Objective:To observe the clinical effects of acupuncture at Jiaji (EX-B 2) points plus tuina manipulation for thoracic facet joint disorder, and get new clinical evidence for treatment of thoracic facet joint disorder. Methods:Totally 106 patients with thoracic facet joint disorder were randomly allocated into an observation group and a control group based on the random number table. Patients in the control group were treated by tuina manipulation, while those in the observation group were treated by acupuncture at Jiaji (EX-B 2) points before tuina manipulation. Patients in the two groups were treated once a day. The improvements of signs and symptoms and the efficacy were observed after 3 treatments. Results:After treatment, there were intra-group statistical differences in scores of 8 signs and symptoms in both groups (P<0.01); the score of each item in the observation group was lower than that in the control group, and there was statistical significance in the inter-group difference (P<0.05). The cure rate of the observation group was 50.9%, versus 26.4% in the control group, and there was a significant difference between the two groups (P<0.05). Conclusion:In treating thoracic facet joint disorder, acupuncture plus tuina manipulation can restore the biomechanical balance of thoracic vertebrae, fully maximized the combined effect, and significantly improve the clinical efficacy.

Objective To observe the expression of matrix metalloproteinase(MMP-2, MMP-9 and vascular endothelial growth factor (VEGF) in retinoblastoma (RB) and its relationship with the differentiation and optic nerve infiltration of RB. Methods Forty paraffin specimens of pathological confirmed RB were studied. They were divided into differentiated group (15 cases) and undifferentiated group (25 cases) , optic nerve infiltration group( 13 cases) and without optic nerve infiltration group(27cases). The expression of MMP-2, MMP-9 and VEGF were detected by immunohistochemistry, their relationships with the differentiation and optic nerve infiltration were also analyzed. Results The positive rate of MMP-2, MMP-9 and VEGF expression in 40 RB cases were 52.5%, 57.5% and 72.5%respectively. The expression of MMP-2, MMP-9 and VEGF in the undifferentiated group were significantly higher than those in the differentiated group (χ2= 9. 037, 9. 253, 8. 095;P＜0. 05). The expression ofMMP-2, MMP-9 and VEGF in RB with optic nerve infiltration group were significantly higher than those in RB without optic nerve infiltration group (χ2=11.045,10. 243, 8. 956;P＜0. 05). The expression of MMP-2,MMP-9 had a positive correlation with the expression of VEGF in RB (r= 0. 126, 0. 314;P ＜ 0. 05).Conclusions MMP-2, MMP-9 and VEGF expressed in RB tumor tissues. The expression of MMP-2,MMP-9 has a positive correlation with the expression of VEGF. The levels of MMP-2, MMP-9 and VEGF expression are related to optic nerve infiltration of RB cells.

Objective To investigate the safety and feasibility of carotid artery stenting (CAS) and evaluate its clinical outcomes. Methods From July 1998 to December 2003, 30 consecutive patients with 32 lesions underwent extracranial CAS procedures. Thirteen patients had a history of stroke or TIAs, 22 were hypertensive, 11 were diabetic and 8 had history of MI. Neurological assessment, Carotid duplex ultrasound, carotid and intracranial angiography were done before CAS in all patients. All the cases were done percutanously from femoral arteries and stenting was applied in all procedures. Carotid duplex ultrasound, cardiac and neurological elevation were performed post procedure. Results 30 patients (26 male and 4 female) underwent a total of 32 CAS procedures. Total 32 self-expandable stents and 1 tubular stent were implanted in all the cases. Direct stenting technique was applied in 9 cases. The other 21 procedures were performed with distal filtration supporting devices. The device can not be delivered due to tortuous target vessel in one case (success rate 95%) and CAS success rate was 97%. The particles were found in all filter baskets. Four patients underwent coronary artery bypass grafting 1 month later post CAS without perioperative neurological and cardiac events. One patient had contralateral cerebral hemorrhage during CAS and died three days later. Another patient died three days after CAS due to acute pulmonary edema. No restenosis was found by means of carotid duplex ultrasound during the follow-up (3-60 months) study. Conclusion CAS is safe and feasible in preventing ischemic stroke. This new alternative has satisfied clinical outcomes in managing cardiac and neurological ischemic diseases. Operative embolic complication can be potentially prevented by neurological protective device.

Objective The purpose of this study was to assess the effectiveness and safety of distal filtration protection devices during carotid artery stenting.Methods Between June and July in 2002, carotid artery stenting (CAS) were performed in 4 patients with asymptomatic severe carotid artery stenosis. Self-expendable stents (Smart, Cordis) and distal filtration devices (Angioguard XP, Cordis) were used in all patients. Primary endpoints were perioperative neurologic complications and mortality. Data were collected prospectively.Results All patients were male, their age were 59?4 years old. One patient had history of transient ischemic attacks(TIAs); and another had inferior myocardial infarction, and right coronary intervention was performed before CAS, and another one underwent CABG 6 months ago. There were two right internal carotid and two left internal carotid severe lesions (82.5?6.0)% were treated. The filtration devices and stents were delivered and deployed successfully in all target artery (technical success rate was 100%) and

0. 05), and were closely correlated to the clinical stage and the curative effect(P 0. 05). Conclusions: VEGF expression was contributes to the tumor neovascularization and tumor metastasis. Local control rate of VEGF positive tumor patients can be increased but their survival can not be prolonged by mono-chemotherapyonly only.

