Objective To investigate whether the splenic metastasis of B16 melanoma cells in C57BL/6 mice occur through the blood.Methods Cultured B16 melanoma cells were collected,adjusted into 1?10~7/ml and inoculated into the left armpit of C57BL/6 mice,200 ?l each.After 15 days,the mice were sac- rificed and metastasis was examined in various tissues and organs by HE stain.1.5 % Evan Blue(EB),2 ml/kg, was administered to normal C57BL/6 mice through the tail vein,and the region stained blue was compared with that of metastasis.Results All of 12 mice inoculated were burdened with melanoma.Splenic metastasis occurred in 9 of them,among which 3 were accompanied by lung metastasis,and all of 9 mice showed differ- ent degree of local lymph node metastasis.The dorsal 1/4 part of the positive spleen was stained black by the metastatic tumor cells.Under the microscope,distribution of the metastatic tumor cells in the spleen was scat- tered,their growth inhibited,and maturation accelerated.The region stained blue by EB also located at the dorsal 1/4 part of the spleen,which was almost the same as the positive part of metastasis.Conclusions Metastasis of B16 melanoma cells in C57BL/6 miee occurs earlier in the spleen than in other organs,and this metastasis is achieved most possibly through the blood.

Objective To assess the contractility of levator ani muscle in postpartum female using pelvic floor three-dimensional ultrasound and to provide an effective imaging basis for the change of levator ani muscle contractility in postpartum female.Methods Totally 75 postpartum women (55 underwent spontaneous vaginal delivery and 20 underwent selective cesarean delivery) and 40 nulliparas were examined by pelvic floor three-dimensional ultrasound.The images were obtained at rest and at maximal levator ani muscle contraction.The sagittal hiatal length (L) on the two-dimensional sagittal images,the area of levator hiatus (A) and the circumference of levator hiatus (C) were measured on the three-dimensional images,and the difference value between rest and contraction were calculated to get the △L,△A,△C.Then the △L,△A,△C between different groups were compared.Results The △L,△A,△C in spontaneous vaginal delivery group and selective cesarean delivery group were smaller than those in nulliparas group (P ＜0.05),and there was no statistical difference between spontaneous vaginal delivery and selective cesarean delivery group (P ＞ 0.05).Conclusions Three-dimensional ultrasound can effectively assess the the contractility of levator ani muscle,the levator ani muscle contractility of postpartum female was poorer than nulliparas,and between spontaneous vaginal delivery and selective cesarean delivery women there is no obvious difference.

Objective To observe the effects of thymosin alpha-1 on T cell subsets and gastricintestinal reac-tion of patients with gastricintestinal tumor. Methods Twenty patients with gastricintestinal tumor were divided into two groups, observe group(n = 10) received thymosin alpha-1 during chemotherapy, and control group(n = 10) only received chemotherapy, T cell subsets and gastricintestinal reaction were observed. Results Before chemotherapy, CD+4,CD+8 and CD+4/CD+8 of observe group were(43.7±7.5),(27.3±2.8) and (1.5±0.1), and control group were (39.4±6.3), (30.9±2.5) and (1.2±0.6). There was significant difference between them (P < 0.05). After chemotherapy, CD+4, CD+8 and NK cell were (40.6±6.8)、( 29.7±2.6) and (19.1±2.7), control group (35.9±5.7), (33.4±2.4) and (18.6±2.3). There was significant difference between them (P < 0.05). Effec-tive ratio of nausea and vomit of observe group were 72.5% and 60.0%, control group 40.0% and 33.3%, There were significant differences between them(P <0.05). Conclusion Thymosin alpha-1 may ameliorate the function of T cell subsets and gastricintestinal reaction of patients with gastricintestinal tumor.

ObjectiveTo observe the morphology and structure of the urogenital hiatus in late pregnancy women using transperineal three-dimensional (3D) pelvic floor ultrasonography.Methods Twenty five nulliparas and fifty late pregnancy women were examined by transperineal 3D pelvic floor ultrasonography.The images of urogenital hiatus were obtained and compared to study their differences between nulliparas and late pregnancy women.Results Compared to nulliparas,urogenital hiatus in late pregnancy women tended to be circular and with an offset position.The puborectalis were bended and could be avulsed.Pelvic floor connective tissue could be loose.The morphology of vaginal might be abnormal and pelvic organ might prolapsed slightly.ConclusionsThe morphology and structure of urogenital hiatus in late pregnancy women are different to those in nulliparas,the hiatus tend to be slackness.3D pelvic floor ultrasonography is an effective imaging method to observe the morphology and structure of urogenital hiatus in pregnancy women.

