The effect of anthocyanin from Dioscorea alata L. after purification, identification on antioxidant capacity in mice.

Increasing findings devote to searching for natural active compositions as additives to ameliorate health status. Anthocyanin, water-soluble natural pigment, has been concerned due to its favorable antioxidant activity. In this study, we purified anthocyanin from Dioscorea alata L., identified its compounds, and evaluated its antioxidant properties. The results indicated that the purity of anthocyanin increased to 39.59 ± 1.56%, 60.18 ± 1.97%, and 81.08 ± 1.97% after purification via AB-8 macroporous resin, Sep-Pak C18 solid phase, and LH-20 Sephadex stepwise. Ultra-performance liquid chromatography tandem mass spectrometer results indicated that paeoniflorin-3,5-O-dihexoside, petunin-3-O-feruloyl-glucoside-5-O-glucoside, cyanidin-3-O-feruloyl glucoside-5-O-glucoside, cyanidin-3-O-sophoroside, and petunin-3,5-O-dihexoside were the major compounds. The purified anthocyanin exhibited stronger antioxidant activity than the unpurified extract and ascorbic acid, whereas weaker than that of cyanidin-3-O-glucoside in general, which was assessed using DPPH, ABTS, and Fe3+ reducing capacity methods. Moreover, the purified anthocyanin increased GSH-Px, total antioxidant capacity, and superoxide dismutase activities and decreased malondialdehyde concentration on serum in mice after administering lipopolysaccharide for 24 h (p < .05). To summarize, the purified anthocyanin boasts more outstanding antioxidant properties than that of crude extracts. These results provide a reference with source of anthocyanin and it is conducive to use Dioscorea alata L. resources.

Microfluidic Preparation of pH-Responsive Microsphere Fibers and Their Controlled Drug Release Properties.

In this study, a capillary microfluidic device was constructed, and sodium alginate solution and a pH-sensitive hydrophobic polymer (p(BMA-co-DAMA-co-MMA)) solution were introduced into the device for the preparation of hydrogel fibers loaded with polymer microspheres. The structure of the microsphere fiber, including the size and spacing of the microspheres, could be controlled by flow rate, and the microspheres were able to degrade and release cargo responding to acidic pH conditions. By modification with carboxymethylcellulose (CMC), alginate hydrogel exhibited enhanced pH sensitivity (shrunk in acidic while swollen in basic condition). This led to an impact on the diffusion rate of the molecules released from the inner microspheres. The microsphere fiber showed dramatic and negligible degradation and drug release in tumor cell (i.e., A431 and A549 cells) and normal cell environments, respectively. These results indicated that the microsphere fiber prepared in this study showed selective drug release in acidic environments, such as tumor and inflammation sites, which could be applied as a smart surgical dressing with normal tissue protective properties.

CDCA5 is negatively regulated by miR-326 and boosts ovarian cancer progression.

PURPOSE: Ovarian cancer (OC) is the fifth leading cause of cancer death in women and one of the most prevalent malignancies in humans. Therefore, improved methods for OC early detection are urgently needed. Research has demonstrated that microRNAs (miRs) are strongly linked to OC tumorigenesis. The potential regulatory mechanism regarding miR-326 in OC is undefined. The present study was aimed to explore miR-326 expression in OC and its roles in OC progression. METHODS: Qualitative (q)RT-PCR was adopted to examine miR-326 expression in OC tissues and cell lines in clinical samples. qRT-PCR and Western blot were applied to detect division cell cycle associated 5 (CDCA5) expression at the messenger RNA (mRNA) and protein levels. CCK-8 and Transwell invasive experiments were utilized to examine cell proliferation and invasion. Cell cycle and apoptosis were analyzed by flow cytometry. Online bioinformatics analysis was employed to predict the target genes of miR-326 and luciferase reporter genes were applied for validation. RESULTS: MiR-326 was remarkably down-regulated in OC tissues and cell lines relative to the corresponding adjacent normal tissues and normal human ovarian surface epithelial cells (HOSE). MiR-326 overexpression markedly restrained the proliferation and invasion of OC cells, whereas miR-326 inhibitors exerted the contrary effect. Additionally, miR-326 up-regulation in OC cells prevented cells from entering S-phase and enhanced apoptosis, a phenomenon that may be due to CDCA5 down-regulation. Furthermore, we proved that CDCA5 was a downstream target of miR-326. CONCLUSIONS: MiR-326 represses the proliferation and invasion of OC cells and enhances apoptosis by specifically modulating CDCA5 expression.

