One-pot synthesis and enzyme-responsiveness of amphiphilic doxorubicin prodrug nanomicelles for cancer therapeutics.

In this study, we report a one-pot synthesis and enzyme-responsiveness of polyethylene glycol (PEG) and glutamic acid (Glu)-based amphiphilic doxorubicin (DOX) prodrug nanomicelles for cancer therapeutics. The nanomicelles were accomplished by esterification and amidation reactions. The nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR) data confirmed the structure of nanomicelles. The DOX-loaded nanomicelles showed a DLS-measured average size of 107 nm and excellent stability in phosphate-buffered saline (PBS) for 7 days. The drug loading and cumulative release rates were measured by ultraviolet-visible (UV-vis) spectrophotometry at 481 nm. The cumulative release rate could reach 100% in an enzyme-rich environment. Further, the therapeutic efficiency of nanomicelles to cancer cells was determined by cell viability and cellular uptake and distribution using HeLa cells. The cell viability study showed that the DOX-loaded nanomicelles could effectively inhibit the HeLa cell proliferation. The cellular uptake study confirmed that the nanomicelles could be effectively ingested by HeLa cells and distributed into cell nuclei. Based on the collective experimental data, this study demonstrated that the synthesized nanomicellar prodrug of DOX is a potential candidate for cancer therapeutics.

microRNA-4270-5p inhibits cancer cell proliferation and metastasis in hepatocellular carcinoma by targeting SATB2.

Hepatocellular carcinoma (HCC) remains a lethal cancer type for both males and females. MicroRNAs (miRNAs) contribute to the initiation, development and metastasis of cancer. Although several miRNAs have been identified as drivers or suppressors of HCC, the molecular mechanisms of many miRNAs have not been investigated. Currently, we discovered that miR-4270-5p was a significantly downregulated miRNA in HCC. We revealed that miR-4270-5p overexpression inhibited cell proliferation and invasion of HCC cells. The data manifested that miR-4270-5p directly targeted SATB2, a key regulator of epithelial mesenchymal transition (EMT), in HCC cells and reversed the EMT process. The rescue experiments suggested that SATB2 overexpression reversed the biological function of miR-4270-5p in HCC cells. Clinical data indicated that SATB2 expression was negatively correlated with miR-4270-5p levels in HCC patients. Our findings provided potential targets for prognosis and treatment of patients with HCC.

Comparisons of prebiotic activity of polysaccharides from shoot residues of bamboo (Chimonobambusa quadrangularis) via different ethanol concentrations.

Three polysaccharide fractions from bamboo shoot (Chimonobambusa quadrangularis), CPS70, CPS75, and CPS80, were prepared using a final ethanol concentration of 70%, 75%, and 80% in the precipitation process. In vitro digestibility and the prebiotic activity of CPS70, CPS75, and CPS80 were evaluated and compared. The results indicated that all three of the CPS fractions exhibit a high degree of nondigestibility to human gastric juice (>98.5%) or α-amylase hydrolysis (>94.5%). Compared with the blank control, the three CPS fractions could not only significantly (p < .05) stimulate the proliferation of B. adolescentis, B. infantis, B. bifidum, and L. acidophilus, but also significantly (p < .05) enhance the production of lactic, acetic, propionic, and butyric acids when these polysaccharides were added as alternative carbon sources to glucose during the in vitro fermentation of four probiotics. Furthermore, when comparing the three CPS fractions, CPS75 displayed the strongest prebiotic potential, as this polysaccharide had the strongest effect on the proliferation of probiotic bacteria as well as the greatest effect on SCFAs production. These results demonstrated that the concentration of ethanol used during the precipitation process has a significant impact on the prebiotic activity of CPS. PRACTICAL APPLICATIONS: Ethanol precipitation is the first step when extracting polysaccharides from aqueous extracts as it is simple, rapid, and easy to carry out. This study focuses on how different concentrations of ethanol used in the precipitation process affect the prebiotic potential of bamboo shoot (Chimonobambusa quadrangularis) polysaccharides (CPS). The result indicated that the concentration of ethanol used during the precipitation process has a significant impact on the prebiotic activity of CPS. To our knowledge, it is the first to evaluate the effects of the concentration of ethanol during the process of precipitation on prebiotic potential of polysaccharides, which can subsequently be applied to the optimization of ethanol concentration when precipitating natural polysaccharides for the purpose of in vitro fermentation.

