Two new iridoid glycosides from the whole plant of Rehmania piasezkii.

Two new iridoid glycosides, piasezkiiosides A (1) and B (2), were isolated from aqueous extract of the whole plant of Rehmannia piasezkii. Their structures were established from the spectroscopic data, chemical transformation, and X-ray diffraction analysis. Compound 1 exhibited weak hepatoprotective activity against APAP-induced HepG2 cell damage.

Seven new pentasaccharides from the roots of Rehmannia glutinosa.

Seven new pentasaccharides (1-7), rehmaglupentasaccharides A-G, were isolated from the air-dried roots of Rehmannia glutinosa. Their structures were established from the spectroscopic data obtained and by chemical evidence. The known verbascose (8) and stachyose (9) were also obtained in the current investigation, and the structure of stachyose was unequivocally defined using X-ray diffraction data. Compounds 1-9 were tested for their cytotoxicity against five human tumor cell lines, influence on dopamine receptor activation, and proliferation effects against Lactobacillus reuteri.

Four new iridoid glycosides from the roots of Rehmannia glutinosa.

Four new iridoid glycosides (1-4), rehmaglutosides L-O, were isolated from the air-dried roots of Rehmannia glutinosa. Their structures were established from the spectroscopic data obtained and by chemical evidence. The known mellittoside (5) and ajugol (6) were also obtained in the current investigation, and the structure of mellittoside was unequivocally defined using X-ray diffraction data. Compounds 1-6 were tested for their cytotoxicity against five human tumor cell lines and proliferation effects on Lactobacillus Reuteri.

Four ionones and ionone glycosides from the whole plant of Rehmannia piasezkii.

Four new ionones and ionone glycosides (1-4) were isolated from the whole plant of Rehmannia piasezkii Maxim. Their planar structures as well as absolute configuration were confirmed via spectroscopic analysis, ECD calculation, and X-ray crystallography. Compounds 1-4 were tested for their cytotoxicity against five human tumor cell lines and ability to inhibit LPS-activated NO production in the BV2 cell line.

The Effect of Posaconazole and Isavuconazole on the Pharmacokinetics of Erdafitinib in Beagle Dogs by UPLC-MS/MS.

Objective: A robust, quick, and reliable ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the quantification of erdafitinib in beagle dog plasma was developed and validated to evaluate the changes of posaconazole and isavuconazole on the pharmacokinetics of erdafitinib in beagle dogs, respectively. Methods: This experiment adopted a three-period self-control experimental design. In the first period (group A), erdafitinib was orally administered to six beagle dogs at a dose of 4 mg/kg. In the second period (group B), the same six beagle dogs were orally given posaconazole at a dose of 7 mg/kg, and after 30 min, erdafitinib was orally given. In the third period (group C), isavuconazole at a dose of 7 mg/kg was given orally, and then, erdafitinib was orally given. At the different time points after erdafitinib was given in the three periods, the blood samples were collected. The concentration of erdafitinib was detected by the developed UPLC-MS/MS method. DAS 2.0 was used to calculate the pharmacokinetic parameters of erdafitinib. Results: Erdafitinib had a good linear relationship in the range of 1-500 ng/ml, and the lower limit of quantification was 1 ng/ml. The precision, accuracy, extraction recovery, matrix effect, and stability meet the requirements of the guiding principles. After erdafitinib was combined with posaconazole, the Cmax and AUC0→t of erdafitinib increased by 27.19% and 47.62%, respectively, and the t1/2 was prolonged to 6.33 h. After erdafitinib was combined with isavuconazole, the Cmax and AUC0→t of erdafitinib increased by 23.13% and 54.46%, respectively, and the t1/2 was prolonged to 6.31 h. Conclusion: A robust and reliable UPLC-MS/MS method was fully optimized and developed to detect the plasma concentration of erdafitinib in beagle dogs. Posaconazole and isaconazole could inhibit the metabolism of erdafitinib in beagle dogs and increase the plasma exposure of erdafitinib.

Susceptibility and Resistance Mechanisms During Photodynamic Therapy of Melanoma.

