Objective To assess the effects of 670nm LED (light-emitting diode) to protect the photoreceptor from the light-induced damage in a rat model. Methods 32 SD rats were randomly assigned to one of eight groups: untreated control group, the LED-treated control group, three groups of light-induced damage,and three groups of light-induced damage treated with LED. Light-induced damage result from exposing to constant light for 3 hours of different illuminations of 900,1800 and 2700 lx, respectively. The LED treatment (50 mW) was delivered for 30 minutes at 3 hours before the light damage and 0,24 and 48 hours after the light damage. Retinal function and morphology were measured by electroretinogram (ERG) and histopathology assay. Results The illumination of 900 lx for 3 hours did not damage the rat retina. The illumination of 1800 lx for 3 hours resulted in thinner ONL and no OS and IS. The ratio of damaged area/total retinal area was 0.48±0.12, the damaged thickness of ONL/normal ONL (L5) was 0.39±0.07,and the amplitude of ERG b wave was (431±120) μV. With the LED treatment the ratio of damaged area decreased (M6=0.17±0.12, P5/6=0.002), and the ratio of the damaged thickness of ONL also decreased (L6=0.22±0.09, P5/6<0.01), and the amplitude of ERG b wave increased to (1011±83) μV(P5/6 <0.001). The illumination of 2700 lx for 3 hours caused severed damage to the rat retina and the LED could not protect them significantly. Conclusions 670 nm LED treatment has an evident protective effect on retinal cells against light-induced damage, which may be a simple and effective therapy to prevent or to delay age-related maeular degeneration.

Objective To establish an early diagnostic test for tuberculous meningitis (TBM) with good sensitivity and specificity. Methods Twenty-five patients with a clinical diagnosis of TBM and 49 controls, including 27 patients with other infectious diseases of central nervous system and 22 patients with noninfectious neurological diseases, were enrolled in our research. We simultaneously detected antimycobacterium bovis BCG IgG secreting cells in both cerebral spinal fluid (CSF) and peripheral blood (PBL)by enzyme linked immunospot assay(ELISPOT),repeated insertion sequence IS6110 specific for mycobacterium tuberculosis in CSF by PCR and anti-BCG IgG titre in both CSF and PBL by enzyme linked immunosorbent assay(ELISA).Results The sensitivity of ELISPOT was 84.0%,much higher than that of PCR(75.0%)and ELISA(52.3%).The specificities of the three tests were 91.8%,93.7%and 91.8%respectively.The numbers of CSF cells secreting anti-BCG IgG tested by ELISPOT were even higher in the early phase of TBM, but declined along with the disease progressing(t=-3.183,P=0.008),which allowed an early diagnosis to be made. Conclusion ELISPOT technique is proved to be the most valuable test for the early diagnosis of TBM.

In this paper, Fourier transform infrared spectroscopy fingerprint analysis of Marsdenia tenacissima samples was used to develop a reliable method of tracing the geographical origins. Forty-eight samples from four provinces of China were analyzed by FTIR. We analyzed and characterized the fingerprints in both the full spectrum peaks and characteristic peaks, then the principal component analysis and the cluster analysis were carried out. The results of fingerprint analysis, correlation analysis, principal component analysis and cluster analysis can identify the geographic origins correctly, which verified and supplemented each other; the identification results and the actual location showed a high degree of consistency, namely the lower the space distance, the greater the similarity of different samples. These results revealed the obvious superiority and practical value in comparison to the more tedious and time-consuming wet chemistry method normally used. Using appropriate metrology methods can trace the geographical source correctly. The M. tenacissima materials from the region of Maguan should be considered as genuine medicinal materials taking into account the good quality.
China ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; classification ; standards ; Geography ; Marsdenia ; chemistry ; classification ; Medicine, Chinese Traditional ; Principal Component Analysis ; Quality Control ; Reproducibility of Results ; Spectroscopy, Fourier Transform Infrared ; methods