Improving the Metabolic and Mental Health of Children with Obesity: A School-Based Nutrition Education and Physical Activity Intervention in Wuhan, China.

This study aimed to evaluate the effectiveness of a school-based nutrition education and physical activity intervention on cardiovascular risk profile and mental health outcomes among Chinese children with obesity. Two primary schools were randomly allocated to the control group (CG) and the intervention group (IG). We selected children with obesity from 1340 students in the third and fourth grades as participants. The IG received 8 months of nutrition education and physical activity intervention, while the CG was waitlisted. A generalized estimating equation model was applied to assess repeated variables over time. A total of 171 children with obesity (99 IG and 72 CG) aged 9.8 ± 0.7 years completed the post-intervention stage. Compared with baseline, significant reductions were observed within the IG for depression and fasting plasma glucose at post-intervention. After adjusting for confounders, group and time interaction effects showed that the IG achieved improvements in the risk of poor well-being (p = 0.051) and social anxiety (p = 0.029), had decreased diastolic blood pressure (p = 0.020) and fasting plasma glucose (p < 0.001), and had significantly increased high-density lipoprotein (p < 0.001) from baseline to post-intervention relative to the CG. The effects of school-based nutrition education and physical activity intervention on children with obesity are diverse, including not only the improvement of metabolic health but also mental health promotion.

The Effect of LncRNA H19/miR-194-5p Axis on the Epithelial-Mesenchymal Transition of Colorectal Adenocarcinoma.

BACKGROUND/AIMS: Since the combined actions of lncRNAs and miRNAs have been considered to be involved in the occurrence and development of various neoplasms, the main purpose of this study was to discover whether and how lncRNA H19 and miR-194 influenced the epithelial-mesenchymal transition (EMT) process of colorectal adenocarcinoma (CRA). METHODS: Totally 214 pairs of CRA and adjacent normal tissues were collected, and 5 human CRA cell lines (i.e. HCT116, HT-29, RKO SW280 and Lovo) were purchased. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was adopted to quantify the H19 and miR-194-5p expressions in cells and tissues. The expressions of FoxM1, E-cadherin, vimentin, N-cadherin were determined using western blot. On the side, si-H19, si-NC, miR-194-5p mimic, miR-194-5p inhibitor and negative control (NC) were transfected into CRA cell lines. Meanwhile, the invasive, migratory and proliferative conditions of the cells were assessed through transwell, wound healing and colony-forming experiments, with final verification of the relationship between H19 and miR-194-5p employing dual-luciferase reporter gene assay. RESULTS: Highly-expressed H19, lowly-expressed miR-194-5p, low-grade differentiation and lymph node metastasis appeared as the independent predictors of unfavorable prognosis in CRA patients' (all P< 0.05). It indicated that FoxM1 expression displayed positive correlations with H19 expression, yet negative associations with miR-194-5p expression within CRA tissues (P< 0.05). In addition, transfection of H19-siRNA and miR-145-5p mimic triggered a conspicuous increase in E-cadherin expression, as well as an evidently down-regulation in vimentin and N-cadherin expressions within HT29 and RKO cells (P< 0.05). On the other hand, the invasive and migratory capacities of CRA cells were significantly hindered (P< 0.05). Moreover, the luciferase reporter gene assay confirmed that H19 modified miR-194-5p expression through directly targeting at it (P< 0.05). Ultimately, FoxM1 could reverse the role of miR-194-5p in inhibiting invasion, migration and EMT of CRA cells (P< 0.05). CONCLUSION: LncRNA H19/miR-194/FoxM1 axis could serve as a profound target for the diagnosis and treatment of CRA.

Associations of physical activity and fruit and vegetable intake with well-being and depressive symptoms among obese schoolchildren in Wuhan, China: a cross-sectional study.