Melanoma is the most aggressive malignant skin tumor and arises from melanocytes. The resistance of melanoma cells to various treatments results in rapid tumor growth and high mortality. As a local therapeutic modality, photodynamic therapy has been successfully applied for clinical treatment of skin diseases. Photodynamic therapy is a relatively new treatment method for various types of malignant tumors in humans and, compared to conventional treatment methods, has fewer side effects, and is more accurate and non-invasive. Although several in vivo and in vitro studies have shown encouraging results regarding the potential benefits of photodynamic therapy as an adjuvant treatment for melanoma, its clinical application remains limited owing to its relative inefficiency. This review article discusses the use of photodynamic therapy in melanoma treatment as well as the latest progress made in deciphering the mechanism of tolerance. Lastly, potential targets are identified that may improve photodynamic therapy against melanoma cells.

[The change of NOS in pulmonary oxygen toxicity induced by different oxygen pressure].

OBJECTIVE: Long time exhaled oxygen will induced oxygen toxicity. Some studies had found that different pathology may exised in normobaric and hyperbaric pulmonary oxygen toxicity, and nitric oxide synthase (NOS) may play a role. In this study, we discussed the change of NOS in normobaric and hyperbaric pulmonary oxygen toxicity. METHODS: Sixty male SD rats were randomly divided into 6 groups (n = 10), exposed to 1 ATA (atmosphere absolute), 1.5 ATA, 2 ATA, 2.5 ATA and 3 ATA, 100% oxygen for 56, 20, 10, 8, 6 hours respectively. Rats were exposed to air as control. After exposure, the protein in bronchoalveolar lavage fluid (BALF), the wet/dry weight of lung and the expression of eNOS, nNOS in lung were defined. RESULTS: As compared to air group, the protein in BALF, the wet/dry of lung were significantly elevated in 1.0 ATA group, while these changes were not so obviously in the other groups, and these changes in hyperbaric oxygen group (approximately 1.0 ATA) were significantly decreased as compared with nonnrmobaric oxygen group (1.0 ATA). The expression of nNOS were not changed in normobaric and hyperbaric pulmonary oxygen toxicity, while the expression of eNOS was significantly decreased in 2 ATA group, and significantly elevated in 2.5 ATA and 3 ATA group. CONCLUSION: The expression of eNOS can change when exposed to different pressures of oxygen.

Cyanidin-3-O-galactoside and blueberry extracts supplementation improves spatial memory and regulates hippocampal ERK expression in senescence-accelerated mice.

OBJECTIVE: To investigate whether the antioxidation and the regulation on the Extracellular Regulated Protein Kinases (ERK) signaling pathway are involved in the protective effects of blueberry on central nervous system. METHODS: 30 Senescence-accelerated mice prone 8 (SAMP8) mice were divided into three groups and treated with normal diet, blueberry extracts (200 mg/kg•bw/day) and cyaniding-3-O-galactoside (Cy-3-GAL) (50 mg/kg•bw/day) from blueberry for 8 weeks. 10 SAMR1 mice were set as control group. The capacity of spatial memory was assessed by Passive avoidance task and Morris water maze. Histological analyses on hippocampus were completed. Malondialdehyde (MDA) levels, Superoxide Dismutase (SOD) activity and the expression of ERK were detected. RESULTS: Both Cy-3-GAL and blueberry extracts were shown effective functions to relieve cellular injury, improve hippocampal neurons survival and inhibit the pyramidal cell layer damage. Cy-3-GAL and blueberry extracts also increased SOD activity and reduced MDA content in brain tissues and plasma, and increased hippocampal phosphorylated ERK (p-ERK) expression in SAMP8 mice. Further more, the passive avoidance task test showed that both the latency time and the number of errors were improved by Cy-3-GAL treatment, and the Morris Water Maze test showed significant decreases of latency were detected by Cy-3-GAL and blueberry extracts treatment on day 4. CONCLUSION: Blueberry extracts may reverse the declines of cognitive and behavioral function in the ageing process through several pathways, including enhancing the capacity of antioxidation, altering stress signaling. Cy-3-GAL may be an important active ingredient for these biological effects.

[Study on the iodine status of pregnant women and thyroid function of neonate in high iodine area].

