Objective:To establish an HPLC method for the simultaneous determination of narirutin, limonin, honokiol and mag-nolol in Zhishi Xiaopi pills. Methods:The separation was performed on a Shim-pack VP-ODS C18(250 mm ×4.6 mm,5 μm)column with the mobile phase consisting of acetonitrile–methanol(1 ∶2) (A) and water (B) with gradient elution. The flow rate was 1. 0 ml ·min-1 . The column temperature was 30℃. All the injection volume was 20 μl. Narirutin, limonin, honokiol and magnolol was de-tected at 283 nm, 210 nm, 294 nm and 294 nm, respectively. Results:Narirutin, limonin, honokiol and magnolol had good linearity within the concentration range of 5. 26-105. 20 μg·ml-1(r=0. 999 8), 7. 65-153. 00 μg·ml-1(r=0. 999 4), 6. 21-124. 20 μg· ml-1(r=0.999 3)and 6.45-129.00 μg·ml-1(r=0.999 6), respectively; the average recovery was 99.00%(RSD=0.77%), 98. 17%(RSD=1. 19%), 98. 78%(RSD=0. 86%) and 97. 90%(RSD=0. 99%), respectively. Conclusion: The method is sim-ple, rapid and reliable, which can be used for the quality control of Zhishi Xiaopi pills.

Adult ; Humans ; Laryngeal Neoplasms ; Male ; Neurofibroma ; Vocal Cords ; pathology

Objective To estimate the degree of oxidative stress and atherosclerosis praecox in patients with systemic lupus erythematosus (SLE), and emphasize on exploring AOPPs' clinical significance in premature atherosclerosis in SLE. Methods The levels of AOPPs, Hey, MDA, SOD, baPMV and CIMT were detected by ELISA and spectropholometry in 44 non-menopausal female SLE patients and 31 healthy middle-aged women respectively, and baPWV. The results were compared with AOPPs of the two groups. Then each group was stratified based on disease duration (≥5 years or <5 years) and the disease activity(active and inactive) in SLE patients. The patients' TC, TG, LDL were analyzed. T test, t' test and Pearson correla-tion were selected. Results The levels of AOPPs, Hcy, MDA in SLE patients were significantly higher than those of the healthy controls (P<0.05). The activities of SOD were lower than controls (P<0.05). The levels of AOPPs, Hcy, MDA, SOD had statistical significance between SLE patients disease duration ≥5 years or <5 years, active or inactive groups. There were two cases with CAP in patients with SLE (more than 5 years disease duration),while there wasnone in healthy controls. The levels of baPWV and CIMT in patients with SLE were higher than healthy controls (P<0.05), and had statistical significance in SLE patients disease duration more than or less than 5 years (P<0.05). The oxidative stress targets (AOPPs, Hey, MDA, SOD) had significant correlation with the level of baPWV and CIMT (P<0.01). The level of serum AOPPs had significant positive correlation with the levels of Hcy, TC, TG and LDL (P<0.01～0.05). Conclusion SLE patients have increased oxidative stress , and significantly higher prevalence of atherosclerosis than healthy controls. The disease duration and oxidative stress play important roles in the duration of atherosclerosis. AOPPs probably involves in the accelerated atherosclerosis of SLE patients. It may be a predictor for SLE complicated with atherosclerosis praecox.

Objective To compare the clinical effect of different doses of methylprednisolone in the treatment of children with severe hand foot mouth disease research and analysis. Methods 100 patients in our hospital from January 2014 to August 2016 were selected and randomly divided into the control group and the experimental group, with 50 patients in each group. The control group were treated with small doses of methylprednisolone, 2 mg/ (kg?d), intravenous injection. The experimental group was treated with large dose of methylprednisolone, 10～15 mg/(kg?d) intravenous infusion. The therapeutic effects of the experimental group and the control group were compared and analyzed. Results after the corresponding treatment, the number of adverse reactions in the experimental group was 31, the adverse reaction rate was 62.0％, and the adverse reaction rate in the control group was 60.0％. There was no significant difference in the incidence of adverse reactions between the two groups. The upper respiratory rate and critical illness in the experimental group were 26.0％ and 32.0％, significantly lower than that of the control group (P<0.05). The average remission time of the patients in the experimental group was (2.19±1.01) days. The symptomatic remission time in the control group was (4.19±1.89) days. The remission time of the control group was significantly longer than that of the experimental group, with statistical difference (P<0.05). Conclusion The short term large dose of methylprednisolone in treatment of children with severe HFMD clinical effect is ideal, can be reduced to critical illness rate, less symptom remission time, with the further promotion of the clinical significance.