BACKGROUND: The prevalence of childhood obesity is increasing and psychological disorder is a common comorbidity of obesity. We investigated the associations of physical activity (PA) and fruit and vegetable (FV) intake with well-being and depressive symptoms among obese schoolchildren. METHODS: Participants included 188 obese children aged 9.8 ± 0.7 years living in Wuhan, China. Self-administered questionnaires were used to collect the children's PA and FV intake information. PA was considered to be high if the child participated in sport and/or vigorous free play at least 3 days per week with 60 min per day, while sufficient FV intake was defined as consuming FV 5 times per day. Children's well-being and depressive symptoms were assessed by standard questionnaires. Multiple logistic regression was performed to determine the odds ratios (ORs) and 95% confidence intervals (CIs) of the relationships of PA and FV intake with well-being and depressive symptoms. RESULTS: High PA and sufficient FV intake were independently associated with significantly decreased risks for depressive symptoms (for PA, OR: 0.39, 95% CI: 0.16-0.92; for FV, OR: 0.21, 95% CI: 0.08-0.55) and poor well-being (for PA, OR: 0.35, 95% CI: 0.16-0.74), respectively. Furthermore, interactive inverse associations were observed between combined high PA and sufficient FV intake with poor well-being and depressive symptoms. Compared to their counterparts, children with high PA and sufficient FV intake had significantly reduced risk for poor well-being (OR: 0.16, 95%CI: 0.05-0.55) and depressive symptoms (OR: 0.12, 95% CI: 0.03-0.48). CONCLUSIONS: High PA and sufficient FV intake are inversely associated with the risks of poor well-being and depressive symptoms among obese Chinese schoolchildren.

Procaine is a specific DNA methylation inhibitor with anti-tumor effect for human gastric cancer.

DNA hypermethylation and the silencing of tumor suppressor genes caused by DNA hypermethylation is considered as a molecular hallmark of many kinds of cancers. Procaine, a local anesthetic, has been shown as a potential DNA methylation inhibitor in some types of cancers. However, the influence of procaine on DNA methylation regulation as well as the biological function in gastric cancer is still unknown. We report here that procaine represses the DNA-methylation level and promotes the proliferation arrest and apoptosis of gastric cancer cells. Global DNA methylation measurement demonstrates that procaine significantly reduces the global DNA methylation level. Analyses of the DNMTs expression and activity show procaine represses the activity, but not the expression, of DNMT1/DNMT3A. Further evidence on specific genes shows that procaine reduces the DNA methylation level in the promoter regions of CDKN2A and RARß genes through abrogating the binding of DNMT1/DNMT3A toward these regions. This repression would not be reversed by the overexpression of DNMT1/DNMT3A. Moreover, RT-qPCR and luciferase report assays demonstrate that procaine leads to the upregulation of CDKN2A and RARß due to the activation of the promoter of these genes. In the end, we test the function of procaine toward gastric cancer cells and find that procaine has the growth inhibitory and apoptosis inducement effect toward gastric cancer cells. Collectively, our data not only uncovers the regulation mechanisms of procaine to DNA methylation but also suggests an anti-tumor potential of procaine specific to the gastric carcinoma and provides a new therapeutic strategy for gastric carcinoma.

miR-938 promotes colorectal cancer cell proliferation via targeting tumor suppressor PHLPP2.

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. Although the development of therapy approaches, the outcome of CRC patients still is poor, understanding the biological mechanism of CRC progression is critical to improve the treatment strategies. miRNAs regulate CRC progression, we found miR-938 was upregulated in CRC tissues and cells, MTT assay, colony formation assay and soft agar growth assay suggested miR-938 overexpression promoted CRC cell proliferation, miR-938 knockdown inhibited CRC cell proliferation. Tumor suppressor PH domain Leucine-rich-repeats Protein Phosphatase 2 (PHLPP2) was a target of miR-938, miR-938 inhibited PHLPP2, luciferase activity assay suggested miR-938 directly bound to the 3'UTR of PHLPP2, meanwhile, we found miR-938 promoted c-Myc and Cyclin D1 expression, confirming miR-938 promoted CRC cell proliferation. Double knockdown of miR-938 and PHLPP2 promoted CRC cell proliferation, suggesting miR-938 promoted CRC cell proliferation by inhibiting PHLPP2.

Desmin detection by facile prepared carbon quantum dots for early screening of colorectal cancer.