Objective:To detect the interleukin-22(IL-22) levels in peripheral blood of the asthma patients and analyze the relationship between the IL-22 levels and IgE,pulmonary function,fractional exhaled nitric oxide(FENO),and to investigate the role of IL-22 in the pathogenesis of bronchial asthma. Methods:Seventy-eight patients with asthma were selected as experiment group.The asthma patients were divided into increased IgE group (n=46)and normal IgE group(n=32)according to the IgE levels.The asthma patients were also divided into acute attack stage and clinical control stage according to patients' clinical symptoms.In addition, twenty healthy controls were chosen as healthy control group.The plasma IL-22 levels, IgE,forced expiratory volume in one second (FEV1) and FENO levels of the subjects in various groups were detected and compared between various groups.The correlations between the plasma IL-22 levels and IgE, FEV1,FENO were analyzed.Results:The plasma IL-22 level and FENO level of the patients in asthma group were higher than those in healthy control group(P<0.05).The IL-22 level and FENO level of the patients in clinical control stage were lower than those in acute attack stage,but they were still higher than those in healthy control group(P<0.05).The plasma IL-22 level of the patients in increased IgE group was higher than that in normal IgE group(P<0.05).There were significant negative correlations between the IL-22 levels and FEV1 in the asthma patients in acute attack group and clinical control group(r=-0.365 3,P<0.05;r=-0.267 2,P<0.05),but there was no significant correlation between the IL-22 levels and FENO(r=0.011 9,P>0.05;r=0.059 8,P>0.05).Conclusion:The increased expression of IL-22 in peripheral blood of the patients with asthma may participate in the pathogenesis of asthma,and may aggravate the deterioration of pulmonary function of the asthma patients.

Objective To study the synergism effect of 131 I-Herceptin and high-energy X-ray on HER2 overexpressed breast cancer SK-BR-3 cells.Methods The protein expression and gene amplification of human epidermal growth factor receptor-2 ( HER2 ) in SK-BR-3 cells were identified by immunohistochemistry and fluorescence in situ hybridization ( FISH ) method, 131 I-Herceptin was prepared by iodogen method, and the IC15 concentration of 131 I-Herceptin on SK-BR-3 cell were selected by MTT method.The cells were divided into control group and drug group according to 131 I-Herceptin used or not, and were delivered five different doses of external irradiation (0,2,4 and 6Gy), and the synergism effect was detected by colonogenic assay.The cells were divided into blank group, drug group(131I-Herceptin), X-ray group(2 Gy external irradiation) and combination group (131I-Herceptin+2 Gy external irradiation), the apoptosis rate and death rate were detected by AO/EB method and cell cycle were detected by flow cytometry.Results The labling rate, radiochemical purity and specific radioactivity of 131 I-Herceptin were 86.8%, 93.9% and 868.3 μci/mg, respectively.The IC15 of 131 I-Herceptin was 15.625μci/mL.131 I-Herceptin and high-energy X-ray significantly reduced surviving fraction ( SF) ( F=628.888,F=964.97,P<0.05) and there were interactions between them (F=113.046,P<0.05).There were significant differences in apoptosis rate and death rate among blank group, drug group, X-ray group and combination group(F=103.324,F=13.33,all P<0.05),and there were significant differences of pairwise comparison (P<0.05).After irradiation and 131I-Herceptin administration, the cell cycle changed obviously from G1-phase to G2-and S-phase.Conclusion 131 I-Herceptin combined with high-energy X-ray has the synergism effect on HER2 overexpressed breast cancer SK-BR-3 cells.

