Objective Tumor vessels were foundation of tumors’formation,growth and metastasis,and the destruction of a spot of them would lead to wide avascular necrosis of tumor cells,so the specific killing effect on tumor vasculature endothelial cells had become one of the most important strategies for tumor therapy. Endostar,a novel recombinant human endostatin expressed and purified with an additional nine-amino acid sequence,is a new anti-drug. This study aimed to determine how endostar to affect the behavior of endothelial cells and tumor endothelial cells in vitro and in vivo. Methods Human umbilical vein endothelial cells ( HUVEC) was isolated and cultured followed by identification with Ⅷ factor related antigens. The cells differentiated into tumor-derived endothelial cells ( Td-ECs) by co-culturing with supernatants of A549 cells. The anti-effect of endostar on Td-ECs was detected with MTT assay,cell migration assay,flow cytometry and TUNEL assay. Results Endostar had a significant effect on the proliferation of Td-EC in a time-and concentration-dependent manner. At the same time,though Endostar at a certain concentration inhibited cell migration process significantly,arrested the cells at G0 /G1 and S phases and induced cell apoptosis ( P 0. 05) . Conclusion Our study indicates a novel function of endostar,to inhibit and Td-ECs proliferation in vitro. So,using endostar for cancer therapy is worthwhile for further study in clinical trials.

Objective To observe the change of diffusion upper limit of macromolecules through pathological retina and the difference between the layers of retina. Methods Retinal edema was emulated by establishing branch retinal vein occlusion(RVO)model in miniature pig eyes under photodynamic method.Two days later,the retinas of both eyeballs were peeled off.The diffusion test apparatus was designed by ourselves.FITC-dextrans of various molecular weights(4.4,9.3,19.6,38.9,71.2 and 150 kDa)and Carboxyfluorescein(376 Da)were dissolved in RPMI-1640 solutions and diffused through inner or outer surface of retina.The rate of transretinal diffusion was determined with a spectrophotometer.Theoretical maximum size of molecule(MSM)was calculated by extrapolating the trend-linear relationship with the diffusion rate.In separate experiments to determine the sites of barrier tO diffusion,FITC-dextrans were applied to either the inner or outer retinal surface,processed as frozen sections.and viewed with a fluorescence microscope. Results FITC-dextrans applying tO inner retinal surface,4.4 kDa dextrans were largely blocked by inner nuclear layer(INL);19.6-71.2 kDa dextrans were blocked by the nerve fiber layer(NFL)and inner plexiform layer;150 kDa dextrans were blocked by NFL.FITC-dextrans applying to outer retinal surface,most dextrans with various molecular weights were blocked before outer nuclear layer(ONL).No matter applying to the inner or outer surface,Carboxyfluorescein can diffuse through the whole retina and aggregate at INL and ONL.After RVO,the inner part of retina became edema and cystoid,loosing the barrier function.Compared with the normal retina,the MSM in RVO tissues increased(6.5±0.39nm Vs 6.18±0.54nm,t=4.143,P=0.0001). Conclusions After RVO,the barrier function of inner part of retinal is destroyed and the upper limit of diffusion macromolecule size increased.which is nevertheless limited.ONL acts as bottle-neck barriers to diffusion,if the outer part of retina is damaged,the change of the diffusion upper limit will be prominent.

The acute and chronic respiratory tract inflammation models were made to investigate the effect and mechanism of sterol extracts from Begonia Sinensis Rhizome (BSR). The first model of acute lung injury was made with Kunming mice by inhaling cigarette smoke, then the mice were treated with different concentrations of BSR sterol extracts. Lung tissue morphology was detected by HE staining, TNF-alpha/MPO were detected by Elisa, and cPLA2 protein were, detected by Western blotting respectively. Results showed that in model group, lung sheet became real, alveolar space shrank or disappeared, alveolar septum was thickened, plenty of inflammatory cells were infiltrated, capillary blood vessels were congestive and the expression of TNF-α, MPO, cPLA2 increased; after administration, a small amount of inflammatory cells were infiltrated, alveolar septum became obvious, capillary congestion status was significantly relieved and the expression of TNF-α, MPO, cPLA2 decreased (P < 0.05). The second model of chronic respiratory tract inflammation in BALB/c mice with bronchial asthma was induced by OVA, then the mice were treated with different concentrations of BSR sterol extracts. Lung tissue morphology was detected by HE staining, indexes such as IL-4, IL-5, IL-13 were detected by Elisa, and the cPLA2 protein expression was detected by Western blotting respectively. Results showed that in model group, a lot of inflammatory cells around lung vessels and bronchi exuded, bronchial goblet cells proliferated and the expression of IL-4, IL-5, IL-13, cPLA2 increased; after administration, inflammatory and goblet cell hyperplasia reduced, the expression of IL-4, IL-5, IL-13, cPLA2 also decreased (P < 0.05). The above results showed BSR sterol extracts could resist against respiratory inflammation by inhibiting cPLA2 in a dose-dependent manner.
Animals ; Asthma ; drug therapy ; genetics ; immunology ; Begoniaceae ; chemistry ; Cytokines ; genetics ; immunology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Interleukin-13 ; genetics ; immunology ; Interleukin-4 ; genetics ; immunology ; Interleukin-5 ; genetics ; immunology ; Lung ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Rhizome ; chemistry ; Sterols ; administration & dosage

