AIM: To prepare urokinase-loaded immunoliposomes, with anti-D-dimer mouse monoclonal antibody that can selectively target to thrombotic site and test the thrombolytic effect in the rabbit model of acute pulmonary embolism (PE) in early phase. METHODS: 40 New Zealand white rabbits were randomly divided into five groups: TBS group (TBS buffer, negative control group), urokinase (UK) group (150 000 IU/kg UK, positive control group), Lip group (Lip-UK, containing 50 000 IU/kg UK), Ab group (Ab-Lip-UK, containing 50 000 IU/kg UK) and 2 Ab group (2 Ab-Lip-UK, containing 50 000 IU/kg UK but double DDmAb of Ab group). PE model was established. Five minutes later, five solutions (TBS, UK, Lip-UK, Ab-Lip-UK, 2 Ab-Lip-UK) were transfused through femoral vein respectively. Right ventricular systolic pressure (RVSP) and right ventricular diastolic pressure (RVDP) were estimated within one hour. The pathological changes of lung, heart, liver and kidney were all observed. RESULTS: RVSP in TBS group had no significant changes within one hour after administration. However, RVSP had respective notable decline at 30 min, 40 min, 30 min, 20 min in UK, Lip, Ab and 2 Ab group, respectively. The means of residual emboli were as follows: 4.0±0 in TBS group, 2.4±0.9 in UK group, 3.1±0.6 in Lip group, 2.4±0.9 in Ab group and 1.9±0.6 in 2 Ab group. The lungs in each group showed scattering local congestion, effusion and swelling on the surface. Obvious hemorrhage of heart, kidney and liver was found with HE staining in UK group, no pathologic change was observed in other groups. CONCLUSION: 2 Ab-Lip-UK with early thrombolytic effect and security may be an ideal thrombolytic agent for treating pulmonary embolism.

Objective To observe the clinical efficacy of skin needling plus flash cupping for hemifacial spasm. Method Forty patients were randomized into a treatment group and a control group, 20 in each group. For both groups, the following acupoints were selected: Baihui (GV20), Sishencong (EX-HN1), Yifeng (TE17), Xiaguan (ST7), Cuanzhu (BL2), Yangbai (GB14), Taiyang (EX-HN5), Quanliao (SI18), Jiache (ST6), Dicang (ST4), Hegu (LI4), and Waiguan (TE5). Skin needling plus flash cupping was used for the treatment group, while even reducing-reinforcing needling method was used for the control group. Seven-day treatments were considered as a treatment course, and the therapeutic efficacy was analyzed after 3 courses. Result The cure rate was 65.0%in the treatment group versus 45.0% in the control group, and the difference was statistically significant (P<0.05). Conclusion Skin needling plus flash cupping can produce a significant efficacy in treating hemifacial spasm, better than ordinary needling method.

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AIM To study the changes of IgE, IL-4 and IFN-y in serum and pulmonary tissue homogenate of asthmatic guinea pigs and the effects of scoparone on them. METHODS To divide animals into three groups: control, asthma and scoparone treatment groups. Choose the model guinea pigs of asthma sensitized with OA, and observe the changes of IgE, IL-4 and IFN-r in serum and pulmonary tissue homogenate of asthmatic guinea pigs and the effects of scoparone on them by means of chemolumi nescence, radio immunoassay, enzyme-linked immunoabsordent assay. RESULTS IgE and IL-4 in serum and pulmonary tissue homoge-nate of asthmatic guinea pigs obviously increase (P

Objective To determine the level of toll-like receptor4 (TLR4) in the kidney of rats with the Multiple Organ Dysfunction Syndrome (MODS),and to investigate the early phase of kidney damage in MODS.Methods The experiment was done at experimental center of medical college of Three Gorges University.Forty Adult male Sprague-Dawley (SD) rats were randomly (random number) divided into two groups,namely the normal control group and the MODS model group.The rats of model group were further divided into four sub-groups as per different intervals (6 h,12 h,24 h and 48 h),and there were 8 rats in each groups.The animal models of MODS were established by two hits,the left eyeball of each model rat was removed to bleed to 2 mL/100 g,and four hours later,Lipopolysaccharide ( LPS,5 mg/kg) was injected into intraperitoneal cavity of model rats.The same volume of saline was injected intraperitoneally into rats of control group.All rats were sacrificed at various intervals.The histological changes in the kidney tissue were observed by naked eye and under light microscope.The lever of TLR4 proteins in serum and kidney tissue were detected by using flow cytometry (FCM).One-way ANOVA was used for comparison among multiple groups.Results (1) There were no histopatholagical changes in kidney of rats in control group,and the kidney injury was serious in rats with MODS.(2) Compared with the rots of control group,the level of TLR4 in kidney tissue of rats with MODS increased at 6 hours and reached peak 12 hours later ( P ＜ 0.01 ),and then decreased 24 hours ( P ＜ 0.01 ).There was no significant difference in levd of TLR4 between two groups 6 and 48 hours.Compared with the rats of control group,the level of TLR4 in peripheral blood leucocyte of rats with MODS increased significantly 6 ～ 48 hours after LPS ( P ＜ 0.01 ).The concentrations of serum and kidney tissue TLR4 proteins were positively correlated with each other ( r =0.893,P ＜ 0.05 ).Conclusions The level of TLR4 markedly increased in the kidney tissue at early stage of MODS,and the TLR4 may play an important role in the pathogenesis of kidney injury in MODS.