A new kind of curcin (curcin 2), induced by several kinds of stresses from Jatropha curcas leaves, under the control of the 35S CaMV (cauliflower mosaic virus) promoter, was introduced into tobacco genome by Agrobacterium tumefaciens-mediated transformation method. Curcin 2 protein was only detected in the transgenic tobacco plants transformed with the cur2p fragment (coding premature curcin 2 protein), but not in the plants transformed with cur2m fragment (coding mature curcin 2 protein). The transgenic lines expressing curcin 2 showed increased tolerance to tobacco mosaic virus (TMV).

BACKGROUND: The dynamic changes of pneumocyte apoptosis and aspartate-specific cysteine proteases-3 (caspase-3) expression in lung tissue of rats during the process of lung ischemia/reperfusion (I/R) injury and the possible action mechanisms remain unclear.OBJECTIVE: This study was to observe the dynamic changes of pneumocyte apoptosis and caspase-3 expression in the rat lung tissue during the process of lung I/R injury, and to analyze the role of pneumocyte apoptosis and the possible action mechanism.DESIGN: A randomized controlled animal experiment.SETTING: Emergency Center, First Hospital, Nanjing Medical University.MATERIALS: This study was carried out in the Animal Laboratory of the First Hospital of Nanjing Medcial University and Nanjing Center for Radioimmunity between April 2006 and September 2006. Twenty-eight male healthy SD rats of clean grade, with body weight of 250 to 350 g, aged 49 to 76 days, were provided by the Experimental Animal Center of Nanjing Medical University. The involved rats were randomized into experimental group and control group, with 14 rats in each.METHODS: ①Experimental intervention: Rats in the experimental group were created into models of lung I/R injury according to the method of Eppinger et al. They were occluded for 45 minutes at the porta of lung (no systolic and diastolic reactions in lung tissue being considered as successful occlusion), and then they were reperfused (recovery of systolic and diastolic function being considered as successful reperfusion); After that, lung tissues were harvested at 3 and 6 hours after lung I/R injury, 7 rats at each time point. Each rat in the control group was subjected to a thoracotony only, but lung tissues were isolated at the same time point by the same method. ②Experimental evaluation: Apoptotic cells in the lung tissue were detected with a flow cytometer by Annexin-V-PI staining, and apoptosis rate was calculated. Caspase-3 expression in the lung tissue was observed by immunohistochemical method and image analysis. Wet to dry weight ratio(W/D) of lung tissue of rats in the two groups was calculated; the number of injured pulmonary alveoli at I/R 3 hours/that at I/R 6 hours was calculated for quantitative evaluation of injured lung tissue; Patho-morphological changes of lung tissue were observed by haematoxylin & eosin staining under an optical microscope.MAIN OUTCOME MEASURES: ①Pneumocyte apoptosis rate and caspase-3 expression in the lung tissue. ②W/D of lung tissue and quantitative evaluation of injured lung tissue. ③Patho-morphological changes of lung tissue.RESULTS: Twenty-eight rats were involved in the final analysis, without deletion. ①Pneumocyte apoptosis rates in the experimental group at I/R 3 and 6 hours were significantly increased as compared with control group (P＜0.01). In the experimental group, pneumocyte apoptosis rate was decreased a little at I/R 6 hours than at I/R 3 hours (P＜0.05). ②Caspase-3 expression in the lung tissue of rats of experimental group reached its top at I/R 3 hours, and was decreased a little at I/R 6 hours. At each time point, caspase-3 expression in the experimental group was increased as compared with control group (P＜0.01). ③In the experimental group, the number of injured pulmonary alveoli at I/R 3 hours/that at I/R 6 hours and W/D ratios of lung tissues were significantly increased as compared with control group (P＜0.01). In the experimental group, two ratios at I/R 6 hours were higher than those at I/R 3 hours (P＜0.05).④In the experimental group, the structure of pulmonary alveoli was destructed, collapsed and disappeared; lots of inflammatory cell infiltration was found; Patho-morphological changes of injured lung tissue at I/R 6 hours were severer than those at I/R 3 hours. No obvious changes were found in the control group.CONCLUSION: At the early stage of lung I/R injury, the alteration of caspase-3 maybe activate pneumocyte apoptosis and induce the apoptosis of lung tissue, and thereby leads to lung injury.

