Chemical profiling of Fritillariae thunbergii Miq prepared by different processing methods reveals two new quality markers: Zhebeininoside and imperialine-3-ß-D-glucoside.

ETHNOPHARMACOLOGICAL RELEVANCE: Fritillariae thunbergii Miq (FTM)exhibit versatile biological activities including the significant antitussive and expectorant activities. As a herbal medicine, the therapeutic effects of FTM may be expressed by multi-components which have complicated integration effects on multi-targets. With the time going, the different processing methods of FTM has been changed a lot. Thus，the study described the effect of processing methods to FTM and its quality. MATERIAL AND METHOD: Studies were undertaken by using UHPLC-LTQ Orbitrap MS and pharmacodynamic models. All reagents were involved of analytical grade. While a HPLC-ELSD's method has been developed and validated, a certified Quality System is conformed to ICH requirements. The experimental animals followed the animal welfare guidelines. AIM OF THE STUDY: We aimed to found the differences after the different processing methods of FTM, and to demonstrate the changes could be selected as quality control indicators, and established a method for simultaneous determination of these for quality control. RESULTS: we have previously found two new steroidal alkaloids: zhebeininoside and imperialine-3-ß-D-glucoside from the different processing methods of FTM, which is the difference between the different processing methods of FTM, mainly on the steroidal alkaloids. The activity analysis of zhebeininoside, imperialine-3-ß-D-glucoside, verticine and verticinone showed that the mouse model of cough expectorant has antitussive effect. The positive drug selected was dextromethorphan syrup. The positive group showed biological activity, but the blank group showed nothing. The model group showed illness which means that the model was effective. There are two ways of the mechanism of action of the expectorant action which can make sputum thin, reduce its viscosity, and be easy to cough up, or can accelerate the movement of mucous cilia in the respiratory tract and promote the discharge of sputum. In our study, the content of phenol red was significantly reduced in the administration group. CONCLUSIONS: To sum up, our results suggest that zhebeininoside and other three components cloud be selected as quality control indicators, and a method for simultaneous determination of zhebeininoside and other three components was established for quality control.

Validation, Optimization, and Application of the Zebrafish Developmental Toxicity Assay for Pharmaceuticals Under the ICH S5(R3) Guideline.

The zebrafish as an alternative animal model for developmental toxicity testing has been extensively investigated, but its assay protocol was not harmonized yet. This study has validated and optimized the zebrafish developmental toxicity assay previously reported by multiple inter-laboratory studies in the United States and Europe. In this study, using this classical protocol, of 31 ICH-positive compounds, 23 compounds (74.2%) were teratogenic in zebrafish, five had false-negative results, and three were neither teratogenic nor non-teratogenic according to the protocol standard; of 14 ICH-negative compounds, 12 compounds (85.7%) were non-teratogenic in zebrafish and two had false-positive results. After we added an additional TI value in the zebrafish treated with testing compounds at 2 dpf along with the original 5 dpf, proposed a new category as the uncategorized compounds for those TI values smaller than the cutoff both at 2 dpf and 5 dpf but inducing toxic phenotypes, refined the testing concentration ranges, and optimized the TI cut-off value from ≥ 10 to ≥ 3 for compounds with refined testing concentrations, this optimized zebrafish developmental assay reached 90.3% sensitivity (28/31 positive compounds were teratogenic in zebrafish) and 88.9% (40/45) overall predictability. Our results from this study strongly support the use of zebrafish as an alternative in vivo method for screening and assessing the teratogenicity of candidate drugs for regulatory acceptance.

Flexible Thermo-Optic Variable Attenuator based on Long-Range Surface Plasmon-Polariton Waveguides.

