OBJECTIVETo investigate the effect of allicin in inhibiting insulin-induced vascular smooth muscle cell (VSMC) proliferation and migration.

METHODThe tissue explant method was adopted to culture rat's aVSMCs, and the immunofluorescence method was used to identify α-SMA. Cells from the third to fifth generations were selected in the experiment The insulin-induced VSMC model was established. The experiment was carried out in five groups: the control group, the insulin group, the allicin group, the ERK inhibitors PD98059 group (20 μmol · L(-1)) and the PD98059 + allicin group. VSMCs' proliferation was determined by CCK8 colorimetric method, while its migration was detected by cell counting; The western blotting was used to detect total ERK, Phospho-ERK, PCNA protein's expression.

RESULTPrimary cultured VSMCs grew well in the spindle shape under the lightmicroscope, with peak and valley. α-SMA immunofluorescence results showed that the cultured cells had typical VSMCs' features. Insulin could stimulate VSMCs' proliferation and migration, with the best effect at the concentration of 100 nmol · L(-1). The pretreatment with allicin could significantly inhibit VSMCs' proliferation and migration induced by insulin in a dose-dependent manner. The pretreatment with PD98059 and allicin + PD98059 could inhibit VSMCs' proliferation and migration induced by insulin remarkably as well. Insulin could significantly accelerate VSMCs' expression of such proteins as p-ERK, PCNA. Contrarily, allicin could notably inhibit VSMCs' expression of such proteins as p-ERK, PCNA in a dose-dependent manner.

CONCLUSIONAllicin could significantly inhibit VSMCs' proliferation and migration induced by insulin, which may be related to the inhibition of the activation of ERK signal path.

Animals ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Insulin ; metabolism ; Male ; Muscle, Smooth, Vascular ; cytology ; drug effects ; metabolism ; Myocytes, Smooth Muscle ; cytology ; drug effects ; Plant Extracts ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Sulfinic Acids ; pharmacology

Objective To establish a method for simultaneous determination of icariin and naringin content in bushen huayu extract by HPLC. Methods Reverse-phase high performance liquid chromatography ( RP-HPLC ) separation was performed one C18 column (4.6 mmí250 mm,5 μm).The mobile phase was acetonitrile containing 0.2%H4PO3(pH=2.9). Gradient elution with a flow rate of 0. 8 mL·min-1 was applied to achieve the separation. The detection wavelength was set at 269 nm,and the column temperature was 30℃. Results Icariin had a good linear range from 0. 3-3. 0μg (r=0. 999 9),the recovery rate was 105. 6%,and RSD was 0. 95%. Naringin was linear within the range of 0. 12-1. 20 μg (r=0. 999 9). The recovery rate was 94. 0%and RSD was 0. 52%. Conclusion The method is simple,stable,accurate,and reproducible,which can be used as the quality control standard for bushen huayue extract.

Humans ; Leukemia, Lymphocytic, Chronic, B-Cell ; Societies, Medical ; United States

BACKGROUND:How does the peripheral nerve block work to inhibit the functions? What effects does it do to the spinal cord and the cerebral center? How does it regulate the peripheral nerve and how does it change anatomy of brain center? Al above are stil unknown.
OBJECTIVE:To observe the effect of peripheral nerve block on the biology of spinal cord nerve cel s.
METHODS: Sixty New Zealand rabbits were selected and randomly divided into three groups: ischiadic nerve block group, subarachnoid block group and extradural nerve block group. Each group had 20 rabbits which were sub-divided into experimental group and control group with 10 rabbits in each subgroup. The rabbits in the three experimental groups were injected with lidocaine, bupivacaine and lidocaine at the middle point between femoral head and ischiadic tuberosity in ischiadic nerve block group, subarachnoid block group and extradural nerve block group, respectively. The rabbits in each control group were injected with normal saline at the same position.
RESULTS AND CONCLUSION:After ischiadic nerve block, extradural nerve block or subarachnoid block for 30 minutes, the Nigeria’s bodies of the little round cel s in the poliomyelia posterior horn laminaeⅢandⅣand the polyhedral cel s in the anterior horn laminaeⅨwere less than those in the control groups. Nuclei leaned towards one side. c-Fos protein expression was decreased or showed no expression, suggesting that the cord nerve cel function of corresponding spinal segments was inhibited. The spinal pia mater of spinal cord had a hierarchical or fracture phenomenon after subarachnoid block.

