Objective:The effects of IL-13 ( Interleukin-13 ) on SDF-1 ( Stromal cell derived factor 1 ) and EGF ( Epidermal growth factor) expression in fibroblasts co-cultured with breast cancer cells were investigated to explore the mechanism for IL-13 in the development of breast cancer.Methods:The co-culture of human breast cancer cell line MDA-MB-231 with the human skin fibroblast line CCC-ESF-1 ( ESF) was used in vitro and in tumor-burdened nude mice.The effects of IL-13 on SDF-1 and EGF expression in the co-cultured fibroblasts in vitro were analyzed using reverse transcription quantitative polymerase chain reaction ( RT-qPCR ) , flow cytometry and Western blot assay.The proliferation of the co-cultured human breast cancer cells in vitro was detected by Cell counting kit-8(CCK-8).The effects of IL-13 on SDF-1 and EGF expression in the fibroblasts of tumor tissue of tumor-burdened nude mice were analyzed using immunofluorescence and laser confocal microscope, and the tumor volumes were examined.Results: IL-13 could up-regulate SDF-1 and EGF expression in the fibroblasts co-cultured with breast cancer cells in vitro,and promoted the proliferation of the co-cultured breast cancer cells.In tumor-burdened nude mice,IL-13 enhanced SDF-1 and EGF expression of fibroblasts in tumor tissue, and accelerated tumor growth.Conclusion:IL-13 up-regulates SDF-1 and EGF expression of fibroblasts co-cultured with breast cancer cells.The molecular mechanism of promoting effect of IL-13 on breast cancer relates to SDF-1 and EGF of fibroblasts in breast cancer stroma.

Objective To investigate the effect of tadpole extract(T871 3) on tumor cells and its mechanism. Methods We studied the effects of T871 3 on proliferation, differentiation and apoptosis of HL 60 cells by cytomorphological observation, cytochemistry and TUNEL method. We also examined gene expression during the induction of apoptosis and differentiation in tadpole extract treated HL 60 cells by in situ hybridization and intact cell mRNA dot blot techniques. Results 1 T871 3 was able to inhibite HL 60 cells proliferation. 2 T871 3 was able to induce HL 60 cells to differentiate along monocyte macrophage lineage at low concentration, and apoptosis at higher concentration. 3 The differentiation of HL 60 cells was accompanied by downregulations of c myc,c myb gene expression, The apoptosis of HL 60 cells was accompanied by downregulations of c\|myc bcl 2 gene expression, suggesting that these genes may be involved in the apoptosis and differentiation process. Conclusion Tadpole extract may have effects on HL 60 cells through changing the oncogene expression. [

Objective To explore the protective mechanisms of Buyanghuanwu decoction (BHD) on heart function by observing its effect on MIRI rabbits. Methods The rabbits were randomly divided into three groups, including sham group, model group, and BHD group. After pretreatment for one week, the model of myocardial ischemia reperfusion injury was established by ligating the branch of left coronary arter-y. The changes of electrocardiography were observed, and the values of LVSP, + dp/dtmax, - dp/dtmax were recorded and compared. Results Compared with the model group, the values of LVSP, + dp/dtmax, - dp/dtmax in BHD group were significantly increased (P<0.01 ). Conclusion BHD can improve heart function of MIRI rabbits.

Objective To induce skin cancer in BALB/c mice using DMBA as initiator and TPA as tumor promot-er. Through optimizing the doses and frequencies of DMBA administration to establish a stable skin cancer model with less time and causing less skin damage. Methods Shaving the back of mice to expose a piece of skin around 2 cm × 2 cm. The mice were divided into a blank control group and four treatment groups randomly. These four groups were given 1, 2, 4, 7 times 100μg/100μL DMBA/acetone, respectively, in the first week, and twice 4μg/100μL TPA/acetone per week in the next 11 weeks. The body weight changes, time of tumor formation and number of tumors formed were recorded during the experiment. The mice were sacrificed at 12th week and samples of tumor tissue and adjacent normal skin tissue were taken for pathological examination using HE staining. Results Tumors were observed at the 7th week in the group with once DMBA administration in the first week and at the 4th week in the group with twice DMBA administration in the first week. Skin cancers were formed also in the group with 4?time DMBA administration in the first week, however, with signif?icantly more severe skin damages. The mice receiving DMBA everyday in the first week died at the 3th week. Conclusions The best induction protocol for skin cancer in BALB/c mice should be twice DMBA in the first week followed by twice TPA