A flexible thermo-optic variable attenuator based on long-range surface plasmon-polariton (LRSPP) waveguide for microwave photonic application was investigated. Low-loss polymer materials and high-quality silver strip were served as cladding layers and core layer of the LRSPP waveguide, respectively. By using finite element method (FEM), the thermal distribution and the optical field distribution have been carefully optimized. The fabricated device was characterized by end-fire excitation with a 1550 nm laser. The transmission performance of high-speed data and microwave modulated optical signal was measured while using a broadband microwave photonics link. The results indicated that the propagation loss of the LRSPP waveguide was about 1.92 dB/cm. The maximum attenuation of optical signal was about 28 dB at a driving voltage of 4.17 V, and the variable attenuation of microwave signals was obviously observed by applying different driving voltage to the heater. This flexible plasmonic variable attenuator is promising for chip-scale interconnection in high-density photonic integrated circuits and data transmission and amplitude control in microwave photonic systems.

Investigation of simultaneously existed Raman scattering enhancement and inhibiting fluorescence using surface modified gold nanostars as SERS probes.

One of the main challenges for highly sensitive surface-enhanced Raman scattering (SERS) detection is the noise interference of fluorescence signals arising from the analyte molecules. Here we used three types of gold nanostars (GNSs) SERS probes treated by different surface modification methods to reveal the simultaneously existed Raman scattering enhancement and inhibiting fluorescence behaviors during the SERS detection process. As the distance between the metal nanostructures and the analyte molecules can be well controlled by these three surface modification methods, we demonstrated that the fluorescence signals can be either quenched or enhanced during the detection. We found that fluorescence quenching will occur when analyte molecules are closely contacted to the surface of GNSs, leading to a ~100 fold enhancement of the SERS sensitivity. An optimized Raman signal detection limit, as low as the level of 10-11 M, were achieved when Rhodamine 6 G were used as the analyte. The presented fluorescence-free GNSs SERS substrates with plentiful hot spots and controllable surface plasmon resonance wavelengths, fabricated using a cost-effective self-assembling method, can be very competitive candidates for high-sensitive SERS applications.

Hispidin induces autophagic and necrotic death in SGC-7901 gastric cancer cells through lysosomal membrane permeabilization by inhibiting tubulin polymerization.

Hispidin and its derivatives are widely distributed in edible mushrooms. Hispidin is more cytotoxic to A549, SCL-1, Bel7402 and Capan-1 cancer cells than to MRC5 normal cells; by contrast, hispidin protects H9c2 cardiomyoblast cells from hydrogen peroxide-induced or doxorubicin-induced apoptosis. Consequently, further research on how hispidin affects normal and cancer cells may help treat cancer and reduce chemotherapy-induced side effects. This study showed that hispidin caused caspase-independent death in SGC-7901 cancer cells but not in GES-1 normal cells. Hispidin-induced increases in LC3-II occurred in SGC-7901 cells in a time independent manner. Cell death can be partially inhibited by treatment with ATG5 siRNA but not by autophagy or necroptosis inhibitors. Ultrastructural evidence indicated that hispidin-induced necrotic cell death involved autophagy. Hispidin-induced lysosomal membrane permeabilization (LMP) related to complex cell death occurred more drastically in SGC-7901 cells than in GES-1 cells. Ca2+ rather than cathepsins from LMP contributed more to cell death. Hispidin induced microtubule depolymerization, which can cause LMP, more drastically in SGC-7901 cells than in GES-1 cells. At 4.1 µM, hispidin promoted cell-free tubulin polymerization but at concentrations higher than 41 µM, hispidin inhibited polymerization. Hispidin did not bind to tubulin. Alterations in microtubule regulatory proteins, such as stathmin phosphorylation at Ser16, contributed to hispidin-induced SGC-7901 cell death. In conclusion, hispidin at concentrations higher than 41 µM may inhibit tubulin polymerization by modulating microtubule regulatory proteins, such as stathmin, causing LMP and complex SGC-7901 cell death. This mechanism suggests a promising novel treatment for human cancer.

Hybrid fiber resonator employing LRSPP waveguide coupler for gyroscope.