Objective To explore the histopathological changes of the pigs′ eyes in vivo after radial optic neurotomy (RON), and provide the experimental foundation for the safety of RON. Methods A total of 12 healthy miniature pigs were used in the experiment, in whom 8 were executed at the 1~st , 3~rd , 7~th , and 48~th day respectively after underwent RON in both eyes, and 4 were executed at the 120~th day after underwent RON unilaterally (the other eye was as the control in 2 and underwent single vitrectomy in 2). All the enucleated eyes were cut in sections routinely and embeded in paraffin. The sections were stained by HE, Masson trichrome staining or Luxol fast blue staining and the different sections of optic nerve were observed by light microscope. Results No damages of the major vessel wall were found and the cerebral pia mater of orbital optic nerves kept integrated. At the 1~st day after the operation, the incisions came into being and local hemorrhages infiltrated into the circumambience and backside. The vacuole-like change induced by the demyelination of optic nerve fiber located at the incisions. At the 3~rd day, the vacuole-like changes widened. At the 7~th day, the fibroblasts aggregated at the incision, with hyperplastic neuroglia cells and dispersed pigmented granules. Lymphocytes and monocytes were the major infiltrated inflammatory cells. At the 48~th day, collagen filled in the incisions and aggregated neuroglia cells of the rear optic nerves behind the incision were found, which showed weak staining with obvious boundary which was somewhat beyond the midline of optic nerves. At the 120~th day, localized atrophy of optic nerve occurred under the incision. No abnormal pathological findings were found in the normal eyes and the eyes undergo vitrectomy. Conclusions Localized atrophy of optic nerves comes into being after the normal pig eyes in vivo underwent RON. The surgery is safe to some extent.

Objective To investigate the visual impairment and its rehabilitation after stroke.Methods After diagnosis of stroke by the neurologists, the patients received examinations of ocular pathology and visual function, especially in vision correction and rehabilitation. Results Five hundred and forty-seven stroke patients were recruited. 47.0% of the patients had eye alignment/movement impairment, 34.0% had visual field impairment, 19. 0% had visual perceptual difficulties and 19.9% had low vision blind. After using telescope, 35.7% blind patient's distance vision was ≥0.05; 58.2% low vision patient's distance vision was ≥0.3. After refraction (average vision:0.36±0.34 vs. 0.48 ±0.36,t =- 3.949,P=0.03) or using telescope(average vision of patients had low vision:0. 13±0. 07 vs. 0. 46±0.26,t=-5. 208,P=0.01 ;average vision of blind patients:0. 01 ±0.01 vs. 0. 06±0. 05,t = -3. 628, P= 0. 05), the improvement of vision for patients was significant.Conclusions The 72% of stroke patients has some kinds of visual impairment, which will affect not only the patients' independent living but also the clinical rehabilitation. Refraction and using telescope are effective for vision rehabilitation. Doctors should pay much attention to the visual therapy and rehabilitation for the stroke patients.

Objective To evaluate the long-term results of vitreoretinal surgery without use of intraocular silicone oil or gas in patients with diabetic tractional retinal detachment (DTRD).Methods The clinical interventional case series study included 104 patients (112 eyes) with DTRD,who were consecutively treated by pars plana vitrectomy without use of intraocular silicone oil or gas.Among the eyes,there were 6 eyes with iris neovascularization (INV),1 eye with neovascular glaucoma (NVG) and 50 eyes with macular retinal detachment.There were no pre-existing retinal holes or breaks prior to surgery nor any iatrogenie retinal breaks developed during vitrectomy.Cataract removal combined with intraoeular lens implant surgeries were performed on 15 eyes.Follow-up duration varied from 12 to 65 months (mean: 29 months).Results Subretinal fluid was completely absorbed within 2 months after surgery.In 107 eyes (95.54%),the retina reattached after surgery and remained attached till the end of follow-up period.Best corrected visual acuity (BCVA) improved in 79 eyes (70.53%),remained unchanged in 14 eyes (12.500%) and got worse in 19 eyes (16.79%).The BCVA improving rate was lower in the macular detached group (33 eyes/50 eyes,66.00% Vs 46 eyes/62 eyes,74.19%,χ2=0.89,P=0.344).No obviously aggravated opacity of lens was observed after vitreoretinal surgeries in the eyes without cataract surgeries.Seven (6.25%) eyes showed INV (5 new onset eyes),and none of them developed into NVG.In multivariate logistic regression,factors associated with postoperative rubeosis iridis were pre-existing rubeosis iridis I-adjusted odds ratio (OR)= 10.2],low preoperative BCVA (OR= 11.1) and low postoperative BCVA (OR = 16.7).Conclusions Vitreoretinal surgery for DTRD may not necessarily be combined with silicone oil or gas tamponade if there are no preoperative or intraoperative retinal breaks,and only using irrigation fluid could access a good long-term prognosis result.