Th aim of this study was to develop a new facile chemical method for early screening of colorectal cancer.The -C(O)OH groups modified Carbon Quantum Dots (CQDs) were prepared by an facile innovative route of acid attacking on carbon nanotubes (CNTs). The -C(O)OH groups were further transported into -C(O)Cl groups by SOCl2 treating. The obtained ClCQDs were conjugated onto the anti-Desmin, which were applied for testing the Desmin concentration in serum by using linearly fitted relationship with photoluminescence (PL) intensity.The obtained carbon quantum dots are quasispherical graphite nanocrystals with photoluminescence at about 455 nm. The Desmin with concentration of 1 ng/mL can lead to a decrease of PL intensity for anti-Desmin conjugated CQDs with good linearity. This assay had good specificity for Desmin with in interferential substances of immunoglobulin G (IgG), alpha fetoprotein (AFP), and carcinoembryoic antigen (CEA).A new facile acid attack method was developed to prepare ClCQDs, which could conjugate onto the anti-Desmin for detection of Desmin in serum with high sensitivity and specificity. As the detection limit is lower than 1 ng/ mL, this work provides a promising strategy for the evaluation of colorectal cancer risk with low cost and excellent sensing performance.

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In order to rapidly monitor chlorophyll content in cotton functional leaf, and establish the quantitative relationship between chlorophyll content and spectral characteristic parameter of single cotton leaf, cotton was pot cultivated in a rain shelter and subjected to waterlogging at squaring stage. Cotton leaf samples were taken and measured every 3 days after waterlogging. The correlation between chlorophyll content and spectral characteristic parameter was synthetically analyzed, and then the estimation model of chlorophyll content was established and verified. The results showed that the chlorophyll content decreased with increasing waterlogging stress. The original spectral reflectance and first order differential spectral reflectance was negatively correlated with the chlorophyll content in the band near 580 and 697 nm. The estimation model established by difference vegetation index and normalized difference vegetation index performed better than that established by linear model of single band. Furthermore, the estimation model with (DR697-DR738)/(DR697+DR738) as the independent variable fitted the best with the correlation coefficient of 0.814, which could be utilized to estimate chlorophyll content of single leaf under waterlogging stress.

MicroRNA-224 aggrevates tumor growth and progression by targeting mTOR in gastric cancer.

Growing evidence suggests that microRNA plays an essential role in the development and metastasis of many tumors, including gastric cancer. Aberrant miR-224 expression has been indicated in tumor growth, the mechanism of miR-224 promoting the proliferation and metastatic ability for gastric cancer remains unclear. Accumulating evidence reports that mTOR signal pathway plays an important role in the cellular process, such as apoptosis, cell growth and proliferation. The goal of the present study was to identify whether miR-224 could inhibit the growth, migration, invasion, proliferation and metastasis of gastric cancer through targeting mTOR expression. Real-time PCR (RT-PCR) was used to quantify miR-224 expression in vitro and in vivo experiments. Luciferase reporter assays were performed to confirm the activity of mTOR pathway, and immunofluorescence staining assay was conducted to observe apoptosis and cell proliferation ability. Bioinformatics as well as cell luciferase function studies distinguished the direct modulation of miR-224 on the 3'-UTR of the mTOR, which leads to the inactivation of apoptosis signaling and the activation of cell proliferation. In addition, inhibition of miR-224 significantly reduced the expression of mTOR and improved caspase-9/3 expression while decreased cyclin D1/2 levels, attenuating gastric cancer cell proliferation. Therefore, the present study revealed the mechanistic links between miR-224 and mTOR in the pathogenesis of gastric cancer through modulation of caspase-9/3 and cyclin D1/2. In addition, targeting miR-224 could serve as a novel strategy for future gastric cancer therapy.

MicroRNA-766 targeting regulation of SOX6 expression promoted cell proliferation of human colorectal cancer.

