Animals ; Male ; Prostate-Specific Antigen ; analysis ; metabolism ; Prostatectomy ; adverse effects ; standards ; Specimen Handling ; methods ; standards

Objective To develop a method for determination of catechins and alkaloids in brick-tea by high performance liquid chromatography(HPLC) with diode array detection(DAD),and to explore optimum extraction conditions for catechins and alkaloids components in brick-tea.Methods Catechins and alkaloids were separated and detected by HPLC.Orthogonal experiment and paired t test was carried out to compare the effect of water and alcohol as extraction solvents and to obtain optimum extraction conditions for extracting catechins and alkaloids components in brick-tea.Results Six kinds of catechins of catechin,epicatechin,gallocatechin,epigallocatechin,epicatechin gallate,epigallocatechin gallate and two kinds of alkaloids of caffeine and theobromine were separated and detected simultaneously.Calibration curves between peak areas and concentration of each component in bricktea were linear within a suitable concentration range,and coefficients of determination (R2) were between 0.9990-0.9999; spiked recoveries were from 83.78％ to 106.35％,and relative standard deviations(RSD) were between 0.50％-1.51％.The optimum extraction condition for catechins and alkaloids was 80％ ethanol,solid-liquid ratio of 1:10,temperature 80 ℃ and extraction time 30 min.Conclusion Alcohol as extraction solvents with optimal combination on HPLC,six kinds of catechins and two kinds of alkaloids are separated accurately,qualitatively,rapidly and sensitively.

Aged ; Anthracosis ; complications ; pathology ; Bronchoscopy ; Fiber Optic Technology ; Humans ; Lung Diseases, Obstructive ; etiology ; pathology ; Male ; Pneumonia ; etiology ; pathology ; Retrospective Studies

p21-Activated kinases including p21-activated kinase 2 contributed to the regulation of actin cytoskeleton and cell dynamics. In order to investigate the function of PAK2 on the maturation of Xenopus oocyte, PAK2-NT(PAK2-N-terminal,PAK2-NT) and PAK2-NTm (PAK2-N-terminal mutation) mRNA were microinjected into Xenopus oocyte respectively. Under fluorescent microscopy germinal vesicle breakdown was observed during cytokinesis. To further observe the relationship of oocyte cytokinesis, polar body formation and Cdc42 activity, confocal microscopy with time-lapse was employed . As a result, occurrences of germinal vesicle breakdown in oocytes were similar to those oocytes injected with PAK2-NT mRNA or injected with PAK2-NTm mRNA,but no cytokinesis and polar body formation were observed in oocytes injected with PAK2-NT mRNA or PAK2-NTm mRNA. These results indicated that PAK2 involved in Xenopus oocytes cytokinesis and polar body formation independent of Cdc42 activity.

Objective To observe the effects of intravitreal injection of conbercept for aggressive posterior retinopathy of prematurity (AP-ROP).Methods It is a retrospective case study.Twenty-one patients (40 eyes) with AP-ROP were enrolled in this study.There were 9 males (18 eyes) and 12 females (22 eyes),with the mean gestational age of (28.30±1.79) weeks and the mean birth weight of (1 021.40±316.70) g.All the lesions of 40 eyes were located in posterior zone,with 24 eyes in zone Ⅰ and 16 eyes in zone Ⅱ.All the eyes were treated with intravitreal injection of conbercept 0.025 ml (0.25 mg).During follow-up,nonresponders or patients with deterioration were retreated with intravitreal injection of conbercept or photocoagulation;patients with progressive deterioration to stage 4 had received vitrectomy.At the 1,2,4,8,12,16,20,24 weeks after treatments,the disappearance or decrease of retinal vessel tortuosity and neovascularization,and the growth of the normal retinal vessels toward the peripheral retina were evaluated.Results Thirty-six eyes were cured for only one injection,the cured rate was 90.00％.However,2 eyes (5.00％) had progressed to stage 4 with contractive retinal detachment,which underwent vitrectomy.Two eyes (5.00％) had received twice injections,whose remaining avascular zone area treated by photocoagulation.No major systemic or ocular complications after injection appeared.All lens remained transparent and no iatrogenic retinal hole was occurred during the follow-up.Conclusion Intravitreal injection ofconbercept is effective in the treatment of AP-ROP.

OBJECTIVETo evaluate the effects of Attractin (Atrn) silence on the anti-oxidative and anti-apoptotic abilities of TM4 Sertoli cells and its influence on the expressions of superoxide dismutase (SOD) and caspase6 in the cells.

METHODSWe observed the apoptotic indexes of TM4 Sertoli cells with normal expression (control), partial deletion, and complete deletion of the Atrn gene (psiRNA-TM4, psiAtrn-TM4, and mu-SC). We determined the mRNA and protein expressions of SOD and caspase6 by Q-PCR and Western blot, measured the SOD activity and malondialdehyde (MDA) contentby spectrophotometry, and detected the apoptotic index of the cells by TUNEL.