Objective To observe the changes of the intercellular spaces of squamous epithelium of lower esophagus in gastroesophageal reflux disease(GERD).Methods Eleven outpatients with GERD symptoms more than 3 months [6 with nonerosive reflex disease(NERD)and 5 with erosive esophagitis(EE)]and 5 healthy volunteers were recruited.All of them underwent endoscopy and 24-hr ambulatory pH monitoring.Biopsies were taken in lower esophagus(2 cm above Z-line)for electron microscope examination.Results Intercellular spaces of esophageal epithelial cell in volun teers,NERD patients and EE patients were (0.374?0.073)?m,(1.308?0.079)?m and (1.332?0.144)?m respectively,with significant differences between the control group and the NERD or EE group.There was no difference between NERD group and EE group.Conclusions Dilated intercellular spaces were seen in both NERD and EE cases,which was significantly different from the control cases.

Aim To purify L-amino acid oxidase(LAAO) from the venom of Naja atra and study its effect on endothelial cells.Methods The NAV-LAAO was purified by ion-exchange chromatography and affinity chromatography.The MTT assay and Western blot were used to detect the viability and apoptosis of HUVEC.The tubule-forming was used to study the angiogenesis of cells.Results The NAV-LAAO was purified successfully from the venom of Naja atra.The molecular weight of NAV-LAAO was determined to be 58 ku by SDS-PAGE.NAV-LAAO effectively inhibited the growth and tubule-forming of HUVEC,and the 50% inhibitory concentration(IC50) was 21.42 mg?L-1.Compared with control,the levels of caspase-3 and caspase-8 increased in HUVEC treated with NAV-LAAO.Conclusions The NAV-LAAO is purified successfully from Naja atra venom by ion-exchange chromatography and affinity chromatography.The NAV-LAAO inhibits the growth and tubule-forming capacity of HUVEC in a dose-dependent manner.

BACKGROUND:In previous experiments, a polymer-shel ed microbubble contrast agent was successful y prepared using L-polylactic acid-polyethylene glycol methyl-covered liquid fluorocarbon perfluoropentane, which developed wel in vivo and in vitro under a second harmonic contrast-enhanced ultrasound with a low mechanical index OBJECTIVE:To optimize the conditions for preparing polymer-shel ed microbubble contrast agents with high yield and suitable size. METHODS:The polymer-shel ed microbubble contrast agent containing perfluoropentane was prepared by single emulsion method. Three preparation conditions, including polymer mass/perfluoropentane volume ratio (4/1, 2/1, 1/1, 1/2), hemogenate speed (18 000, 26 000, 35 000 r/min) and time (15, 30, 60, 120 seconds), were optimized, and the microbubble yield and size were set as judge standard. The contrast-enhanced ultrasonography of rabbit kidney was performed using the optimized microbubbles, and the arrival time, time to peak, half time of descending and peak intensity were analyzed by TCA software. RESULTS AND CONCLUSION:The preparation conditions were optimized as a polymer mass/perfluoropentane volume ratio of 2/1, homogenate speed of 26 000 r/min and homogenate time of 60 seconds, which presented a high microbubble yield of (1.8±0.4)×109/mL and mean size of (3.7±1.3)μm with uniform distribution. The arrival time, time to peak, half time of descending and peak intensity were detected as (3.1±0.6) seconds, (2.9±0.5) seconds, (4.0±0.7) seconds and (4.7±1.1)×10-5 AU, respectively. The polymeric microbubbles prepared by optimized conditions were of high yield, with suitable size and good contrast-enhanced effect in vivo.

