Expression of lncRNA TUG1 in hypertensive patients and its relationship with change state of an illness.

OBJECTIVE: To investigate the expression of LncRNA TUG1 in hypertensive patients and its relationship with the change state of illness. PATIENTS AND METHODS: A prospective analysis was carried out. Eighty-two patients with hypertension admitted to our hospital from March 2016 to February 2019 were regarded as a research group, and 79 healthy people admitted to our hospital during the same period were regarded as a control group. The serum LncRNA TUG1, platelet activating factor (PAF), endothelin-1 (ET-1), serum tumor necrosis factor-α (TNF-α), high sensitive C-reactive protein (hsCRP), and the relationship between the expression of LncRNA TUG1 and the clinicopathological characteristics of patients with hypertension in the two groups were detected, and finally, the risk factors of hypertension were analyzed. RESULTS: The expression levels of LncRNA TUG1, PAF, ET-1, TNF-α, and hsCRP in the serum of patients in research group were significantly higher than those in control group (p<0.05). LncRNA TUG1 has a positive correlation with the severity of an illness of hypertensive patients (r=0.881, p<0.001), and also has a positive correlation with the expression levels of PAF, ET-1, TNF-α, and hsCRP (r=0.735, p<0.001; r=0.756, p<0.001; r=0.712, p<0.05; r=0.723, p<0.05). The expression of LncRNA TUG1 in the serum of patients with hypertension was related to hypertension mode, severity of disease, hyperlipidemia, and obesity (p<0.05). Obesity (OR: 3.469, 95% CI: 2.175-4.095), drinking (OR: 3.677, 95% CI: 1.695-4.892), hyperlipidemia (OR: 3.374, 95% CI: 1.759-6.988), and the high expression of TUG1 (OR: 2.693, 95% CI: 1.679-7.472) are independent risk factors for the attack of hypertension. CONCLUSIONS: LncRNA TUG1 is highly expressed in the serum of hypertensive patients and it is closely related to the progression of hypertension. Also, it may be one of the independent risk factors for hypertension and a new molecular target for hypertension treatment.

Short communication: Relationship between lysine/methionine ratios and glucose levels and their effects on casein synthesis via activation of the mechanistic target of rapamycin signaling pathway in bovine mammary epithelial cells.

The synthesis of protein requires the availability of specific AA and a large supply of energy in bovine mammary epithelial cells (BMEC). Whether an interaction exists between Lys/Met ratio and glucose level on milk protein synthesis and its potential regulatory mechanism is unclear. We investigated the effects of different Lys/Met ratios and glucose levels on casein synthesis-related gene expression in BMEC to elucidate the underlying molecular mechanisms. Primary BMEC were subjected to 4 treatments for 36 h, arranged in a 2 × 2 factorial design with Lys/Met ratios of 3:1 (1.2:0.4 mM, LM3.0; total AA = 8.24 mM) and 2.3:1 (1.4:0.6 mM, LM2.3; total AA = 8.64 mM) and glucose levels of 17.5 mM (high glucose level) and 2.5 mM (low glucose level). No interactions between Lys/Met ratio and glucose level on cell viability, cell cycle progression, mRNA, or protein expression levels were found. High glucose level increased cell proliferation and promoted cell cycle transition from intermediate phase (G1 phase) to synthesis (S phase) by approximately 50%, whereas Lys/Met ratio had no effect. Both mRNA and protein abundance of αS1-casein and ß-casein were positively affected by LM3.0, whereas a high glucose level increased protein abundance of αS1-casein and ß-casein and increased gene expression of CSN1S1 but not of CSN2. Furthermore, high glucose increased the mRNA abundance of ELF5 and decreased that of GLUT8, enhanced protein expression of total and phosphorylated mechanistic target of rapamycin (mTOR), and decreased phosphorylated AMP-activated protein kinase (AMPK) levels. Treatment LM3.0 had a stimulatory effect on total and phosphorylated mTOR but did not affect AMPK phosphorylation. The mRNA levels of JAK2, ELF5, and RPS6KB1 were upregulated and mRNA levels of EIF4EBP1 were downregulated with LM3.0 compared with LM2.3. Our results indicate that casein synthesis was regulated by Lys/Met ratio via JAK2/ELF5, mTOR, and its downstream RPS6KB1 and EIF4EBP1 signaling. In contrast, glucose regulated casein synthesis through promoting cell proliferation, accelerating cell cycle progression, and activating the ELF5 and AMPK/mTOR signaling pathways. Within the range of substrate levels in the present study, a change in Lys/Met ratio had a stronger effect on abundance of αS1-casein and ß-casein than a change in glucose level.

