ECV304 was reported first in 1990 as a spont aneously-transformed and immortalized cell line derived from a Japanese HUVEC. S ubsequently, many studies validated that the ECV304 is a permanent endothelial cell line. It has been used widely as an endothelial cell model and an useful re search tool in biomedicine and pharmacology. However, several distinct differenc es exist between ECV304 and HUVEC. Some studies even pointed out that ECV304 is not of HUVEC origin. According to the research data including ours, this reporte dly endothelial-derived permanent human cell line ECV304 may be dedifferentiated towards an epithelial phenotype. It is therefore not an appropriate cell line t o study endothelial cell biology. But cultured ECV304 cells can still be used as a model, tool or target in the pathophysiological and pharmacological studies, depending on whether or not their functional expression or markers are suitable for the research work.

The change of myocatdial protein kinase C (PKC) activity during ische-mia and reperfusion was studied in isolated Langendorff perfused rat hearts. The enzymeactivity was determined by measuring the incorporation of ~(32)P from (r-~(32)P) ATP intohistone. The cytosolic PKC activity was similar in control, ischemic and reperfused hearts;however, there were significant increases in the membrane PKC activity during ischemia and reperfusion There were 1.68, 1.88, 2.18 and 1.34 fold increases of the membrane PKC activity at 15, 30, 45 and 60 minutes after ischemia respetively Following15 minutes of ischemia, repetfusion of heart only caused 1.37 fold increase in the mem-brane PKC activity, compared with that at 15 minutes after ischemta no siginificant differ-ence was found. These results suggested, that the signal transduction mediated by PKC wasimpaired during the development of ischemic and post-ischemic reperfusion injuty of theheart. Panaxadiol saponin decreased the enhanced membrane PKC avtivity induced by is-chemia by 62.5%.

Objective To understand the patients of congenital microtia malformation families knowledge of skin expander and influencing factors. Methods Self-made questionnaire to sample survey of 500 cases of our department (Plastic Surgery Hospital, Chinese Academy of Medical Sciences, the second microtia concer) patients′ families. Results 47.8％(239/500) of 500 patients of expander knowledge level is high, 41.2％(206/500) pass the exam, 11.0％(55/500) fall the exam, only 13.4％(67/500) really have a comprehensive understanding on expander achieve excellent. Scores of male and female were (16.06 ± 1.99) points and (16.39 ± 2.16) points, t = 1.752, P > 0.05, there was no statistically significant difference comparing the 2 group. Patients′ families score of different cultural levels, respectively (14.06 ± 2.36), (14.98 ± 2.02), (16.54 ± 2.00), (16.73 ± 1.88) points, F = 21.736, P < 0.01, difference of four groups was statistically significant. Different age patients′families score ( 16.21 ± 1.96), (16.62 ± 2.14), (14.86 ± 2.11), (13.98 ± 2.02), (13.73 ± 1.88) points, F = 15.685, P > 0.05, there was no statistically significant difference comparing the 5 groups. Patients with different professional families score (13.25 ± 2.19), (13.79±2.27), (16.08±1.89), (14.10±2.08), (14.13±2.35), (14.45±2.09), (14.56±1.75), (16.84± 1.81) points, F = 2.737, P < 0.01, difference of eight groups was statistically significant. Conclusions Congenital microtia patients′families skin expander knowledge needs to be improved, it is necessary to take various forms, conduct for families of expander knowledge through propaganda and education.

