Dan Huang Ming Mu Recipe Suppresses the Progression of Streptozotocin-induced Diabetic Retinopathy After Retinal Laser Photocoagulation in Brown Norway Rats via Down-regulating Vascular Endothelial Growth Factor and Up-regulating Pigment Epithelium-derived

Objective To analyze the effects of Dan Huang Ming Mu Recipe(DHMMR)(a pharmaceutical preparation from herbs and having the function of replenishing vital essence,removing heat,promoting blood circulation and excreting pathogenic water) on diabetic retinopathy(DR) after retinal laser photocoagulation through regulating the expression of vascular endothelial growth factor(VEGF) and pigment epithelium-derived factor(PEDF).Methods Forty male Brown Norway(BN) rats were randomly divided into blank group(group A,10 BN rats) and model group(30 BN rats).DR models were induced by 40 mg/kg streptozotocin(STZ) and the body weight and blood glucose of rats were monitored.After 12-week injection,fluorescein fundus angiography(FFA) and histopathological examination were detected to confirm the successful establishment of DR models.Subsequently,the right eyes of model group rats were conducted retinal laser photocoagulation with the left eyes having no retinal laser photocoagulation and rats of the model group were randomly divided into group B(the model control group),group C(the positive control group),and group D(the DHMMR group),with 10 rats in each group.Rats of the group A and the group B were given vehicle,and the group C were given calcium dobesilate suspension by gavage,and the group D were given DHMMR by gavage.After 4-week gavage,FFA was carried out to observe the fundus microvascular change.Then,the rats were sacrificed to do histopathological examinations and the serum levels of P-selectin,cAMP,cGMP,and the relative expression of the protein of VEGF and PEDF in retina were detected.Results After 12-week STZ injection,the blood glucose of the model group were pronouncedly higher than the blank group(P < 0.01).Fundus micro-hemangioma changes were observed in the rats of the model group through FFA,and the rats of the blank group did not see any changes.The pictures of HE staining showed that the retinal structure of the model group was more disordered than the blank group.After 4-week gavage,treatments with DHMMR showed dramatic reduction of serum levels of Pselectin,cAMP and cGMP compared with the group B(P < 0.01).FFA and histopathological examinations of DHMMR-treated rats revealed significantly suppression of retinal edema,fundus microvascular destruction and retinal destruction distinguishing from the group B.And the relative expression of VEGF protein of the group D and the group A was markedly lower than that of the group B(P < 0.01).The relative expression of PEDF protein of the group D and the group A was dramatically higher than that of the group B to the contrary(P < 0.01).Conclusions DHMMR demonstrates pronounced suppressive effects on the progression of DR after retinal laser photocoagulation,through down-regulating VEGF and upregulating PEDF.

2型糖尿病患者房水中PEDF，VEGF，Leptin水平测定的临床意义

目的检测2型糖尿病患者眼房水中色素上皮衍生因子（pigment epithelium-derived factor，PEDF）、血管内皮生长因子（vascular endothelial growth factor，VEGF）和瘦素（Leptin）的含量，并探讨其临床意义。方法对70例2型糖尿病患者房水中PEDF和VEGF的含量采用双抗体夹心酶联免疫吸附（ELISA）法检测，Leptin含量采用放射免疫法（RIA）检测。根据散瞳眼底检查和眼底荧光素血管造影检查后，实验组分为：无糖尿病视网膜病变组（NDR）22例，单纯型糖尿病性视网膜病变组（BDR）28例，增生型糖尿病性视网膜病变组（PDR）20例，对照组为健康的老年性白内障患者20例。结果NDR组，BDR组，PDR组的房水VEGF含量分别为250．32±26．77，300．11±58．89，496．23±91．06pg／ml；PEDF含量分别为0．41±0．55，0．37±0．42，0．29±0．19ng／ml；Leptin含量分别为69．80±21．37，155．08±32．76，230．27±56．92pg／ml。对照组房水VEGF含量为144．69±26．55pg／ml，PEDF含量为0．52±0．47ng／ml，Leptin含量为43．62±20．02pg／ml，对照组与实验组比较差异均有统计学意义（F=118．62，P〈0．01；F=-110．53，P〈0．01；F=101．22，P〈0．01）。4组房水VEGF，PEDF，Leptin含量有明显增加的趋势。房水中的VEGF与PEDF，Leptin含量有相关性（r=0．995，P〈0．01；r=0．776，P〈0．01）；房水中VEGF，PEDF，Leptin含量与糖尿病患者的病程及糖尿病视网膜病变的严重程度有相关性（r=0．722，P〈0．01；r=0．716，P〈0．01），（r=-0．869，P〈0．01；r=-0．865，P〈0．01），（r=0．776，P〈0．01；r=0．765，P〈0．01）。结论VEGF，PEDF，Leptin在糖尿病视网膜病变的形成过程中有重要作用，且VEGF与PEDF，VEGF与Leptin之间有相关性。