OBJECTIVE: To reveal the iodine status of pregnant women and its impact on thyroid function of neonates in high iodine area. METHODS: A total of 210 pregnant women were chosen from a hospital in Haixing county, Hebei province. Pregnant women's random urinary and neonates' cord blood were collected. The urinary iodine concentration (UIC) was examined by arsenic-cerium catalytic spectrophotometry. The levels of free triiodothyronine (FT3), free thyroxine (FT4) and sensitive thyroid-stimulating hormone (sTSH) in serum were measured by chemiluminescence. RESULTS: The age of 210 pregnant women was (27.69 +/- 4.73) years old, whose urinary iodine median (inter-quartile range ) was 1240.70 (1292.68) microg/L. 84.3% (177/210) of the pregnant women had excessive iodine intake (UIC > or = 500 microg/L), 13.8% (29/210) had more than adequate iodine intake( UIC within 250 - 499 microg/L), 0.5% (1/210) had adequate iodine intake (UIC within 150 - 249 microg/L) and 1.4% (3/210) had insufficient iodine intake (UIC < 150 microg/L). The average serum level of FT3, FT4 in neonates were (2.93 +/- 0.59), (15.03 +/- 1.92) pmol/L, respectively. The median (inter-quartile range) of serum sTSH in neonates were 7.33 (5.59) mU/L 78.1% (164/210) of the neonates' serum TSH were beyond 5 mU/L. There were no correlation between pregnant women's urinary iodine level and neonates' serum FT3 and sTSH level (P > 0.05), but there was a positive correlation between pregnant women's urinary iodine level and neonates' serum FT4 level (P < 0.01). Serum FT4 level of the neonates with whose mothers had insufficient iodine intake ((12.99 +/- 1.10) pmol/L) were statistically lower than those with their mothers had excessive iodine intake (15.16 +/- 1.83) pmol/L) (P < 0.05). CONCLUSION: Most of the pregnant women in high iodine area were excessive for iodine nutrition, but still a few pregnant women had insufficient iodine nutrition. The level of neonates' serum sTSH were relatively high and monitoring of risk of hypothyroidism and subclinical hypothyroidism should be strengthened.

[Investigation of thyroid function abnormalities in children in high water iodine areas of Hebei province].

OBJECTIVE: To understand the level of thyroid function and the prevalence of the thyroid diseases of children in high water iodine areas. METHOD: Two primary schools were selected by purposive sampling from Haixing county, Hebei province in April 2010. A total of 371 children were selected from six classes by cluster sampling in the schools. Morning-urine and venous blood (5 ml) were collected to measure the levels of urinary iodine, free triiodothyronine (FT(3)), free thyroxine (FT(4)) and sensitive thyroid-stimulating hormone (sTSH) in serum and to evaluate the iodine status and thyroid function of children. RESULTS: The median urinary iodine of the children was 1032.08 µg/L. Overall, 96.2% (357/371) of the children's urinary iodine were ≥ 300 µg/L and 68.5% (254/371) were ≥ 800 µg/L. The level of FT(3) and FT(4) were (6.28 ± 0.81) pmol/L and (16.37 ± 2.72) pmol/L, respectively. The median of sTSH was 4.01 mU/L. The FT(4) among boys ((16.63 ± 2.60) pmol/L) was significantly higher than that among girls ((15.99 ± 2.85) pmol/L) (P < 0.05). Overall, 44 subjects were diagnosed as thyroid disease (11.9%), 25 were subclinical hypothyroidism (57% of the total patients), 4 were hypothyroidism (9%), 10 were hyperthyroidism (23%) and 5 were subclinical hyperthyroidism (11%). In the urinary iodine of 200 - 799 µg/L, the prevalence of thyroid disease and hypothyroidism were 6.3% (7/111) and 14.6% (27/254), respectively. When urinary iodine ≥ 800 µg/L, the prevalence of thyroid diseases and hypothyroidism were 3.6% (4/111) and 8.3% (21/254), respectively. CONCLUSION: Most of the children in high water iodine areas had excessive iodine intake. Levels of the children's thyroid hormone were within the normal range. The prevalence of thyroid disease were high, in these areas and the thyroid disease were mainly the subclinical hypothyroidism.

[Construction of the dioxin bioassay method based on the clonal expressed aryl hydrocarbon receptor system].

