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.

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 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.

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 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.

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.

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).

Objective To explore the effect of Bailing capsule on epithelial-mesenchymal transition( EMT) in rats with adenine-in-duced tubulointerstitial fibrosis. Methods Tubulointerstitial fibrosis animal models were established and SD rats were divided into mo-del group ( n = 30), treatment group ( n = 30) and control group( n = 30), randomly. Experimental rats were harvested at 7 w, 12 w,17 w after onset of experiment and functional evaluations were performed. Histology, immunohistology were examined to investigateboth histolopathology changes and the expression of bone morphogenic protein-7 (BMP-7), transforming growth factor-β1 (TGF-β1 )and a-smooth muscle actin (α-SMA) in kidneys at three time points mentioned above, respectively. Results Compared with controlgroup, 24 h urinary protein in model group lost increasingly and significantly difference appeared at three time points relative to controlgroup ( P ＜ 0.01 ). Urinary NAG in model group was markedly higher than that in control group from 7 w after onset (P ＜ 0.01 ) andwas increasingly raised at 12 w and 17 w (P＜0.01). The value of blood BUN and Cr in model group increased at 7 w (P＞0.05) rel-ative to control group. There was significant difference at 12 w and 17.w (P ＜ 0.01 ). Histologically, kidneys in model group, at 7 w,exhibited tubular casts and gently tubular dilation, granuloma in cortex, mononuclear cells infiltration in tubulointerstitial areas, andmild interstitial fibrosis. At 12 w, the degree of tubular injury and tubulointerstitial fibrosis gradually aggravated. Up to 17 w, diffusetubular dilation or atrophy was observed and focal tubules disappear. Diffuse interstitial fibrosis was exhibited. In normal kidneys, im-munohistochemistry suggested that the light expression of BMP-7 was detected in proximal renal tubular epithelial cells and marked ex-pression was identified in distal tubule, collecting duct, and renal tubular epithelial in junction area between cortex and medulla. How-ever, the expression of BMP-7 in kidneys of model group significantly decreased with increasing tubulointerstitial fibrosis and was nega-tive correlation with the expression of TGF-β1(r = -0. 981 P＜0.01) and α-SMA (r= -0.975 P＜0.01). Bailing capsule ad-ministration protected the expression of BMP-7 and reduced TGF-β1 and α-SMA expression before 12 w(P＜ 0.01 ). Conclusions Ourstudy shows an anti-fibrotic reno-protective function of Bailing capsule in rats with tubulointerstitial fibrosis via prevention of epithelial-mesenchymal transition at early stage. However, the beneficial effect lost with increasing tubulointerstitial fibrosis.

Objective To evaluate the safety limits of amygdalin in the Chinese patent medicines and Chinese herbal medi-cines,so as to provide reference for establishing the standard in the safety limits of endogenous cyanide glycosides as toxic ingredients. Methods The toxicity differences between D-amygdalin and L-amygdalin were explored by CCK-8 assay. The daily safe dose of amyg-dalin was deduced by the no-observed-adverse-effect level(NOAEL)of cyanide. Based on the deduced results,the safety limits of amygdalin in thirty-six Chinese patent medicines and three different kinds of Chinese herbal medicine were calculated. Results The half maximal inhibitory concentration of D-amygdalin and L-amygdalin was(3.5±2.0)and(6.6±0.4)mmol/L,respectively,and the daily safe dose of amygdalin was 104.30 mg. Conclusion The toxicity of D-amygdalin was approximately two times that of L-amygda-lin. To establish the safety limits of amygdalin,the sum content of the epimer could be regarded as the D-amygdalin itself to enlarge the safety scope. The content of amygdalin of three Chinese patent medicines was determined to exceed the maximum safety limit for all measured batches,indicating some kind of safety risk. Moreover,in the researches on safety limit establishment,the toxicity-alleviat-ed processing effect should be included.

Objective To study the effects of fluid loading during fluid resuscitation on hemodynamics and cerebral perfusion(CPP)under different levels of intra-abdominal pressure(IAP).Methods Forty swine were employed in the study.Hypovolemia was made by blood withdrawal of 30% of estimated blood volume from each animal through the carotid artery line.All swine were randomized(random number) into four groups, namely IAPL0, IAPL15, IAPL25 and IAPL35.Then N2 gas was used to inflate in the abdomen slowly for elevating the IAP to 0,15, 25 and 35 mmHg.Fluid loading were performed with 500 mL hydroxyethyl starch within 30 minutes.Hemodynamic variables were evaluated by PiCCO.Heart rate (HR),mean arterial pressure(MAP), central venous pressure(CVP), cardiac output(CO), global end-diastolic volume index(GEDVI) and stroke volume index(SVI)were measured 30 minutes before fluid loading and 30 minutes after fluid loading.After placement of intracranial pressure optical fiber probe in the ventricle connected to intracranial pressure monitor for continuous monitoring of ICP changes, cerebral perfusion pressure (CPP) could be calculated.Results (1)HR decrease, MAP increased, SVI, CI and GEDVI significantly increased after fluid loading in all four groups (IAPL0,IAPL15,IAPL25 and IAPL35)(P<0.05),and there were not significant changes in CVP and EVLWI(P>0.05).SVRI decreased significantly in group IAPL15,IAPL25 and IAPL35 respectively(P<0.05).(2) ICP increased significantly in group IAPL15, IAPL25 and IAPL35(P<0.05), and CPP increased significantly in group IAPL25 and IAPL35(P<0.05) while CPP only slightly increased in group IAPL15(P>0.05).Conclusion When combined with intra-abdominal hypertension,fluid loading could improve the circulation of swine due to increase in CO and improve CPP.