To offer the reference and method for salt damage in the cultivation of Marsdenia tenacissima, the seeds of M. tenacissima collected from Maguan city ( Yunnan province) were taken as the test materials to study the effects of different priming materials on improving germination and growth under high-level salt stress condition. Four different treatments, which were GA3, KNO3-KH2PO4, PEG-6000, NaCl, combined with ANOVA were applied to test the performance of germination energy, germination percentage, germination index, MDA, SOD, and CAT. The results showed that the seed germination was obviously inhibited under salt stress and the soaked seeds with different priming materials could alleviate the damage of salt stress. Under these treatments, the activities of SOD, CAT the content of soluble protein significantly increased. While the content of MDA significantly decreased. The maximum index was obtained when treated with 1.20% KNO3-KH2PO4, the germination percentage increased from 52.67% to 87.33% and the activity of SOD increased from 138.01 to 219.44 respectively. Comparing with the treatment of 1.20% KNO3-KH2PO4, the germination percentage of treating with 300 mg x L(-1) GA3 increased from 52.67% to 80.67%, while the activity of SOD increased from 138.01 to 444.61.
Germination ; drug effects ; physiology ; Marsdenia ; drug effects ; growth & development ; Nitrates ; pharmacology ; Polyethylene Glycols ; pharmacology ; Potassium Compounds ; pharmacology ; Seeds ; drug effects ; growth & development ; Sodium Chloride ; pharmacology ; Stress, Physiological ; Xanthones ; pharmacology

Objective To detect the synergetic effect and mechanism of arsenic trioxide(As2O3)and Trichostatin A(TSA)during inducing apoptosis of HL-60 cells.Methods MTT method was used to test the proliferation of HL-60 cells.Cell cycle and apoptosis were detected by FCM.Semi-quantitative RT-PCR was used to detect the mRNA expression of Bax and Bcl-2 in the cells treated by As2O3 and(or)TSA.Results As2O3 combined with TSA could inhibit proliferation and induce cell cycle arrest at G0 and G1.The percent of apoptosis induced by combination of As2O3 and TSA was obviously higher than that of either As2O3 or TSA.Bax gene expression was increased,while Bcl-2 gene expression was decreased,Bax/Bel-2 ratio was up-regulated.Conclusion Synergetic effect by As2O3 and TSA is remarkable in inducing apoptosis of HL-60 cells.Cell cycte arrest and Bax/Bcl-2 ratio play an important role in apoptosis of HL-60 cells induced by As2O3,TSA or their combination.

Objective To explore the dynamic expressions of interleukin-1 (IL-1)receptor associated kinase (IRAK)-M and IRAK-4 in rats with or without endotoxin tolerance (ETT)in acute liver failure (ALF).Methods Sixty-six male SD rats were divided into three groups:ALF group,ETT group and control group.The rats in ETT group received daily lipopolysaccharide (LPS)intraperitoneal injection for 5 days,while the rats in ALF group received daily injection with same volume of 0.75% NaCl solution.Both ETT group and ALF group received intraperitoneal injection with D-galactosamine (D-GalN)and LPS at 24 hours after the 5th injection of LPS or NaCl solution.At 2,6,12,24 and 48 h after D-GalN and LPS injection,the serum levels of tumor necrosis factor-α (TNF-α)and interleukin-6 (IL-6)were detected by enzyme-linked immunosorbent assay (ELISA).The liver pathologic changes were observed with HE staining by microscope.The mRNA expressions of IRAK-4,IRAK-M and NF-κB (p65)were detected by reverse transcriptase polymerase chain reaction (RT-PCR).The pairwise comparison between groups was done by lease significant difference (LSD)and Dunnet's t test.Results The liver pathologic changes in ETT group were much milder than those of ALF group.In ALF group,IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24and 48 h after D-GalN and LPS challenge were 0.711 ±0.074,0.904±0.118,1.012 ±0.098,1.534±0.279 and 1.451±0.290,respectively,while the IRAK-M mRNA/β-actin absorbance ratios were 0.496±0.018,0.516±0.089,0.503±0.023,0.503±0.057 and 0.469±0.142,respectively.In ETT group,the IRAK-4 mRNA/β-actin absorbance ratios at 2,6,12,24 and 48 h after D-GalN and LPS challenge were 0.619±0.083,0.587±0.033,0.623±0.034,0.720±0.044 and 0.654±0.041,respectively,while the IRA'K-M mRNA/β-actin absorbance ratios were 0.929 ± 0.064,1.111±0.138,1.113±0.027,1.891±0.315 and 1.710±0.303,respectively.The IRAK-M mRNA expression level was significantly increased and IRAK-4 mRNA expression level was relatively decreased in ETT group compared to ALF group.The differences were all statistically significant at 2,6,12,24 and 48 h after D-GalN and LPS challenge (F= 17.305,54.921,121.031,67.607,55.279,respectively; F=19.506,43.777,110.823,302.681,202.822,respectively; F=172.003,59.519,987.055,68.463,96.601,respectively; all P＜0.05).Conclusion LPS pretreatment may induce ETT in rat model by downregulating the expression of IRAK-4 and upregulating the expression of IRAK-M.