OBJECTIVE: To study the specific binding of the artificial clonal aryl hydrocarbon receptor translocator (ARNT) with the natural aryl hydrocarbon receptor (AhR) and the recolonization by polyclonal antibody. The dose-response relationship with tetrachlo-rodibenzo-dioxin (TCDD) was also studied to develop TCDD detection method and the binding degree related to dose response. METHODS: (1) The target genes including AhR-PAS, AhR-C and ARNT-PAS were amplified by RT-PCR by using the total RNA purified from the liver cells of C57BL/6J mice as templates to construct pGEX-5X1 recombinants. The recombinant plasmids were expressed in E. coli. (2) The rabbits were immuned by the clonal fusion proteins: AhR-PAS, AhR-C to prepare the polyclonal antibody. (3) The natural AhR from the hepatic cytosol of C57BL/6J mice was extracted. The artificial cloning expressed fusion protein:GST-ARNT-PAS and the natural AhR were incubated in different dose of TCDD. The quantity of the heterodimer through affinity adsorption and Western blots were measured. RESULTS: (1) The target proteins including AhR-PAS, AhR-C and ARNT-PAS were successfully cloned and expressed in E. coli. (2) The detection limit of polyclonal antibody AhR-PAS and AhR-C were 5 ng and 1 ng, respectively. (3) The total protein concentration prepared from the liver cells was 60.5 mg/ml. The artificial clonal protein ARNT-PAS could specifically bind to the natural AhR complex with the existence of TCDD. The detection limit of TCDD was 0.25 pmol which was 80 pg approximately. CONCLUSION: A TCDD detection method based on the aryl hydrocarbon receptor system was established and the detection limit might reach pg grade.

[Role of mitogen-activated protein kinase pathway in chloracne].

OBJECTIVE: To investigate the role of mitogen-activated protein kinase (MAPK) signal transduction pathway in chloracne. METHODS: Immunohistochemical technique was used to detect the expression of phosphorylated epidermal growth factor receptor (p-EGFR) and p-MAPK proteins in the epithelium of chloracne group and control group. RESULTS: p-EGFR and p-MAPK was found in all chloracne tissues, whereas no expression of p-EGFR and p-MAPK protein was found in control group. In the skin of chloracne patients, p-EGFR was mainly distributed in the membrane and the cytoplasm, especially in the vicinity of membrane; major positive signal of p-MAPK was in core and serosity. CONCLUSION: EGFR and MAPK phosphorylation is found in chloracne tissues. MAPK signal transduction pathway is one important molecular mechanism of chloracne.

[Effect of 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin on the expression of AhR and TGF-alpha].

OBJECTIVE: To study HaCaT-keratinocyte cell lines, a chosen model of human epidermis in an attempt to analyze the mRNA expression of AhR and TGF-alpha induced by TCDD METHODS: Semi-quantitative reverse transcription PCR-technique was used for assaying the relative levels of AhR and TGF-alpha mRNA of HaCaT-cells during the proliferation period when the cells were cultured for 24 hours. RESULTS: Relative level of the AhR-transcripts (corrected with beta-actin) decreased with the elevated concentration of TCDD and the relevant coefficient between the proliferation rate and concentration was -0.548, and the differences among all groups were significant (F =4.124, P =0.021). The vehicle control was respectively compared with 7 x 10(-10) mol/L (0.0620 +/- 0.0085) and 7 x 10(-9) mol/L (0.0518 +/- 0.0194) group, significantly different from the control group (0.1138 +/- 0.0227) (t = -3.48, P <0.05; t = -4.17, P <0.01), the expression amount being 55% and 45% of the control. Relative levels of TGF-alpha mRNA tended to increase with the elevated concentration with the significant coefficient of 0.695 (P < 0.01), and the differences among all groups were significant (F = 15.789, P =0.000). In two higher concentration group 7 x 10(-10) mol/L (0.1474 +/- 0.0390) and 7 x 10 (-9) mol/L (0.2133 +/- 0.0364), their relative expression amount of TGF-alpha mRNA was 2.6-fold, 3.8-fold of the control group (0.0561 +/- 0.0100) respectively. Further analysis for the relevant relationship between the amounts of the AhR mRNA and TGF- alpha mRNA showed a highly negative correlation, the coefficient being - 0.561 (P <0.05). CONCLUSIONS: TCDD down-regulate the expression of AhR and up-regulate the expression of TGF-alpha. A strong negative correlation between AhR and TGF-alpha expression is found.

[The effect of selenium on the expression of Fas/FasL in experimental autoimmune thyroiditis rats' thyroid with adequate iodine].