MicroRNAs (miRNAs) have emerged as important regulators of cancer-cell biological processes. Previous studies have shown that miR-766 plays an important role in a variety of biological processes in various human cancers. However, the underlying mechanism of miR-766 in colorectal cancer (CRC) cells remains unclear. In this study, we investigated miR-766's role in CRC cell proliferation. Polymerase chain reaction results showed that miR-766 expression was significantly upregulated in CRC tissues and cells. Ectopic expression of miR-766 promoted cell growth and anchorage-independent growth in CRC cells. Bioinformatic analysis predicted SOX6, a potential target of miR-766, acting as a tumor suppressor. Luciferase reporter assay results demonstrated that miR-766 directly bound to the 3'-untranslated region of SOX6. Overexpression of miR-766 suppressed SOX6 expression, resulting in the downregulation of p21 and upregulation of cyclin D1. In a further experiment, SOX6-silenced SW480 cells transfected with miR-766 promoted cell growth, suggesting that downregulation of SOX6 was required for miR-766-induced CRC cell proliferation. Taken together, these results suggested that miR-766 represents an onco-miRNA and participates in the development of CRC by modulating SOX6 expression.

Simple framework for understanding the universality of the maximum drag reduction asymptote in turbulent flow of polymer solutions.

Self-consistent direct numerical simulations of turbulent channel flows of dilute polymer solutions exhibiting friction drag reduction (DR) show that an effective Deborah number defined as the ratio of polymer relaxation time to the time scale of fluctuations in the vorticity in the mean flow direction remains O(1) from the onset of DR to the maximum drag reduction (MDR) asymptote. However, the ratio of the convective time scale associated with streamwise vorticity fluctuations to the vortex rotation time decreases with increasing DR, and the maximum drag reduction asymptote is achieved when these two time scales become nearly equal. Based on these observations, a simple framework is proposed that adequately describes the influence of polymer additives on the extent of DR from the onset of DR to MDR as well as the universality of the MDR in wall-bounded turbulent flows with polymer additives.

A competitive strategy based on cucurbit[7]uril supramolecular interaction for simple and sensitive detection of dibucaine.

In this work, the competitive interaction between dibucaine and three fluorescent probes (i.e., berberine, palmatine, and coptisine) for occupancy of the cucurbit[7]uril (CB[7]) cavity was studied by fluorescence spectra, UV-visible absorption spectra, (1)H NMR spectra, and theoretical calculations in acidic aqueous solution. Based on the fluorescence enhancement of berberine, palmatine, and coptisine upon binding with CB[7], respectively, a series of fluorescence detection methods for dibucaine were proposed. At the optimized conditions, the fluorescence intensity of berberine-CB[7], palmatine-CB[7], and coptisine-CB[7] complexes showed negative correlation to the concentration of dibucaine, which led to a series of simple and sensitive fluorescence methods for the determination of dibucaine for the first time. Linear ranges obtained in the detection of the dibucaine were 0.018-3.34 µmol L(-1), 0.032-4.47 µmol L(-1), and 0.079-4.42 µmol L(-1) with detection limits of 6.0 nmol L(-1), 12.0 nmol L(-1), and 25.0 nmol L(-1), respectively. Moreover, the proposed method was successfully applied for the determination of the drug in biological fluids. The competitive mode based on CB[7] superstructure provided a promising assay strategy for fluorescence detection in various potential applications.

Dual­sensitive HRE/Egr1 promoter regulates Smac overexpression and enhances radiation­induced A549 human lung adenocarcinoma cell death under hypoxia.

The aim of this study was to construct an expression vector carrying the hypoxia/radiation dual­sensitive chimeric hypoxia response element (HRE)/early growth response 1 (Egr­1) promoter in order to overexpress the therapeutic second mitochondria­derived activator of caspases (Smac). Using this expression vector, the present study aimed to explore the molecular mechanism underlying radiotherapy­induced A549 human lung adenocarcinoma cell death and apoptosis under hypoxia. The plasmids, pcDNA3.1­Egr1­Smac (pE­Smac) and pcDNA3.1­HRE/Egr-1­Smac (pH/E­Smac), were constructed and transfected into A549 human lung adenocarcinoma cells using the liposome method. CoCl2 was used to chemically simulate hypoxia, followed by the administration of 2 Gy X­ray irradiation. An MTT assay was performed to detect cell proliferation and an Annexin V­fluorescein isothiocyanate apoptosis detection kit was used to detect apoptosis. Quantitative polymerase chain reaction and western blot analyses were used for the detection of mRNA and protein expression, respectively. Infection with the pE­Smac and pH/E­Smac plasmids in combination with radiation and/or hypoxia was observed to enhance the expression of Smac. Furthermore, Smac overexpression was found to enhance the radiation­induced inhibition of cell proliferation and promotion of cycle arrest and apoptosis. The cytochrome c/caspase­9/caspase­3 pathway was identified to be involved in this regulation of apoptosis. Plasmid infection in combination with X­ray irradiation was found to markedly induce cell death under hypoxia. In conclusion, the hypoxia/radiation dual­sensitive chimeric HRE/Egr­1 promoter was observed to enhance the expression of the therapeutic Smac, as well as enhance the radiation­induced inhibition of cell proliferation and promotion of cycle arrest and apoptosis under hypoxia. This apoptosis was found to involve the mitochondrial pathway.