Long non-coding RNA TUG1 promotes endometrial cancer development via inhibiting miR-299 and miR-34a-5p.

It is generally known that the human genome makes a large amount of noncoding RNAs compared with coding genes. Long non-coding RNAs (lncRNAs) which composed of more than 200 nucleotides have been described as the largest subclass of the non-coding transcriptome in human noncoding RNAs. Existing research shows that lncRNAs exerted biological functions in various tumors via participating in both oncogenic and tumor suppressing pathways. The previous studies indicated that lncRNA taurine upregulated 1 (TUG1) play important roles in the initiation and progression of malignancies. In this study,based on previous research, we investigated the expression and biological role of the lncRNA-TUG1. We analyzed the relationship between lncRNA-TUG1and endometrial carcinoma (EC) in a total 104 EC carcinoma specimens, compared with that in normal tissues. We found that lncRNA-TUG1 expression in cancer tissues was significantly higher than that in adjacent tissues. Through a series of experiments, the results demonstrated that lncRNA-TUG1 enhances the evolution and progression of EC through inhibiting miR-299 and miR-34a-5p.

[Sequence analysis of coding regions of KCNJ5 gene in unilateral adrenal hyperplasia].

OBJECTIVE: To investigate the prevalence of KCNJ5 gene missense mutations and their role in patients with unilateral adrenal hyperplasia (UAH). METHODS: Fourteen UAH tissues were collected through surgical resection, and all the tissues were confirmed by pathology. Peripheral blood samples of the same patients were collected as control. The coding regions of the KCNJ5 were detected by direct DNA sequencing. Protein structure and function were predicted with specific software. RESULTS: Three missense mutations were detected among the 14 patients with UAH, which included c.451G>C/A (p.G151R) (2/14), c.503T>G (p.L168R) (1/14), c.830T>A (p.S209T) (9/14). Among these, c.830T>A is a newly identified somatic mutation. Protein structure prediction showed that S209T lied in the second transmembrane domain, a conservation region of KCNJ5. S209 was also the phosphorylation site of PKC that is located in intracellular area. CONCLUSION: Missense mutations of KCNJ5 gene may be associated with UAH. Protein structure prediction has suggested that KCNJ5 mutations may be associated with UAH.

[Association of GIRK4 gene polymorphisms with essential hypertension in obese ethnics Uygur from southern Xinjiang].

OBJECTIVE: To assess the association of polymorphisms of G protein-coupled inwardly-rectifying potassium channels 4 (GIRK4) gene with essential hypertension in ethnic Uygurs from southern Xinjiang. METHODS: A total of 1194 (461 males and 733 females) Uygur residents aged 30 to 70 and with a body mass index (BMI) over 18.5 kg/m(2) were selected from Hetian region. All of the subjects have received questionnaire survey, physical examination, biochemical analysis and blood pressure measurement. They were divided into hypertensive group and normotensive group. Genotyping by the TaqMan polymerase chain reaction method was performed for 4 common single nucleotide polymorphisms (rs4937391, rs2604204, rs6590357 and rs1122149), and a case-control study was carried out. RESULTS: Genotype distributions of rs4937391, rs2604204, rs6590357 and rs1122149 in both groups were in Hardy-Weinberg equilibrium (P> 0.05). The average systolic blood pressure of CC genotype of rs11221497 single nucleotide polymorphism (SNP)[(132.69± 26.9) mmHg)] was higher than the CG genotype [(127.4± 22.7) mmHg] and GG genotype [(121.1± 26.3) mmHg]. There has a significantly difference in average systolic and diastolic blood pressures between CC and GG genotypes (P< 0.05). A case-control association analysis revealed that the rs11221497 SNP was in association with essential hypertension with the dominant model [P< 0.05, OR= 0.67 (0.49-0.93)]. Haplotype analysis indicated that H6(C-G-C-G) was significantly more common in normotensive group than hypertensive group (P= 0.001). CONCLUSION: The rs11221497 SNP of the GIRK4 gene is associated with essential hypertension in ethnic Uygur population in Xinjiang.