Chlorpyrifos-induced hormesis in insecticide-resistant and -susceptible Plutella xylostella under normal and high temperatures.

Hormesis induced by insecticides at the dosage lower than what ostensibly directly causes death on insects was studied. This paper reports the effects of the in vivo application of varied concentrations of chlorpyrifos (CPF) on Plutella xylostella (DBM). The insecticide concentrations applied included 0.000025-2.5 mg l-1, which are far lower than LC1 (7.2 mg l-1), for the CPF-susceptable (Si) DBM, and 250 mg l-1 which is far below LC1 (1286 mg l-1), for the CPF-resistant (Rc) DBM, as well as LC10- and LC50-doses for both strains. Significant hormesis was found with the 'hermetic-CPFs', i.e., 0.0025 mg l-1 for Si DBM and 2.5 mg l-1 for Rc DBM, at the normal or high temperature either in a 24 h or under a long-term treatment. These doses of CPF significantly stimulated the development and increased the fecundity of Si and Rc DBM at 25°C with approximately 23.5-29.8% activity increase on acetylcholinesterase (AChE) and 30.5-91.3% increase on glutathione S-transferases (GSTs) at 25 or 38°C in 4-24 h. The enzymatic activities were significantly reduced by LC50-CPF at 25°C in vivo, but the inhibition was relieved significantly, if the insects were first subjected to a hormetic-CPF pretreatment. It was remarkable that the average rates of enzymatic activity increase were 67.5-76.6% for AChE and 366-546% for GSTs. Consequently, it was concluded that the hormesis on Si and Rc DBM could be induced by CPF doses far below LC1 at normal or high temperature in short- or long-term treatment. These findings might help to improve the current insect control practices in the field.

Technical note: Isolation and characterization of porcine mammary epithelial cells.

Within the mammary gland, functional synthesis of milk is performed by its epithelial (alveolar) cells. The availability of a stable mammary epithelial cell line is essential for biochemical studies to elucidate cellular and molecular mechanisms responsible for nutritional regulation of lactation. Therefore, porcine mammary epithelial cells (PMEC) were isolated from mammary glands of a 9-mo-old nonpregnant and nonlactating gilt and cultured to establish a nonimmortalized cell line. These cells were characterized by expression of cytokeratin-18 (an intermediate filament specific for epithelial cells), ß-casein (a specific marker for mammary epithelial cells), and α-lactalbumin. In culture, the PMEC doubled in number every 24 h and maintained a cobblestone morphology, typical for cultured epithelial cells, for at least 15 passages. Addition of 0.2 to 2 µg/mL prolactin to culture medium for 3 d induced the production of ß-casein and α-lactalbumin by PMEC in a dose-dependent manner. Thus, we have successfully developed a useful PMEC line for future studies of cellular and molecular regulation of milk synthesis by mammary epithelial cells of the sow.

High nuclear expression of STAT3 is associated with unfavorable prognosis in diffuse large B-cell lymphoma.

BACKGROUND: The purpose of the study was to investigate the expression and prognostic value of STAT3 in diffuse large B-cell lymphoma (DLBCL). METHODS: Seventy-four DLBCL patients from 2001 to 2007 were reviewed in the study. The STAT3 expression in their tumor tissues was examined using the immunohistochemistry (IHC) method, and evaluated for its association with clinicopathological parameters. RESULTS: Strong nuclear staining of STAT3 and phosphorylated-STAT3tyr705 (P-STAT3) were observed in 19 cases (25.7%) and 24 cases (32.4%), respectively, and the expression levels were highly consistent between them (P = 0.001). The high nuclear expression of STAT3 was more frequent in the non-germinal center B cell-like (non-GCB) DLBCL than that in the GCB subtype, but not reaching significance (P < 0.061). The high nuclear expression of STAT3 was found to be correlated with poor overall survival (OS) (P = 0.005). Multivariate Cox regression analysis showed that the STAT3 expression was an independent prognostic factor for DLBCL patients regardless of CHOP or R-CHOP regimen used as the first-line therapy. CONCLUSION: STAT3 is more frequently expressed in non-GCB DLBCL than that in GCB subtype, and its strong nuclear expression is correlated with poor OS in DLBCL.

