AIM: To investigate the changes of plasma glutathione peroxidase (GSH-PX) and catalase (CAT) activities in nude mice (NM) bearing human nasopharyngeal carcinoma (NPC) and observe the effect of naja naja atra venom (NNAV) on them. METHODS: Plasma GSH-PX and CAT activities in human NPC bearing NM treated ( i.p. ) by low, middle or high concentration NNAV solution (1 mg/L, 5 mg/L, 10 mg/L) were determined by colorimetry. RESULTS: Plasma CAT activity (16 450 U/L) in NM bearing tumor group decreased significantly in comparison with the control group (20 680 U/L)(P0.05). Treated by low, middle or high concentration NNAV solution, CAT activities of three NM bearing tumor groups (20 570 U/L, 23 090 U/L, 21 280 U/L ) were higher than that of the NM bearing tumor group without NNAV treatment (16 450 U/L) (P0.05). GSH-PX activities of the three groups (especially high concentration group) were higher than that of the group without NNAV treatment (P

AIM: To investigate the angiogenesis in the process of sarcoma 180 (S_(180)) tumor transplantation and changes of regulator factors, and explore the possible mechanism. METHODS: The S_(180) transplanted tumor in the Km mouse was used to detect the tumor angiogenesis by immunohistochemical examination of FⅧ. The levels of VEGF (V) and endostatin (E) in serum and the homogenate of tumor tissue were measured by ELISA and EIA, and the correlation between tumor weight and microvessel count (MVC) and morphology in tumor was also analyzed by multiple ANNOVA method. RESULTS: MVC, the relative count of total vessels and relative total vessel area increased with the development of transplanted S_ 180 . VEGF level in tumor tissue were higher at the 10th and 15th day than the 5th day after tumor transplantation. Endostatin in the tumor tissue and serum both reached the highest level at the 15th day, V/E ratio did not changed in this process. Furthermore, MVC, average vessel area and relative total area had a significant correlation with tumor weight. CONCLUSION: MVC increases in the development of S_(180) transplantation tumor and is related with the tumor weight; the positive regulator of angiogenesis in the tumor tissue is up-regulated during tumor growth, and the regulators in the tumor tissue maintains a relative balance.

AIM:To establish a HPLC method for determining contents of tetrahydropalmatine and pepper alkali in Zhitong Capsule(Rhizoma Corydalis,Fructus Piperis,Radix Paeoniae alba,etc.)to control its quality.METHODS:The samples were extracted in water by ultrasonic wave,and then extracted by aether to refine after being acidized by 1 mol/mL HCl.Following that the pH of water solution was adjusted to 9.0-10.0 by NH_3?H_2O,and then extracted by aether again.After that,the aether solution was collected to evaporation to dryness and metered by methanol.The sample solution was determined by high performance liquid chromatography on a Hypersil ODS 2 C_ 18 column(4.6 mm?250 mm,5 ?m)with mobile phase composed methanol-0.1% phosphoric acid water solution(the pH was adjusted to 6.0 by triethylamine)(55∶45).The flow rate was 1.0 mL/min,the column temperature was at 30 ℃ and the signal from 0 to 14.5 minutes were acquired at 280 nm,and that from 14.5 to 22 minutes were detected at 328 nm.RESULTS:The resolution of tetrahydropalmatine and pepper alkali was good,with no miscellaneous peak.The linear range was at 0.196-1.96 ?g(r=0.999 6)for tetrahydropalmatine,0.03-0.3 ?g(r=0.999 7)for pepper alkali.The average rate of recovery of tetrahydropalmatine was 97.74%(RSD= 0.42%,n=6),and that of pepper alkali was 104.51%(RSD=2.01%,n=6).CONCLUSION:The method is simple,reliable,accurate and can be applied to the quality control of the preparation.

Objective To Screen out incidence of vitamin K deficiecy and complicated with hemorrhage in newborn patients, infant patients and normal neonates, and also study on the treatment of vitamin K deficiency. Methods Using emzymoimmunoelectrophoresis to test PIVKA Ⅱ in umbilical and vein blood. Results The incidence of vitamin K deficiency in normal neonates, newborn patients (≤ 5 days) and infants patients (25~60 days) are 31.2%,47.6% and 31.8%. The incidence of hemorrhage in newborn patients (≤5 days) is 26.0%, infant patients (25~60 days) is 66.6%. Intramuscular injection of vitamin K 1 1 mg is the proper dosage to prevent and treat vitamin K deficiency. Conclusion The neonates right after birth or about 25 days after birth, especially those of breast feeding and who are getting lievr and gall diseases should receive vitamin K 1 to prevent vitamin K deficiency.