This study examined the neuroprotective effect of cluster of differentiation molecule 200 (CD200) against methamphetamine (METH)-induced neurotoxicity. In the in vitro experiment, neuron-microglia cultures were treated with METH (20 μmol/L), METH (20 μmol/L)+CD200-Fc (10 μg/mL) or CD200-Fc (10 μg/mL). Those untreated served as control. Microglia activation expressed as the ratio of MHC-II/CD11b was assessed by flow cytometry. The cytokines (IL-1β, TNF-α) secreted by activated microglia were detected by enzyme-linked immunosorbent assay (ELISA). In the in vivo experiment, 40 SD rats were divided into control, METH, METH+CD200-Fc and CD200-Fc groups at random. Rats were intraperitoneally injected with METH (15 mg/kg 8 times at 12 h interval) in METH group, with METH (administered as the same dose and time as the METH group) and CD200-Fc (1 mg/kg at day 0, 2, 4 after METH injection) in METH+CD200-Fc group, with CD200-Fc (1 mg/kg injected as the same time as the METH+CD200-Fc group) or with physiological saline solution in the control group. The level of striatal dopamine (DA) in rats was measured by high-performance liquid chromatography (HPLC). The microglial cells were immunohistochemically detected for the expression of Iba-1, a marker for microglial activation. The results showed that METH could increase the microglia activation in the neuron-microglia cultures and elevate the secretion of IL-1β and TNF-α, which could be attenuated by CD200-Fc. Moreover, CD200-Fc could partially reverse the striatal DA depletion induced by METH and reduce the number of activated microglia, i.e. Iba-1-positive cells. It was concluded that CD200 may have neuroprotective effects against METH-induced neurotoxicity by inhibiting microglial activation and reversing DA depletion in striatum.

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.

Objective To investigate the influence of Ethyl pyruvate(EP) on the level of high mobility group box 1 (HMGB1) in brain injury of infant rats induced by lipopolysaccharide (LPS) and its significance.Methods Two hundred and forty normal healthy 1-month-old Spragne-Dawley (SD) rats were randomly divided into 3 groups:9 g/L sodium chloride (NS) group(n=80),LPS group (n=80),and EP group (n=80).LPS (1 mg/kg) was injected via internal carotid.In EP group,after injecting LPS,each rat was immediately administrated 4 mL EP(40 mg/kg) intraperitoneally; in control group,4 mL Ringer's solution was given instead of EP.Rats were decapitated at 6,12,24,48 and 72 h following drug injection.The evan's blue (EB) content of brain tissues was meteraged by the formamide methods.Immunohistochemistry technology was used to detect the expression of HMGB1,neuron specific enolase(NSE) and glial fibrillary acidic protein(GFAP) protein,and reverse transcribe polymerase chain reaction technology was applied to study the expression level of HMGB1 mRNA in brain tissue.Meanwhile,the pathological changes of brain tissues were observed under the light microscope.Results Six hours after LPS was given,brain EB content,the levels of NSE,GFAP protein started to rise,reaching the peak at 24 h,and still higher than those in control group at 48 h and 72 h.The expression pattern of HMGB1 protein and mRNA in brain tissue was consistent with the severity of brain injury,increased at 6 h after LPS was given and reached the peak at 24 h,still higher than those in control group at 48 h and 72 h.Positive correlation was found among HMGB1 protein,HMGB1 mRNA and EB content,NSE protein,GFAP protein,respectively.In EP group,the levels of HMGB1 protein and mRNA,the levels of brain EB content,NSE,GFAP protein were found,positive correlation was still gotten between HMGB1 protein,EB content and NSE,GFAP protein,HMGB1 mRNA in LPS group and EP group.Conclusion EP has neuroprotective effect on brain injury induced by LPS,which may be relevant to decreasing of the expression of HMGB1 and suppressing inflammation action.