Polycomb protein EZH2 regulates E2F1-dependent apoptosis through epigenetically modulating Bim expression.

Deregulation of the pRB/E2F pathway, which occurs frequently in human malignancy, is often associated with inappropriate proliferation and/or apoptosis. While the role of E2F1 in apoptosis induction has been well-established, it remains unclear how this pro-apoptotic activity is regulated in cancer. Here we describe EZH2, an oncogenic polycomb histone methyltransferase and an E2F1 target, as an important regulator of E2F1-dependent apoptosis. We show that E2F1 induces EZH2 expression, which in turn antagonizes the induction of E2F1 pro-apoptotic target Bim expression. RNAi-mediated gene depletion of EZH2 enhances E2F1-dependent Bim expression, thereby promoting the pro-apoptotic activity of E2F1. Hence, the concomitant induction of EZH2 and Bim by E2F1 constitutes a fail-safe mechanism to allow tumor cells with aberrant E2F1 activity to evade apoptosis. These findings reveal a novel mechanism by which the apoptotic activity of E2F1 is restrained in human cancer and also provide the first evidence that EZH2 directly regulates apoptotic process in cancer cells.

Sampling survey on intellectual disability in 0 approximately 6-year-old children in China.

OBJECTIVES: To understand the current status of intellectually disabled children and the prevalence of intellectual disability (ID) in children aged 0 approximately 6 years and its risk factors, and to provide scientific evidence to formulate relevant policies for helping intellectually disabled children. METHODS: Multiphase, stratified, unequal proportional and cluster sampling was adopted to investigate 60 124 children aged 0 approximately 6 years. All the children investigated were screened for ID using the Denver Developmental Screening Test, and those with positive screening test would be further diagnosed by varied specialists using the Gesell Developmental Inventory. RESULTS: In total, 560 of 60 124 children were diagnosed as intellectually disabled with an overall prevalence of 0.93%. Prevalence of ID was highest in children living in medium-developed areas with a prevalence of 1.20%, higher than in those living in developed areas (0.75%) and in underdeveloped areas (0.84%). It was higher in rural areas (1.03%) than in urban areas (0.83%), and higher in boys (1.01%) than in girls (0.84%). Prevalence of ID increased with the age of children and decreased with the educational level of their parents. CONCLUSIONS: The study suggested that ID is still prevalent in the children of China, and rehabilitation for them is lagging behind current needs. Early prevention of ID in children and pre-school education for them should be strengthened.

Optical coherence tomography for noninvasive diagnosis of epithelial cancers.

We summarize our recent progress in the development of the optical coherence tomography (OCT) systems suitable for clinical diagnosis and the preliminary results for in vivo diagnosis of epithelial cancers (e.g., bladder cancers). The endoscopic spectral-domain OCT system allows simultaneous, real-time, cross-sectional OCT images of tissue structure and functions (i.e., local Doppler blood flow) of biological tissue for enhanced diagnosis. A new approach to use spectral demodulation of elastic scattering is discussed for potential cancer grading. The transverse and axial resolutions of the OCT scopes are 12 microm and 10 microm, respectively. Results of the preliminary clinical studies show that unlike animal carcinogenesis models, bladder cancers in humans are more complicated in terms of epithelial backscattering changes: some lesions exhibit enhanced backscattering; some show reduced scattering owing to complex surface condition changes such as asperities or invaginations induced by tumorigenesis (e.g., papillary transitional cell cancers). Nevertheless, promising results can be provided by incorporating other diagnostic parameters such as changes in local vasculature and urothelial heterogeneity.