OBJECTIVE: To study the effect of different selenium intake on the expression of apoptosis protein Fas/FasL in experimental autoimmune thyroiditis (EAT) rats' thyroid with adequate iodine. METHODS: Thirty-two female lewis rats were divided stochastically into 4 groups as C group, M group, Se(+) + M group, Se(-) + M group, respectively, and pretreated with feedstuffs containing different concentrations of selenium (Se(+) + M group 2 mg/kg, C and M group 0.20 mg/kg, Se(-) + M group 0.02 mg/kg, respectively) for two weeks, and immunized the rats with porcine thyroglobulin (pTg) to establish an EAT model. The thyroid gland was sampled, embedded in mineral wax and sliced, and the expression of Fas/FasL was measured with immunohistochemistry. RESULTS: Both the expressions of Fas and FasL of EAT rats were significantly increased as compared with control group. The expression of Fas in rats' thyroid follicular cells with EAT was down-regulated as the increased selenium intake (optical density: 0.059 +/- 0.006), the expression of Fas of Se(+) + M group (0.036 +/- 0.004) was significantly inhibited (q = 11.591, P = 0.000), and expression of Fas was lower in the Se(+) + M group than Se(-) + M group (0.050 +/- 0.005) (q = 7.055 , P = 0.000). Effect of selenium on FasL was not identified. CONCLUSION: Increasing the intake of selenium might decrease the expression of Fas on thyroid follicular cells and restrain the development of EAT.

[Study on content of iodine in food in Tianjin market and iodine nutrition conditions among college students].

OBJECTIVE: To know about content of iodine in foods sold in Tianjing markets presently, and the iodine nutrition conditions in college students. It was also aimed to probe the functions of the iodized salt complement with the dietary iodine intake, and whether the urine iodine could reflect dietary iodine intake. METHODS: 278 food samples in markets were collected by a randomly stratified sampling method, while the arsenic-cerium catalytic contact method was used to determine the content in food. The dietary information of students for seven days was recorded, and the urine iodine was determined through the arsenic-cerium catalytic spectrophotometry. RESULTS: The determination of 47 kinds and 278 food samples indicated that the content of iodine within animal foods (7.8 microg/100 g - 30.8 microg/100 g) was higher than that within plant foods (1.8 microg/100 g - 16.1 microg/100 g). The investigation also showed that students who regarded vegetarian food as principle accounted for 70. 19%. The amount of dietary iodine intake among those students, based on the dietary survey, was (111.67 +/- 53.18) microg/d, while supplementary iodine from iodized salt was about (230.27 +/- 45.55) microg/d. Therefore, the total iodine provided from diet would be (341.95 +/- 89.58) microg/d. Modified by urine creatinine, the median of urine iodine was 271.28 microg/gCr, and the urine iodine and dietary iodine intake was found positively related (r(s) = 0.463, P < 0.01). CONCLUSIONS: Regarding the vegetarian food as the principle, most of students investigated are not rich. The dietary iodine intake is lower than RDA (150 microg), but it can be obtained the iodized salt by 230. 27 microg, which is the possible supplement to the shortage from foods.

[Cloning, identification and tissue expression of novel genes differentially expressed in diabetic rats supplemented with zinc].

OBJECTIVE: To clone novel gene fragments differentially expressed from diabetic rats supplemented with zinc and to detect their expression distribution in various tissues. METHODS: cDNA fragments from former research project were cloned, sequenced and BLASTn analysed. The RT-PCR of the two novel genes were made using the primers designed according to the sequence of cDNA to observe the expression changes in liver of various groups and their expression distributions in various tissues. RESULTS: 2 # cDNA and 6 # cDNA were shown to be the novel gene fragments for no matched gene with them in GenBank. The expression level of DM group and DM + Zn group was obviously lower than those of NC group. The expression level of DM + Zn group was higher than those of DM group ( P < 0.05) . The expression level of 2 # cDNA was higher in brain and pancreas, lower in kidney, and none in heart, muscle and thymus. The expression of 6 # cDNA can be found in heart, muscle, brain, kidney and pancreas, but thymus. 2 # cDNA and 6 # cDNA were registered to GenBank with the numbers of AY952968 and AY952970 respectively. CONCLUSION: Two novel genes differentially expressed in diabetic rats supplemented with zinc were cloned and expressed in various tissues, and were successfully submitted to GenBank.