Objective To analyze the mammographic features of ductal carcinoma in situ (DCIS) of the breast and the correlation between the mammographic and pathologic findings, and try to provide clinical criteria for selecting patients for appropriate local treatment. Methods A retrospective study was performed to analyze the mammographic features and to correlate the mammographic and pathologic findings in 29 consecutive cases of DCIS including 8 cases of DCIS associated with small invasive foci. Results (1)There were various features in mammograms of DCIS, including cluster microcalcifications (20 cases), ill defined mass with calcifications (3 cases), mass (4 cases), nipple retraction associated with big ductal dilatation (1 case), and normal mammogram (1 case). (2)The shape of the calcification cluster distributed as V shaped in 7 cases, round in 8 cases, irregular in 5 cases, and scattered as many small areas in one quadrant in 3 cases.(3)1 lesion appeared as strip from the deep aspect of the breast to the nipple, and 3 masses were round. (4)The comedo subtype (7/9) and the high grade nuclei of DCIS (6/9), correlating with poor prognosis, were more likely to be accompanied by linear and branching calcification. Noncomedo DCIS (11/12) and low grade nuclei (11/12) were more likely to be associated with granular and punctate calcification when microcalcification were seen on mammogram. There was statistically significant difference between the two groups with P =0.002 and P =0.009 respectively (Chi square test, Fisher′s exact method). Conclusion The mammographic findings of DCIS were characteristic. They were closely associated with the pathologic features that were correlated with the biomolecular findings. To some extent, the choice of treatment could be based on these mammographic findings.

Background Reasearches showed that α-melanocyte-stimulating hormone (α-MSH) inhibits inflammation and ameliorates the ocular surface abnormalities in a scopolamine-induced dry eye rat model,and the managing effect of sodium carboxymethylcellulose (CMC) on dry eyes also has been determined.However,whether α-MSH can enhance the therapeutic effects of CMC remains to be investigated.Objective This study was to investigate the protective effects of α-MSH combined with CMC on ocular surface in a scopolamine-induced dry eye rat model.Methods Sixty clean female Wistar rats were randomly divided into normal control group,model control group,NaCl group,CMC group,α-MSH group and α-MSH+CMC group,and 10 rats for each group.The dry eye models were established by subcutaneous injection of scopolamine hydrobromide at 9:00,12:00,15:00 and 18:00 per day for 28 days.0.9％ NaCl solution,1×10 3 mg/ml α-MSH solution,0.5％ CMC eye drop,and 1 ×10-3 mg/ml α-MSH+0.5％ CMC solution were topically administered twice a day (8:00,17:00) since the initial day of modeling according to grouping.Shirmer Ⅰ test (S Ⅰ t),breakup time of tear film (BUT) and corneal fluorescence staining were performed before and 7,14,21,28 days after the application of drugs.At 28 days following the administration of drugs,the eyeballs of the rats were collected.Hemotoxylin and eosin staining was employed to examine the morphology of corneas,and periodic acid schiff (PAS) staining was used to count the conjunctival goblet cells.This study protocol was approved by Experimental Animal Ethic Committee of Tianjin Medical University (SYXK 2009-0001),and the use and care of the rats complied with ARVO Statement.Results The S Ⅰ t and BUT values were significantly reduced,and the corneal fluorescence staining scores were significantly increased over time following modeling in the model control group (all at P＜0.01).No significant differences were found in the S Ⅰ t,BUT and corneal fluorescence staining scores between model control group and NaCl group at various time points (all at P＞0.05).At 7,14 and 21 days after intervention,the S Ⅰ t values were (4.800±0.789),(4.100±0.516) and (4.300±0.856) mm in the α-MSH+CMC group,which were considerably higher than (2.875 ±0.719),(2.375 ±0.619) and (2.532±0.957)mm in the NaCl group (all at P＜0.01).At 7 days after intervention,the BUT values were (4.938± 1.843) seconds and (5.000±1.491) seconds in the α-MSH group and α-MSH+CMC group,which were significantly higher than (3.250±1.000) seconds in the NaCl group (both at P＜0.01).The corneal fluorescence staining scores in the CMC group,α-MSH group and α-MSH+CMC group were significantly lower than that in the NaCl group,with the lowest score in the α-MSH +CMC group (all at P＜0.05).The thickening of corneal epithelial layer,corneal edema and arrangement disorder of corneal stroma were found in the model control group and NaCl group;while slight corneal edema and epithelial cell proliferation were exhibited in the α-MSH+CMC group by hemotoxylin and eosin staining.PAS staining showed that the number of goblet cells was much more in the CMC group,α-MSH group and α-MSH+ CMC group than that in the model control group and NaCl group (all at P ＜ 0.01).Conclusions The sole application of α-MSH or CMC alleviates ocular surface damage and morphological abnormality to certain extent,and the combination of α-MSH and CMC generates more effective protection in comparison with sole administration of α-MSH or CMC.The early application of the drugs plays an improvement role in tear secretion and tear film stability in dry eyes.