Objective To evaluate curative effects of percutaneous laser disc decompression(PLDD) in the treatment of discogenic low back pain.Methods A total of 36 patients with discogenic low back pain was included in the study.The PLDD was performed by using the DIOMED semiconductor laser treatment,with a power of 15 W,1 s-pulse duration,1 s-interval,and an energy of radiation of 800~1 200 J.Curative outcomes were evaluated based on the Visual Analog Scale(VAS) criteria. Results The operation time was 15~60 min(mean,30 min).The pain provocation was observed in 32 patients.Follow-up examinations were conducted in the 36 patients for 6~36 months(mean,11 months).The postoperative VAS scores were improved by ≥ 3 points in 18 patients and ≥ 2 points in 14 patients.Results revealed that 32 patients(88.9%) reported a good response and 4(11.1%) reported no response. Conclusions Percutaneous laser disc decompression is a safe,effective,and minimally invasive procedure that can be used for patients with discogenic low back pain.

Objective To set up an antibody-capture ELISA method to detect the Japanese encephalitis virus(JEV)antibody.Methods ELISA plate was coated with the monoclonal antibody which was specific to the envelope protein epitope E39 of JEV,JEV SA14-14-2 strain as the source of antigen was used to absorb the monoclonal antibody,the absorbed virus used to capture the JEV'S antibody.The antibody that captured ELISA was established.The indirect ELISA method using the virus particles from cell culture was compared with coating ELISA plate,105 clinical serum were checked.Results The background in indirect ELISA assay could not be abscised,positive and negative serum diluted in a ratio of 1:10,1:100,1:1000,the relative value of A posative/A negative were 1.02,0.99,1.13,all<2.1.But the antibody-captured ELISA method when the serum dilution was 1:10,1:100,the A posative/A negative were 3.57,2.94,all>2.1;when the dilution was 1:1000,the A posative/A negative was 1.42,<2.1,it meant the method could distinguish the positive and negative serum efficiently when the dilution Was 1:100,the background problem in indirect ELISA assay could be solved.Antibody-capture method was used to check 105 serum samples,the A posative/A negative over a range of 0.257～0.321(0.262±0.050),all<2.1,no positive sample found.Conclusion The antibody-capture ELISA method has been preliminary set up with a high specificity,capable of quickly identifying JEV from other virus.

Objective To observe the characteristics of coronary hemodynamics in acute hibernating myocardium (AHM) model by intracoronary doppler guidewire. Methods 13 little domestic Chinese swine were established to the model of AHM by interventional method (closed chest),intracoronary Doppler guidewire were used to measure average peak velocity (APV),diastolic systolic velocity ratio (DSVR) and coronary flow velocity reserve (CFVR) distal to the stenosis in the period of pre model,post model and reperfusion.Then animals were executed,the histomography changes of AHM cells were observed.Results 10 animals (76 92%) were established to the model of AHM successfully,myocardium necrosis were not found under microscopical examination.APV and CFVR distal to the stenosis decreased significantly in AHM model ( P