Detection of DNA strand breaks, DNA-protein crosslinks, and telomerase activity in nickel-transformed BALB/c-3T3 cells.

Although nickel compounds are known carcinogens, the underlying carcinogenic mechanisms are not fully understood. The objective of this research was to determine if the genotoxic lesions of DNA strand breaks and DNA-protein crosslinks are present in nickel-transformed BALB/c-3T3 cells, and to further elucidate the potential carcinogenesis of insoluble and soluble nickel compounds through telomerase activity in nickel-transformed BALB/c-3T3 cell lines. DNA strand breaks, DNA-protein crosslinks and telomerase activity were investigated by single cell gel electrophoresis (comet assay), (125)I-postlabelling techniques, and the TRAP-silver staining assay, respectively. Results showed that both DNA strand breaks and DNA-protein crosslinks were present in nickel-transformed BALB/c-3T3 cells. However, the highest levels of DNA strand breaks and DNA-protein crosslinks were found in insoluble crystalline NiS-transformed cells and high levels of DNA strand breaks and DNA-protein crosslinks were also found in the transformed cells induced by two water-soluble NiCl(2) and NiSO(4) at moderate concentrations of cytotoxicity. These data suggest that these two genetic endpoints are useful biomarkers and are associated with cell transformation and carcinogensis of insoluble and soluble nickel compounds. Also, we found that the crystalline NiS- and NiCl(2)-transformed cells possessed a high telomerase activity. A weak telomerase was found in NiSO(4)-transformed cells. The results seem to indicate that in addition to crystalline NiS, some water-soluble nickel compounds such as NiCl(2) are also highly carcinogenic. These results may partly explain the cell transformation and relative carcinogenic potency of insoluble crystalline NiS, soluble NiCl(2), and NiSO(4).

[Study on taxol release in the two-liquid-phase cultures of Taxus cuspidata].

Effects of rare earth compound (ammonium sulphate), organic solvents(oleic acid and dibutylphthalate) and the integrated function of the rare earth compound and organic solvents were studied on taxol release in the Taxus cuspidata suspension cultures. And then effects of different organic solvents(paraffin, organic acid, alcohol and ester), their volumetric fraction and phase toxicity were studied on taxol release in the two-liquid-phase cultures of Taxus cuspidata. The results showed that the addition of the rare earth compound or the organic solvents could strengthen obviously taxol release, especially the organic solvents. But the addition of the rare earth compound could not strengthen further taxol release in the twoliquid-phase cultures of Taxus cuspidata. Therefore the organic solvents were very good permeabilizing reagents, which could enhance obviously secondary metabolite in the twoliquid-phase cultures of plant cells. Release percentage of taxol was increased into more than 75% from 40% of the control.

Effects of metabolites of benzo(a)pyrene on unschedule DNA synthesis in BALB/3T3 cell line.

In order to explore the damage from metabolites of benzo(a)pyrene on DNA of mammalian cells, the effects of four metabolites of benzo(a)pyrene (anti-BPDE, syn-BPDE, 3-OH-BP and 9-OH-BP) on synthesis of DNA and unschedule DNA synthesis (UDS) in BALB/3T3 cells were assayed, by methods of single-labeling and double-labeling. The results showed that all of the four agents were able to increase the synthesis of DNA, but only three of them (apart from syn-BPDE) induced UDS in BALB/3T3 cells. The above indicates that the metabolites of benzo(a)pyrene are able to damage DNA in BALB/3T3 cells, and that this effect may be relative to the sterical structure of metabolites of benzo(a)pyrene.

Promotive effect of diethylstilbestrol on urethan-induced mouse lung tumorigenesis.

The effect of diethylstilbestrol (DES) on urethan-induced mouse lung tumorigenesis was assessed by a single intraperitoneal injecting of urethan (50 mg/kg) or/and multi intramuscular injecting of DES (5 or 50 mg/kg). All mice were sacrificed 18 weeks after administration, and the lung tumors were examined histopathologically. DES did not produce an elevated lung tumor response when administered alone, but it produced a statistically significant enhancement of incidence of tumors, average number of tumors, incidence of cancers and constituent ratio of malignant tumors when given in conjunction with urethan. The results indicated that DES may be a promoter in lung tumor formation.

