VEGF水平及其受体分型和角膜细胞增殖能力的相关性

目的探讨血管内皮生长因子(VEGF)水平及其受体分型(VEGFR1、VEGFR2)与角膜细胞增殖能力的相关性。方法受试者为A、B、C 3组,每组20例。A组为新生血管性青光眼组,B组为原发性青光眼组,C组为年龄相关性白内障组。酶联免疫吸附试验(ELISA)检测血清和房水中VEGF、VEGFR1、VEGFR2水平和促红细胞生成素(EPO)水平。结果3组血清VEGF水平差异有统计学意义(F=680.667,P<0.001),3组血清EPO水平无显著差异(F=0.773,P>0.05)。3组房水VEGF水平差异具有统计学意义(F=552.247,P<0.001)。3组房水中EPO水平差异有统计学意义(F=1757.436,P<0.001)。新生血管性青光眼房水VEGF、EPO水平相关性不显著(r=-0.182,P>0.05)。A、B组血清VEGF、VEGFR2含量与C组相比有显著差异(均P<0.01)。3组血清VEGFR1含量未见明显差异(P>0.05)。结论血管生成性青光眼患者体内VEGF、VEGFR2和EPO局部水平升高,但EPO与虹膜血管生成无明显相关性。

Cloning and Sequence Analysis on cDNA of Growth Hormone Releasing Hormone Receptor Gene from Wuzhishan Miniature Pig

[Objective] cDNA of growth hormone releasing hormone （GHRH） receptor gene from Wuzhishan miniature pig was cloned and its sequence was also analyzed. [Method] Using genomic DNA extracted from porcine ear tissues of Wuzhishan miniature pig as the template, three pairs of primers were designed by the reported cDNA sequence of porcine GHRH, and cDNA was also amplified by RT-PCR. After being recovered and purified, PCR products were ligated to pMD18-T and then transformed into Escherichia coli DH5a. The transformation products were analyzed by PCR and double enzyme digestion to screen positive clones, and the positive clones were sequenced after identification in LB liquid medium. [ Result] cDNA of Wuzhishan miniature pig GHRH receptor gene was obtained successfully, and its length was 1 577 bp coding 423 amino acids. BLAST analysis showed that there were only 23 nuoleotides in difference between this fragment and pomine GHRH receptor gene, and its homology was 98%. However, both GHRH receptor genes were constituted by 423 amino acids with the sequence homology of 96%. [ Conclusion] This study provides theoretical basis for further studies on the dwarf mechanism of Wuzhishan miniature pig.

子宫内膜孕激素抵抗在多囊卵巢综合征发病机制中的研究进展

多囊卵巢综合征（PCOS）是一种妇科内分泌失调性疾病,在世界女性中发病率约为4%,在育龄期女性中占18%。尽管PCOS常在生育早期诊断,但其确切病因及发病机制仍不清楚。PCOS的主要临床病发症状为高雄激素血症、不排卵或稀发排卵、卵巢多囊样改变,常常伴有胰岛素抵抗,远期发生Ⅱ型糖尿病、心血管疾病、子宫内膜病变的风险增大。临床研究发现,75%的PCOS患者出现不排卵、不孕,50%发生复发流产。

Recombinant adenovirus-mediated expression of GHS-R1a in HEK 293 cells

Objective To construct the recombinant adenovirus vector carrying human growth hormone secretagogue receptor type 1a （GHS-R1a） ,for genetic transfection.Methods The full-length human GHS-R1a gene was obtained by PCR amplification and then cloned into the shuttle plasmid pAdTrack-CMV.The linearized plasmid pAdTrack-CMV-GHS-R1a was co-transformed into Escherichia coli （E.coli） BJ5183 cells along with an adenoviral backbone plasmid pAdEasy1.The HEK293 cells were then infected with adenoviruses.The expression of GHS-R1a was indicated by green fluorescent protein （GFP） ,and confirmed by Reverse Transcription Polymerase Chain Reaction （RT-PCR） and Western blot.Results Enzymatic digestion of pAdGHS-R1a yielded a large fragment （approximately 30 kb） and a small fragment （4.5 kb） ,indicating the success-ful construction of recombinant adenovirus expression vector.Expression of GFP was observed by confocal laser scanning microscopy at 24 h after infection.RT-PCR and Western blot further confirmed that GHS-R1a was efficiently expressed in 293 cells.Conclusion Recombinant adenovirus （AdGHS-R1a） is successfully constructed,and the target gene can be expressed efficiently in 293 cells,which provide a valuable tool for further studying the function of GHS-R1a.

Effects of electro-acupuncture on embryo implanted potential of patients with infertility of different syndromes

