Inhibition efficacy of CFB-siRNA on laser-induced choroidal neovascularization in rat / 中华实验眼科杂志

BackgroundChoriodal neovascularization is an important ocular manifestation of angiogenesis in eyes,which derives from the choroid capillaries.Recent studies have found that complement activation is playing a key role in the laser-induced CNV.Because of the key position of CFB in the alternative pathway,bytargeting CFB and blocking the alternative pathway may provide an approach to observe the role of this alternative pathway in the generation of CNV.Objective This study was to investigate the inhibitory effect of reconstructed complement factor B (CFB)-small interfering ribonucleicacid(siRNA)on choroidal neovascularization (CNV)and its mechanism. Methods Experimental CNV was induced by laser photocoagulation in 96 eyes of 48 clean Brown Norway rats.The rats were randomly divided into 4 groups.25,50 and 75 μg B factor siRNA were injected via caudal vein on 1 day,3,5 days after photocoagulation in different dose groups,and normal saline solution was injected at the same way in experimental control group.Other 12 normal rats were used as blank control group.Fundus fluorescein angiography(FFA) was performed on 3,7,14,21,28 days after injection of CFB-siRNA and CNV was scored.The expressions of vascular endothelial growth factor(VEGF) and factor Ⅷ in choroid were detected by immunochemistry.The expressions of CFB-siRNA,VEGF,transforming growth factor β2( TGF-β2 )proteins in choroid were determined using immunochemistry in 7,14,21,28 days,and the expressions of mRNA of CFB-siRNA,VEGF,TGF-β2 were examined by reverse transcription polymerase chain reaction(RT-PCR). ResultsFFA revealed that the CNV rates in various doses of CFB-siRNA groups were significant lower than those of experimental control group in various time points(P＜0.05),and those in 75 μg B factor siRNA were decreased in comparison with 25 μg B factor siRNA (P＜0.05).Immunochemistry showed that the intensities of the VEGF and factor Ⅶ expression in various doses of CFB-siRNA groups were weaker than the blank control group ( P ＜ 0.05 ).Compared with the control group,the expression of CFB reduced in 7 days,and then approached to the level near the control group.Fourteen to twenty-one days after injection of CFB-siRNA,VEGF and TGF-β2 depressions in different doses of CFB-siRNA groups were lower than blank control group( P＜0.05 ).CFB expression in choroid showed the lower levels in CFB-siRNA injection group compared with blank control group in from 7 through 21 days (P＜0.05).RT-PCR displayed the gradual increase of CFB mRNA and curve-like changes of VEGF and TGF-β2 with time prolong. Conclusions Recombinated CFB-siRNA can effectively inhibit laser-induced CNV by down-regulating the expression of VEGF and factor Ⅷ.Alternative pathway of complement plays an important role in the production of CNV.

Expression of inhibin B betaB subunits in the testicular tissues of azoospermia patients with different pathological alterations / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the expression of inhibin B betaB subunits in human testicular tissues.</p><p><b>METHODS</b>Eighty-three cases of the azoospermia underwent testicular biopsy. In accordance with the pathological alterations of spermatogenesis, the samples were divided into four groups: Sertoli-cell-only syndrome (n = 21); hypospermatogenesis (n = 20), maturation arrest (n = 24) and almost normal spermatogenesis (n = 18). Immunohistochemical staining for inhibin B betaB subunits was conducted on the paraffin-embedded sections of different spermatogenesis to localize inhibin B betaB subunits in the seminiferous tubules.</p><p><b>RESULTS</b>Immunohistochemically, positive products of inhibin B betaB subunits were found in both the seminiferous tubules and interstitial tissues of the testis as brown or yellow particles in the cytoplasm. Leydig cells and early intermediate spermatogenic cells showed a very strong positivity; Sertoli cells in the seminiferous tubules were mostly positive; peritubular myoid cells showed a weak positive staining; but no positive expression of inhibin B betaB subunits was found in late spermatids and mature sperm.</p><p><b>CONCLUSION</b>Inhibin B may be produced by both Sertoli cells and early spermatogenic cells in the seminiferous tubules.</p>

Variation of nestin expression in SD rat eyes in different stages of postnatal development / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the position and quantity of nestin expression in SD rat eyes in different stages of postnatal development.</p><p><b>METHODS</b>Immunocytochemical method was used to identify nestin expression in the eyes of SD rats of 1 to 30 days old.</p><p><b>RESULTS</b>Nestin expression was detected in the retina and extraocular muscles of SD rats. The expression varied with the time of postnatal development, distributing in the entire retina layers in earlier stages and confined in the nerve fiber layer in later stages. The quantities of nestin expression in the extraocular muscles decreased gradually with growth.</p><p><b>CONCLUSION</b>Nestin expression in the retinas and extraocular muscles of SD rats decreases during the postnatal development.</p>

Establishment of an undifferential orchitis model in rats and changes of spermatogenic epithelium / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the changes of rat testicular spermatogenic epithelium stimulated by bacterial lipopolysaccharide (LPS) in vivo.</p><p><b>METHODS</b>Twenty Wistar rats were divided into two groups: control group and experimental group. The control group was treated with pyrogen-free saline (1 ml/kg) and the experimental group was injected ip with saline containing LPS (1 mg/kg) once every two days. Two groups were operated after ten days in order to investigate the testicular pathological changes by HE staining and the expression of proliferating cell nuclear antigen( PCNA), alpha-catenin in spermatogenic epithelium by immunohistochemistry assay.</p><p><b>RESULTS</b>The testes of the experimental group showed inflammatory changes. The positive expression of PCNA in seminiferous epithelium was significantly lower than that of control group. The number of positive cells in every seminiferous, in which only spermatogonia were stained in experimental group were 59 +/- 5 and it showed significant decrease compared with the control (P < 0.01). Furthermore, the percentage of such seminiferous tubules was 0.673 +/- 0.054 and increased apparently (P < 0.01). The expression of alpha-catenin in testicular tissue of the experimental group declined (P < 0.01), and cellular positive granular light density was 0.150 +/- 0.014.</p><p><b>CONCLUSION</b>The ability of spermatogonium proliferation and the function of conglutination of cells under inflammatory condition of the testes declined, which may be one of the etiologies of male infertility.</p>

The establishment of testicular fibrosis model in Wistar rats / 中华男科学杂志

<p><b>OBJECTIVES</b>To establish the testicular fibrosis model in rats.</p><p><b>METHODS</b>Wistar rats were divided into control group(n = 12) and model group(n = 40) randomly. Testicular fibrosis model was built with the classical method of establishing experimental autoimmune orchitis (EAO) combined with injecting Bacille Calmette-Guerin (BCG) into left testis.</p><p><b>RESULTS</b>The incidence rate of EAO and the rate of testicular fibrosis were 100%, 11.1% and 100%, 81.5% at 80, 140 days after the first infection, respectively.</p><p><b>CONCLUSIONS</b>The model of rat testicular fibrosis was established successfully.</p>