To establish an HPLC-MS/MS method for the analysis of quercetin, kaempferid and isorhamnetin in rats plasma and study its pharmamacokinetics after an intragastrical administration of Hippophae rhamnoides extracts. Five healthy male Sprague-Dawley (SD) rats were given single doses of H. rhamnoides extracts (quercetin 26.35 mg x kg(-1), kaempferid 4.040 mg x kg(-1), isorhamnetin 31.37 mg x kg(-1)), and then their orbital sinus blood samples were collected at different time points. The drug plasma concentration of the three flavonoids was determined by HPLC-MS/MS method. After that, the main pharmacokinetics parameters were calculated by using Kinetica 5. 0. 11 software. The methodological test showed that the linear concentration ranges of quercetin, kaempferid and isorhamnetin were 7.500-600.0 μg x L(-1) (R2 = 0.998 5), 1.000-80.00 μg x L(-1) (R2 = 0.998 5 ) and 10.00-800.0 μg x L(-1) (R2 = 0.998 0), respectively. The inner and inter-days precisions were both less than 14.0%. The plasma samples showed a good stability and consistency with the requirement of biological sample analysis after the samples were frozen once and placed at - 20 degrees C for 15 d and room temperature for 6 h and the treated analytes were placed at -20 degrees C for 24 h. For quercetin, the pharmacokinetic parameter t(½β), AUC(0-∞), MRT(0.∞), C.(max) and T(max) were (113.3 ± 19.37) min, (12 542.14 ± 3 504.05) μg x h x L(-1), (119.6 ± 13.29) h, (164.6 ± 27.33) μg x L(-1) and (5.199 ± 0.840 3) h, respectively. For kaempferid, the pharmacokinetic parameters t(½β), AUC(0-t), MRT(0-∞), C(max) and T(max) were (79.85 ± 17.15) min, (934.51 ± 94.59) μg x h x L(-1), (81.50 ± 13.75) h, (80.15 ± 14.24) μg x L(-1) and (3.827 ± 0.902 7) h, respectively. For isorhamnetin, the pharmacokinetic parameters t1,2,, AUC(0-t), MRT(0-∞), C(max) and T(max) were (118.3 ± 20.73) min, (26 067.77 ± 4 124.60) μg x h x L(-1), (129.0 ± 16.30) h, (269.6 ± 29.32) μg x L(-1) and (6.513 ± 1.450) h, respectively. The HPLC-MS/MS analysis method established in this study was proved to be sensitive and accurate and could be applied in the pharmacokinetic study of quercetin, kaempferid and isorhamnetin in rat plasma.
Animals ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; pharmacokinetics ; Hippophae ; chemistry ; Kaempferols ; blood ; pharmacokinetics ; Male ; Quercetin ; analogs & derivatives ; blood ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry ; methods

Objective To determine the prevalence of metabolic syndrome ( MS) in women who had gone through natural menopause, and to provide basic data for developing effective prevention and intervention of metabolic syndrome in Taixing area Jiangsu province. Methods A cluster sample of natural postmenopausal women aged 45 to 64 years were sampled and assessed by questionnaires, physical measurements, and biochemical tests; and analyzed according to the International Diabetes Federation ( IDF, 2005) diagnostic criteria for MS. Results The crude prevalence of metabolic syndrome was 35. 25% and the standardized prevalence rate was 31. 47% after age being adjusted. The prevalence of metabolic syndrome was increased with age ( P < 0. 01). The constituent ratio of the combinations of 3, 4, and 5 components in MS was 21. 45% , 10. 73% , and 3. 07% , respectively. Conclusion The prevalence of the metabolic syndrome among older women after menopause is high and increasing with aging. Public education should be strengthened to improve the quality of life in postmenopausal women and prevention and intervention should be adopted earlier as well.

Adolescent ; Child ; Child, Preschool ; Female ; Gastric Mucosa ; microbiology ; Gastrointestinal Diseases ; diagnosis ; microbiology ; Helicobacter Infections ; diagnosis ; microbiology ; Helicobacter pylori ; growth & development ; isolation & purification ; Humans ; Infant ; Male ; Mouth Mucosa ; microbiology

OBJECTIVE Neuroinflammation plays a critical role in neurodegenerative disorders, although the inflammation may not the initiating factor. Parkinson disease (PD) is characterized patho?logically by the accumulation of alpha synuclein (α-syn) and the loss of the dopamine (DA) neurons in the substantia nigra (SN), which has been reported to be induced by the stereotaxic injection of lipopolysaccharide (LPS) to the SN region in rodents. This study is to investigate the therapeutic benefit of the inhibition of miR-873 in PD. METHODS Rats received the right-unilaterally injection with concentrated LV-sponge or LV-EGFP 3 d before LPS treatment, 7 or 14 d after LPS treatment. The animals were tested for rotational behavior with the dopaminergic agonist apomorphine dissolved in sterile saline at 21 d after LPS injection. The regulation of miR-873 on the genes related with cholesterol transport and inflammation was assayed in SH-SY5Y cells and U251 cells. RESULTS TLR4-MyD88 signaling pathway was involved the regulation of miR-873 by LPS. The luciferase assay showed that HMGCR, ABCA1 and A20 were down- stream genes of miR- 873. The transfection of miR- 873 decreased the cholesterol levels in cell membrane, but increased in lysosome in SH-SY5Y cells. Compared with the control SH-SY5Y cells, cholesterol levels were higher in lysosome with α-synuclein overexpression or LPS treatment. The transfection of miR-873 increased the α-syn levels in lysosome in cells with α-synuclein overexpression. The loss of dopaminergic neuorns induced by LPS was significantly respectively decreased by 22.8%, 35.6% and 57% after the inhibition of miR-873 at 3 d before LPS treatment, 7 or 14 d after LPS treatment. Compared with LPS-treated group, the number of the rotation of rats was decreased by 60.4%, 33.5% and 13.2% after the inhibition of miR-873 at 3 d before LPS treatment, 7 or 14 d after LPS treatment. The inhibition of miR-873 significantly decreased accumulation of α-syn. The mRNA levels of HMGCR, ABCA1 and A20 in SN were decreased by LPS treatment, which was attenuated by the injection of LV- sponge. CONCLUSION The selective regulation of miR- 873 can protect the dopaminergic neurons from the LPS-induced damage. The inhibition of miR-873 can attenuate the relocation of cholesterol in lysosome and the accumulation of α-syn in neurons induced by LPS via the regulation of HMGCR, ABCA1 and A20.

