Chemical constituents of sesquiterpenes from Chloranthus multistachys / 中国中药杂志

With repeated silica gel, octadecyl silica(ODS), and Sephadex LH-20 column chromatography, normal-phase and reverse-phase high performance liquid chromatography(HPLC), etc., a pair of new enantiomers and 5 known compounds were separated from the 95% ethanol extract of Chloranthus multistachys. These compounds were identified by the nuclear magnetic resonance spectroscopy(including 1 D-NMR and 2 D-NMR), single-crystal X-ray diffraction, circular dichroism(CD) spectroscopy, mass spectrometry(MS), and some other methods as(1R,4R,5R,8S,10R)-chloraeudolide H(1 a),(1S,4S,5S,8R,10S)-chloraeudolide H(1 b), hydroxyisogermafurenolide(2), 4α-hydroxy-5α,8β(H)-eudesm-7(11)-en-8,12-olide(3), chloraniolide A(4), chlorantene D(5), 4α,8β-dihydroxy-5α(H)-eudesm-7(11)-en-8,12-olide(6). Compounds 1 a and 1 b are a pair of new eudesmane-type sesquiterpene enantiomers, and compounds 2-4 were isolated from C. multistachys for the first time.

Simultaneous determination of six bioactive constituents in Reduning Injection by UPLC-QqQ-MS/MS / 中草药

Objective: An ultra high performance liquid chromatography coupled to triple quadrupole mass spectrometry (UPLC- QqQ-MS/MS) method has been established to simultaneously determine the contents of six bioactive constituents [japonicaside A (JA), L-phenylalanion secologanin B (PSB), luteolin, scopolamine, (1S,6R,7R,10R)-6-carboxy-10-methyl-α-methylene-1-(1- oxobutyl)-cyclohexane acrylic acid (CMCA), and 3α,5α-tetrahydrodeoxycordifoline lactam (TL)] from Reduning Injection. Methods: This chromatographic separation was performed on an Agilent Zorbax SB-Aq C18 (150 mm × 2.1 mm, 3.5 μm) column with the mobile phase consisting of 0.1% formic acid water (A) and methanol (B) in a gradient elution (0.01-2.00 min, 5% B; 2.00-4.00 min, 5%-40% B; 4.00-11.00 min, 40%-95% B; 11.00-13.00 min, 95% B; 13.00-13.10 min, 95%-5% B; 13.10-14.00 min, 5% B) at a flow rate of 0.5 mL/min and the colunm temperature was set at 20℃. The analytes were detected using electrospray ionization (ESI) source by positive and negative ion monitoring mode. A triple quadrupole mass spectrometer was operated by ESI source in positive and negative ionization mode with multiple reaction monitoring for the detection of the six compounds. In the positive ion monitoring mode, the flow rate of Collision Gas (CAD) was 8 mL/min, the flow rate of Curtain Gas (CUR) was 20 mL/min, the temperature (TEM) was 500℃ and the Ion Spray Voltage (IS) was 4 500 V. In the negative ion monitoring mode, the flow rate of Collision Gas (CAD) was 8 mL/min, the flow rate of Curtain Gas (CUR) was 20 mL/min, the temperature (TEM) was 500℃ and the Ion Spray Voltage (IS) was -4 500 V. Results: All calibration curves of the six components showed excellent linear regressions (r ≥ 0.999 0) within the test range. The average recoveries were 78.93%, 114.65%, 101.99%, 90.98%, 98.08%, and 115.58%, and the average contents of six bioactive constituents in 16 batches of Reduning Injection were 2.00, 26.63, 52.63, 5.29, 34.64, and 9.69 μg/mL, respectively. Conclusion: The established method is rapid, accurate, and has high repeatability, which could provide scientific evidences for the quality control of Reduning Injection.

