Hotair mediates tumorigenesis through recruiting EZH2 in colorectal cancer.

Long noncoding RNAs (lncRNA)  have been demonstrated to extensively participate in a wide spectrum of biological activities ranging from embryogenesis and cancer progression. HOX transcript antisense RNA (Hotair), an lncRNA located in the HOXC locus, has been reported to play an important role in carcinogenesis. As a well-known oncogene, it potentiates cancer metastasis and tumor progression. And it also serves as a biomarker for poor prognosis and tumor recurrence. In this study, Hotair was found to be upregulated in colorectal cancer (CRC) cells and clinical specimens. Further investigation showed that knockdown of Hotair dramatically suppressed cell proliferation and colony formation, suggesting that Hotair may stimulate tumorigenesis of CRC. The enhancer of zeste homolog 2 (EZH2), a regulator of epigenetic modification, was upregulated in CRC cells and clinical samples. And the silence of EZH2 significantly suppressed cell viability and colony formation. Furthermore, the RNA immunoprecipitation assay revealed that Hotair directly bound EZH2 in CRC cells. In conclusion, Hotair mediated tumorigenesis via recruiting EZH2, which might shed light on the development of a novel therapeutic approach for patients with CRC.

[Nuclear magnetic resonance-based metabonomics for sedative and hypnotic effect of Banxia Houpo decoction].

Insomnia was a common disease, which might be correlated with γ-aminobutyric acid A (GABAA) receptor mechanism, cytokine regulatory mechanism, excitatory amino acid mechanism and hydroxytryptamine (5-HT) receptor mechanism, but the correlations between these independent mechanisms and pathological characterization were still unclear. To further explore the effect of Banxia Houpo decoction on known or unknown biological pathways during treatment of insomnia, the metabonomics method based on ¹H-NMR was developed for detecting the significant changes in metabolomics after the administration with Banxia Houpo decoction in pentobarbital sodium-induced rat sleeping experiment. Serum and urine samples were analyzed by ¹H-NMR. Principal component analysis (PCA) was carried out for endogenous small molecule metabolites in urine and serum. H-NMR spectroscopies and relevant metabolites were found and identified by Simca-p 17.0 (Umet-rics, Umea, Sweden) and Chenomx NMR Suite 7.1 (Chenomx, Inc., Edmonton, Alberta, Canada) software. The result suggests that Banxia Houpo decoction group and indiplon group had significant differences. The load diagram showed the biggest variation metabolites and intergroup significant differences among 10 metabolic substances. According to the experiment, Banxia Houpo decoction group and indiplon group can prolonge the sleeping time of pentobarbital sodium-induced sprague-dawley rats, with a synergistic effect. The significant changes of these biomarkers indicated that the Banxia Houpo decoction could aid sleep by adjusting the content of glutamine, creatine phosphate, 2-oxoglutarate, and reducing the activity of brain nerves.

Modulation of metallothionein isoforms is associated with collagen deposition in proliferating keloid fibroblasts in vitro.

The keloid fibroblast (KF) is known to have higher proliferative capacity than normal dermal fibroblast (NF). Metallothionein (MT), a metal-binding protein, has been reported to promote cell proliferation. In this study, we evaluated the expression of MT isoforms at the mRNA level in fetal bovine serum (FBS)-stimulated proliferating KF. Although the morphological appearance of NF and KF was similar when viewed under light, confocal and transmission electron microscopy, there was surprisingly a generally lower expression of MT isoforms in KF when compared with NF and also reduced MT staining in dermal fibroblasts of keloids as opposed to normal skin. Primary cultures of KF grown in 5% FBS or 10% FBS compared to without FBS demonstrated significantly higher proliferative activity and more abundant deposition of collagen. Contrary to expectation, MT-1A, -1F, -1G, -1X and -2A isoforms were significantly down-regulated in proliferating KF. Moreover, stimulating KF with TGF ß1, which is known to promote collagen synthesis and keloid formation, increased expression of Collagen 1A and 3A genes accompanied by reduction in MT-2A gene expression. Furthermore, down-regulation of the MT-2A gene in proliferating KF by siRNA-mediated silencing enhanced cell proliferation with concomitant up-regulation of the NF-κB gene and 10 of 13 other NF-κB pathway-related genes analysed but no alteration of the Collagen 1 and Collagen 3 gene expression. It would appear that down-regulation of MT isoforms in proliferating KF, in particular MT-2A, enhances keloidogenesis with the possible involvement of the NF-κB signalling pathway.