Systemic evaluation of hypoxic-ischemic brain injury in neonatal rats.

The objective of the study was to evaluate the systematically rat model of neonatal hypoxic-ischemic brain damage. The right carotid arteries of 7-day-old healthy Wistar rats were ligated, and then, the rats were subjected to an environment with 8 % of oxygen. Four weeks after the birth, neurobehavioral test, water maze test, and motor-evoked potential and neuropathologic examinations were performed. The footprint analysis showed significantly larger and instable paces in the hypoxic-ischemic group (P < 0.05); the time that rats crossed the balance beam in the hypoxic-ischemic group was longer than the control group (P < 0.05). The water maze test showed that the escape latency of hypoxic-ischemic group was significantly longer than that of control group (P < 0.05). The hindlimb quadriceps compound muscle-evoked potential CMEP of rats in hypoxic-ischemic group showed that the wave amplitude was lower than that of control group (P < 0.05). HE staining showed visible periventricular leukomalacia in hypoxic-ischemic groups; disrupted nuclear membrane was detected in the IH group with transelectronmicroscopy; Immunohistochemistry: compared with control group, MBP-positive neurocytes decreased, glial fibrillary acidic protein positive neurocytes increased in the periventricular zone (P < 0.05). Carotid artery ligation combining the hypoxic chamber created a reliable and stable rat model of neonatal hypoxic-ischemic brain damage and can be used for experimental research related to management of cerebral palsy.

[Expression of GIRK4 gene in kidney tissues of obese rat].

OBJECTIVE: To investigate the expression of GIRK4 gene in the kidney tissues of obese rats. METHODS: Obese rat models were established using diet-induced method. The GIRK4 protein expression in kidney tissues was determined in 20 obese rats and 10 normal rats using Western blot analysis. RESULTS: The relative expression level of GIRK4 protein in the kidney tissues of obese rat (1.75±0.42) was significantly lower than that in normal rats (3.37±0.68, P<0.05). CONCLUSION: GIRK4 has a low protein expression in the kidney tissues of obese rat.

[Relationship between the G protein gated inward rectifier potassium channel 4 gene polymorphism and dyslipidemia of Uyghur residents].

OBJECTIVE: To investigate the relationship between the G protein-gated inward rectifier K+ channel subunit 4 (GIRK4) gene polymorphism and the dyslipidemia among Uyghur residents in Xinjiang. METHODS: The polymorphisms of rs2604204, rs4937391, rs6590357, and rs11221497 among the Uyghur residents were genotyped using Taqman polymerase chain reaction (PCR). Lipid levels were measured by conventional methods and were analyzed. RESULTS: In the less-than-50-years population, the genotype distributions of the rs6590357 was statistically significant different in subjects with or without abnormal triglycerides (P=0.005). Aslo, the the genotype distributions of the rs11221497 also significantly differed in subjects with normal compared or abnormal TG (P=0.011). Logistic regression analysis suggested that rs6590357 still had positive association with TG abnormalities in subjects under 50 years (P=0.014). rs11221497 also had positive association with TC abnormalities. The TG levels of CT+TT genotypes were significantly higher than the CC group (P=0.006). Haplotype analysis found that the differences of H3 haplotype frequencies between the TG abnormal and normal groups were statistically significant (P=0.007). CONCLUSION: The polymorphisms of rs11221497 and rs6590357 of GIRK4 gene may play a role in the development of dyslipidemia in Uygur population.