Objective To study the potential efficacy of transplanted bone marrow-derived mesenchymal stem cells (MSCs) in treating and repairing the acute lung injury in animal models. Methods MSCs were isolated from mouse bone marrow, cultrued and amplified in vitro. The lipopolysaccharide (LPS) was inhaled through postnasal tract to cause acute lung injury in mice and the MSCs labeled by Brdu were administrated via vein into the mice. The migration and differention of the cells were identified by immunostaining and double immunostaining. The pathological changes, pulmonary edema index and the content of IL-1β in lung homogenate were used to accese the therapeutical effect of MSCs. Results The cultured MSCs dispalyed a positive CD44 and a negative CD34. The Brdu-labeled cells were detected in the lungs of the recipient 4 days after transplantation, indicating its origin of MSCs. Theses cells also exhibited characteristics of aveolar epithelials, expressing the cytokeratin-the marker of epithelium. Compared with the injuried ones, the mice treated with MSCs showed a decreased pulmonary edema in-dex and IL-1β content in the lung homogenate. Conclusion These data suggest a therapeutical effects of MSCs in treating and repairing the mouse acute lung injury.

Objective To discuss the design of pedicled thoracodorsal artery perforator flap (TDAP flap),and to evaluate the aesthetic results and donor-site complications for immediate partial breast reconstruction (IPBR) after breast conserving surgery (BCS) for breast cancer patients.Method Clinical data of 13 breast cancer cases treated with BCS + IPBR using TDAP flap from November 2004 to November 2010 were retrospectively analyzed.Perforators were identified with Doppler preoperatively in all patients.Results All perforators originated within a median distance of 8.0 cm ( range,7.5 to 9.5 cm) from axillary plica at the posterior line of axilla.Median area of the flaps was 6.0 × 8.0 cm ( range,5.0 × 7.0 cm to 8.0 × 10.0 cm).One flap was muscle-sparing,while a small muscle strip was left embedding the perforators in other twelve flaps to increase the reliability of the vascular pedicle.Postoperatively patients were followedup from 4 to 71 months.Median follow-up time was 41 months.Flap necrosis and seroma in the donor-site were not found in all patients.Aesthetic results were graded as excellent or good in 9 patients,fair in 3,and poor in one.Conclusions TDAP flap is a good choice for IPBR after BCS for breast cancer patients whether lesions in outer quadrants or inner quadrants,especially for those patients with excisional biopsy.Preoperative mini-Doppler is helpful for determining the precise location of the main perforators,and decreasing the risk of vessels injury.

AIM and METHODS: The effects of angiotensin II(AngII) on the activation of three transcription factors were investigated by using EMSA and western-blot methods in endothelial cells respectively. RESULTS: The EMSA results showed that AngII stimulation could increase NF-κB, SP-1 and AP-1 activation in ECV304, which suggested that activity changes in these three transcription factors could partly contribute to the dysfunction of endothelial cells.The binding affinity of NF-κB, SP-1 and AP-1 with corresponding oligonucleotides in AngII-treated ECV-304 were respectively 10.98,3.89,1.33 times as large as in control. The nuclear appearance of AngII-activated NF-κB was examined by western-blot, which corroborates our results from EMSA analysis. While as the protein appearance of AP-1 and SP-1 in nucleus, were little higher than the control group. The result of western-blot suggested that AngII-induced EC activation of these three transcription factors maybe mainly due to the enhanced binding ability to its corresponding cis-acting factors. CONCLUSION: NF-κB, a ubiquitously exposed nuclear transcription factor, is involved, together with SP-1,AP-1, in the regulation of endothelial cell dysfunction related to cardiovascular diseases such as atherosclerosis and hypertension.

AIM: To determine whether the high mobility group protein I (HMGI) is able to bind to the upstream sequence of platelet-derived growth factor B-chain gene and to characterize the HMGI-binding AT-rich regions. METHODS: Recombinant human HMGI (rhHMGI) protein was prepared and electrophoresis mobility shift assay (EMSA) was used. RESULTS: The binding of rhHMGI to PDGF-B (-1 758 / +43 bp) was observed in vitro. Two major HMGI-binding fragments -1 392 / -1 180 bp and -188 / +43 bp were identified, which contained the same AT-rich sequence TTTATAAA (-1 333 / -1 326 bp, -1 314 / -1 307 bp and -30 / -23 bp). An oligonucleotide bound to the TTTATAAA and the GAGACC, the core sequence of the shear stress response element of the PDGF-B, could also bind to the HMGI. Furthermore, HMGI facilitated the binding of NF-?B to the GAGACC in the oligonucleotide. CONCLUSION: The HMGI could bind to the upstream sequence of the PDGF-B gene via the AT-rich sequence TTTATAAA, which may play a role in the transcriptional regulation of the PDGF-B gene.