Objective To analyze the effect of combined detection of serum tumor markers, Ki-67 and P-glycoprotein (PGP) in lymph node metastasis and postoperative recurrence of colorectal cancerpatients. Methods The clinical data of 517 colorectal cancer patients who had underwent surgicaltreatment were retrospectively analyzed. Among them, lymph node metastasis was found in 165 cases(lymph metastasis group), and 352 cases did not have lymph node metastasis (non lymphoid metastasis);postoperative recurrence was found in 224 cases (postoperative recurrence group), and 293 cases did havepostoperative recurrence (non postoperative recurrence group). The serum tumor markers levels of cancerantigen 19- 9 (CA19- 9), carcinoembryonic antigen (CEA), cancer antigen 125 (CA125) and theexpressions of Ki-67, PGP were compared. The correlation between the serum tumor markers levels andthe expressions of Ki-67, PGP was analyzed. Results The serum levels of CEA, CA19-9 and CA125 and the positive expression rates of Ki-67 and PGP in lymph metastasis group were significantly higher than those in non lymph metastasis group:(21.39 ± 3.15)μg/L vs. (10.12 ± 2.48)μg/L, (68.48 ± 5.82) U/L vs. (35.26 ± 3.51) U/L, (82.16 ± 7.53) U/L vs. (32.46 ± 6.24) U/L, 69.70% (115/165) vs. 13.64% (48/352) and 72.73%(120/165) vs. 14.77%(52/352), and there were statistical differences (P<0.01). The serum levels of CEA, CA19-9 and CA125 and the positive expression rates of Ki-67 and PGP in postoperative recurrence group were significantly higher than those in non postoperative recurrence group:(18.26 ± 2.34)μg/L vs. (9.18 ± 1.26)μg/L, (47.52 ± 4.85) U/L vs. (21.43 ± 2.18) U/L, (59.16 ± 4.25) U/L vs. (33.17 ± 3.46) U/L, 60.27% (135/224) vs. 9.56% (28/293) and 6.70% (15/224) vs. 7.51% (22/293), and there were statistical differences (P<0.01). The correlation analysis results showed that the positive expression rates of Ki-67 and PGP were positive correlated with the serum levels of CEA, CA19-9 and CA125 (P<0.05). Conclusions The levels of serum tumor markers and the positive expression rates of PGP and Ki-67 in colorectal cancer patients with lymph metastasis and postoperative recurrence are high. The levels of tumor markers are closely related to the positive expression rates of Ki-67 and PGP, which indicates that the combined detection of these indexes has a good effect on evaluating lymph metastasis and postoperative recurrence of colorectal cancer patients.

Objective To investigate the effect of ischemic preconditioning (IPC) on coronary microcirculation function in short-term hibernating myocardium(SHM) animal model.Methods Twelve little domestic Chinese pigs were established as the model of SHM by interventional method (closed-chest) and divided into 2 groups at random: the control group (CON group, n =6) and ischemic preconditioning group (IPC group, n =6).IPC was elicited by 2 cycle of 5 min of ischemia followed by 5 min of reperfusion before the establishment of model.Intracoronary Doppler guidewire was used to measure average peak velocity (APV),diastolic systolic velocity ratio(DSVR) and coronary flow reserve(CFR) of the distal to the stenosis at the baseline and 10 min,30 min,60 min,120 min after the establishment of model.Results APV,DSVR and CFR all decreased significantly in SHM model in both groups( P 0.05 ),at 60 min CFR of the CON group decreased significantly than before( 0.96 ? 0.27 vs 1.74 ? 0.49 , P 0.05 ).Conclusions The coronary microcirculation dysfunction happened at the time point of 60 min after coronary stenosis in SHM model,IPC can protect the coronary microcirculation function in SHM model.

BACKGROUND:Patients with spinal metastases may appear different degrees of pain and abnormal spinal stability, and can be treated with percutaneous baloon kyphoplasty combined with bone cement implantation. OBJECTIVE:To explore the effect of percutaneous baloon kyphoplasty with bone cement implantation on spinal stability and pain in patients with spinal metastases. METHODS:Twenty-three patients with metastatic spinal tumor were treated with percutaneous baloon dilatation kyphoplasty with polymethyl methacrylate bone cement. There were 10 females and 13 males, and their age ranged from 23 to 71 years. The visual analogue scale score, anterior and posterior edge height of vertebral body, quality of life, and motor function score of patients were observed before and after treatment. RESULTS AND CONCLUSION: Compared with before treatment, the visual analogue scale score and motor function score were significantly decreased, while anterior and posterior edge height of vertebral body were significantly increased in 23 patients at 24 hours after treatment (P < 0.05). After 12 months of folow-up, no case appeared to have spinal nerve root damage, serious adverse reactions and bone cement embolism. The patient'squality of life was significantly improved, compared with before treatment (P < 0.05). Experimental findings indicate that percutaneous baloon dilatation kyphoplasty with polymethyl methacrylate bone cement can significantly improve the spinal stability, relieve the degree of pain, and exert exact effects in treatment of spinal metastases.