Objective To establish a HPLC/MS/MS method for the determinati on of gentiopicroside in human urine.Methods The urine sample was treated by solid-phase extraction with internal standard of caffeine;The RESCEK C8 colum n(150mm?2.1mm,5 ?m) was used as the analytical column with a mobilep hase consisting of methanol-10mmol? L-1 NH4AC buffer(pH=6.5)-acetonit rile(50∶40∶10,V/V),the flow rate was 0.2 mL? min-1;A triple quadruple tandem mass spectrometer was used as the detector,Electrospray ioniza tion source was applied and operated in positiveion mode.Gentiopicroside and caffeine were detected by monitoring the ion transition of m/z 374.1→ 195.2 and m/z 195.2→ 138.2 respectively.Results The linear range was 30～ 9000 ng?mL-1(r=0.9980) for gentiopicroside in human urine.The recovery was 91.10% ～ 9 6.21 %,The absolute recovery was 100.52% ～ 103.83%,The within-day and between-day precisions were less than 10 %.Conclusion The method is proved to be sensitive,accurate,rapid,specific.

Objective To investigate the expression of transforming growth factor?1(TGF-?1)in human colorectal carcinoma and its value for predicting the prognosis.Methods The expression of TGF-?1 and vascular endothelial growth factor(VEGF)was measured in specimens of 52 coloreetal cancers by immunohistoehemistry.The features of clinical pathology were analyzed and the follow-up of all patients were conducted.The correlation between the expression of TGF-?1 and the survival time was studied with Log-rank test.Results Of 52 patients,no expression of TGF-?1 and VEGF was observed in 11 and 14 patients,and the expression was noticed in 41 and 38 patients,respectively.There was a signifi- cant positive correlation between expression of TGF-?1 and expression of VEGF(x~2＝0.633,P＜0.01). Furthermore,the expression of TGF-?1 was significantly correlated with Dukes staging(x~2＝19.866,P＜0.01)and metastasis of lymph nodes(x~2＝13.152,P＜0.01).The 3-year overall survival rates(OSR)in all patients was 49.1% and the 3-year OSR of patients with and without expression of TGF-?1 were 20.5% and 69.2% respectively(x~2＝11.64,P＝0.0006).Conclusion The expression of TGF-?1 could be served as an important predicator for prognosis of coloreetal carcinoma.

Background Sweep pattern visual evoked potential (SVEP) is an objective method of visual test.There is a clear correlation between SVEP acuity and subjective vision,but they are not identical.Recent studies showed that new regression method can improve the accuracy of SVEP acuity. Objective This trial was to investigate and compare the outcome between amplitude-spatial frequency (A-SP) regression method and amplitudelogVA (A-logVA) regression method in extrapolating the SVEP acuity.Methods SVEP was recorded in 113 eyes of 64 subjects using GT-2000 ( Guo Te,China) with the gratings of 10 different spatial frequency from 0.99 to 12.89 cpd as stimulus.The 1 13 eyes included cataract,glaucoma,corneal disease,optical neuropathy,retinal disease,ocular trauma,refractive error and normal eyes.The correlation were analyzed of SVEP acuity,decimal visual acuity and LogMAR visual acuity.The response were averaged and DFT on the monitor display.SVEP acuity was calculated by extrapolating 0 response amplitude.Results The correlation indices of decimal visual acuity curves obtained by the A-logVA function was 0.663,and that obtained by the A-SP function was 0.705.The positive correlation was seen between subjective decimal visual acuity and A-logVA decimal visual acuity (r =0.540,P＜ 0.01 ) and between subjective decimal acuity and decimal acuity calculated by the A-SP regression method (r=0.620,P＜0.01 ).SVEP decimal acuity calculated by the A-SP function regression method was significantly different from the that calculated by the A-logVA function regression method (Z =-8.688,P＜0.01 ).And the correlation indices of LogMAR visual acuity curves obtained by the A-logVA function was 0.733 and that obtained by the A-SP function was 0.715.The positive correlation was found between the subjective LogMAR acuity and that calculated by the A-SP regression method (r=0.700,P＜ 0.01 ) and between the subjective LogMAR acuity and LogMAR acuity calculated by the A-logVA regression method (r=0.710,P＜0.01 ).SVEP LogMAR acuity from A-SP function regression method was significantly different from the LogMAR acuity from A-logVA function regression method (Z=-8.748,P＜0.01 ).No significant differences of VA LogMAR were found in gender,eyes,type of disease and age(x2 =2.171,P=0.338;x2 =0.976,P=0.614;x2 =6.032,P=0.420;x2 =14.720,P=0.257 ).Conclusions SVEP can obtain the visual outcome in human.The amplitude-logVA function regression method is more accurate in extrapolating SVEP acuity.