Objective To study the inhibitory effects of siRNA targeting survivin on the expression of survivin,as well as the apoptosis,proliferation and invasion of a human melanoma cell line,M14.Methods Two siRNAs targeting survivin were designed,chemically synthesized,and used to construct the recombinant plasmids,pRAT-H1.1/neo-survivin-siRNA1 and pRNAT-H1.1/neo-survivin-siRNA2.Then,recombinant plasmids were transfected into M14 cells mediated by Lipofectamine 2000 reagent.Those cells untransfected or transfected with empty vector served as the control.After culture over various periods of time.cells were collected for the detection of mRNA and protein expression of survivin with RT-PCR and Western blotting,respectively,and for the examination of apoptosis and proliferation of M14 cells by flow cytometry and MTT methods,respectively.Also,Transwell assay was performed to detect the invasive capability of M14 cells.Results A statistical decrease in the mRNA and protein expressions of survivin was observed along with an increase in apoptotic rate(x2=31.55,P＜0.01)in M14 cells transfectcd with siRNA-containing plasmid compared with untransfected and empty vector-transfected cells.As MTT assay indicated,on day 4 after the transfcorion,the proliferation of M14 cells was inhibited by(55.4±4.3)%,(34.5±4.3)%and(13.3±4.6)%,with pRNAT-H1.1/neo-survivin-siRNA1,pRNAT-H1.1/neo-survivin-siRNA2 and empty vector,respectively:there was a significant difference among the three groups(P＜0.05).Decreased invasive capability was noticed in M14 cells transfected with siRNA-containing plasmid compared with untransfected cells(all P＜0.05).Conclusions The plasmid containing siRNA against survivin can specifically inhibit the expression of sarvivin,proliferation and invasion of tumor cells,and induce cell apoptosis.The inhibition of survivin expression by siRNA may be a rational approach to the gene therapy for malignant melanoma.

Objective To measure the changes in levels of tumor necrosis factor α(TNF-α) secreted by human endothelial cells EC-304 after hematoporphyrin monomethyl ether-photodynamic treatment (HMME-PDT),and to explore the relationship between cytokines and inflammation initiation after management of nevus flammeus with photodynamic therapy.Methods EC-304 cells were cultured in 6-well plates,and classified into 4 groups:HMME-PDT group pretreated with HMME followed by irradiation with laser,HMME control group treated with HMME only,laser control group irradiated with laser only,and blank control group without any treatment.Culture supematants of EC-304 cells were collected from HMME-PDT group at 12,24 and 48 hours after the irradiation,and from the other three groups at the same time points.The supernatant TNF-α level was measured with enzyme linked immunosorbent assay (ELISA).Results The difference was statistically significant in the supernatant TNF-α level between different time points in each group (F=62.276.P<0.01) and between the 4 groups at each time point (F=11.538,P<0.01).Multiple comparison analysis showed that HMME-PDT group differed significantly from the other 3 control groups in the supernatant TNF-αlevel at each time point (all P<0.01),while no significant difierence was observed among the other three control groups at any time point (all P>0.05).Conclusion HMME-PDT promotes the secretion of TNF-α by EC-304 cells.