Objective To investigate the effects of Shenfu injection ( SF,a Chinese herbal medicine preparation made of Codonopsis pilosula and Aconitum carmichaeli) on the cell apoptosis of focal cerebral ischemic-reperfusion injured rats and the expression of heme oxygenase-1 (HO-1). Methods Forty-two male Sprague-Dawley rats used for producing unilateral brain ischemia reperfusion model were randomly divided into three groups:sham operation group ( Sham group),ischemia reperfusion group ( IR group),and SF Injection group (SF group).The model of focal cerebral ischemia-reperfusion injury was induced by transient occlusion of middle cerebral artery (ischemia for 2 h,and reperfusion for 3,6 h respectively).In SF group,SF ( 10 mg/kg) was intraperitoneally injected duri(n)g reperfusion.Cell apoptosis rate in brain tissue was detected by the technique of Annexin-V-PI double staining and was counted in flow cytometer.Expression of HO-1 in brain was measured by RT-PCR,while the pathological and ultra structure changes of cerebral tissue were also observed.Results Cell apoptosis rate of brain tissue were significantly higher in IR group than that in Sham group (P ＜0.01 ),while SF group had less significant changes in cell apoptosis rate, HO-1 level of brain tissue than IR group (P ＜ O.01 ).The ultra structure change of brain tissue was less in SF group than that in IR group.Conclusions During early stage of brain IR injury,SF inhibits cellular apoptosis and in turn protects the brain from injury which is attributed to the increase in HO-1 expression induced by SF.

Objective To evaluate the inhibitory effect of photocatalytic titanium dioxide (TiO2)on the growth of a human epidermal squamous cell carcinoma cell line A431 and its mechanism.Methods Cultured A431 cells were classified into various groups to remain untreated (blank control group),be treated with different concentrations (100,200,300,400,500,600 mg/L) of TiO2 nanoparticles alone or in combination with ultraviolet (UV,main wavelength 253.7 nm,power 30 W,distance 30 cm,exposure duration 15 min) irradiation.After additional culture for different durations,methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate cell growth,annexin V-fluorescein isothiocyanate/propidium iodide (PI) double staining to observe cell apoptosis,and Rho123 staining to determine mitochondrial transmembrane potential.Statistical analysis was carried out using SPSS 13.0 software.Analysis of variance (AOV),t test and Student-Newman-Keuls (SNK) test were performed to assess the differences in these parameters between these groups.Results The growth of A431 cells was inhibited by pretreatment with TiO2 nanoparticles followed by UV irradiation,and the inhibitory effect was enhanced as the dose of TiO2 nanoparticles increased.As AOV and SNK test showed,there were significant differences in the growth inhibition rate among A431 cells treated with different concentrations of TiO2 nanoparticles at the three time points (24,48 and 72 hours) after UV irradiation (n =6,F =21.54,77.56,20.27,respectively,all P ＜ 0.05).No statistical inhibition was observed in the growth of A431 cells treated with TiO2 nanoparticles alone compared with untreated A431 cells (all P ＞ 0.05).Photocatalytic TiO2 nanoparticles also induced the apoptosis but decreased the mitochondrial transmembrane potential in A431 cells.In detail,the apoptosis rate was 8.86％ ± 0.22％,11.72％ ± 0.29％ and 31.24％ ± 0.78％ in A431 cells treated with TiO2nanoparticles of 100,200,400 mg/L followed by UV irradiation,respectively,compared to 2.69％ ± 0.28％ in the blank control group (n =3,F =256.61,P ＜ 0.05).Decreased mitochondrial transmembrane potential (expressed as total fluorescence intensity) was observed in A431 cells treated with TiO2 nanoparticles of 100,200,400 mg/L followed by UV irradiation compared with blank control group (758.48 ± 15.42,676.60 ± 14.35,557.71 ± 13.12vs.2943.65 ± 70.26,F =208.57,P ＜ 0.05,n =3),and SNK test also revealed statistical differences between these groups.Conclusions TiO2 nanoparticles combined with UV can inhibit the growth of but induce the apoptosis in A431 cells,which may be associated with the reduction in mitochondrial transmembrane potential in A431 cells,while TiO2 nanoparticles alone show no inhibitory effect on the growth of A431 cells.