Objective To explore the impacts of electro-acupuncture on embryo implanted potential and its molecular mechanism in patients with infertility of different syndromes. Methods Among infertile patients treated with in vitro fertilization and embryo transplantation （IVF-ET） combined with electro-acupuncture, 82 cases of kidney deficiency syndrome （group A）, 74 cases of liver qi stagnation syndrome （group B） and 54 cases of phlegm dampness syndrome （group C） were selected. Patients in three groups all received a long program of ovarian hyperstimulation. Additionally, electro-acupuncture was applied before controlled ovarian hyperstimulation （COH） and in the process of ovarian hyperstimulation. Contents of human leukocyte antigen G （HLA-G） in the serum were determined in three groups respectively on the 2nd day of menstruation （M2） and the day of human Chorionic Gonadotropin （hCG） injection. Content of HLA-G in the culture solution of three groups was examined separately on the day of embryo transplantation. The fertilization rate, implantation rate and clinical pregnancy rate were observed for patients in three groups. Results Contents of HLA-G in the serum on the day of hCG injection and in the culture solution on the day of embryo transplantation in group A and B were significantly higher than those in group C （all P0.05）. However, there was no significant difference with contents of HLA-G in serum on M2 day among three groups. The high quality embryo rate in either group A （73.6%, 352/478） or group B （70.6%, 379/537） was significantly higher than that in group C （54.2%, 208/384） with significantly statistical difference （all P0.01）. But there were no significant differences in clinical pregnancy rate, fertilization rate and cleavage rate among three groups. Conclusion Electro-acupuncture combined could increase contents of HLA-G in the body and the level of HLA-G secreted from embryos of patients in the process of IVF-ET. Eventually, the pregnancy outcome and the pregnancy rate are improved. It showed better clinical effects of electro-acupuncture for patients with kidney deficiency and liver qi stagnation syndromes than those of phlegm dampness syndome.

Three-dimensional common-feature hypotheses for Toll-like receptor 7 agonists

Toll-like receptor 7 （TLR7）, the best known TLRs, has been demonstrated to be useful in fighting against infectious disease. In our study, three-dimensional （3D） pharmacophore models were constructed from a set of 5 TLR7 agonists. Among the 10 common-featured models generated by program Discovery Studio/HipHop, a hypothesis （Hypo2） including one hydrogen-bond donor （D）, one hydrogen-bond acceptor （A）, and two hydrophobic （H） features was considered to be important in evaluating the ligands with TLR7 agonistic activity. The obtained pharmacophore model was further validated using a set of test molecules and the Catalyst TLR7-agonist-subset database. Hypo2 has been shown to identify a range of highly potent TLR7 agonists. Finally, the obtained pharmacophore was further validated using docking studies. Taken together, this model can be utilized as a guide for future studies to design the structurally novel TLR7 agonists.

Characterization of subtype selection properties of R-(-)-DM-phencynonate hydrochloride and its racemate on muscarinic receptors

In order to compare the potential selectivity of R-（-）-DM-phencynonate hydrochloride with its racemate （±）-DM- phencynonate hydrochloride on acetylcholine muscarinic receptor subtypes, the five human acetylcholine muscarinic receptor subtypes （M1- M5） （CHO-hml-5R） were cloned and expressed in Chinese hamster ovary （CHO-K1） cell line. The specific mRNAs of the five acetylcholine muscarinic receptor subtypes were detected by the reverse transcription-polymerase chain reaction （RT-PCR） method, demonstrating the definite expression of muscarinic receptor subtype genes （CHO-hml-5R）. The affinity and saturability of different muscarinic receptor subtypes to [^3H] N-methylscopolamine （[^3H]-NMS） were obtained by radioligand binding assay. Equilibrium binding assay revealed that the maximum binding capacity of [^3H]-NMS （Bmax value） to CHO-hml-5R were 40.22±3.23, 24.53±4.11, 29.65±2.65, 25.41±2.46, 32.78±4.81 pmol/mg·protein, respectively. Kd values of [^3H]-NMS to muscarinic receptors M1 to M5 were 0.97±0.22, 1.16±0.14, 0.99±0.06, 0.56±0.08, 1.12±0.06 nM, respectively. R-（-）-DM- phencynonate hydrochloride was found to block the M4 receptor with a much higher potency （pD2 = 7.48） than those displayed on M1 （pD2 = 6.20）, M2 （pD2 = 5.99）, M3 （pD2 = 5.99） and M5 （pD2 = 6.70） subtypes. However, for （±）-DM-phencynonate hydrochloride, no significant subtype receptor selectivity was found. Both （±）-DM- and R-（-）-DM-phencynonate hydrochloride showed allosteric effects on muscarinic receptors, the Hill coefficient （nH） of five receptor subtypes was less than 1, respectively. The results revealed that R-（-）-DM-phencynonate hydrochloride showed selectivity torwards M4 subtype, and there were allosteric effects for both R-（-）-DM-phencynonate hydrochloride and （±）-DM-phencynonate hydrochloride on muscarinic receptors.

Electron Transfer Between Molecules: Study with Langevin Equations

In this paper, we investigate the electron transfer(ET) in donor-acceptor model. The Langevin equation with random forces is used. The oscillations of the primary states observed in experimental data have been shown with this approach. And other features on the dependence of the rate of ET on temperature, free energy, and reorganization energy have also been clearly shown.

Mathematical analysis on a multidimensional model of morphogen transport with receptor synthesis

In biological development, morphogens are locally produced and spread to other regions in organs, forming gradients that control the inter-related pattern and growth of developing organs. Mechanisms of morphogen transport were built and investigated by numerical simulations in [A. D. Lander, Q. Nie and F. Y. M. Wan, Do morphogen gradients arise by diffusion？ Developmental Cell 2 （2002） 785-796]. In that paper, model C, which considers endocytosis, exocytosis and receptor synthesis and degradation, is in a one-dimensional spatial region and couples a partial differential equation with ordinary differential equations. Here, this model is promoted to an arbitrary dimension bounded region. We prove existence, uniqueness and non-negativity of a global solution for this advanced model, of its steady-state solution and linear stability of steady state by operator semigroup, the Schauder theorem and local perturbation method. Our results improve previous results for this model in a one dimension region.