[Treatment of pediatric malnutrition by acupuncture of Sifeng (EX-UE10) combined with oral administration of Wang's Baochi Pill].

OBJECTIVE: To investigate the clinical effect of acupuncture at Sifeng (EX-UE10) combined with Wang's Baochi Pills in the treatment of pediatric malnutrition, so as to provide a more effective method for pediatric malnutrition. METHODS: A total of 201 children with malnutrition were randomly divided into combined treatment group (n＝102) and control (Baochi Pill) group (n＝99). The children in the combined treatment group were treated by acupuncture stimulation of Sifeng (EX-UE10, till no more yellowish-white effusion out) and oral administration of Wang's Baochi Pills, and those in the control group treated by oral administration of Wang's Baochi Pills only. The course of treatment was one month for both groups. The integral score of symptom was assessed according to the main symptoms as body weight and height and food-intake, and to the secondary symptoms including mentality, agitation, sleep, hair gloss, susceptibility to cold, hydrosis, abdominal distension, and susceptibility to diarrhea or constipation. The therapeutic effect was assessed by consulting the "Criteria for Diagnosis and Therapeutic Effect Evaluation of Syndromes/Illnesses of Traditional Chinese Medicine (TCM)" and "TCM Professional Criteria of the People's Republic of China for Diagnosis and Therapeutic Effect Evaluation of Syndromes/Diseases of TCM Pediatric". RESULTS: After the treatment, the total symptom scores of both groups were significantly decreased in comparison with their own pre-treatment (P<0.05), and the scores of total symptom after the treatment and the 1st, 2nd and 3rd follow-up surveys were obviously lower in the combined treatment group than in the control group (P<0.05). Of the 99 and 102 cases in the control and combined treatment groups, 47 and 59 were cured, 39 and 37 experienced improvement in their symptoms, and 13 and 6 failed, with the effective rate being 86.87% (86/99) and 94.11%(96/102), respectively. The effective rate of the combined treatment was evidently higher than that of the simple medication (P<0.05). CONCLUSION: Acupuncture of Sifeng (EX-UE10) combined with Wang's Baochi Pills is better than administration of Wang's Baochi Pills alone in the therapeutic effect for pediatric malnutrition.

Comparison of the effects of differentially sulphated bovine kidney- and porcine intestine-derived heparan sulphate on breast carcinoma cellular behaviour.

Heparan sulphate is a sulphated glycosaminoglycan and is able to bind to and regulate the activity of many growth and signalling factors. We have previously shown that its expression is correlated with tumour grade and cell proliferation in breast phyllodes tumours. In this study, we examined the use of heparan sulphate as a biomarker of invasive ductal carcinoma and the effects of differentially sulphated heparan species on breast cancer cell behaviour. Immunohistochemistry using the 10E4 monoclonal antibody was carried out on 32 paraffin-embedded breast cancer specimens and paired non-cancerous breast tissues to compare the expression patterns of heparan sulphate. Upregulated expression of the sulphated 10E4 epitope in heparan sulphate was detected in both epithelial and stromal compartments of breast cancer compared with normal mammary tissues, with a 2.8X increase in immunoreactivity score. To determine the effects of differentially sulphated heparan sulphate molecules on breast cancer behaviour, cultured breast carcinoma cells were treated with chlorate, a competitive inhibitor of glycosaminoglycan sulphation, and two different heparan sulphate species. Inhibition of glycosaminoglycan sulphation resulted in a significant increase in cancer cell adhesion and a reduction in cell migration, together with upregulated expression of focal adhesion kinase and paxillin. Both porcine intestine- and bovine kidney-derived heparan sulphate species could block the change in cell adhesion. However, the former heparan sulphate species completely abolished, while the latter exacerbated, the chlorate-induced decrease in cell migration. The results show that heparan sulphate is a useful biomarker of breast invasive ductal carcinoma. Different sulphation patterns of heparan sulphate residues have differential effects in regulating breast cancer cellular behaviour, and this may be exploited to develop heparan sulphate into a useful target for treatment of breast carcinoma.