BACKGROUND: In recent years, there are many studies designed to explain the protective effect of lidocaine on brain, but few about the therapeutic effect on traumatic cerebral edema. The content of aquaporin-4(AQP-4) in brain tissue is the highest and it has been proved that AQP-4participants in the formation of cerebral edema induced by cerebral trauma, cerebral infarction, eerebrai tumor and other reasons.OBJECTIVE: To observe the effect of lidocaine on the expression of AQP-4 of experimental rats following brain injury and analyze the therapeutic effect of lidocaine on brain edema.DESIGN: A randomized and control animal experiment.SETTING: Department of Anesthesiology, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: The experiment was carried out in the Institute of Brain Disease, the Medical School Hospital of Qingdao University between January and July 2004. Totally 65 three-month-old healthy male Wistar rats,were enrolled in the experiment and randomly divided into 3 groups: normal control group (n=5), model group (n=30) and treatment group (n=30). Thirty rats in the model group and treatment group were respectively assigned into 6 subgroups according to 6 different time points: 1,4,6,12,24 and 48 hours following brain injury, with 5 rats at each time point.METHODS: Animal models of brain injury at right parietal lobe were created according to the method from Feeney et al. As for the rats of normal control group, they only underwent operation to be injured at the corresponding part. Rats recovered the access to food and water 2 to 8 hours after operation. For the rats in the treatment group, they were intraperitoneally injected of lidocaine at the 1st, 4th,6th, 12th, 24th, and 48th hours following injury, 5 rats at each time point. The initial dosage was 30 mg/kg, then 15 mg/kg was maintained . Administration was conducted every 6 hours in 3 days; For the rats in the model group, they were intraperitoneally injected of 30 mg/kg normal saline and rats in the normal control group were given no special treatments. Water content of brain was calculated 5 days following brain injury with dry and wet weight method:water content of brain=[brain mass (wet)-brain mass (dry)]/brain mass(wet)×100%. Expression of AQP-4 of brain tissue of rats was detected with immunohistochemical method and cytomorphological change of brain tissue was observed under optical microscope.MAIN OUTCOME MEASURES: ① Water content of brain of rats in each group. ② Expression of AQP-4 of brain tissue of rats in each group.③ Pathological results of brain tissue of rats in each group.RESULTS: No rats died accidentally or for other factors in the process of experiment, finally, all the 65 rats entered the stage of result analysis. ①Compared with model group, administration of lidocaine within 6 hours following brain injury could significantly decrease the water content of brain tissue [1 hour after brain injury: (81.09±0.29)%, (83.04±0.25)% ,P ＜ 0.05];[4 hours after brain injury: (81.34±0.35)%, (83.31±0.48)%,P ＜ 0.05] ;[6 hours after brain injury: (82.01±0.21)%, (83.25±0.37)% ,P ＜ 0.05]. Compared with model group, administration of lidocaine could significantly decrease the expression of AQP-4 [1 hour after brain injury:(0.19±0.02), (0.24±0.03),P ＜ 0.05]; [4 hours after brain injury: (0.21±0.05 ), (0.25±0.05) ,P ＜ 0.05]; [6 hours after brain injury: (0.21±0.03 ),(0.24±0.02) ,P ＜ 0.05]. There were no significant differences of AQP-4expression and water content of brain tissue when administration was conducted at the 12th, 24th and 48th hours following brain injury (P ＞ 0.05). ②Under the microscope, AQP-4 positive cells presented vacuolus, they mainly lay in the edema area of peripheral part of trauma, cortex of traumatic side and around the blood vessel as well as astrocyte of white substance, choroid plexus and ependymal layer. ③ Necrosis was found in most cells in the central area of trauma and apoptosis in most cells in the peripheral area. Compared with model group, necrotic and apoptotic cells were significantly less within 6 hours following trauma, but not at the 12th,24th and 48th hours following trauma in the treatment group.CONCLUSION: High dosage of lidocaine can decrease the expression of AQP-4 and lighten cerebral edema following brain injury, but administration should be given as early as possible.

Objective To study the effects of triptolide on the apoptosis in and proliferation of a human melanoma cell line M14.Methods M14 cells were cultured with the presence of 5 concentrations (12.5,25,50,100,200 nmol/L) of triptolide for 24,48 and 72 hours respectively,and cell counting kit-8 (CCK-8) was used for the detection of cell proliferation.Some M14 cells were treated with triptolide at 10 nmol/L,20 nmol/L and 30 nmol/L for 48 hours followed by the analysis of cell cycle by flow cytometry and detection of cell apoptosis by flow cytometry following annexin V-fluorescein isothiocyanate (FITC)/propidium iodide double staining.The morphological changes of M14 cells treated by triptolide at 30 nmol/L for 48 hours were observed by Hoechest 33258 staining.Results Compared with untreated M14 cells,an increase of cell population in S phase was observed in triptolide-treated cells,along with a decline in cell population in G2/M phase.The apoptosis rate was (2.92 ± 0.17)％,(20.99 ± 0.40)％,(34.28 ± 2.04)％ and (63.38 ± 0.71) ％ respectively in M14 cells treated with triptolide at 0,10,20 and 30 nmol/L for 48 hours,suggesting that triptolide enhanced the proliferation of M14 cells in a dose-dependent manner.After treatment with triptolide of 30 nmol/L,M14 cells showed morphological changes characteristic of apoptosis.Conclusion Triptolide could inhibit the proliferation of and induce the apoptosis in M14 human melanoma cells.

Objective To compare the absorption characteristics of HMME by a human umbilical vein endothelial cell line ECV 304 versus a human keratinocyte cell line HaCaT.Methods Exponentially growing ECV304 and HaCaT cells were incubated with various concentrations (50,100,150,200 and 250 mg/L) of HMME for 16 h or HMME of 150 mg/L for various durations (15 min,30 min,1 h,3 h,8 h,12 h and 24 h).The quantity of HMME absorbed by the cells were determined by laser scanning confocal microscopy (LSCM).Results The fluorescence intensity was 74.00,125.57,135.24,141.99 and 132.09 for ECV304 cells,93.88,102.45,112.59,108.23 and 104.70 for HaCaT cells,after incubation with HMME of 50,100,150,200 and 250 mg/L,respectively.After treated with HMME of 150 mg/L for 15 min,30 min,1 h,3 h,8 h,12 h and 24 h,ECV304 cells showed a fluorescence intensity of 95.07,103.97,105.96,108.99,112.93,115.36 and 122.91,respectively,and HaCaT cells displayed a fluorescence intensity of 104.25,106.60,108.72,113.75,117.66,114.90 and 118.14,respectively.Conculsions Within a defined range of concentration and duration,the absorption of HMME by both ECV304 and HaCaT cells is,to some extent,concentration- and time-dependent.