Objective:To establish an HPLC method for the determination of chlorogenic acid in honeysuckle stem and honeysuck-le from different sources in different harvest periods. Methods:A Waters C18 column(250 mm × 4. 6 mm,5 μm)was used. The mo-bile phase consisted of acetonitrile-0. 4% H3 PO4(10:90)with the flow rate of 1. 0 ml·min-1 . The detection wavelength was 327 nm and the column temperature was 30℃. An external standard method was established for the determination of chlorogenic acid in honey-suckle stem and honeysuckle in six different harvest periods from three sources. Results:There was a good linear relationship within the range of 0. 065-1. 300 μg for chlorogenic acid(r=0. 999 8). The content of chlorogenic acid in honeysuckle was the highest in Sep-tember and October. The content of chlorogenic acid in Lonicera acuminata was the highest among the honeysuckle stem from three dif-ferent sources. Conclusion:The content of chlorogenic acid in honeysuckle has a certain relationship with the harvest time,which can provide theoretical basis for the choice of harvest time for honeysuckle stem.

OBJECTIVETo study the effect of ethyl acetate fraction of Blumea balsamifera (BBE) residue on treating rats of adjuvant arthritis (AA) and its mechanism.

METHODThe rats were immunized with the Freund's complete adjuvant (FCA). After modeling, 28 days' treatment with BBE was performed. During the experimental process, rat mass, toe girth, arthritic index (AI), proliferation of immune organs and pathological section were measured. After treatment, blood samples were collected through fossa orbitalis vein for detection of serum SOD, MDA, GSH, NO, OH*, ALP, AST, ALT, NAG and SA content using colorimetric method and IL-1, IL-6, TNF-α content using ELISA method.

RESULTAdministration with BBE (high dose) could significantly ameliorate joint swelling and arthritis index, effectively inhibit synovial hyperplasia, down-regulate the levels of MDA, NO, OH*, ALP, AST, ALT, NAG, SA, IL-1, IL-6, TNF-α and up-regulate the SOD and GSH levels in serum.

CONCLUSIONThe results suggested BBE possesses substantial anti-arthritis and antioxidant activities.

Acetates ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; administration & dosage ; isolation & purification ; Arthritis, Experimental ; blood ; drug therapy ; metabolism ; Asteraceae ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; Humans ; Interleukin-1 ; blood ; Interleukin-6 ; blood ; Male ; Malondialdehyde ; blood ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; blood

AlM:To observe the changes of corneal endothelium after phacoemulsification cataract surgery in different types of cataract patients.
METHODS: Randomly selected age-related cataract, diabetic cataract and cataract of high myopia 30 eyes of 30 cases, respectively, in our hospital. All patients underwent phacoemulsification combined with intraocular lens implantation, corneal endothelial density and the percentage of hexagonal cells were measured by corneal endothelial cell instrument without touching before surgery and one week after surgery.
RESULTS: The difference of the preoperative corneal endothelial cell density and the percentage of hexagonal cells among three groups were not statistically significant (P>0. 05). One week after surgery, the cell density in three groups were respectively 2 496. 86 ± 298. 96/mm2 , 2 379. 51 ± 375. 13/mm2 , 2 425. 38 ± 312. 68/mm2 , the percentage of hexagonal cells were respectively ( 46. 20 ± 12. 03)%, (43. 44±13. 99)%, (44. 35±8. 13)%. Both the cell density and the percentage of hexagonal cells one week after surgery were lower than those before operation. There were significant difference in three groups ( P <0. 05). Both the measurements in diabetic cataract group and cataract of high myopia group after surgery were lower than those in age-related cataract group, the cell density and the percentage of hexagonal cells in diabetic cataract group were lower obviously compared with those in age- related cataract group and the difference was significant (P<0. 05).
CONCLUSlON:The tolerance of corneal endothelial cell to phacoemulsification cataract surgery is lower in cataract with diabetes and high myopia. Corneal endothelium should be assessed preoperatively and protected intraoperatively.

AIM: To prepare Nano-Liposome encapsulated MAGE3/HSP70(NL M3H) and study its character and antitumor immunity in mouse. METHODS: NL M3H was prepared by the thin film-dispersion ultrasonic. The shape and size of NL M3H were detected by electron microscope. The encapsulation rate, drug-carrying capacity, stability and the releasing character were tested by Sephedex-G100 gel filtration. The mouse was immunized by NL M3H, and the antitumor immunity was detected by ELISPOT and LDH release assay. RESULTS: The mean size of NL M3H was lower than 100 nm. The encapsulation rate was 38％.The drug content was 0.038 g/L. NL M3H has good stability after stored in 4℃ for 6 months. The releasing profile showed that 74 percent of proteins was released during the first 24 hours in saline. The results of ELISPOT and LDH release assay showed that NL M3H generated tumor specific cytotoxic T lymphocyte(CTL)to damage tumor cel1. CONCLUSION: NL M3H has novel characters, it can generate specific CTL to kill tumor cell, and can be used as new kind of vaccine agsinst tumor.