Objective To explore the possibility of the inner vertical outer spiral complex tubular urethra scaffold vascularization in repairing long segment of urethral defect.Methods From August 2014 to October 2015,27 clean male New Zealand white rabbits were divided into 3 groups,S1 group was transfected recombinant vascular endothelial growth factor(VEGF) gene lentiviral vector group.S2 group was vascular pedicle transfer tube group.C group was simple stent group.A 3.0 cm inner vertical outer spiral complex scaffold was constructed by using the small intestine acellular matrix (SIS) and polylactic acid copolymer (PLGA) modified by type Ⅰ collagen surface,and adipose-derived mesenchymal stem cells (ADSC) and smooth muscle cells after transformation from New Zealand white rabbits.In S1 group,the seed cells were transfected by recombinant vascular endothelial growth factor (VEGF) gene lentivirus,which express VEGF protein.The complex scaffold was used to repair 3.0 cm rabbit urethral defect In S2 group,the untransfected cells were seeded into the scaffold and embedded in the skin near the groin artery 3 weeks for repairing urethral defect with vascular pedicle transfer tube.In group C,the unseeded scaffold was used to repair the urethral defect alone.Postoperative observation and urethrography were followed 4,8 and 24 weeks after implantation.The HE staining,fluorescence tracing,immunohistochemical and scanning electron microscopy were evaluated at the same phase.Results In S1 group,there were one urinary fistula and one urethral stricture-related death,respectively.The urethra was smooth and patent,histological examination showed active hyperplasia of urethral capillary.In S2 group,there were one urinary fistula and two urethral stricture-related deaths,respectively.The urethral was rough,local thinning or dilated.Fat accumulation and mucosal contraction were found in the urethral submucosal,respctively.In C group,there were one urinary fistula,three hypospadias,and three urethral stricture-related deaths.The thickness of the urethra was uneven and stricture bending.The urethral mucosa was poorly repaired and the scar was narrow.HE and CD31 staining showed that S1 and S2 groups were active in the proliferation of urethral capillaries,and the angiogenesis was abundant.VEGF staining showed that the cytoplasm of endothelial cell layer,smooth muscle layer of vascular wall and the urothelial epithelial cell layer were fully expressed at 24 weeks,especially in epithelial cell layer.CKpan staining showed that the epithelium of S1 and S2 group developed to stratified epithelium,and the morphology of urethra was similar to normal urethra at 24 weeks.The urethral epithelial in C group of grew poor as single-level,irregular arrangement,24 weeks is still a lack of effective stratified epithelium.HE and oα-SMA staining showed that the smooth muscle and actin gradually increased in group S1 and S2,α-SMA staining in group C was scarce and increased at 24 weeks.PLGA was encapsulated by the surrounding tissue and the structure of electrospinning was clear after 4 weeks,absorbed and degraded after 8 weeks and absorbed after 24 weeks.Conclusions The inner vertical outer spiral comnplex tubular urethra scaffold maybe a reasonable method in repairing long segment urethral defects,and the methods of tubular urethra scaffold vascularization by transfected VEGF gene recombinant lentiviral vector and vascular flap deserve more research.