ObjectiveTo determine the levels of type Ⅳ collagen and matrix metalloproteinase-9 ( MMP-9 ) in the serum and kidney of rats with the multiple organ dysfunction syndrome ( MODS ) and investigate the mechanism of extracellular matrix damage in renal failure of MODS.MethodsForty adult male Sprague- Dawley (SD) rats were randomly (ramdom number) divided into two groups,namely the normal control group ( n =8 ) and the MODS model group ( n =32 ).The rats of model group were further divided into four sub-groups as per different intervals,6 h,12 h,24 h and 48 h,after modeling (n =8 in each).The animal models of MODS were established by two hits,the left eyeball of each model rat was removed to bleed to 2 ml bloocd/100 g body weight and lipopolysaccharide ( LPS,5 mg/kg) was injected into intraperitoneal cavity of model rats four hours later. The same volume of saline instead was injected intraperitoneally into rats of control group.All rats were sacrificed at different intervals.Creatinine (Cr)and blood urea nitrogen (BUN) were determined by using a Hitachi Automatic Biochemical Analyzer.The histological changes in renal tissue were observed under light microscope and transmission electronic microscope.The levels of serum type Ⅳ collagen and MMP - 9 protein were measured by using enzyme linked immunosorbent assay (ELISA).Type Ⅳ collagen and MMP- 9 protein levels in renal tissue were detected by western blot.One - way ANOVA was used for comparison among multiple groups.Results Compared with the control group,Cr and BUN were significantly higher in MODS group ( P ＜0.05 ).There were no histopathological changes in kidney of rats in control group,and the renal injury was serious in rats with MODS.The remarkable edema of basement membrane and defect of collagen fibers in renal tissue were observed in MODS group.Compared with the control group,the levels of MMP-9 in serum increased 6-48 hours after modeling and peaking at interval of 12 hours after modeling ( P ＜ 0.05 ).The protein levels of type Ⅳ collagen increased not significantly in 6 h group and 12 h group in comparison with control group (P ＞ 0.05 ),while those in 24 h group and 48 h group significantly increased ( P ＜ 0.01 ).The levels of MMP -9 in renal tissue of rats with MODS increased in 6 h group and 12 h group (P＜0.05),peaked in 24 h group ( P ＜ 0.05 ),and decreased in 48 hours.However,the level of Ⅳ collagen in serum of rats with MODS decreased significantly 6-48 hours after modeling (P ＜ 0.05 ) Conclusions The injury of extracellular matrix is an important factor to the kidney damage in MODS and it may he used for early diagnose and as a treatment target for kidney injury in MODS.

Lactic acid production of twelve strains of LAB isolated from broiler intestine and tolerance property of three strains were investigated. The results of lactic acid production showed that among all strains K6 exhibited the most rapid production during the first twelve hours, the seconds were K9 and C1; D17 exhibited the highest production of lactic acid by twenty-four hours, C1 exhibited the highest production of lactic acid by forty-eight hours. The pH values in three strains of K9、D17 and C1 culture showed the fast decline during the first twelve hours, with the final values significantly lower than those of other strains cultures. The results of tolerance property showed that the survival counts of C1could be detected when pH value was at 2 after three hours, but the survival counts of D17 and K9 could not be detected after one hour. When pH value was at 2.5 after three hours ,the survival counts of C1 declined from 10~ 8.2 /mL to 10~ 4.8 /mL, K9 from 10~ 8.2 /mL to 10~ 4.6 /mL, the survival counts of D17 could not be detected. 0.08% bile had few effects on the survival counts of three strains; when incubated in the medium with 0.40% bile, the survival counts of C1 declined from 10~ 8.4 /mL to 10~ 6.5 /mL,D17 from 10~ 10.3 /mL to 10~ 7.5 /mL, and K9 from 10~ 9.8 /mL to 10~ 7.7 /mL. When the group treated with 37℃ for 20 minutes was served as the control, the survival counts of C1 and K9 was not detected when treated with 80℃, but the survival counts of D17 were 10~ 4.9 /mL, when treatment with 65℃ the survival counts of C1 and K9 decreased significantly .