7,8-Dihydroxycoumarin inhibits A549 human lung adenocarcinoma cell proliferation by inducing apoptosis via suppression of Akt/NF-κB signaling.

The Akt/NF-κB pathways are involved in numerous anti-apoptotic and drug-resistance events that occur in non-small cell lung cancer (NSCLC). In the present study, the role of 7,8-dihydroxycoumarin in the regulation of the anti-apoptotic Akt and NF-κBp65 signaling pathways was explored. A549 human lung adenocarcinoma cells were exposed to 7,8-dihydroxycoumarin with a final concentration of 25, 50 and 100 µmol/l for 48 h. Quantitative polymerase chain reaction (PCR) and western blotting were performed to detect mRNA and protein expression, respectively. The MTT assay was performed to detect cell proliferation. The results demonstrated that anti-apoptotic phospho-Akt1 (pAkt1), phospho-IκBα (pIκBα), NF-κBp65 and Bcl-2 were inhibited and pro-apoptotic caspase-3 was upregulated in a concentration-dependent manner. At a concentration of 100 µmol/l, the anti-apoptotic NF-κBp65 and Bcl-2 mRNA expression levels decreased 0.12 (5.82/48.5, treated/control)-fold and 0.17 (6.7/39.4, treated/control)-fold, respectively. The pro-apoptotic caspase-3 mRNA was upregulated 4.43 (39.4/8.9, treated/control)-fold. The anti-apoptotic pAkt1, pIκBα, NF-κBp65 and Bcl-2 proteins were downregulated, with blot grayscale values of 7.3 (vs. 52.4 control), 4.3 (vs. 42.2 control), 5.08 (vs. 44.5 control) and 5.92 (vs. 38.5 control), respectively. The proapoptotic caspase-3 was upregulated to a blot grayscale value of 27.8 (vs. 5.8 control). The proliferative activity of A549 cells was reduced significantly compared with that of the control cells (83.7, 27.2 and 9.5 vs. 100%, respectively; P<0.05 for each). 7,8-Dihydroxycoumarin plays an important role in the induction of apoptosis via suppression of Akt/NF-κB signaling in A549 human lung adenocarcinoma cells in a concentration-dependent manner. 7,8-Dihydroxycoumarin may be a candidate naturally-occurring drug for the treatment and prevention of lung adenocarcinoma.

Differences in HER2 over-expression between proximal and distal gastric cancers in the Chinese population.

AIM: To investigate HER2 expression and its correlation with clinicopathological variables between proximal and distal gastric cancers (GC) in the Chinese population. METHODS: Immunostaining of HER2 was performed and scored on a scale of 0-3 in 957 consecutive GC cases, according to the revised scoring criteria of HercepTest(TM) as used in the ToGA trial. Correlations between HER2 expression and clinicopathologic variables of proximal (n = 513) and distal (n = 444) GC were investigated. RESULTS: Our results showed that HER2 expression was significantly higher in the proximal than in distal GC (P < 0.05). Overall, HER2 expression was significantly higher in male patients (P < 0.01), the Lauren intestinal type (P < 0.001), low-grade (P < 0.001) and pM1 (P < 0.01) diseases, respectively. There was a significant difference in HER2 expression among some pTNM stages (P < 0.05). In contrast, HER2 expression in the distal GC was significantly higher in male patients (P < 0.001), low-grade histology (P < 0.001), the Lauren intestinal type (P < 0.001), and pM1 (P < 0.001). In the proximal GC, however, higher HER2 expression scores were observed only in tumors with low-grade histology (P < 0.001) and the Lauren intestinal type (P < 0.001). CONCLUSION: HER2 over-expression in GC of Chinese patients was significantly more common in proximal than in distal GC, and significantly correlated with the Lauren intestinal type and low-grade histology in both proximal and distal GC, and with pM1 disease and male gender in distal GC.