Objective To observe the formation of asymmetric dimethylarginine (ADMA)and the expression of dimethylarginine dimethylaminohydrolase 2 (DDAH-2) of human umbilical vein endothelial cells (HUVECs) stimulated by uric acid (UA), and to explore the role of ADMADDAH axis in the vascular endothelial dysfunction induced by uric acid. Methods HUVECs were cultured in M199 medium supplemented with 10% FBS. Cells were exposed to different concentrations of UA (0, 60, 120 mg/L) for 6 h and 24 h. Under different concentrations and times, the level of ADMA in cell suspension was detected by high performance liquid chromatography (HPLC) technique; the gene and protein expressions of DDAH-2 were detected by RT-PCR and Western blotting; the fluorescence intensity of intracellular 2',7'-dichlorofluorescein (DCF) which represented the productions of ROS was detected by the flow cytometry (FCM). The activity of DDAH-2 in HUVCEs which were exposed to different concentrations of UA (0, 60, 120mg/L) or UA (120 mg/L) +NAC (10 mmol/L) for 24 h was estimated by directly measuring the amount of ADMA metabolized by the enzyme and the role of NAC in the activity was studied.Results The expression of ADMA induced by urid acid was dose-depent and higher at 24 h than that at 6 h in the same dosage (all P＜0.05). The dosage and stimulation time of UA did not have any influence on the expression of intracellular DDAH-2 (all P＞0.05). When HUVECs exposed to UA (120 mg/L) for 24 h, the production of intracellular ROS was significantly increased while the activity of DDAH-2 was decreasesd (all P＜0.05) as compared to 60 mg/L stimulation. This effect could be inhibited by the intervention of anti-oxidant NAC. Conclusions The high UA stimulation on HUVECs can increase the expression of intracellular ROS and inhibit the activity of DDAH-2 which increases the concentration of ADMA by decreasing the degradation of ADMA as well as the formation of NO. DDAH-ADMA axis may participate in the vascular endothelial dysfunction induced by UA.