Polarization error and temperature noise are two main limits to the performance of resonant fiber optic gyroscope (RFOG). To overcome these limits, we demonstrated a hybrid resonator consisting of a polymer-based long-range surface plasmon polariton (LRSPP) waveguide coupler and a silica fiber. Single-polarization property of LRSPP waveguide and the offsetting of the opposite thermo-optical characteristics between the polymer-based LRSPP waveguide and the silica fiber can effectively inhibit both the polarization error and the temperature noise of RFOG. The measured resonance spectrum of the hybrid resonator shows the absence of polarization noise. The temperature dependence of wavelength shift (TDWS) of resonator dropped to about 2 pm/°C, or even to 0 pm/°C with optimal structure, which dramatically improves the temperature stability of gyroscope system. In addition, the hybrid resonator also shows tremendous application potential in rate-grade and tactical-grade gyroscopes.

[Study of animal model of hypertensive disorder complicating pregnancy in pregnant rats stimulated by homocysteine and monosodium glutamate].

OBJECTIVE: To determine whether homocysteine (Hcy) and monosodium glutamate (MSG) could lead to animal model of hypertensive disorder complicating pregnancy and its mechanism. METHODS: Female adult Wistar rats were randomly divided into 4 groups: pregnant control group (PN), pregnant Hcy group (PH), pregnant glutamic acid group (PG) and pregnant Hcy and glutamic acid group (PHG). The rats of each group were injected with Hcy 200 mg/kg or physiological saline every day intraperitoneally and with MSG or 0.9% saline every other day via Hcy injection from the 10th day to the 20th day of pregnancy. The blood pressure, urine protein, function of liver and kidney, weight of placenta, length and weight of fetus were all measured. The histological change of the pallium and the change of behavior of pregnant rats were also observed. RESULTS: (1) The blood pressure in PH [(107 +/- 8) mm Hg, 1 mm Hg = 0.133 kPa], and PHG group [(109 +/- 10) mm Hg] after the treatment increased significantly compared with those in other groups from the 12 th day after pregnancy (P < 0.01). (2) The level of urine protein [(1.42 +/- 0.53) g/L, (1.53 +/- 0.24) g/L] in PH and PHG groups after the treatment was significantly higher than the other groups (P < 0.01). (3) Obvious changes of the function of liver and kidney [alanine aminotransferase (ALT) (57 +/- 15) U/L, (69 +/- 24) U/L, aspartate aminotransferase (AST) (265 +/- 61) U/L, (293 +/- 118) U/L, blood urea nitrogen (BUN) (9.5 +/- 0.8) mmol/L, (9.5 +/- 1.6) mmol/L, creatinine (Cr) (54 +/- 10) micromol/L, (54 +/- 10) micromol/L], were found in PHG group and PH group (P < 0.01). (4) The weight of placenta [(0.49 +/- 0.28) g, (0.45 +/- 0.03) g], length and weight of fetus [(3.6 +/- 1.5) cm, (3.5 +/- 1.5) cm] in PH group and PHG group were significantly lower than that in PN group and PG group (P < 0.01). Obvious histological changes in pallium and kidney were found in PH group and PHG group. Changes of behavior were found in PHG group. CONCLUSION: Hcy and MSG could induce the symptoms of hypertensive disorder complicating pregnancy in pregnant rats, especially pre-eclampsia, possibly through injuring vascular endothelial cell and nerve cell of the cerebral cortex.

[Relationship between changes of insulin like growth factor-1 and insulin like growth factor binding protein-1 in maternal serum and placenta and pathogenesis of hypertensive disorder complicating pregnancy].