A new sesquiterpene from Acorus calamus (Ⅱ) / 中国中药杂志

A new quaiane-tgpe sesquiterpene was isolated from the 95% ethanol extract of the rhizomes of Acorus calamus by silica gel and sephadex LH-20 column chromatographic methods. Structure and absolute configuration of the sesquiterpene were elucidated by spectroscopic data and X-ray crystallographic analysis, and named as 1R,5R,7S-guaiane-4R,10R-diol-6-one.

Langerhans cell protein 1 (LCP1) binds to PNUTS in the nucleus: implications for this complex in transcriptional regulation

Protein phosphatase-1 (PP1) nuclear targeting subunit (PNUTS), also called PP1R10, p99, or CAT 53 was originally isolated as a mammalian nuclear PP1-binding protein. In this study, we performed yeast two-hybrid screens to identify PNUTS-interacting proteins. Here, we report that LCP1 (epidermal Langerhans cell protein 1), a novel member of the HMG-box protein family, binds tightly to PNUTS. Co-immunoprecipitation of deletion constructs revealed that the C-terminus of LCP1 is sufficient for the interaction with an N-terminal region of PNUTS that is distinct from its PP1-binding domain. Furthermore, immunofluorescence studies showed that a subpopulation of LCP1 co-localizes with PNUTS in nuclear speckles. Importantly, we found that the N-terminus of LCP1 has a strong trans-activation activity in a GAL4-based heterologous transcription assay. The transcriptional activity of LCP1 is markedly suppressed by its interaction with PNUTS, in a PP1-independent manner. These findings suggest that the coordinated spatial and temporal regulation of LCP1 and PNUTS may be a novel mechanism to control the expression of genes that are critical for certain physiological and pathological processes.

The Utility of Hand Held Autorefractometer for Refraction in Children

We studied the utility of hand held autorefractometer on the cycloplegic refraction for poor coorperated children by analyzing the data taken from hand held autorefractometer(Retinomax) and Canon R-10 autorefractometer. Seventy patients(140 eyes) were studied. We divided them into two groups according to the age: Group I included patients less than 4 years, Group II over 4 years. There were no statistic significant differences in the spherical and cylindrical values regardless of age and cooperability in cases of less than 6 diopter but statistic significant difference in cases of high refractive errors (>6 diopter) or cylindrical axis values. In Conclusion, this study revealed that hand held autorefractometer was useful for checking refraction of the poor coorperated children and the spherical and cylindrical values of Retinomax have no differences from the values of R-10 in the cases of less than 6 diopter.

A Study about the Accuracy of Automated Refraction

We analysed the differences between the values of noncycloplegicautomated refraction and manifest refraction by ophthalmologist 150 patients(274 eyes)were studied. We divided these patients into five groups according to the age; Group 1 included patients under 10 yea(n=50 eyes), Group 2 from 10 yrs to 19 yrs(n=74 eyes), Group 3 from 20 yrs to 39 yrs(n=66 eyes), Group 4 from 40 yrs to 49yrs(n=39 eyes), Group 5 over 50 yrs(n=46 eyes); We used Canon Auto Ref R-10 autorefractor and ophthalmologist's refractions were the results of both subjective refraction by retinoscope and objective refraction used by cross cylinder. Between the two examinations, the percentages of eyes of disagreement beyond +/-0.5D were 22%(group 1), 39.2%(group 2), 14.1%(group 3), 15.8%(group 4), 17.4%(group 5) in spherical power; 18%, 23%, 27.3%, 29%, 15.2% in cylindrical power respectively; 20%, 31.9%, 18.2%, 15.8%, 19.6% in spherical equivalent respectively and the percentages of eyes of disagreement beyond +/-10 degrees in cylindrical axis were 25%, 42.6%, 39.2%, 40%, 52.9% respectively. There was no statistic significance between the age groups when we compared the absolute differences of values between two examinations. Conclusively, this study revealed that there were large differences between the values of noncycloplegic automated refraction and manifest refraction by ophthalmologist in the large amout of patients and then our results indicated that automated refraction was dangerous to patient's eyes and subjective refinement was needed by ophthalmologist if patients wanted the spectacle or contact lens.