[Single nucleotide polymorphism analysis on the 5' regulatory region of IGF2 gene in Wuzhishan pig].

Single nucleotide polymorphisms (SNPs) in partial 5' regulatory region of the insulin-like growth factor 2 (IGF2) gene were studied by DNA sequencing in 60 pigs from the Wuzhishan, Diannan small-ear, Xiang, Meishan and Large White pig breeds. Thirteen SNP sites were detected, including one transversion at T6029A, 4 A<---->G transitions (A5976G, G13520A, G13563A and G13669A) and 8 C<---->T transitions (C5872T, C5888T, C6010T, C6037T, C6043T, C6063T,C6112T, C6164T). These 13 SNPs formed 23 composite genotypes. The gene, genotype and composite genotype frequencies of every SNP site in the whole group and in each breed were calculated. Results showed that the predominant allele in 3 miniature pig breeds was G, T and A at A5976G, C6164T and G13669A sites respectively, but the A-C-G allele was pre-dominant in Meishan and Large White breeds. Moreover, H15 and H19 were the characteristic composite genotype for the large versus the miniature breeds, respectively. In addition, the C5888T SNP was analyzed in 123 Wuzhishan pigs by the PCR-RFLP method. Results showed that the predominant allele was C, and the predominant genotype was CC. chi2-test results indicated that the Wuzhishan pig breed was at Hardy-Weinberg equilibrium with respect to this SNP. These results provide the miniature pig breeds such as the Wuzhishan pig with certain genetic references on the regulation of growth and development, and the mechanism of its dwarfism.

P53 Codon 72 polymorphisms: a case-control study of gastric cancer and potential interactions.

P53 codon 72 polymorphisms have been reported to be associated with cancers of the lung, esophagus and cervix. However, there have been no reports on the interaction of select risk factors and p53 codon 72 polymorphisms in gastric cancer susceptibility. 155 gastric cancer cases and 134 cancer-free controls were enrolled at the Memorial Sloan Kettering Cancer Center (MSKCC) from November 1992 to November 1994. The crude odds ratio (OR1) associated with the (Pro/Pro) polymorphism and the risk of gastric cancer was 1.27 (0.70-2.33). Adjusting for age, sex, race and education (OR2) and further adjusting for BMI, calories, sodium, smoking, vitamin C, fiber, alcohol, fat, and H. pylori status (OR3) did not yield significant results. Significant joint effects were associated with high fat consumption (OR1=2.61 (95% CI:1.13-6.06); OR2=2.85 (95% CI:1.14-7.15) for total cancers and for proximal tumors (OR1=2.56 (95%CI:1.00-6.54)). The low vitamin C intake/high-risk polymorphism group (Pro/Pro) had an OR1 of 4.82 (95% CI: 1.72-13.45) and the OR2 was 6.19 (95% CI: 2.08-18.40) for distal tumors. The point estimates were increased for interaction odds ratios but not statistically significant (OR1=4.25 (95% CI: 0.66-27.50); OR2=4.73 (95% CI: 0.67-33.43); OR3=5.55 (95% CI: 0.66-46.47)). Further studies specifically looking at proximal and distal tumors are required to confirm any potential interaction between the p53 codon 72 polymorphisms and environmental risk, in particular low dietary vitamin C and high fat consumption.

Lazaroid U83836E protects the heart against ischemia reperfusion injury via inhibition of oxidative stress and activation of PKC.

