be robust using different miRNA target data sets.

To investigate the effects of polysaccharide fraction of Cordyceps sinensis (PSCS) on triptolide (TPL)-induced apoptosis in the HL-60 cells and the involved molecular mechanism.The cultured leukemia HL-60 cells were divided into three groups: control group, TPL group (cells were treated with 5 ng/ml TPL only), and PSCS+TPL cells group (cells treated with 5 ng/ml TPL and 100 microg/ml or 200 microg/ml PSCS for 18 h). Cell viability was tested by MTT assay and apoptotic cells were quantitatively measured by flow cytometry with Annexin V/PI double stain.The expressions of Caspase-3, 6, 7, 9 and NF-kappa B proteins were tested by Western blot.MTT assay showed that different concentrations of PSCS inhibited the cell viability. Flow cytometry indicated that TPL markedly increased the apoptosis rate of the HL-60 cells, and PSCS enhanced the apoptosis in a dose-dependent manner. Western blot showed that TPL did not inhibit the expression of the Caspase-3, 6, 7, 9 and NF-kappa B proteins, and when cells were treated with PSCS, the expression of proteins decreased with the PSCS concentration rising.PSCS can enhance TPL-induced apoptosis in HL-60 cells and inhibit the expression of NF-kappa B and Caspase 3,6,7,9,which might be the possible signaling pathway of inducing apoptosis.

Purpose: Persistent activation of signal transducers and activators of transcription 3 (Stat3) and its overexpression contribute to the progression and metastasis of several different tumor types. For this reason, Stat3 is a reasonable target for RNA interference–mediated growth inhibition. Blockade of Stat3 using specific short hairpin RNAs (shRNA) can significantly reduce prostate tumor growth in mice. However, RNA interference does not fully ablate target gene expression in vivo, owing to the idiosyncrasies associated with shRNAs and their targets. To enhance the therapeutic efficacy of Stat3-specific shRNA, we applied a combination treatment involving gene associated with retinoid-IFN–induced mortality 19 (GRIM-19), another inhibitor of STAT3, along with shRNA.Experimental Design: The coding sequences for GRIM-19, a cellular STAT3-specific inhibitor, and Stat3-specific shRNAs were used to create a dual expression plasmid vector and used for prostate cancer therapy in vitro and in mouse xenograft models in vivo.Results: The coexpressed Stat3-specific shRNA and GRIM-19 synergistically and more effectively suppressed prostate tumor growth and metastases when compared with treatment with either single agent alone.Conclusion: The simultaneous use of two specific, but mechanistically different, inhibitors of STAT3 activity exerts enhanced antitumor effects.