OBJECTIVE: To investigate the role of insulin like growth factor-1 (IGF-1) and insulin like growth factor binding protein-1 (IGFBP-1) in the pathogenesis of hypertensive disorder complicating pregnancy. METHODS: The levels of IGF-1 and IGFBP-1 in serum and placenta were detected from 60 patients with hypertensive disorder complicating pregnancy (hypertensive disorder complicating pregnancy group) and 18 normal pregnant women (control group) by enzyme linked immunosorbent assay (ELISA) and immunohistochemistry. RESULTS: (1) Serum level of IGF-1 (229 +/- 100) microg/L in hypertensive disorder complicating pregnancy group was significantly lower than that in control group (336 +/- 120) microg/L, (P < 0.01). There were significant differences among gestational hypertension (303 +/- 80) microg/L, mild preeclampsia (233 +/- 77) microg/L and severe preeclampsia groups (155 +/- 73) microg/L, (P < 0.05). (2) The expression of IGF-1 in the placenta of hypertensive disorder complicating pregnancy group (48%, 29/60) was significantly reduced than that in control group (83%, 15/18; P < 0.01). Its expressions in mild and severe preeclampsia patients were lower than that in control group (P < 0.05, P < 0.01). (3) Serum IGFBP-1 from hypertensive disorder complicating pregnancy group (161 +/- 90) microg/L was significantly higher than that in control group (98 +/- 75) microg/L, (P < 0.01). There were significant differences among gestational hypertension (97 +/- 73) microg/L, mild preeclampsia (157 +/- 69) microg/L and severe preeclampsia groups (225 +/- 81) microg/L (P < 0.05). (4) The expression of IGFBP-1 in hypertensive disorder complicating pregnancy group (77%, 46/60) was higher than that in control group (39%, 5/18), (P < 0.01). Its expression in mild and severe preeclampsia patients were higher than that in control group (P < 0.05). (5) The levels of IGF-1 in serum and placenta had a significant negative relationship with that of IGFBP-1 in hypertensive disorder complicating pregnancy group (r = -0.269, P < 0.05; r = -0.369, P < 0.01). Levels of IGFBP-1 in maternal serum were positively related with that of placenta (P < 0.01). CONCLUSION: The changes of levels of IGF-1 and IGFBP-1 are related to the occurrence and development of hypertensive disorder complicating pregnancy.

[Thyroid functional changes of normal human fetus and newborns].

OBJECTIVE: To study the changes of the thyroid hormones level of human fetus and newborns. METHODS: More than 71 cases of medically indicated cordocentesis have been done in 16 -36 gestational weeks in our hospital during last three years. Among them, 71 fetus who were free of diseases and their maternal thyroid function were normal were included into the study group. The blood samples were sent to analysis of thyroxine (T(4)), triiodothyroxine (T(3)), free thyroxine (FT(4)), free triiodothyroxine (FT(3)) and thyrotropin (TSH). 140 umbilical cord blood samples taken at the time of term delivery were sent to analysis of FT(4), FT(3) and TSH as a control. Normal range of different gestational weeks was calculated. Statistical analysis was done for the changes of all these thyroid hormones before 28 weeks and after. RESULTS: All the thyroid hormones can be detected in 16 weeks of pregnancy, FT(4) already reaches the top level of adults (5.8 +/- 2.6) pmol/L and will continually increase with the increase of gestational age. There was a parallel increment of all the fetal thyroid hormone concentrations with the gestational age. The concentrations of T(4), T(3) and FT(4) have a rapidly increase after 28 weeks and have a statistically significant difference from (2.8 +/- 1.8) nmol/L, (37.2 +/- 27.2) nmol/L and (10.6 +/- 3.1) pmol/L, respectively to (5.8 +/- 2.6) nmol/L, (55.9 +/- 33.3) nmol/L, (13.0 +/- 4.5) pmol/L, respectively. TSH level of fetus was increased gradually along the gestation, reaching the up level of the adults at the 20 weeks and peaking at the birth time. While the T(3) and FT(3) keep in a lower level in gestation. CONCLUSIONS: Fetal thyroid hormones increase with the gestational age. The diagnosis of congenital fetal thyroid hormone malfunction in the second half of the pregnancy should be monitored mainly by the T(4), FT(4) and TSH levels in different gestational age. For this consideration, to set up a reliable data for normal human fetus thyroid hormone concentrations is a very important and essential step to provide a practical guide for doctors to do intra-uterine diagnosis and treatment of associated high-risk groups. The peaking level of TSH at the birth time will surely company the changing of other thyroid hormones, so it might not be the best time to screening the congenital thyroid malfunction at the 72 hours after birth.