AIM:To establish the murine model of oxygen induced retinopathy ( OIR) and to evaluate the inhibition of retinal neovascularization by erythropoietin ( EPO) blockade.
METHODS: Neonates of C57BL/6 mouse ( P7 ) were exposed to 75%±2% oxygen for 5d and return to normal air environment when 12d ( P12 ) to establish oxygen-induced retinal neovascularization model. The neonates were divided into groups, injected with 0. 5μL solution containing 25ng ( group A), 50ng ( group B), 250ng ( group C) of soluble erythropoietin receptor ( EPO-R) or PBS (group D) at P12, P14 and P16 in the right eye. On P17, the litters were sacrificed and their right eyes were enucleated and fixed with 4% paraformaldehyde, made pathological section. The number of breakthrough internal limiting membrane neovascular nuclei was counted with pathological retinal morphology, understanding theproliferative degree of retinal neovascularization.
RESULTS: The pathological sections showed the neovascular cell nuclei which penetrating the inner limiting membrane in intravitreal EPO receptor injection group was reduced statistically than that in PBS injection group, the difference was statistically significant ( P<0.01). And, neovascular nuclei count differences in the
various concentrations of EPO receptor group was statistically significant (P<0. 01). With the EPO receptor concentration increase, neovascular endothelial cells broken through the internal limiting membrane was reduced.
CONCLUSION: Intravitreal injection of soluble EPO receptor can block EPO and improve neovascularization. The new method is expected to become new treatment of ocular neovascular diseases.

AIM: To investigate the therapeutic effect of endothelial progenitor cell-conditioned medium (EPC-CM) on the lung structure of neonatal rat exposed to hyperoxia, and to explore the mechanisms.METHODS: Bone marrow-derived endothelial progenitor cells (EPCs) were collected from new born Sprague-Dawley (SD) rats and the EPCs were identified.The conditioned medium from the passage 3 EPCs was collected.Newborn SD rats (n=40) were randomly divided into 4 groups.The rats in room air group were exposed to the room air (21% O2) for 21 d.The rats in hyperoxia group were exposed to hyperoxia (85% O2) for 21 d.The rats in endothelial cell basal medium (EBM) group were exposed to hyperoxia for 21 d, and received 100 μL EBM on postnatal day 14 (P14) in a single intratracheal (IT) injection.The rats in EPC-CM group were exposed to hyperoxia for 21 d, and received 100 μL EPC-CM on P14 in a singlie IT injection.The rats were sacrified on the 21st day.The left lungs were excised, placed in 4% paraformaldehyde, serially dehydrated in ethanol and embedded by paraffin.Serial sectioning of the paraffin-embedded left lung tissues was prepared for 5 μm thickness, and stained with hematoxylin and eosin.The pulmonary radical alveolar count (RAC) and alveolar mean linear intercept (MLI) were then calculated.The microvascular density was determined by FVIII immunostaining.The mRNA expression of KGF, VEGF, SP-A and SP-C in the right lung tissues was detected by real-time fluorescence quantitative PCR.RESULTS: The cultured cells had typical EPC morphological characteristics, and had the abilities to bind to FITC-UEA-1 and uptake DiI-ac-LDL.The body weight of the rats on day 21, RAC, MLI and microvascular density were significantly lower in hyperoxia group and EBM group than those in room air group (P<0.05).The EPC-CM group had significantly higher RAC and microvascular density than those in hyperoxia group and EBM group (P<0.05), but the body weight and MLI had no significant difference.The mRNA expression levels of KGF, VEGF, SP-A and SP-C in hyperoxia group and EBM group were significantly lower than those in room air group (P<0.05).The mRNA expression levels of KGF, VEGF, SP-A and SP-C in EPC-CM group were significantly higher than those in hyperoxia group and EBM group (P<0.05).CONCLUSION: EPC-CM promotes the lung alveolarization and microvascular formation in neonatal rats exposed to hyperoxia.These benefits may be correlated with the increased KGF and VEGF mRNA expression.

Objective: To investigate the potential role of me latonin in the adrenal cortex of human embryo. Methods:Specifi c melatonin receptors was localized and characterized in the adrenal cortex of h u man embryo by means of immunohistochemistry. Results: mt1 （Me l1a）and MT2 （Mel1b）subtype of melatonin receptors was principally localize d to cytoplasm in zona glomerulosa， fasciculata and reticularis. Conclu sion: It is possible that mt1 and MT2 subtype of melatonin receptors co-exist in the adrenal cortex of human embryo.