Objective To investigate the influence of proline-rich tyrosine kinase 2 (Pyk2)gene RNA interference on proliferation,invasion and migration of Hep3B hepatocellular carcinoma cells.Methods The Pyk2 gene RNA interference vector was transfected in Hep3B hepatocellular carcinoma cells by lipofectamine. The Hep3B cells divided into three groups:siRNA group (the vector with Pyk2 RNAi gene was transfected), negative control group (the vector without Pyk2 RNAi gene was transfected),and blank control group (no vectors was transfected).Pyk2 mRNA and protein were detected using reverse transcription reverse transcription-poly-merase chain reaction (RT-PCR)and Western blotting.The biological behavior including cell proliferation,inva-sion and migration were detected by 3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide (MTT), transwell and wound healing assay,respectively.Results The expression of Pyk2 mRNA of Hep3B cell line in siRNA group (0.1 6 ±0.03)was significantly decreased than those in negative group (0.74 ±0.1 3)and blank control group (0.77 ±0.1 6),with statistically significant differences (t=51 .46,P=0.000;t=53.21 ,P=0.000).The expression of Pyk2 protein of Hep3B cell line in siRNA group (0.24 ±0.06)was significantly decreased than those in negative group (0.83 ±0.05)and blank control group (0.91 ±0.06),with statisti-cally significant differences (t=57.29,P=0.000;t=68.53,P=0.000).The cell proliferation inhibition rate at 48 hours in siRNA group (26.1 7%±0.28%)was significantly raised than those in negative group (9.28%± 0.22%)and blank control group (6.47%±0.31%),with statistically significant differences (t=31 .45,P=0.004;t=34.64,P=0.002).The number of transmembrane cells in siRNA group (32.5 ±8.5)/1 0 HP was significantly declined than those in negative group (98.4 ±1 2.3 )/1 0 HP and blank control group (1 1 2.6 ± 1 1 .3)/1 0 HP,with statistically significant differences (t=95.64,P=0.000;t=1 05.1 7,P=0.000).The wound healing assay in siRNA group (28.1 7%±1 .46%)was significantly lower than those in negative group (77.38%±2.24%)and blank control group (79.41%±3.1 7%),with statistically significant (t=85.86,P=0.000;t=89.37,P=0.000).Conclusion Pyk2 gene involves the proliferation,invasion and migration of Hep3B cells,which has close correction with development and metastasis of hepatocellular carcinoma.Pyk2 gene is very helpful to become a molecular target for the diagnosis and treatment of hepatocellular carcinoma.

Objective To explore the clinical application of ultrasonography on tethered spinal cord syndrome (TCS).Methods Ultrasonic feature of 45 patients with TCS were analyzed retrospectively. Pre-and post-operative blood flow rates of cone were recorded by color doppler.Results Ultrasound was the same as computer tomography not only presented the image of anatomy and pathology of TCS,but also showed the lack of pulsatile motion of distal cord with TCS.Before the operation,blood flow rates of cone were( 0.047?0.012) m/s.After the operation,blood flow rates of cone were(0.158?0.029) m/s.There was significantly different(P

Objective To observe the treatment of ~(99)Te-MDP on typeⅡcollagen induced arthritis (CIA)in rat,and the effect on the expression of synovial MMP-3 and TIMP-1 mRNA.To explore the mech anisms of the ~(99)Te-MDP in the treatment of rheumatoid arthritis.Methods The rats in which C1A(n=24)were divided into three group:the control group(n=8),~(99)Tc-MDP group(n=8)and Methotrexate group(n=8). Arthritis were evaluated by arthritis index and histopathological index and the expressions of MMP-3 and TIMP-1 mRNA in synovium were detected by RT-PCR.Results①The arthritis indexs of the control group, the methotrexate group,the ~(99)Tc-MDP group were increased with time.②The histopathological scnres of the control group were significantly higher than those of methotrexate group and ~(99)Tc-MDP group(P＜0.01).The histopathological scores of cartilage destruction and bone erosion of ~(99)Tc-MDP group were lower than those of methotrexate group(P＜0.05).③The levels of MMP-3 mRNA of the control group,~(99)Tc-MDP group, methotrexate group were notably higher than those of the control group(P＜0.01).The levels of control group was notably higher than that of the ~(99)Tc-MDP group(P＜0.05).There was not significant difference in all groups on the levels of TIMP-1 mRNA(P＞0.05).Conclusions ~(99)Tc-MDP can notably relieve the arthritis symdrome and retard the catilage damage and bone erosion of CIA in rats,and could significantly decrease the MMP-3 mRNA in the synovium.Which may be one of the therapeutic mechanism.~(99)Tc-MDP is better than methotrexate in retarding catilage and bone erosion and decreasing MMP-3 mRNA in CIA rats in a 3-week therapeutic intervention.

Objective To examine the expression of toll-like receptor 2(TLR2)in peripheral blood monocytes and explore its association with disease stages and clinical manifestations and to explore the patho- genesis of rheumatoid arthritis(RA).Methods The expression of toll-like receptor 2 in peripheral blood monocytes from 47 RA patientis(27 in active stage and 20 in stable stage)and 18 normal individuals were de- tected by flowcytometry and RT-PCR.Results The expression of toll-like receptor 2 in peripheral blood monocytes in patients with active disease was significantly increased compared to non-active patients and nor- mal individuals,The expression was found to correlate with the Disease Active Score(DAS),serum C-reactive protein(CRP)level and the erythrocyte sedimentation rate(ESR),but not correlate with rheumatoid factor (RF)and the anti-cyclic citrullinated peptide(CCP)antibody.Conclusion The expression of toll-like recep- tor 2 in peripheral blood monocytes of patients with active RA is significantly increased.And the expression is correlated with disease activity index.The innate immune system is activated in patients with active disease. And the increased expression may promote the activities of monocytes.

Objective To determine the effects of~(99)Tc-MDP on joint inflammation and bone destruc- tion in collagen-induced arthritis(CIA)rats model and its effect on tumor necrosis factor alpha(TNF-?). Methods CIA was induced by immunization of male SD rats with an emulsion of collagen.~(99)Tc-MDP or placebo was intravenous infused to rats for 20 days.Joint inflammation was assessed by arthritis index.Lesions of bone were assessed based on the histological changes in ankle joints,radiographic analysis in hind paw with Larsen score.Systemic TNF-?level was measured by radioimmune assay.Results~(99)Tc-MDP suppressed joint swelling(P