BACKGROUND: Tumor necrosis factor-alpha (TNF-α) in the brain can promote inflammative reaction and thrombosis through inducing the expression of endothelioal adhesion molecule during cerebral ischemic reperfusion.OBJECTIVE: To investigate the good regulation of scalp point through point acupucture with Baihui (GV-20) to Qubin (GB-7) to ischemic cerebral injury, and to inhibition of TNF-α over-expression within the brain as well.DESIGN: A randomized controlled trial.SETTING: Emergency Neurological Department of the Hospital Affiliated to Medical College of Qingdao University.MATERIALS: The experiment was completed from August 2002 to August 2003 at the Institute of Cerebrovascular Disease in Medical College of Qingdao University. A total of 15 healthy female SD rats of clean grade, weighing 230-270 g, provided by Shanghai Experimental Animal Center of Chinese Academy of Sciences, were at random divided as sham operation group,acupuncture group and control group, with 5 rats in each group.for rats in acupuncture group and control groups, the procedur for rats in sham operation group was similar to those in control group, but without obstruction of the middle cerebra artery. After ten minutes ischemia, the rats in acupuncture group were given acupuncture treatment. The hairs in parts correspondent to Baihui (GV-20) and Fengfu (GV-16) points in human were cut down and conventionally sterilized, then a filiform needle in 0.35 mmx40 mm was used to point to point acupucture correspondent to Baihui and Qubin in human to the depth of 0.8 cm. Another needle was used to perpendicularly insert at Fengfu (GV-16) point to the depth of 0.5 cm. A pulse electric machine was applied for 30 minutes, with the positive electrode connecting to Baihui (GV-20) and negative electrode to the Fengfu GV-16), the frequensection and in situ hybridization assay of TNF-α: All rats of every group,after treatment of 2 hours ischemia and 12 hours reperfusion, were deeply anesthetized, quickly given 200 mL normal saline through the catheterized ascending aorta, and continually given 300 mL paraformaldehyde of 40 g/L concentration; then they were decapitated for collecting the brain, the brain was fixed for 1 hour with the same solidification solution. The coronal sec-tion about 7 μm was prepared starting from the part of optic chiasm towards the back. The cytoplasm of TNF-α positive cell was stained brownish yellow, and that of TNF-α negative control cell was not stained. VIDAS-21 image analysing system was used to analyse the image, the result was extissue: After conventional staining with hematoxylin-eosin, the pathological changes of focal areas in both acupuncture group and control group were observed under light microscope.ischemic cerebral tissues in acupuncture group and control group.TNF-α in the cortex and striate body of brain in rats with ischemic reperfusion: Compared with sham operation group, the expressions in acupuncture group and control group were significantly increased (0.302±0.04,0.320±0.02; 0.466±0.08, 0.423±0.02; 0.367±0.03, 0.362±0.02; P ＜ 0.05);compared with acupuncture group, the expression in control group was sigbral tissues: In control group the extent of degeneration and necrosis was more serious, the neuron was degenerative necrosis, the neuroglia cell was in vacuolar degeneration, there was obvious phenomenon of leukocytic aggregation and margination within the vessel, and also leukocytic infiltration around the vessels. In acupuncture group there were only a small nub of neurons being in degenerative degeneration and neuroglia cells in vacuolar degeneration, and less leukocytic aggregation within the vessel.CONCLUSION: The level of TNF-α in the cortex and striate body of brain was increased after ischemic reperfusion. Scalp point through point acupuncture with Baihui (GV-20) to Qubin (GB-7) could markedly inhibit its expression, so that the therapy could effectively check over-expression of inflammatory mediators and their biological effects, hence reducing the inflammatory injury after cerebral ischemia.

Objective Through the DNA barcoding 12s rRNA sequences alignment and analysis of several rhinoceros horns products involved in cases to analysis the application feasibility of 12s rRNA in the rhinoceros horns products’ species identification. Methods Use rhinoceros horns products in 3 cases as materials, total DNA were extracted with improved method, PCR ampliifcation the DNA barcoding. Results The alignment and analysis of sequences show that 12s rRNA could identify rhinoceros horns products at the species level. Conclusion The DNA barcoding 12s rRNA could be used as a new way to identify the rhinoceros horns products which can’t be identiifed with morphological characteristics, provide a reliable basis for the qualitative and sentencing of cases.

Objective To observe the expression of IL-1? and to study the mechanism of acupuncture for focal cerebral ischemia. Methods The ischemia-reperfusion model was established in rats. The immunohistochemistry technique was used to detect the expression of IL-1?in the sham operation group, control group and acupuncture group at 2h, 6h,12h,24h after reperfusion following middle cerebral artery occlusion in rats, TTC staining method was used to observe the infarct volume at 24h after reperfusion. Results The sham operation group showed a baseline expression of IL-1? in cortex and striate body. The cells expressing IL-1? were increased in 2h after reperfusion and peaked at 12h after reperfusion in the control and acupuncture groups. The expression of IL-1? was weaker in acupuncture group as compared to that in the control group at all the time points. Conclusion Acupuncture could downregulate the expression of IL-1?in cortex and striate body and reduce infarct volume. It was indicated in the study that downregulated expression of IL-1? might be one of the mechanisms of acupuncture protecting the brain from ischemic damage.