Phytochemical investigation of the roots of Scrophularia buergeriana Miq. (Scrophulariaceae), resulted in the isolation of a new iridoid derivative named as buergerinin (1). Its structure was elucidated as rel-(1R, 5R, 6R)-(2-oxa-bicyclo[3.3.0]oct-7-en-6, 7-diyl)dimethoxypropane based mainly on MS and 1D and 2D NMR spectroscopic analyses.
Iridoids ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Mass Spectrometry ; Plant Roots ; chemistry ; Scrophularia ; chemistry

To study the sesquiterpenoid constituents in the whole plant of Sarcandra glabra, silical column chromatography, Sephadex LH-20, reverse phase ODS column chromatography and preparative HPLC were used to isolate 70% EtOH extract of Sarcandra glabra. The structures were elucidated based on spectroscopic data (HR-ESI-MS, 1H NMR, 13C NMR, HSQC, HMBC and NOESY). Four sesquiterpenoids were obtained and identified as 4alpha-hydroxy-5alphaH-lindan-8 (9)-en-8, 12-olide (1), chloranthalactone E (2), 8beta, 9alpha-dihydroxylindan-(5), 7 (1)-ieb-8alpha, 12-olide (3) and chloranoside A (4), respectively. Compound 1 is a new sesquiterpene lacone.

Objective To evaluate the changes of the waveform of the photopic negative response in flash-electroretinogram, visual acuity and central retinal thickness in the treatment of intravitreal injections of bevacizumub. Design Retrospective self-comparative case series. Partidpants 8 subjects (9 eyes) with exudative age-related macular degeneration and 3 subjects (3eyes) with proliferative diabetic retinopathy. Method Evaluation protocol included examinations of the Early Treatment Diabetic Retinopathy study visual acu-ity, visual field, intraocular pressure, fundus fluorescein angiography, optical coherence tomography and flash-electroretinogram. Intravit-real injections of bevacizumab, 1.25 mg (0.05ml), were given under an operating microscope and aseptic conditions. All the subjects were followed-up one month later. Main outcome Measure The amplitudes of PhNR, visual acuity and central retinal thickness. Re-sult At 1 months, the mean amplitudes of PhNR and mean visual acuity in all cases had no obvious change (n=12, P>0.05).The central retinal thickness reduced obviously (n=12, P<0.05), but it was neither significantly correlated with PhNR (r=0.294, P=0.145) nor with visual acuity(r=-0.358, P=0.073). Conclusion The single intravitreal injection of bevacizumab is showed promising in absorption of in-traretinal edema and subretinal fluid in patients with exudative age-related macular degeneration and proliferative diabetic retinopathy, but the changes of visual function (including PhNR) might need further investigation. (Ophthalmol CHN, 2009, 18: 243-246)

Background Previous studies showed that after herpes simplex virus-1 (HSV-1) infection of the cornea,matrix metallo proteinase-2 (MMP-2) (produced by corneal cells and corneal epithelial cells) plays an important role in the development of HSK.Focal adhesion kinase (FAK)plays an important role on the expression,release and activation of MMPs.This study explored the expressions of MMP-2 and FAK,which induced by HSV-1 infected human corneal epithelial cells.Objective To investigate MMP-2 and FAK expression in HSV-1 infected human corneal epithelial cells.Methods Human corneal epithelial cells were infected with HSV-1 in vitro to establish cell model of viral infection.The expression of MMP-2 and FAK were detected by reverse transcription PCR (RT-PCR),Western blot,immunohistochemical method and immunofluorescence method at 2 hours,20 hours and 40 hours after infection.Results At 2 hours,20 hours and 40 hours after infection,the expressionis of MMP-2 mRNA and FAK mRNA were significantly increased in comparison with uninfected cells (MMP-2 mRNA:Ftime =0.968,P=0.436 ; Fgroup =47.649,P =0.000 ; Fi ion =0.757,P =0.536.FAK mRNA:Ftime =0.658,P =0.631 ; Fgroup =35.182,P=0.000;Finteraction =1.386,P=0.137).Western blot assays showed that there were no significant differences in p-FAK,FAK or MMP-2 expressions between infected cells and control cells after 2 hours (P＞0.05),but the expression levels of infected cells were significantly increased at 20 hours and 40 hours (both at P ＜ 0.05).Immunohistochemistry results showed that longer infection time was associated with an increased number of cells staining for MMP-2,FAK and p-FAK.Conclusions At the initial stage of HSV-1 infected,p-FAK plays an important role in the process of virus invading and MMP-2 activation.