Objective To construct the expression plasmid of porcine leukemia inhibitory factor(LIF) in the prokaryotic system,and purify the expressed recombinant porcine LIF protein as antigen to immune rabbit for preparation of rabbit anti-pig LIF polyclonal antibody,and immunolocalize of LIF in pigs.Methods The cDNA fragment encoding mature porcine LIF protein was amplified by PCR,then cloned into the prokaryotic expression vector PET-31b at restriction sites NdeⅠand XhoⅠ.After transformed into E.coli BL21 for expression,the recombinant porcine LIF protein was purifed for preparation of anti-pig LIF polyclonal antibody as immunogen.Western blotting and ELISA were used to test the specificity and titer of the purifed IgG.Immunostaining was used to assay the expression of LIF in pigs.Results The constructed expression plasmid was identified by DNA sequencing and restriction enzyme digestion.The expression of the recombinant porcine LIF protein was performed exactly in E.coli BL21.By immuning the rabbit,the polyclonal antibody was successfully prepared.The results of ELISA and Western blotting showed that the antibody had high titer(1∶10000) and specificity.By immunostaining,LIF protein was faintly expressed in porcine myocardial cell,splenic cord and monolayer covering epithelium cell and pulmonary fibroblast.Conclusion The prepared polyclonal antibody has high titer and specificity;LIF has faint positive immunoreactivity in the normal porcine,myocardial cell,splenic cord and monolayer covering epithelium cell and pulmonary fibroblast.

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 explore the feasibility of minimally invasive video-assisted thyroidectomy (M IVAT). Methods Clinical data of 8 consecutive patients with ben ign thyroid nodules treated by MIVAT in this hospital from September 2002 to Sep tember 2004 were retrospectively reviewed. Results All the pro cedures were successfully completed. The operative time was 90~150 min (mean, 12 0 min), and the hemorrhage volume, 10~30 ml (mean, 20 ml). No complications happ ened and no conversions to open surgery were needed. All the patients were compl etely recovered and discharged from hospital 3~4 days after the surgery. Follow- up checkups for 6~12 months (mean, 9 months) in the 8 patients found normally he aled wounds and no paraesthesia. Conclusions MIVAT is safe and effective, with excellent cosmetic outcomes.

Objective: To examine the causal relationships between obesity, peripheral blood lipid indicators and non-small cell lung cancer (NSCLC) using Mendelian randomization (MR) method, so as to provide the basis for developing NSCLC prevention and control strategies. Methods: Genetic variation data of three obesity evaluation indicators, including body mass index (BMI), body fat ratio (BFR) and waist-to-hip ratio (WHR), and seven peripheral blood lipid indicators, including triglyceride (TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), apolipoprotein A1 (ApoA1), apolipoprotein B (ApoB) and lipoprotein a [LP (a)] were collected through genome-wide association studies (GWAS) and related public databases. Potential causal relationships between obesity, peripheral blood lipid indicators and NSCLC were analyzed using inverse-variance weighted (IVW) method and multivariable MR analysis upon a random effect model. Heterogeneity and horizontal pleiotropy of instrumental variables were evaluated using Cochran's Q test and MR-Egger regression. Results: There was statistically association between BMI with NSCLC (OR=1.256, 95%CI: 1.087-1.451); there were no statistically associations between BFR, WHR, seven peripheral blood lipid indicators and NSCLC (all P>0.005). There was heterogeneity in the association between BMI, BFR, WHR, TG, HDL-C and NSCLC (all P<0.05); no horizontal pleiotropy of instrumental variables was found (all P>0.05). There was no statistically association between BMI and NSCLC after adjusting BFR (OR=1.367, 95%CI: 0.878-2.128); there was still statistically association between BMI and NSCLC after adjusting WHR and peripheral blood lipid indicators (both P<0.05). Conclusions The increase of BMI is associated with the increased risk of NSCLC incidence. BFR may be a potential influencing factor for the association between BMI and NSCLC.