The tumor suppressor gene RhoBTB1 is a novel target of miR-31 in human colon cancer.

miRNAs are a class of endogenous non-coding RNA, which can regulate downstream target genes through binding to the 3'UTR of those genes. Numerous studies have indicated that abnormal expression of miRNAs is implicated in tumor development. Aberrant expression of miR-31 has been found in various cancers, including colorectal cancer. Here, we show that miR-31 is upregulated in human colon cancer tissues and cell lines, and that repression of miR-31 inhibited colon cancer cell proliferation and colony formation in soft agarose. To further elucidate the mechanism underlying the role of miR-31 in promoting colon cancer, we used online miRNA target prediction databases and found that the tumor suppressor RhoTBT1 may be a target of miR-31. Imunohistochemistry assay revealed that RhoBTB1 was significantly decreased in HT29 cells. In addition, ectopic expression of miR-31 reduced RhoBTB1 in the colon cancer cell line HT29. The results suggested that suppression of RhoBTB1 may be responsible for colon tumorigenesis, which was inhibited directly by miR-31. The results of MTT and soft agarose colony-formation assays showed that knockdown of RhoBTB1 by RNAi induced cell proliferation, and colony formation in soft agarose, which mimicked the function of miR-31. This further suggested that suppression of RhoBTB1 was responsible for colon tumorigenesis. In conclusion, we found that miR-31 acts as an oncogene in colon cancer and identified RhoBTB1 as a new target of miR-31 further study demonstrated that miR-31 contributed to the development of colon cancer at least partly by targeting RhoBTB1.

Determination of ranitidine, nizatidine, and cimetidine by a sensitive fluorescent probe.

A validated, simple, and sensitive fluorescence quenching method for the determination of ranitidine, nizatidine, and cimetidine in tablets and biological fluids is presented. This is the first single fluorescence method reported for the analysis of all three H(2) antagonists. The competitive reaction between the investigated drug and the palmatine probe for the occupancy of the cucurbit[7]uril (CB[7]) cavity was studied using spectrofluorometry. CB[7] was found to react with the probe to form a stable complex. The fluorescence intensity of the complex was also enhanced greatly. However, the addition of the drug dramatically quenched the fluorescence intensity of the complex. Accordingly, a new fluorescence quenching method for the determination of the studied drugs was established. The different experimental parameters affecting the fluorescence quenching intensity were studied carefully. At optimum reaction conditions, the rectilinear calibration graphs between the fluorescence quenching values (ΔF) and the medicament concentration were obtained in the concentration range of 0.04-1.9 µg mL(-1) for the investigated drugs. The limits of detection ranged from 0.013 to 0.030 µg mL(-1) at 495 nm using an excitation wavelength of 343 nm. The proposed method can be used for the determination of the three H(2) antagonists in raw materials, dosage forms and biological fluids.

Determination of ethambutol by a sensitive fluorescent probe.

The competitive reaction between ethambutol and two fluorescent probes (i.e., berberine and palmatine) for occupancy of the cucurbit[7]uril (CB[7]) cavity was studied by spectrofluorometry. The CB[7] reacts with these probes to form stable complexes, and the fluorescence intensity of the complexes is greatly enhanced. In addition, the excitation and emission wavelengths of their complexes moved to wavelengths of 343 nm and 495 nm, respectively. However, the addition of ethambutol dramatically quenches the fluorescence intensity of the two complexes. Accordingly, a couple of new fluorescence quenching methods for the determination of ethambutol were established. The methods can be applied for quantifying ethambutol. A linear relationship between the fluorescence quenching values (ΔF) and ethambutol concentration exists in the range of 5.0-1000.0 ng mL(-1), with a correlation coefficient (r) of 0.9997. The detection limit is 1.7 ng mL(-1). The fluorescent probe of berberine has higher sensitivity than palmatine. This paper also discusses the mechanism of fluorescence indicator probes.