A rat pup model of cerebral palsy induced by prenatal inflammation and hypoxia.

Animal models of cerebral palsy established by simple infection or the hypoxia/ischemia method cannot effectively simulate the brain injury of a premature infant. Healthy 17-day-pregnant Wistar rats were intraperitoneally injected with lipopolysaccharide then subjected to hypoxia. The pups were used for this study at 4 weeks of age. Simultaneously, a hypoxia/ischemia group and a control group were used for comparison. The results of the footprint test, the balance beam test, the water maze test, neuroelectrophysiological examination and neuropathological examination demonstrated that, at 4 weeks after birth, footprint repeat space became larger between the forelimbs and hindlimbs of the rats, the latency period on the balance beam and in the Morris water maze was longer, place navigation and ability were poorer, and the stimulus intensity that induced the maximal wave amplitude of the compound muscle action potential was greater in the lipopolysaccharide/hypoxia and hypoxia/ischemia groups than in the control group. We observed irregular cells around the periventricular area, periventricular leukomalacia and breakage of the nuclear membrane in the lipopolysaccharide/hypoxia and hypoxia/ischemia groups. These results indicate that we successfully established a Wistar rat pup model of cerebral palsy by intraperitoneal injection of lipopolysaccharide and hypoxia.

[Association between GIRK4 gene polymorphisms and insulin resistance in Xinjiang Uygur population].

OBJECTIVE: To assess the association between polymorphisms of protein-activated inwardly rectifying K+ channel (GIRK4) gene and insulin resistance (IR) in Xinjiang Uygur population. METHODS: A cross-sectional epidemiological survey-based case-control study was carried out, for which 1295 subjects (including 324 IR patients and 971 non-IR controls) were randomly selected. Functional region of the GIRK4 gene was sequenced for 48 randomly selected IR patients. Representative variable sites were chosen, with its association with IR assessed in 1295 Uygur subjects. RESULTS: rs11221497 variant was associated with IR in Uygur subjects under 50 years old (P=0.017 in genotype model, P=0.009 in dominant model). Subjects with dominant model of CC genotype have an OR of 1.833 (95%CI: 1.157-2.905) for IR. CONCLUSION: GIRK4 gene polymorphisms may be associated with IR in Uygur ethnics from Xinjiang. The CC genotype of rs11221497 variant is a risk factor for IR.

Advances in research on G protein-coupled inward rectifier K(+) channel gene.

G protein-coupled inward rectifier K(+) channel 4(GIRK4) is a G protein-coupled inward rectifier potassium channel family member. Encoded by the KCNJ5, it is widely distributed in the mammalian heart, brain, and other tissues and organs. Recent studies have demonstrated that the abnormal expression of GIRK4 gene is associated with atrial fibrillation, and meanwhile may be closely related to obesity, metabolic syndrome, and many other clinical conditions. Further research on the role the GIRK4 gene in the pathophysiology of these clinical conditions will definitely facilitate their clinical diagnosis and treatment.

Functional recovery following traumatic spinal cord injury mediated by a unique polymer scaffold seeded with neural stem cells and Schwann cells.