AIM: To investigate the anticancer effect of manumycin on abdominal metastatic breast cancer cell line-SK-BR-3 and its relationship with p38 MAPK . METHODS: The test of anticancer effect was performed by the method of MTT, apoptosis induced by manumycin and affected by SB203580, a specific p38 MAPK inhibitor, were examined by caspase-3 activity assay kit, and the protein expression was detected by immunoblotting assay. RESULTS: The inhibition rates at 24 h after treatment with manumycin of 6 ?mol/L, 18 ?mol/L, 54 ?mol/L were (7.4?3.9)%, (21.0?4.4)% and (64.7?4.1)%, respectively and showed dosage-effect relationship. Compared with the control group, the survival rates of the last two treatment groups were decreased significantly (P

Objective To observe functional changes of renal tubular epithelial cells stimulated by high mobility group protein box 1 (HMGB1) and associated mechanism.Methods Renal tubular epithelial cells (NRK52E) were divided into control group,HMGB1 group and HMGB1+ lipopolysaccharide from Rhodobactersphaeroides (LPS RS) group.Toll-like receptor 4 (TLR4) expression was detected by immunofluorescence and Western blotting.Apoptosis rate and cell cycle arrest were identified with flow cytometry.The activation of MAPK signaling pathway and NF-κB were detected by Western blotting.The IL-1,IL-6 and tissue inhibitor of metalloproteinases 2 (TIMP2) mRNA levels were measured by real-time PCR.The secretion levels of IL-1,IL-6 and TIMP2 were measured by protein chips assay.Results TLR4 was expressed by NRK52E cells.Compared with the control group,there were increased cell cycle G1 arrest,MAPK signaling pathway and NF-κB activation in HMGB1 group.Furthermore,IL-1,IL-6 and TIMP2 mRNA levels were increased and IL-1,IL-6 and TIMP2 were secreted by NRK52E when stimulated with HMGB1 (all P ＜0.05).However,effects mediated by HMGB1 stimulation could be inhibited by LPS RS (all P＜0.05).Conclusions Inflammatory activation of NRK52E cells can be mediated by the interaction of HMGB1 and TLR4.

BACKGROUND:With increased age,bone marrow mesenchymal stem cells(BMSCs)are influenced with regard to quantity and quality,which will induce great damages to the donors.Many studies have focused on seeking substitute MSC source.In contrast it remains controversial whether umbilical cord blood contains MSCs.OBJECTIVE:To isolate MSCs from human umbilical cord blood,and to detect their biological properties.METHODS:Umbilical cord blood samples were sterilely isolated using Percoll density gradient centrifugation to harvest intermediate layer cells.DMEM medium containing fetal bovine serum,penicillin,streptomycin and L-glutamine was added.Following several adherences and purification,the floating cells were discarded.Thus,many adherent cells with a confluence were collected.When cells were 60% 80% confluent,cells were digested by trypsin for subculture.Cells at passages,1,5 and 9 were obtained and their morphological changes were observed.Cell surface antigens were measured using flow cytometry.Growth curves were drawn,and cell viability was determined utilizing MTT.RESULTS AND CONCLUSION:Isolated umbilical cord blood MSCs presented an even size,showing spindle or star-shape fibroblasts-like cells.Umbilical cord blood MSCs at 1,5,9 passages were greatly positive for CD29,CD105 and CD166,but weakly positive for CD34 and CD45.Following 5 days of incubation,cells entered logarithmic growth phase.The number was decreased at day 9.Population doubling time was(53.5±8.32)hours.Cells grew well.Cells at 1-7 passages showed similar viability(P ＞ 0.05).Till passage 9,cell proliferation viability was decreased,but no significant difference was determined(P ＞0.05).Results verified that MSCs can be successfully isolated from umbilical cord blood in vitro.Cells at passages 1-9 presented a good reproductive activity.