RESULTSCompared with psiRNA-TM4, after inhibition of the Atrn expression, the Sertoli cells in the psiAtrn-TM4 and mu-SCgroups showed significantly decreased expressions ofSOD mRNA (70.76% and 92.58%) and protein (65.11% and 71.0%) (both P < 0.05). The levels of caspase 6 mRNA and protein were increased 5.28 and 3.40 times in the psiAtrn-TM4 and 2.97 and 2.50 times in the mu-SCgroup as compared with the normal control (both P < 0.05). Atrn deletion markedly increased the apoptotic indexes of the cells in the psiAtrn-TM4 and mu-SC groups by 16.22% and 22.03% (P < 0.05) and reduced the activity of SOD by 23.00% and 39.37% (P < 0.05); it also elevated the level of MDA by 155.22% (P < 0.05).

CONCLUSIONThe Atrn gene exerts influence on the function of Sertoli cells in multiple ways, in which antioxidative stress and apoptosis regulation may play an important role.

Animals ; Apoptosis ; Caspase 6 ; metabolism ; Cells, Cultured ; Gene Deletion ; Male ; Membrane Proteins ; genetics ; metabolism ; Mice ; Oxidative Stress ; Sertoli Cells ; metabolism ; pathology ; Superoxide Dismutase ; metabolism

Objective To investigate the efficiency,decision of intra-operative puncture route,treatment of perioperative complications and discuss the other relative problems of the treament for lumbar disc herniation with percutaneous transforaminal endoscopic discectomy.Methods To excise the nucleus pulpesus under percutaneous transforaminal endoscopic discectomy,use the Macnab standard,visual analogue scale and infrared thermal imaging to estimate the efficiency.Results Among followed-up of 208 patients,182 patients were excellent and good outcome,23 patients favorable,2 patients fair,0 patient poor.The leg and back VAS was a significant improvement 1 week post-operation compared with pre-operation (P ＜ 0.05),but no statistical difference among 3 months,6 months,12 months and 1 week post-operation (P ＞ 0.05).The infrared thermal imaging point out that the legs skin temperature of D-value was a significant improvement post-operation compared with pre-operation (P ＜ 0.05).Conclusions The method excised the nucleus pulpesus,and provided the spine maximum protection about the stability and flexibility.Intra-operative puncture route of individuation design can reduce the complications of intervertebral foramen perioperative,and the key to improve the effectiveness.

This study was aimed to establish an effective method for the simultaneous content determination of strychnine and brucine in rat plasma.The Wistar rat plasma samples were processed by liquid-liquid extraction and separated on a UPLC BEH C18 column.The mobile phase was acetonitrile - acid water (17?83).The acid water was 0.02 mol·L-1.The potassium dihydrogen phosphate and 0.014 8 mol·L-1 sodium heptane sulfonate was mixed with equal amount.The 10% phosphonic acid was used to adjust the system to the pH of 2.8.The detection wavelength was set at 260 nm.The results showed that foreign materials in the Wistar rat plasma did not interfere with the content determination of samples.The calibration curve of strychnine was in good linearity within the range of 0.067 84-3.392 00μg·mL-1.The calibration curve of brucine was in good linearity with the range of 0.052 48-2.624 00μg·mL-1.The intra-and inter-day precisions were less than 10%.The mean extraction recoveries were more than 85%.The stability results met the requirements for biopharmaceutical analysis.It was concluded that the UPLC detection method was sensitive and precise,which can be used in the pharmacokinetics study of strychnine and brucine in Tong-Mai(TM) pill.

Objective To investigate the exact protocol eliciting the hippocampal CA1 long-term depression (LTD) of rats in vivo and the effect of amyloid β-protein (Aβ) on the LTD.Method By applying test stimulation to Schaffer collateral in hippocampal CA1 region in rats,recorded the in vivo field excitatory postsynaptic potentials (fEPSPs) ;further,observed the induction of LTD with different low frequency stimulation (LFS) and investigated the effect of Aβ25-35 on the LTD.Results Prolonged LFS (1,5 and 10 Hz) but not paired-pulse stimulus (PPS) effectively elicited the LTD in the hippocampal CA1 region,with significantly decreased amplitude of fEPSPs after LFS ; 1 Hz 900 pulses group induced a stronger LTD,being (63.7 ± 3.8) ％ at 120 min post-LFS,lower (P ＜ 0.05) than (75.1 ± 3.2) ％ in 600 pulses group ; different frequencies (1,5 and 10 Hz) of LFS with same pulses induced similar degree of LTD,the amplitude of fEPSPs were (63.7 ± 3.8) ％,(61.2 ± 3.6) ％ and (59.8 ± 3.9) ％ respectively,without significant differences between any two groups (P ＞ 0.05) ; after applying 12.5 nmol and 25 nmol Aβ25-35,the amplitude of fEPSPs decreased to (63.2 ± 3.8) ％ and (46.8 ± 3.9) ％,respectively,and lower and than that in control ((73.9 ± 3.0) ％,P ＜ 0.05).Conclusion Prolonged LFS effectively induced in vivo hippocampal LTD of rats,which provides an important electrophysiological protocol for the study of synaptic plasticity; Aβ25-35 injection dont affect the baseline synaptic transmission,but dose-dependently enhance the in vivo hippocampal LTD of rats,indicating that Aβ-induced LTD facilitation may be involved the early impairment of learning and memory in Alzheimer's disease.