Objective Objective To discuss the teaching pattern of experimental classes,and to probe the deep development of the functional subject about experimental teaching and the approach of cultivating talents.Methods Student volunteers participated in a series of activities in the summer vocation including experimental design,implementation,results reports and discussion.Results The reflection of students for the learning pattern were as follows: providing a good opportunity of learning(100%);helpful for improving the abilities of finding and dealing with problems(87.50%);different from the past physiological labs including representing special subject characteristics(93.75%),more precise form(68.75%),excellent interaction between teachers and students(75%);improving the manipulation abilities(93.75%).Deficiencies: the experimental conditions were not good enough(68.75);the teachers should give more guidance in theory and techniques(50%).Conclusions The is a meaningful educational form.Under the new experimental teaching pattern,students could use basic medical knowledge and experimental techniques to design and accomplish experiments voluntarily.This pattern is propitious for cultivating integrated capabilities of students,helping them to learn the basic approach and technique means.And it will have much developing space in experimental education in future.

Objective: To investigate the effect of cinobufacini on proliferation, celly cycle distribution, invasion capability of MDA-MB-231 breast cancer cell line in vitro and possible mechanism. Methods: The effect of cinobufacini on cell growth was measured by CCK-8 reagent kit. Cell cycle distribution was determined by flow cytometry. The invasion capability in vitro was detected by Transwell chamber assay. The mRNA expressions of cell cycle related factors (cyclin) and p21 were tested by RT-PCR. Results: Cinobufacini inhibited proliferation of MDA-MB-231 cells. The half inhibition concentration (IC50) was 0.31 mg/mL. The inhibitory effect was timE-dependent (P<0.05). Cinobufacini significantly decreased invasion capability of MDA-MB-231 cells in vitro compared with control group (P<0.05). Cinobufacini induced S-phase arrest of MDA-MB-231 cells in a concentration-dependent manner (P<0.000 1). Cinobufacini down-regulated the expression levels of cyclin A1, cyclin D1, and cyclin E1, while up-regulated that of p21 in MDA-MB-231 cell line. However, there was no marked change in the expression of cyclin B1. Conclusion: Cinobufacini inhibits cell proliferation and influences the cell cycle distribution in vitro by regulating the expression of cyclin A1, cyclin D1, cyclin E1 and p21 in breast can-cer cells.

Objective To study the clinicopathologic characteristics of triple-negative breast cancer (TNBC) and its value in the prediction of prognosis. Method In this study,500 cases of female breast cancers were examined immunohistochcmically for the TNBC. The clinicopathologic characteristics of the 243 TNBC cases were inspected. Results TNBC accounted for 17.6% (88/500) of the 500 breast cancers. The histological types of the TNBC included mainly infihrative ductal carcinoma, metaplastic carcinoma and medullar carcinoma. Among those, histological grade Ⅲ accounted for 72.7% (64/88) of all the TNBC and was more common than that in hormone receptor positive breast cancers (HR+ group ) and Her-2 overexpression breast cancers (Her-2 group)(P=0.000). The positive rates of CK5/6 and EGFR in the TNBC were 30.7% (27/88) and 34.1% (30/88), respectively. The positive rates of ERCC1 and KIT in the TNBC were 28.4% (25/88) and 34.1% (30/88), respectively, Both of which were higher than those in the HR + group and Her-2 group, respectively (P=0.032 and P=0.026). 3-year survival rate of the TNBC was 71.5% and it was lower than that of HR group (P=0.021) and not significantly different from that of Her-2 group (P=0.474). Conclusions TNBC is the breast cancer with high aggressive pathologic futures and poor prognosis. EGFR and ERCC1 expression were positive in a portion of TNBC cases.