An animal model of septicemia-induced hypercatabolic acute renal failure.

A rat model of hypercatabolic acute renal failure (ARF) was developed in order to further investigate the mechanism of this condition. Sprague Dawley rats were separated into three groups: a septicemic group, an ischemic ARF group, and a hypercatabolic ARF group. Septicemia was produced by the i.p. injection of 1 x 10(7) colony-forming units/mL of Escherichia coli. Ischemic ARF was induced by 60 minutes clamping of the left renal artery following a contralateral nephrectomy. Hypercatabolic ARF was produced by combining ischemic ARF with the i.p. injection of 1 x 10(7) colony-forming units/mL of Escherichia coli. The hypercatabolic ARF group exhibited septic clinical features after the surgical procedures. The blood urea nitrogen and the serum creatinine, potassium and carbon dioxide combining power of hypercatabolic ARF were significantly higher than other two groups 24 hours after surgery. In addition, the rats wit hypercatabolic ARF had a greater loss of body weight and a higher mortality rate compared to the other two groups. The features of this form of experimental ARF are similar to the clinical characteristics of hypercatabolic ARF. Consequently, this appears to be a useful model of hypercatabolic ARF.

Differential effects of aneugens and clastogens on incidences of multinucleated cells and of micronucleate cells in Chinese hamster lung (V79) cell line in vitro.

Two aneugens, vinblastine (0.025-0.4 microgram/ml) and colchicine (0.05-0.4 microgram/ml), and two clastogens, mitomycin C (0.05-0.4 microgram/ml) and cyclophosphamide (1-16 micrograms/ml) were applied respectively to the micronucleus test in Chinese hamster lung (V79) cells in vitro, and the frequency of multinucleated cells (Fmu) and that of micronucleate cells (Fmi) in each group were observed. The results showed that at relatively high concentrations, all four agents increased both Fmu and Fmi, but the ratios of Fmu to Fmi in groups of the two aneugens (average of 2.2, 2.8, respectively) were much (10-30 folds) higher than that in groups of the two clastogens (0.09, 0.20). The difference between aneugens and clastogens in the above ratio was much more remarkable than that in areas of micronuclei (only 1.6-2.5 folds for the latter). At relatively low concentrations, the two clastogens increased only Fmi (but not Fmu), while the two aneugens increased only Fmu (but not Fmi). This indicates that the induction of multinuclei by aneugens may be more sensitive than by clastogens, and the induction of micronuclei by clastogens may be more sensitive than by aneugens. So, it is possible for the ratio of Fmu to Fmi to become a simple and sensitive (though indirect) index for distinguishing aneugens from clastogens. Further studies with other mutagens and (or) other cell types will be needed to confirm the deduction. As no difference in frequency of polyploid cells was observed between control group and each treatment, the multinucleation does not seem related to endoreduplication of chromosomes.

Detection of genomic instability in lung cancer tissues by random amplified polymorphic DNA analysis.

Genomic instability resulting in multiple mutations is believed to be a driving force in the carcinogenic process. In this study, the random amplified polymorphic DNA (RAPD) technique, a simple PCR-based DNA polymorphism assay system, was used for detecting genomic instability in lung cancer tissues. DNAs from 20 lung cancer (18 non-small cell lung cancers and two small cell lung cancers) and their corresponding normal tissues were amplified individually by RAPD with seven different 10-base arbitrary primers. PCR products from RAPD were electrophoretically separated in agarose gels and banding profiles were visualized by ethidium bromide staining. The ability to detect genomic instability in 20 cancer tissues by each single primer ranged from 15 to 75%. DNA changes were detected by at least one primer in 19 (95%) cancer tissues. These results seem to indicate that genomic rearrangement is associated with lung carcinogenesis and that RAPD analysis is useful for the detection of genomic instability in lung cancer tissues.

Ethanol alters the subcellular localization of delta- and epsilon protein kinase C in NG108-15 cells.