Objective: To determine protein binding characteri stic and signal transmission pathway of melatonin（Mel） receptor(MR) in human e mbryonic peripheral organ tissues. Methods: MR was measured by radio ligand-binding assay and the effect of GTPγS on melatonin specific bindi ng was studied. Results: Mel specific binding sites were det ermined in 16 kinds of human embryonic tissue and this binding could be inhibit ed by GTPγS, supporting the theory that MR is coupled to inhibitory G-proteins system. Conclusion: MR is measured in human embryo tissue, the se results provide experimental data for elucidating the mechanism of the effect of Mel.

Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

Objective: To make it clear whether there exists m elatonin receptor in the thyroid of human embryo. Methods: Thyr oid was collected and sliced up to be stained with methods of immunohistochemis try and in situ hybridization. Results: The thyroid tissue w as p ositively dyed, melatonin receptor mt1 and MT2 were with both immunohistoche mistry and in situ hybridization while brown granules dep osited in the membrane, plasma and nuclear of the thyroid cell were with the imm unohitochemistry. Conclusion: There exists melatonin rece ptors in human embryo thyroid, either mt1 or MT2, and they exist in the memb rane, plasma and nuclear.

OBJECTIVETo summarize the clinical experience about surgical treatment of aortic dissection.

METHODSThe clinical data of 51 patients with aortic dissection admitted from December 2004 to December 2006 were analyzed retrospectively. There were 35 male and 16 female patients with a mean age of 55.7 years (ranged from 18 to 83-years-old). Twenty-seven patients of type I was performed under deep hypothermic circulatory arrest and selected cerebral perfusion with stent-graft which was implanted into the descending aorta through aorta arch. Five patients of type II was performed including Bentall operation in 3 patients, Wheat operation in 1 patient, ascending aorta replacement in 1 patient. Nineteen patients of type III was performed with stent-graft which was implanted into the descending aorta through aorta arch under deep hypothermic circulatory arrest.

RESULTSThe time of cardiopulmonary bypass (CPB) in type I patients was 250 to 290 min with an average of (274 +/- 53) min, and the arrest time was 40 to 59 min with an average of (53 +/- 14) min. CPB time of type II patients was 130 to 159 min with an average of (146 +/- 43) min, and the cross clamp time was 60 to 79 min with an average of (66 +/- 15) min. CPB time of type III patients was 240 to 280 min with an average of (260 +/- 28) min, and the arrest time was 20 to 27 min with an average of (24 +/- 3) min. The mean hemorrhage volume of the entire group was (500 +/- 250) ml. The mean ICU retention time was (5.0 +/- 1.5) d and the length of stay was (15.0 +/- 2.5) d. Three patients died during perioperative period. Two patients appeared cerebrovascular accident after operation. One patient appeared descending aorta dilation in the follow-up of 2 to 21 months.

CONCLUSIONDifferent clinical manifestations and treatment should be selected according to the different condition of aortic dissection aneurysm.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Aneurysm, Dissecting ; surgery ; Aorta, Thoracic ; surgery ; Aortic Aneurysm, Thoracic ; surgery ; Blood Vessel Prosthesis Implantation ; Female ; Humans ; Male ; Middle Aged ; Stents

OBJECTIVETo observe the effect of Chinese herbs for stasis removing and collaterals dredging (CHSRCD) upon angiotensin-converting enzyme 2-angiotensin-(1-7)-Mas axis in the renal cortex of diabetic nephropathy rats.

METHODSTotally 89 male Sprague-Dawley rats were randomly divided into the blank control group (C group, n=22), the high-glucose high-fat control group (H group, n=10), and the streptozotocin (STZ)-injecting group (n=57). The diabetes rat model (n=50) was induced by feeding high-glucose high-fat diet in combination with intraperitoneal injection of STZ, which were further divided into the model group (M group, n=24), the irbesartan group (I group, n=13), and the CHSRCD (Z group, n=13). Rats in I and Z groups were intragastrically fed with suspension of irbesartan and CHSRCD, once daily for 16 weeks. Equal volume of drinking water was administrated to rats in the rest groups. Blood glucose and 24 h urine protein quantitation were tested at four time points. And the mRNA expression of ACE2 and Mas at various time points was detected by Real-time PCR, immunohistochemical assay, and Western blot. Quantitative analyses of ACE2 and Mas protein expression were performed at the end of week 16.