Objective: To study and discuss the application effect of TCM characteristic nursing combined with individualized health education in vertigo multi-specialty integrated diagnosis and treatment platform, and to provide reference basis for clinical research in the future. Methods: From June 2016 to June 2017, 60 patients with vertigo treated by multi-professional integrated diagnosis and treatment platform in our hospital were studied. The subjects of both groups were treated with acupoint application, and were randomly divided into routine group and combined group. There were 30 cases in each group. The routine standardized nursing method was applied to the vertigo multi-specialty integrated diagnosis and treatment platform, and the combined group was applied to the platform based on the routine group, and the application effect of the two groups was compared. Results: The total effective rate of nursing in the observation group was 96.67% (29/30), which was significantly higher than that in the routine group (86.67% (26/30), and the difference was statistically significant (χ²= 4.172, P < 0.05).There was no significant difference in the quality of life score before treatment between the two groups (t = 0.178, P > 0.05), but the score of the observation group was significantly higher than that of the routine group (t = 4.237, P < 0.05), and the quality of life score of the observation group was significantly higher than that of the routine group (t = 4.237, P < 0.05). At the same time, the scores of quality of life after treatment in the two groups were significantly higher than those before treatment (t = 9.239, 5.217, P < 0.05).The nursing satisfaction of the observation group was 96.67% (29/30), which was significantly better than that of the conventional group (83.33% (25/30), and the difference was statistically significant (x = 5.372, P <0.05). After one year of follow-up, the number of recurrences in the observation group was 2, the recurrence rate was 6.67%, the number of recurrences in the conventional group was 6, the recurrence rate was 20.00%, the observation group was superior to the normal group, and the difference was statistically significant (χ²= 5.137, P<0.05). Conclusion In the integrated diagnosis and treatment platform of vertigo, the application of TCM nursing combined with individualized health education can improve the nursing effect, improve the patients' quality of life and nursing satisfaction, and reduce the recurrence rate. To provide new theoretical thinking and clinical methods for improving vertigo patients.

Neuritin is a new neurotrophic factor found recently. In order to identify the function of Neuritin clearly, the coding sequence of human neuritin was amplified by PCR from neuritin cDNA , this fragment digested by NocI and NotI was inserted into pET32a by T4 ligase and transformed into E. coli BL21 then the recombinant plasmid named pET32a-neuritin was constructed successfully . Neuritin was expressed distinctly after inducing by EPTG. The product was identified as neuritin by SDS-PAGE and Western blot analysis . The expression production was purified on Ni2+-NTA column.

This study was aimed to investigate the effect of curcumin in combination with bortezomib on the proliferation and apoptosis of human MM cell line H929 in vitro, and to explore its mechanisms. MTT assay was applied to detect the inhibitory effects of curcumin and bortezomib either alone or combined at different concentrations on H929 cells, and flow cytometry was employed to assay the apoptosis rate. In addition, RT-PCR was used to analyze the mRNA expression of gene BCL-2, BAX, cyclin D1. Immunofluorescence technique was performed to study the location changes of NF-κB P65 in different groups. The results showed that both curcumin and bortezomib inhibited the proliferation of MM cell line H929 in dose-dependent manner, and combination of these two drugs displayed synergistical effect. A much higher apoptosis rate was determined by flow cytometry in combinative groups than that in single or control group. And RT-PCR showed, as compared with curcumin or bortezomib group, there was mRNA expression decrease of BCL-2, cyclin D1 but increase of BAX in combined group. The expression of NF-κB P65 in nucleus was downregulated in either the curcumin or bortezomib group, however, distribution of NF-κB P65 in cytoplasm was observed in combined group. It is concluded that the combination of curcumin and bortezomib is much more effective for the inhibiting proliferation and promoting apoptosis of H929 cell line, which may function by inhibiting the transcription of NF-κB and apoptosis-related genes.
Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Curcumin ; pharmacology ; Cyclin D1 ; metabolism ; Drug Therapy, Combination ; Humans ; Multiple Myeloma ; metabolism ; NF-kappa B ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Pyrazines ; pharmacology ; Transcription Factor RelA ; metabolism ; bcl-2-Associated X Protein ; metabolism