In order to study the excretion of genistein (GEN) capsule, an estrogen drugs, in human, 30 healthy volunteers were selected and orally administered 50, 100, and 300 mg genistein in an parallel study. Genistein were determined in urine by LC-MS/MS and glucuronidated genistein (GENG) were indirectly determined with enzymatic hydrolysis in urine by LC-MS/MS, and the pharmacokinetic parameters were analyzed by DAS software (ver 2.0). The result showed that the concentrations of genistein in human urine were less than 1% of the GENG, and the cumulative excretion of GEN in 48 h were 0.037, 0.134, and 0.142 mg, separately, and the urinary excretion percentage were only 0.07%, 0.13%, and 0.05%, separately. But the cumulative excretion of GENG in 48 h was 5.3, 13.8, and 15.4 mg, separately, and the urinary excretion percentage were 10.6%, 13.8%, and 5.1%, separately, and the max urinary excretive rate was 0.4, 1.0, and 1.4 mg x h(-1), separately (tmax were 6 h). Studies showed that part of drug excreted through kidney in a form of GENG in human, and the cumulative urinary excretion and the maximum excretion rate of GENG showed a proportional increase conditioned with the dose in the range of 50-100 mg, but showed non-linear increase feature in 300 mg.

AIM: To investigate the effect of ischemia induced by central retinal artery occlusion on retinal microstructure of macula using optical coherence tomography (OCT). METHODS: Fourteen eyes of 14 patients with unilateral central retinal artery occlusion (CRAO) in two to three days without fully recovery of retinal circulation underwent OCT examination with 4.5 mm length horizontal and vertical line scans through foveola to measure the retinal neurosensory layer (RNL) thickness on foveola, 175 ?m (fovea), 750 ?m (macula) to foveola, respectively. The other normal eyes of patients as control group underwent the same examination and measurement. RESULTS: The mean RNL thickness(?m) on foveola, fovea, macula were 169 91?10 96, 176 36?11 74 and 256 45?16 95 respectively in normal control eyes, and 235 64?47 02 , 241 84?49 36 and 401 57?54 53 respectively in CRAO eyes with retinal ischemia. There was a significant difference in thickness between two group ( P

AIM: To investigate the time - effect relationship between the expression of rhodopsin and recoverin and photoreceptor damage induced by N - nethl - N -nitrosourea ( MNU) .
METHODS: Thirty-six 7-week old Sprague-Dawley ( SD ) rats were intraperitoneally injected with MNU ( 60mg/kg ) and were put to death by dislocation of cervical vertebra 6, 12, 24h; 3, 7d after injection ( 6 per group) , respectively. As a control, six rats were injected with phosphate buffer saline (PBS) 5mL/kg and sacrificed on d3 after injection. The degree of photoreceptor apoptosis was detected by HE staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling ( TUNEL ) and transmission electron microscope ( TEM ) in the right eyes. The mRNA expressions of rhodopsin and recoverin were detected different time after injection by Western blot and immunohistochemical method in the left eyes.
RESULTS:The dissolution of photoreceptor nucleus and apoptosis body were first perceived at 12h by TEM; most of cells at outer nuclear layer were presented positive reaction. The apoptotic index reached peak ( 29. 7% ±2.3%) at 24h which was coincided with the observation of TEM. The results of immunohistochemistry displayed that rhodopsin and recoverin were on a declining curve with time extension. Furthermore, the results of Western blot indicated that rhodopsin had dramatic decline at 6h after injection (P<0. 05), and extremely significant difference comparing to control group after 12h ( P<0. 01 ); while recoverin dramatic declined at 12h, and extremely significant difference after 24h (P<0. 01).
CONCLUSION:60mg/kg MNU intraperitoneally injection one - time may specifically induce photoreceptor apoptosis, The mechanism of down - regulation of rhodopsin and recoverin may be related to the selected apoptosis of photoreceptors.