RESULTSCompared with the C group, blood glucose increased in the H and M groups (P < 0.01). It was higher in the H group (P < 0. 01). 24 h urine protein quantitation at different time points increased in the M group, and it was higher than that in the H group (P < 0.05). Compared with the M group, 24 h urine protein quantitation decreased at the end of week 8 in the I group, and at the end of week 8 and 16 in the Z group (P < 0.05). It was lower in the Z group than in the I group at the end of week 16 (P < 0.05). Compared with the C and H groups, the expression of ACE2 mRNA in the renal cortex was lower in the M group at the end of week 16 (P < 0.01). Compared with the M group, it was higher in the Z group (P < 0. 01). There was no statistical difference in the expressions of Mas mRNA at the end of week 16 between the C group and the M group (P > 0.05). It was lower in the M group than in the H group (P < 0.05). It was higher in the Z group than in the M group (P < 0.05), and higher than in the I group (P < 0.05). The expression of ACE2 and Mas protein in the M group decreased as time went by. The expression quantitation of ACE2 and Mas protein at the end of week 16 was lower in the M group than in the C group (P < 0.05). Compared with the M group, ACE2 expression of the Z group and Mas of the I and Z groups increased more significantly (P < 0. 05).

CONCLUSIONCHSRCD could play a role in renal protection for diabetic nephropathy rats by up-regulating the mRNA and protein expression of ACE2 and Mas, promoting the ACE2-Ang-(1-7)-Mas axis, and lowering urinary protein.

Angiotensin I ; metabolism ; Animals ; Diabetes Mellitus, Experimental ; metabolism ; Diabetic Nephropathies ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Kidney Cortex ; metabolism ; Male ; Peptide Fragments ; metabolism ; Peptidyl-Dipeptidase A ; metabolism ; Proto-Oncogene Proteins ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, G-Protein-Coupled ; metabolism

Objective To retrospectively evalute the value and accuracy of adenosine stress and rest SPECT myocardial perfusion imaging. Methods A total of 1858 patients who were suspected or known for coronary artery disease (CAD) underwent 99Tcm-methoxyisobutylisonitrile ( MIBI ) myocardial perfusionSPECT with adenosine infusion using the standard 2-day protocol. Images were interpreted by two or more experienced nuclear medicine physicians . Coronary angiography was carried out in all patients within one month. Kappa test was used to analyze the correlation between the two imaging studies. Results By coronary angiography, there were 957 patients diagnosed of CAD (one-, two-, three-vessel disease: 506,256,195, respectively) and 901 normal. Stenosis was found in 1603 vessels, including left anterior descending coronary artery (LAD): 765, left circumflex coronary artery (LCX): 399 and right coronary artery (RCA): 439. By adenosine induced stress myocardial perfusion imaging, 876 patients were diagnosed of myocardial ischemia ( sensitivity: 876/957, 91.54% ) and 651 patients had negative findings ( specificity:651/901,72.25 % ). The positive and negative predictive values were 77.80% ( 876/1126 ) and 88.93% (651/732), respectively. The correlation coefficient between the two imaging studies was 0.641. The vessel-based sensitivity was 81.31% (622/765) for LAD, 56.64% (226/399) for LCX and 70.62% (310/439) for RCA, respectively. The sensitivity for detection of one-, two-, three-vessel stenosis was 87.55% (443/506), 94.92% (243/256) and 97.44% (190/195), respectively. The side-effects was mild and transient with an incidence rate of 84.12% ( 1563/1858), without major cardiac events. Conclusion Stress myocardial perfusion imaging induced by adenosine is reliable for the evaluation of myocardial blood supply in CAD patients.

Objective To establish an isolated rat pancreas perfusion technique,a method for the precise measurement of insulin secretion in vitro.Methods An isolated rat pancreas perfusion technique was applied in the study of insulin secretion from?-cells in 10 high-fat diet-induced obese Wistar rats.Results For the assessment of the functional integrity of the perfused pancreas,the isolated pancreas of 6 rats met all the criteria: (1)The constancy of perfusion pressure was kept over the whole experiment time[(70?5)mm Hg,1 mm Hg= 0.133 kPa].(2)The duodenal peristaltic activity of isolated pancreas and duodenum block was present after perfusion experiment.(3)Total insulin response to arginine stimulation was significantly increased as compared with glucose stimulation[maximum insulin secretion rate:(987?100)?U/min vs(545?50)?U/min,P