Objective To study the effects of iodine on the proliferation activity of fibroblasts at different doses. Methods The cultured fibroblasts were treated with different dose iodine(100 ?g/L, 500 ?g/L, 1 000 ?g/L, 3 000 ?g/L, 5 000 ?g/L, 7 000 ?g/L, 9 000 ?g/L, 11 000 ?g/L)for 24 h and observe the morphology of fibroblast. MTT colorimetry was used to detect the fibroblast proliferation activity. Results Iodine could significantly increase the fibroblast proliferation activity at certain range of concentration (7 000 ?g/L). The proliferation activity was highest from 5 000 ?g/L to 7 000 ?g/L, the differences were significant compared with the iodine free control. Conclusion Iodine, the exposure level is 7 000 ?g/L.

Objective To study the protective role of nerve growth factor (NGF) on cerebral ischemic reperfusion injury and its effect on Bax expression. Methods The models of focal cerebral ischemic reperfusion in rats were made by occlusion middle cerebral artery using an intraluminal filament method. NGF was given through vena caudalis. The expression of Bax was studied by immunohistochemistry method and the change of brain was observed using pathological method.Results Bax positive cells in the cortex and striatum areas in ischemia-reperfusion group (115.25?15.07) were more than those in NGF treatment group (70.00?12.77) ((P

Objective Some cases of lipid storage myopathy (LSM) are similar to the polymyositis(PM) in clinic.To study the muscle pathology is helpful as to making clear the diagnosis and elucidating the mechanism of the LSM. Methods 14 cases of LSM with high level of CK in serum were analyzed clinically and compared with their muscle morphological and histochemical changes.Results Muscle weakness and fatigue were the main complaints of these patients.Most of them had acute or sub acute onset,some of them had relapse courses.All of the muscle specimens showed typical accumulation of lipid in muscle fibers.Necrotic fibers were found in 9 cases,3 of them showed changes with the courses. Conclusion Muscle biopsies and histochemical studies are necessary to differentiate LMS with PM.The fiber necrosis and CK leakage in LSM may be caused by changes of the inter circumstance and the membrane disturbance of cells.

Objective:To study the ultrastructural changes of the junctional epithelial after dental preparation. Methods: Junctional epithelium (JE) of left premaxillary teeth was detached by diamond bur in 8 rabbits (group 1) and by surgical knife in another 8 rabbits (group 2). JE of the right teeth without treatment was used as the controls. 4 animals in each group were sacrificed 1 and 2 weeks after operation respectively.Samples were prepared for transmission electron microscope observation.Results: In the two tested groups reattachment was observed 2 weeks after operation,degeneration of mitochondria in JE cells and destruction of basement membrane were observed. No significant difference was observed between group 1 and group 2.Conclusion: Dental preparation may result in ultrastructural changes of JE cells.

Objective To summarize the therapeutic efficacy of electrocoagulation for varicose veins of the lower extremities.Methods Totally 168 cases(238 limbs)of varicose veins of the lower extremities were treated with electrocoagulation in our department from April 2000 to April 2007.Through a small ankle incision,electrocoagulation probe was inserted ascendingly into the groin via the great saphenous vein.After high ligation of the great saphenous vein,the varicose veins and great saphenous trunk were electrocoagulated.Results The patients were discharged from hospital in one to two days after the operation.None of them had incisional infection.The ulcers at the ankle were healed in four weeks.Postoperative complications in this series,included Ⅰ degree burn injury in one limb,Ⅱ degree burn injury in one limb,superficial phlebitic pain in two limbs(two cases);and numbness around the ankle in 14 limbs(10 cases).Totally 148 cases with 208 limbs were followed up for 2 to 9 years with a mean of 6.6 years.During the period,the rate of recurrent varicosity was 3.4%(7/208);and healing rates of ache,dermatitis and eczema,ankle pigment,swelling around the calf and ankle were 95.3%(162/170),85.7%(12/14),41.7%(15/36)and 36.4%(16/44)respectively.Conclusion Electrocoagulation is a minimally invasive and effective method for the treatment of varicose veins of the lower extremities,with quick recovery and low rate of recurrence.

Objective To explore the nechanism of the interaction between K-opioid receptor and ? adrenal receptor. Method The effects of u50 488H on L-type calcium currents in the normal and hypoxic rat ventricular myocytes were studied by using whole-cell patch clamp technique. Results The basal as well as Isoproterenol-stimulated I_ Ca,L were inhibited by U50,488H in a dose-dependent manner in normal rat ventricular myocytes. In the hypoxic rat ventricular myocytes,the inhibitory effect of U50,488H was decreased. U50,488H had no significant effect on Forkolin-stimulated I_ Ca,L . Conclusion The results indicated that the negative modulation of ?-opioid receptor on ?-adrenoceptor was attenuated in the hypoxic ventricular myocytes,and the target of U50,488H on ?-adrenergic system might be situated between ?-adrenoceptor and adenylate cyclase.

Objective To screen a cell line which stably suppresses DAPK expression and to observe the growth characters.Methods Four pairs of shRNA were designed,synthesized and inserted into the pDsRed1-N1-U6 vector.The recombinant plasmids were purified and transfected into PC12 cell.Meanwhile,a pDsRed1-N1-U6 vector was transfected as control.The cell clones were screened by G418,and the stable PC12 cell line was established.DAPK expression was detected by Western blot.MTT method and Flow Cytometry(FCM) assay were used to assess the growth characters of the cell line.Results The shRNAs were transfected into PC12 cell and the cell clones were successfully screened out.Of the four recombinant plasmids,the F2 was the best interfering shRNA.Beyond our expectation,the F1 recombinant plasmid had an enhanced effect on DAPK expression.Conclusion A stable PC12 cell line with stable inhibition of DAPK expression by was established using siRNA expression vectors.