Objective To establish analysis methods of two-dimensional(2-DE)gel electrophoresis for human osteosarcoma.Methods A series of methods,including Immobilized pH gradient were used as ID.Some applications,such as sample preparation used as choice of IPG gel,were improved.Coomassie brilliant blue staining,ImageMaster 2D Elite 3.01 analysis software,MALDI-TOF/ TOF MS and SWISS-PROT database serching were used to separate and indentify the proteins of human osteosarcoma.Results The good use pattern including repetitive experiments showed that in three experiments,the amount of protein spots of the same team sample deviates from the relative standard as following,the average of variation coefficient(%) : 23.00?10.11 and 20.33?9.90;and the range of variation coefficient(%) were:3.80-6.89 and 2.706.89 from osteosarcoma and normal group respectively.The isoelectric point and molecular weigh of the same protein spots in three experiments deviated from relative standard as following:(8.93%?1.17)%,(10.16?2.02)%,(10.87?3.86)%,respectively.Therefore,better resolution and repetitive 2-DE atlas were obtained.The proteins from 11 pairs of sample were analysed by mass spectrometry,9 identified proteins(transthyretin precursor, Triosephosphate isomerase,slow skeletal muscle,cardiac muscle Troponin T,Cofilin-1,Myosin light chain 1,Calgranulin B,Heat-shock protein,Annexin A5, Fanconi anemia group D2 proteins) were more abundant in osteosarcoma tisstues and 2 proteins manganese SOD and carbonic dehydratase appeared down-regulation in osteosarcoma tisstues.Conclusion This optimized 2-DE map is an important tool for further study on osteosarcoma,and these identified proteins were related-proteins with osteosarcoma.It is suggested that the changes of the proteome are involved in the pathology processe of osteosarcoma.

At present, nucleic acid testing technology has been widely used in clinical laboratory diagnosis. Conventional detection technique such as real-time PCR is complicated, time consuming, and dependent on specific instruments. Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein (Cas) system is an adaptive immune defense system against viruses in bacteria and archaea, which has been developed into a powerful technology for genome editing. Recently, the leading groups engaged in CRISPR have set up new tools for nucleic acid detection based on Cas13a, Cas12a and newly discovered protein-Cas14, which plays an important role in rapid diagnosis of infectious diseases, detection of gene mutations in cancer and genotyping. Since they are ultrasensitive, specific, rapid and cost-effective, it is expected to bring great potential for molecular diagnosis. In this review, the mechanism of CRISPR/Cas system and the principle, the applications of the newly-developed diagnostic platforms are introduced. What′s more, the advantages, limitations and prospects of the technologies are summarized.

Objective To evaluate the effect of goal-directed fluid therapy (GDFT) guided by different goals on endothelial glycocalyx,inflammatory cytokines and postoperative complications in patients undergoing high-risk abdominal surgery.Methods Eighty patients of both sexes,aged 18-64 yr,with body mass index of 18-30 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective retroperitoneal tumor resection,were divided into 2 groups (n =40 each) by a random number table method:stroke volume variation (SVV) 9％-14％ group (L group,n=40) and SVV＜9％ group (H group,n =40).SVV 9％-14％ and SVV＜9％ were used as the target and GDFT was performed though combing with cardiac index and mean arterial pressure.The concentrations of syndecan 1,hyaluronic acid,heparan sulfate,tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) in serum were determined at 5 min before skin incision (T0),1 h after skin incision (T1),4 h after skin incision (T2) and 24 and 72 h after operation (T3,4).The intraoperative urine volume,blood loss,volume of liquid infused,volume of blood infused,amount of norepinephrine consumed,operation time,extubation time and length of postoperative hospital stay were recorded.Results Compared with the baseline value at T0,the concentrations of syndecan 1,hyaluronic acid,heparan sulfate,TNF-α and IL-6 in serum were significantly increased at T1-4 in H group,and the concentrations of syndecan 1,heparan sulfate,TNF-α and IL-6 in serum at T1-4 and concentrations of hyaluronic acid at T1-3 were significantly increased in L group (P＜0.05).Compared with H group,the volume of liquid infused was significantly reduced,the amount of norepinephrine consumed was increased,the concentrations of syndecan 1,IL-6 and TNF-α in serum were decreased at T1-4,the concentrations of hyaluronic acid were decreased at T2,3,and the concentrations of heparan sulfate were decreased at T1-3 in group L (P＜0.05).Conclusion Compared with GDFT with the target of SVV＜9％,GDFT with the target of SVV 9％-14％ is more helpful in decreasing degradation of endothelial glycocalyx during the perioperative period and in reducing damage to endothelial barrier,and the mechanism may be related to inhibiting inflammatory responses of patients undergoing high-risk abdominal surgery.