Purpose To study the effect of China cobra venom active factor(CCVAF) from China cobra venom on endothelial cells and its mechanism.Methods MTT experiment was adopted to evaluate the effect of CCVAF on bovine arteria pulmonalis vascular endothelial cells(BAVEC).The Eosin-Coomassie brillient blue and rhodamine-phalloidin method was used for actin cytoskeleton.Flow cytometry for [Ca~(2+)]_i and spectrophotometry were used for lactate dehydrogenase(LDH) and nitrogen oxide(NO) levels in cell culture supernatant.Results CCVAF(0.625-20 μg/mL) inhibited the proliferation of BAVEC in dose-dependent manner,and IC50 of CCVAF on BAVEC was 2.45 μg/mL. After CCVAF and BAVEC coincubation, it was showed that regression of intercellular conjunctions and disorder of F-actin distribution occurred. The content of [Ca~(2+)]_i, [LDH] and [NO] increased respectively.Conclusion CCVAF can inhibit BAVEC proliferation and it maybe associated with the change of cytoskeleton and increasing of [Ca~(2+)]_i,[LDH] aod [NO].

OBJECTIVETo explore the effect of Shoutai Pill (STP) containing serum on bioactivity behaviors of trophoblast cells in spontaneous abortion (SA) patients such as cell proliferation, invasion, migration and secretion.

METHODSTrophoblast cells in artificial abortion in normal pregnancy and SA patients were isolated and cultured in vitro, which were then treated with STP containing serum at various concentrations (5%, 10%, 20%, respectively). Blank serum was taken as the normal control group and dydrogesterone containing serum as the dydrogesterone control group. The proliferation, cycle distribution, invasion and migration capacity, and beta human chorionic gonadotropin (p-HCG) level were detected by methyl thiazolyl tetrazolium (MTT) colorimetry, flow cytometry (FCM), Transwell experiments, and ELISA, respectively.

RESULTSCompared with the normal control group, the activity of cell proliferation obviously decreased, ratios of apoptotic cells (SubGO/G1) and G2/M phase were obviously elevated, S phase cell ratio was obviously reduced (all P < 0.05). Transwell experiments indicated invasion and migration capacity obviously decreased, secreted beta-HCG level were obviously reduced after 72-h intervention (P < 0.05). Compared with the SA group, the activity of cell proliferation obviously increased, ratios of apoptotic cells and G2/M phase were obviously reduced, S phase cell ratio was obviously elevated, invasion and migration capacity were obviously enhanced, secreted beta-HCG level were obviously elevated after 72-h intervention in the dydrogesterone control group and each STP containing serum group (all P < 0.05). The activity of trophoblastic cell proliferation, S phase cell ratio, invasion and migration capacity, and secreted beta-HCG level were strengthened along with increased STP containing serum. Besides, the effects of 20% STP containing serum group were significantly superior to those of the dydrogesterone control group (P < 0.05).

CONCLUSIONSTP containing serum could dose-dependently enhance the proliferative activity of trophoblastic cells, invasion and migration capacity, secretion of beta-HCG, and reduce the apoptosis of trophoblast cells, which might be one of mechanisms for STP preventing and treating SA.

Abortion, Spontaneous ; Apoptosis ; Cell Cycle ; Cell Proliferation ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Dydrogesterone ; pharmacology ; Female ; Humans ; Pregnancy ; Trophoblasts ; drug effects

ObjectiveTo investigate the clinical efficacy of acupuncture in treating Tourette's syndrome.MethodEighty patients were allocated, using a random number table, to a treatment group of 40 cases and a control group of 40 cases. The treatment group received acupuncture and the control group, Western drug treatment.ResultThe YGTSS score decreased significantly in the two groups of patients after treatment(P<0.01). After treatment, the range of decrease in the YGTSS tic score was larger in the treatment group than in the control group; there was a statistically significant difference (P<0.05). The total efficacy rate was higher in the treatment group than in the control group (P<0.05). The incidences of adverse reactions were significantly lower in the treatment group than in the control group.ConclusionThe effect and safety of treatment for Tourette's syndrome are higher in the treatment group than in the control group.