Objective To find the effect of alter-expressed PER2 on proliferation,apoptosis and other clock genes expression in human oral squamous cell carcinoma SCC15 cells.Methods Short hairpin RNA interference was used to knockdown PER2 in SCC15 human oral squamous cell carcinoma cells.Flow cytometry analysis was used to testify the cell proliferation and apoptosis.Quantitative real-time PCR was used to testify the mRNA expressions of PER3,BMAL1,DEC1,DEC2,CRY2,TIM,RORα,NPAS2,PER1 and REV-ERBα.Results The proliferation was enhanced and apoptosis was decreased after PER2 knockdown in SCC15 cells (P<0.05).The mRNA expression of PER3,BMAL1,DEC1,DEC2,CRY2,TIM,RORα and NPAS2 was significantly down-regulated,and the mRNA expression of PER1 and REV-ERBα was significantly up-regulated (P<0.05).Conclusions Clock gene PER2 plays an important role in regulating other clock genes of the clock gene network in cancer cells,PER2 knockdown can enhance proliferation and recede apoptosis of cancer cell.

Aim To investigate the effects of YMⅢ,a derivative of naftopidil, on the vascular contractive activities in rabbit aorta and to explore its vasodilative mechanisms.Methods The isotonic contractions of the thoracic aorta strips from rabbits were recorded, and the effects of YMⅢ on the concentration-response curves of noradrenaline(NA),high potassium and 5-hydroxytryptamine(5-HT)were observed.Intracellular free Ca2+([Ca2+]i)was investigated in the pressence of and the absence of YMⅢ in different conditions.Results YMⅢ(10-8,5?10-8,10-7 mol?L-1)shifted the concentration-response curve of NA with a parallel manner to right, the maximum response was unchanged and the pA2 value was 8.00; YMⅢ (10-5 mol?L-1)also shifted the concentration-response curve induced by high potassium to right but with non-parallel manner,the response was depressed and the pD′2 value was 4.26. However, YMⅢ(10-7,10-6,10-5 mol?L-1)had no statistical influence on the concentration-response curve induced by 5-HT, although it tended to depress the response of the curve at 10-5 mol?L-1.In Ca2+-free medium,YMⅢ (10-8,5?10-8 and 10-7mol?L-1) significantly inhibited the transient contraction induced by NA and the long-lasting one induced by addition of Ca2+ with a concentration-dependent manner.But even at 10-5 mol?L-1,it did not inhibit the contraction induced by caffeine.Conclusions YMⅢ may be ?-adrenergic receptor blocker.Its vasodilative mechanism may be related to:blocking ?-adrenergic receptor on cell membrane resulting in the inhibition on the influx of extracellular Ca2+ and the release of intracellular calcium.

Background Acute stress can provoke the apoptosis of retina cells and induce increasing expression of calcitonin gene related peptide(CGRP)in retina.However,the role of CGRP in pathology of the stressinduced apoptosis of the retina ceils is still elusive.Objective The aim of this study was to investigate the effects of endogenous CGRP on retinal cell apoptosis induced by stress of acute myocardial ischemia after coronary artery occlusion in rats. Methods The acute myocardial ischemia model was established by ligating the left anterior descending branch of coronary artery in 12 male Sprague-Dawley rats.The rats were randomized into CGRP8-37 injection group and normal saline injection group,6 rats 12 eyes for every group.CGRP8-37(10-7 mol/L),a specific antagonist of CGRP receptor,was intravenously injected in CGRP8-37 group by caudal vine at 15 minutes prior to the coronary artery occlusion,and the equivalent amount of normal saline was used at the same fashion in normal saline group.The retinal samples of the rats were collected at 3 hours after coronary artery occlusion for TUNEL staining and caspase-3 activity detection respectively. Results The cellular displacement was observed in inner and outer nuclear layer,and vacuolar degeneration of retinal ganglion cells was found in the coronary artery occlusion animals.The total apoptosis index of retinal cells in CGRP8-37 group was significantly higher than that in normal saline group (42.8％±2.8％ vs 37.5％±2.9％,t=-3.244,P<0.01).The retinal capase-3 activity was significantly enhanced in the CGRP8-37 group compared with saline group(11.3±3.1 fold vs 4.9±1.2 fold,t=-4.603,P<0.01)at 3 hours of coronary artery occlusion.Conclusion The results suggest that the endogenous CGRP may play an anti-apoptotic role in the stress.induced retinal cell injury.