Predictive Value of Serum Vascular Endothelial Growth Factor and Pigment Epithelium-derived Factor in Ovarian Hyperstimulation Syndrome

Objective To explore whether the serum concentrations of vascular endothelial growth factor （VEGF） and pigment epithelium-derived factor （PEDF） could serve as the predictors of ovarian hyperstimulation syndrome （OHSS） in patients undergoing in vitro fertilization-embryo transfer （IVF-ET）. Methods Enzyme-linked immunoadsordent assay （ELISA） was employed to measure the serum concentrations of VEGF and PEDF on the day of hCG administration, oocyte retrieval and embryo transfer, respectively. Based on OHSS classification of the criteria of Golan, 85 patients were divided into three groups. Patients in group A （n=10） showed symptoms of severe OHSS and patients in group B （n=13） suffered from moderate OHSS. The control group （group C, n=62） contained patients without symptoms of OHSS as well as patients with mild OHSS.Results In groups A, B and C, serum concentrations of PEDF on the day of hCG administration （h-PEDF）（166.54 ± 102.81 pg/ml, 159.45 ±136. 77 pg/ml, 172.05±170.95 pg/ml, P=0.48）, oocyte retrieval （o-PEDF）（176.91 ± 103.37 pg/ml, 122.52± 92.54 pg/ml, 179.82±177.47 pg/ml, P=0.27） and embryo transfer （e-PEDF）（169.02± 240.08 pg/ml, 136.80 ±139.21pg/ml, 157.38 ±222.54 pg/ml, P=0.95）, h-VEGF （175.55 ± 103.54 pg/ml, 218.84 ±179.70pg/ml, 153.39±145.06 pg/ml, P=0.36） and o- VEGF （171.93 ± 128.55 pg/ml, 220.36±149.82 pg/ml, 138. 74 ±% 139.30 pg/ml, P=0. 15） showed no significant differences. There was a statistical difference in serum concentration of e-VEGF between group A （197.04±156.63 pg/ml） and group C （110.69±49.55 pg/ml）（P=0.008）. The serum level of estradiol showed a positive correlation with the count of large follicles （r=0. 744）. The ratios of h-VEGF/h-PEDF, o-VEGF/o-PEDF and e-VEGF/e-PEDF were calculated and showed a clear difference among groups A, B and C （4.04±3.39, 2.10±2.14, 1.05± 4.80, P〈0.001; 4.54 5.69, 2.29 ±1.67, 0.94 ±0.59, P〈0.001; 5.43±6.16, 1.81±1.36, 2.42±2.60, P=0.04）. Conclusion While neither serum concentrations of VEGF nor PEDF can be used as an OHSS predictor, the ratios of h-VEGF/h-PEDF, o-VEGF/o-PEDF and e-VEGF/e-PEDF may have great predictive value.

小鼠黑色素细胞沉默及过表达色素上皮衍生因子对黑色素合成的影响

旨在研究色素上皮衍生因子(pigment epithelium-derived factor,PEDF)与黑色素合成调控因子之间的互作关系,探索PEDF对黑色素细胞活力和黑色素合成的调控作用,为研究动物毛色形成机制提供基础。本研究将PEDF siRNA及PEDF过表达载体分别转染小鼠黑色素细胞,对黑色素细胞活力、黑色素含量进行检测,采用qRT-PCR、Western blotting及细胞免疫荧光对PEDF、Wnt1、Wnt3α、MITF和β-catenin的表达和定位进行检测。黑色素细胞PEDF沉默后,黑色素细胞的细胞活力增强,黑色素含量增加,Wnt1、Wnt3α、MITF和β-catenin的mRNA和蛋白表达量升高;黑色素细胞PEDF过表达后,黑色素细胞活力减弱,黑色素含量减少,Wnt1、Wnt3α、MITF和β-catenin的mRNA和蛋白表达量降低。细胞免疫荧光结果显示:PEDF、Wnt1、MITF、β-catenin表达于黑色素细胞的细胞质;Wnt3α表达于黑色素细胞的细胞质和细胞膜。本研究证实:PEDF使黑色素细胞活力减弱,可通过下调促黑色素生成相关的因子Wnt1、Wnt3α、MITF和β-catenin,从而抑制黑色素的生成。

Contemporary Approaches for Identifying Rare Bone Disease Causing Genes

Recent improvements in the speed and accuracy of DNA sequencing, together with increasingly sophisti- cated mathematical approaches for annotating gene networks, have revolutionized the field of human genetics and made these once time consuming approaches assessable to most investigators. In the field of bone research, a particularly active area of gene discovery has occurred in patients with rare bone disorders such as osteogenesis imperfecta （OI） that are caused by mutations in single genes. In this perspective, we highlight some of these technological advances and describe how they have been used to identify the genetic determinants underlying two previously unexplained cases of OI. The widespread availability of advanced methods for DNA sequencing and bioinformatics analysis can be expected to greatly facilitate identification of novel gene networks that normally function to control bone formation and maintenance.