BACKGROUND: The most important objective of transplant studies in the injured spinal cord has been to provide a favorable environment for axonal growth. Moreover, the continuing discovery of new grafts is providing new potentially interesting transplant candidates. Our purpose was to observe the morphological and functional repair effects of the co-transplantation of neural stem cell (NSC), Schwann cells (SCs) and poly lactide-co-glycolide acid (PLGA) on the spinal cord injury of rats. METHODS: A scaffold of PLGA was fabricated. NSCs and SCs were cultured, with the NSCs labeled with 5-bromodeoxyuridine, and the complex of NSC/PLGA or NSC + SCs/PLGA were constructed. Thirty-six Wistar rats were randomly divided into three groups: group A (transplantation of PLGA), group B (transplantation of NSC/PLGA) and group C (transplantation of NSC + SCs/PLGA). The 3 mm length of the right hemicord was removed under the microscope in all rats. The PLGA or the complex of PLGA-cells were implanted into the injury site. Basso-Beattie-Bresnahan (BBB) locomotion scores, motor and somatosensory evoked potential of lower limbs were examined to learn the rehabilitation of sensory and motor function at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury. All the recovered spinal cord injury (SCI) tissues were observed with HE staining, immunohistochemistry, and transelectronmicroscopy to identify the survival, migration and differentiation of the transplanted cells and the regeneration of neural fibres at 4 weeks, 8 weeks, 12 weeks and 24 weeks after injury. RESULTS: (1) From 4 weeks to 24 weeks after injury, the BBB locomotion scores of cell-transplanted groups were better than those of the non-cell-transplanted group, especially group C (P < 0.05). The amplitudes of the somatosensory evoked potential (SEP) and motor-evoked potential (MEP) were improved after injury in groups B and C, but the amplitude of SEP and MEP at 4 weeks was lower than that at 12 weeks and 24 weeks after injury. Compared with group B, the amplitude of SEP and MEP in group C was improved. The amplitude of SEP and MEP was not improved after injury in group A. (2) HE staining revealed the volume of the scaffold decreased and the number of cells in the scaffold increased. Newly-grown capillaries also could be seen. Immunohistochemistry staining showed the transplanted NSCs could survive and migrate until 24 weeks and they could differentiate into neurons and oligodendrocytes. The regenerated axons were observed in the scaffold-cell complex with transelectronmicroscopy. The above manifestations were more extensive in group C. CONCLUSIONS: The transplanted NSC can survive and migrate in the spinal cord of rats up to 24 weeks after injury, and they can differentiate into various neural cells. Co-transplantation of cells/PLGA can promote the functional recovery of the injured spinal cord. The effect of co-transplanting NSC + SCs/PLGA is better than transplanting NSC/PLGA alone.

[Effects of fuzheng quxie granule on immune cells and cytokines in populations with respiratory viral infection].

OBJECTIVE: To investigate the effect of fuzheng quxie granule (FQG) on immune cells and cytokines in populations with respiratory viral infection. METHODS: Fifty-nine patients were randomly divided into 3 groups, that is, 19 patients treated with conventional western medicine (WM) plus FQG in the treated group, 19 patients treated with conventional western medicine alone in the WM group, and 21 patients treated with FQG alone in the TCM group. The levels of T lymphocyte subsets, interleukine-2,4,6,10 (IL-2, IL-4, IL-6, IL-10), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (INF-gamma) and Th1/Th2 were determined before treatment, and at the end of 1st and 2nd week of treatment respectively. RESULTS: Before treatment, levels of TNF-alpha, IL-2, IL-6, IL-10 and INF-gamma in all patients were significantly higher than normal range (P < 0.05). After being treated for 1 week, the levels of serum TNF-alpha, IL-6, and IL-10 were significantly decreased in all groups (P < 0.05), serum IL-2 and INF-gamma decreased to the normal level in the WM group, but in the treated and the FQG group by the end of the 2nd week, the two indexes still remained at the rather higher level (P < 0.05). The ratio of Th1 and Th2 in the treated group and the FQG group increased significantly by the end of 2nd week, reached the level higher than that in the WM group and that before treatment (P < 0.05). No significant difference in, T lymphocytes subsets (CD3+ , CD4+ , CD8+) and percentage of B and NK cells before and after treatment was found in all the 3 groups. CONCLUSION: FQG can positively regulate the immune function of patients with respiratory tract viral infection in certain degree.