Oxidative stress has been demonstrated to be important during myocardial ischemia/reperfusion injury (MIRI). The lazaroid U83836E, which combines the amino functionalities of the 21­aminosteroids with the antioxidant ring portion of vitamin E, is a reactive oxygen species scavenger. The aim of the current study was to investigate the effect of U83836E on MIRI and its mechanisms of action. Rat hearts were subjected to 30 min ligation of the left anterior descending coronary artery, followed by 2 h reperfusion. The results demonstrated that at 5 mg/kg, U83836E markedly protected cardiac function in ischemia/reperfusion rat models, decreased the malondialdehyde content and creatinine kinase activity, while increasing superoxide dismutase and glutathione peroxidase activity. Additionally, U83836E significantly decreased the histological damage to the myocardium, reduced the area of myocardial infarction in the left ventricle and modified the mitochondrial dysfunction. Furthermore, U83836E enhanced the translocation of protein kinase Cε (PKCε) from the cytoplasm to the membrane. However, the cardioprotective effects of U83836E were reduced in the presence of the PKC inhibitor, chelerythrine (1 mg/kg). Therefore, the results of the present study suggest that U83836E has a potent protective effect against MIRI in rat models through the direct anti­oxidative stress mechanisms and the activation of PKC signaling.

[Genetic analysis of 32 microsatellite loci in 13 families of Wuzhishan pig by multiplex PCR and gene scanning technique].

Wuzhishan pig is one of the rare and endangered breeds in china. They have the following characteristics such as :light body weight and small size, early sexually maturity, high meat quality and genetic identification with 6 approximately 8 pares litter size,body weight of born 0.3 approximately 0.4 kg, 15 approximately 16 kg at 6 month old, 35 kg at 2 years old, and so on. They may be used for laboratory utilization, comparative studies on human medical model, embryonic engineering, nutrition metabolism, sensitivity test on virus and bacteria, skin brut and tranfer, removing lipid, teeth and mouth cavity diseases, studies on cardiovascular model and evaluation of new medicine products. The polymorphisms of 32 microsatellites in 13 families of Wuzhishan pig in Hainan were Analysed. Number of alleles in each family was counted, mean heterozygosity and polymorphism Information content(PIC) were calculated. The results showed that number of alleles was 13.66, mean heterozygosity was 0.559 while polymorphism information content was 0.731. This revealed that genetic diversity is abundant in Wuzhishan pig in Hainan. These results have instructional significance for preserving breeds, selection and breeding, development and utilization of Wuzhishan pig in Hainan.

[The synthesis and spectral study of multicomponent heteropoly compounds with Keggin structure. II].

The multicomponent heteropoly compounds [Me4N]4 + x[SiVxMoyW12 - x - yO40].n H2O with Keggin structure have been synthesized and characterized by FTIR. The vibrational frequencies of containing-oxygen bonds (Si-Oa,M=Od, M-Ob-M and M-Oc-M) in these heteropoly compounds have been studied. The vibrational frequencies nu as(M=Od) and nu as(M-Ob-M) are demonstrated to explain why the acidity of the heteropoly compounds becomes weaker and the oxidative ability becomes stronger when W atoms are substituted by V atoms. The rule of the change is similar with [PVxMoyW12 - x - yO40](3 + x)-. In addition, the acidity of P series is stronger than the acidity of the corresponding Si series.

Developmental competence of embryos derived from reciprocal in vitro fertilization between Yak (Bos grunniens) and cattle (Bos taurus).