Protein kinase C (PKC) has been shown to regulate the ethanol sensitivity of membrane-bound receptors and transporters, but little is known about the molecular mechanisms underlying this regulation. PKC is a family of isozymes that translocate to new intracellular sites on activation. Here we present immunochemical data showing that ethanol causes translocation of delta- and epsilon-PKC to new intracellular sites. Ethanol causes translocation of delta-PKC from the Golgi to the perinucleus; this translocation is similar to that induced by activation of PKC with phorbol esters. In contrast, epsilon-PKC translocation caused by ethanol is different from that induced by phorbol esters; ethanol causes translocation of epsilon-PKC from the perinucleus to the cytoplasm, whereas phorbol ester activation causes translocation of epsilon-PKC to the nucleus. Because the substrate specificity of these kinases is determined by their site of localization, ethanol-induced translocation of delta- and epsilon-PKC to new intracellular sites may explain some of the pleiotropic effects of ethanol on cellular functions.

Altered behavior and long-term potentiation in type I adenylyl cyclase mutant mice.

The murine Ca(2+)-stimulated adenylyl cyclase (type I) (EC 4.6.1.1), which is expressed predominantly in brain, was inactivated by targeted mutagenesis. Ca(2+)-stimulated adenylyl cyclase activity was reduced 40-60% in the hippocampus, neocortex, and cerebellum. Long-term potentiation in the CA1 region of the hippocampus from mutants was perturbed relative to controls. Both the initial slope and maximum extent of changes in synaptic response were reduced. Although mutant mice learned to find a hidden platform in the Morris water task normally, they did not display a preference for the region where the platform had been when it was removed. These results indicate that disruption of the gene for the type I adenylyl cyclase produces changes in behavior and that the cAMP signal transduction pathway may play an important role in synaptic plasticity.

Antitransforming activity of chlorophyllin against selected carcinogens and complex mixtures.

Chlorophyllin, a derivative of chlorophyll, is known to be an antimutagenic agent. Studies were performed to determine whether chlorophyllin can also inhibit morphological transformation of BALB/3T3 cells induced by carcinogens and complex mixtures. Chlorophyllin was added to the cultures simultaneously with carcinogens or complex mixtures while the transformation assay was conducted. At concentrations that did not significantly affect cell growth, chlorophyllin was found to inhibit morphological transformation induced by N-methyl-N'-nitro-N-nitrosoguanidine, 3-methylcholanthrene, 7,12-dimethylbenz(a)anthracene, benzo(a)pyrene, aflatoxin B1, and extracts of coal dust, tobacco snuff, and chewing tobacco. In all cases, the mean number of transformed foci per flask treated with chlorophyllin was significantly lower than that of untreated cultures. The reduction in the number of transformed foci was dependent on the concentration of chlorophyllin tested. These results indicate that chlorophyllin is an antitransforming agent.

Iron loading enhances susceptibility to renal ischemia in rats.

Because chronic iron overload can cause organ injury in hemochromatosis and because iron participates in injury during renal ischemia-reperfusion, the effect of mild subacute renal iron loading on the susceptibility to ischemic acute renal failure was evaluated. Male Sprague-Dawley rats were injected with iron nitrilotriacetate (1 mg iron/kg BW i.p. daily) for 5 days. Controls were instead injected with nitrilotriacetate. Seventy-two hours later animals were subjected to 40-min renal artery ischemia. Iron loading produced a 28% increase in kidney iron content without any change in baseline renal function (plasma creatinine) or histology. Ischemic renal injury was far more severe in iron-loaded animals. Plasma creatinine 24 and 48 h after ischemia was significantly higher in iron-loaded rats (3.3 and 3.4 vs. 2.2 and 0.8 mg/dL) and GFR was significantly lower in iron-loaded rats (0.30 vs. 0.78 mL/min). In addition, iron-loaded rats showed a dramatically greater extent of damage by histologic evaluation using a semiquantitative scoring method. Therefore, a small increase in renal iron content greatly increased renal injury after an ischemic insult. These findings may be relevant to human renal disease because there is accumulating evidence of renal iron accumulation in a variety of proteinuric and chronic renal diseases.