Objective To explore the ultrasound contrast agent (SonoVue) effects upon rabbit testis which were injured after the acute unilateral complete torsion-detorsion and its related mechanisms,on the condition of the diagnostic ultrasound radiation with different doses and different imaging modes.Methods 35 healthy New Zealand white adult rabbits were randomly divided into 5 groups(S,A,B,C,and D),with 7 in each group.Left testis needed ischemia reperfusion after complete 3 hours.Group S:reperfusion without SonoVue group;group A:small-dose (0.1 ml/kg) SonoVue conventional imaging group;group B:high-dose (1 ml/kg) SonoVue conventional imaging group ; group C:small-dose (0.1 ml/kg) SonoVue blasting group; group D:high dose(1 ml/kg) SonoVue blasting group.Groups needed ultrasonic imaging before operation and 24 hour reperfusion.The changes for each group's contrast parameters were analyzed.The contents of malondialdehyde(MDA) and superoxide dismutase (SOD) of testicular tissue were detected,and the expressions of Caspase 3 were examined after 24 hours to make a comparison study among these groups.Results In group D,the time to peak(TTP) and half of declining time(DT/2) were significantly prolonged (P ＜ 0.05),peak-base indensity (PBD) was significantly descended (P ＜ 0.05),and the time-intensity curve manifested that the peak was lower and delayed.SOD in C was lower slightly than in group S.Compared with other groups,SOD in group D decreased,and MDA increased (P ＜ 0.05).Under light microscope,the Johnen's scores in groups A,B,C showed no difference with group S2 (P ＞0.05),while were lower compared with group D (P ＜0.05).Compared with others groups,apoptosis index(AI) and Caspase-3 in group D increased with significance (P ＜0.05).Conclusions Under the diagnostic ultrasound radiation,when conventional imaging and small-does imaging blasted,they did not aggravate the ischemia testicular,while for high-dose,it aggravated.

Objective To study the impact of ultrasound contrast agent with different doses or imaging modes on rabbit spermatogenic cell in the diagnosis of radiation dose.Methods Thirty-three New Zealand white rabbits were randomly divided into 3 groups.In group Ⅰ ,three minutes gray scale ultrasound radiation was performed for three rabbits.In group Ⅱ, SonoVue was bolus injected from ear vein for eighteen rabbits.The rabbits in this group were divided into 6 sub-group according to different doses and different sampling times, the single injections of SonoVue with dosage of 0.1 ml/kg(A1, A2) or 1 ml/kg (C1 ,C2) and repeated injections with dosage of 0.1 ml/kg interval 15 min (B1 ,B2) were taken.In group Ⅲ ,twelve rabbits were bolus injected with SonoVue and then the agent bubbles were blasted when the SonoVue suffused the whole testis.The rabbits in this group were divided into 4 sub-groups according to different doses and different sampling times, and the dosages of SonoVue were 0.1 ml/kg(D1, D2) and 1 ml/kg(E1,E2).Testis were drew immediately or 24 hours later for the observation under light microscope and transmission electron microscope.Results Apoptotic cells in group Ⅲ were more than those in group Ⅰ and Ⅱ ( P ＜0.05) ,while no significant difference of the total number of apoptotic cells between group Ⅰ and Ⅱ ( P ＞0.05).The appearances of focal ultra-structural damage were observed by transmission electron microscope in group C1, such as intercellular space widening and mitochondria swelling, and no distinct damage were observed in the other groups.The appearances of intercellular space widening, mitochondria swelling and cellular edema were observed by transmission electron microscope in group Ⅲ, and spotty necrosisetc was also observed in group D1 and E1.Conclusions Large dose of contrast agent could cause germ cells slight instantaneous effects, while conventional dose had no effect.Contrast agent blasting may cause germ cells unrecoverable damage.

Objective To probe the differential diagnostic value of color Doppler ultrasonography(CDU)in cystitis glandularis.Methods Thirty-nine cystitis glandularis and 192 bladder cancer were examined with CDU.The location,shape,boundary,internal echo and relationship with wall of urinary bladder of lesion were observed.Internal blood flow disposition of lesion were detection.The results of CDU were compared with those of cistoscope biopsy and pathologic diagnosis after operation.Results 59.0%(23/39)cystitis glandularis and 52.9%(74/140)bladder cancer lesions located in trigone of bladder and surrounded the orifice of ureter.53.8%(21/39)cystitis glandularis showed part wall thickening and 72.9%(140/192)bladder cancer showed single nodus or bolus.Sacculiform anechoic area were observed in 69.2%(27/39)cystitis glandularis and were not observed in bladder cancer.Punctiform hyperechogenicity appeared in periphery of 48.4%(93/192)bladder cancer and 7.7%(3/39)cystitis glandularis.The detection rate of interior blood flow signal of bladder cancer was 84.9%(163/1 92),which was obviously higher than that of cystitis glandularis.The diagnostic accurate rates of CDU in cystitis glandularis and bladder cancer were 84.6%(33/39)and 95.3%(183/192),respectively.Conclusions CDU can clear display the location,shape,boundary,internal echo and relationship with wall of urinary bladder of cystitis glandularis and is of important value in diagnosis,differential diagnosis and clinical fllow-up of cystitis glandularis.