The purpose of this study was to investigate fertilization ability and embryo development to the blastocyst stage after reciprocal in vitro fertilization (IVF) between yak and cattle in an attempt to clarify the problem of low conception rate after mating yak females with cattle bulls. In vitro-matured (IVM) cattle and yak oocytes were inseminated with either Holstein or yak spermatozoa, and after an 18-h of coincubation period, a proportion of the oocytes was fixed and examined for sperm penetration, polyspermy and male pronuclear formation. The remaining oocytes were cultured in vitro and evaluated for cleavage and blastocyst formation rates. The percentage of IVM oocytes penetrated by spermatozoa ranged from 78.5 to 90.5%, and the formation of one or two pronuclei and the incidence of polyspermy did not differ among the different combinations. The cleavage and blastocyst rates were not affected by the species of the sperm, but they were affected by the species of the oocytes (P<0.05), with cattle oocytes having a higher (P<0.05) cleavage and blastocyst rates (69.9 and 31.3%) than yak oocytes (62.7 and 11.5%). The blastocyst formation rate was calculated from the cleaved zygotes. The interaction between sire and oocytes species (P<0.05) influenced blastocyst formation rate, with the highest blastocyst rate occurring in cattle oocytes fertilized with yak spermatozoa (36.5%) and the lowest rate occurring in yak oocytes fertilized with yak spermatozoa (9.4%). The effect of heterosis was apparent at the blastocyst stage, but there was a large reciprocal difference in blastocyst production between crosses. It was concluded that the low conception rate that results from crossing yaks with cattle is not due to either a species-specific block of fertilization or the developmental competence of the early stage embryo.

Transient expression of endothelins in the amoeboid microglial cells in the developing rat brain.

Amoeboid microglial cells (AMC) which transiently exist in the corpus callosum in the postnatal rat brain expressed endothelins (ETs), specifically endothelin-1 (ET-1) and ET3 as revealed by real time RT-PCR. ET immunoreactive AMC occurred in large numbers at birth, but were progressively reduced with age and were undetected in 14 days. In rats subjected to hypoxia exposure, ET immunoexpression in AMC was reduced but the incidence of apoptotic cells was not increased when compared with the control suggesting that this was due to its downregulation that may help regulate the constriction of blood vessels bearing ET-A receptor. AMC were endowed ET-B receptor indicating that ET released by the cells may also act via an autocrine manner. In microglia activated by lipopolysaccharide (LPS), ET-1 mNA expression coupled with that of monocyte chemoattractant protein (MCP-1) and stromal derived factor-1 (SDF-1) was markedly increased; ET-3 mRNA, however, remained unaffected. AMC exposed to oxygen glucose deprivation (OGD) in vitro resulted in increase in both ET-1 and ET-3 mRNA expression. It is suggested that the downregulated ETs expression in vivo of AMC subjected to hypoxia as opposed to its upregulated expression in vitro may be due to the complexity of the brain tissue. Furthermore, the differential ET-1 and ET-3 mRNA expression in LPS and OGD treatments may be due to different signaling pathways independently regulating the two isoforms. The present novel finding has added microglia as a new cellular source of ET that may take part in multiple functions including regulating vascular constriction and chemokines release.

Gene regulation profile reveals consistent anticancer properties of progesterone in hormone-independent breast cancer cells transfected with progesterone receptor.

Absence of estrogen receptor (ER) and progesterone receptor (PR) is the hallmark of most hormone-independent breast cancers. Previous studies demonstrated that reactivation of PR expression in hormone-independent MDA-MB-231 breast cancer cells enabled progesterone to suppress cell growth both in vitro and in vivo. We determined the whole genomic effect of progesterone in PR-transfected MDA-MB-231 cells. We identified 151 progesterone-regulated genes with expression changes > 3-fold after 24 hr treatment. Most are novel progesterone target genes. Real-time RT-PCR analysis of 55 genes showed a 100% confirmation rate. Twenty-six genes were regulated at both 3 and 24 hr. Studies using translation inhibitor suggest that most of the 26 genes are primary progesterone target genes. Progesterone consistently suppressed the expression of genes required for cell proliferation and metastasis and increased the expression of many tumor-suppressor genes. Progesterone also consistently decreased the expression of DNA repair and chromosome maintenance genes, which may be part of the mechanism leading to cell cycle arrest. These data suggest potential usefulness of progestin in combating ER-negative but PR-positive breast cancer and indicate that progesterone can exert a strong anticancer effect in hormone-independent breast cancer following PR reactivation. The identification of many novel progesterone target genes open up new avenues for in-depth elucidation of progesterone-mediated molecular networks.