2'-Methyl-pyrazolo[4',3':16,17]androst-5-en-3ß-ol.

In the title compound, C(21)H(30)N(2)O, there are five fused rings. The A and C rings adopt chair conformations, ring B adopts an 8ß,9α-half-chair conformation and ring D adopts a 14α-envelope conformation. The pyrazole ring is planar. Inter-molecular O-H⋯N hydrogen bonds [H⋯N = 1.88 (5) Å] help to stabilize the crystal structure. The absolute structure was deduced from those of the starting materials.

Bioptic significance of incarcerated contents at sclerotomy sites during vitrectomy.

PURPOSE: To study the clinical significance of incarcerated materials at sclerotomy sites during vitrectomy. METHODS: Fifty samples prolapsed from the entries to standard three-port pars plana vitrectomies were collected along the scleral surface. Samples from rhegmatogenous retinal detachments (RD) (n = 28), traumatic RD (n = 4), miscellaneous vitreous hemorrhages (n = 12), and intraocular foreign bodies (n = 6) were stained with hematoxylineosin and examined under light microscopy after being fixed in paraffin sections or smeared on slides. RESULTS: The specimens collected after sclerotomy contained vitreous tissue mixed with dispersive and sheet nonpigmented ciliary epithelia, scattered pigment granules, and small pigment gobbets. Specimens collected during vitrectomy contained pigment granules and various cells with a jellylike appearance. Fibrous tissue and remnants of ciliary body and retina were found in some specimens. The incarcerated tissues caused 12 cases of iatrogenic retinal breaks, among which RD recurred in six cases postoperatively due to anterior proliferative vitreoretinopathy. In the noniatrogenic retinal break group, only 4 RD recurred (6/12 versus 4/38, chi2 = 6.586, P = 0.01). CONCLUSION: The incarcerated contents at sclerotomy sites were mainly adjacent tissues and cells. Prolapsed and incarcerated ciliary body and retina fragments were common and might cause iatrogenic retinal breaks. Attention should be paid to this material intraoperatively.

[Effect of endotoxin-induced uveitis on GATA-3 expression and ACAID development].

OBJECTIVE: To investigate the changes of the GATA-3 expression and development of anterior chamber associated immune deviation (ACAID) after anterior chamber (AC) injection of interphotoreceptor retinoid binding protein (IRBP) under ocular inflammation. METHODS: ACAID was induced by injection of IRBP into the AC of 30 Spar-Dawley (SD) rats. Then the animals were divided into -4 days group, -24 hours group, 0 hour group, 3 days group, and 7 days group according to the lipopolysaccharide (LPS) injection 4 days and 24 hours before, or 0 hour, 3 days and 7 days after IRBP inoculation respectively. 6 rats were used as controls 8 days after IRBP injection, Serum interleukin-4 (IL-4) was evaluated to count the development of ACAID; Western blot and RT-PCR were used to determine the protein and mRNA levels of GATA-3 expression. RESULTS: In -24 hours group and 0 hour group, the ocular inflammation reached a maximum 24 hours after LPS injection; on 8 days after IRBP inoculation, serum IL-4 couldn't be detected and GATA-3 expression were not changed both on mRNA and protein levels compared with control group. In -4 days group, the ocular inflammation was subsided gradually 24 hours and disappeared 96 hours after LPS injection; serum IL-4 and GATA-3 expression were significantly elevated 8 days after IRBP injection. In 7 days group, the serum IL-4 and GATA-3 expression in spleen increased 8 days after IRBP inoculation. CONCLUSION: In ocular inflammation, the up-regulation of GATA-3 expression is inhibited and ACAID development is blocked after antigen was injected into anterior chamber. Once GATA-3 is up-regulated, LPS injection cannot affect ACAID development.

[A study on the effects of rAd-gfp-bcl-X(L) on retinal degeneration in mice].

OBJECTIVE: To observe the therapeutic effect of rAd-gfp-bcl-X(L) on retinal degeneration (RD) in mice. METHODS: rAd-gfp-bcl-X(L) granules were injected into the subretinal space of right eyes of RD mice in Group A (postnatal days 10, p10) and Group B (postnatal days 22, p22) (n = 20). The morphological changes were observed by light microscopy and transmission electron microscopy at 15 - 30 days post-injection. Frozen section was used to observe the expression of GFP. The level of bcl-X(L) mRNA and bcl-X(L) protein were assayed by RT-PCR and immunohistochemistry assay. RESULTS: In frozen section, GFP was observed in the treated eyes under fluorescence microscopy, indicating that rAd-gfp-bcl-X(L) had transfected into the retinal cells successfully and that bcl-X(L) was expressed. In Group A, an increased expression of bcl-X(L) mRNA and Bcl-X(L) protein were observed in the treated eyes. The outer nuclear layer of the treated eyes in Group A was thicker than that of the control eyes at 30 days post-injection. The inner segment of treated eyes in Group A was healthier than that of control eyes, as indicated by transmission electron microscopy. No difference was found in the thickness of retina between treated eyes and control eyes in Group B. CONCLUSION: The bcl-X(L) can be expressed and have an anti-apoptosis effect when the rAd-gfp-bcl-X(L) particles are injected into the subretinal space of RD mice's eyes at an early stage. But the expression and protective effects were not observed in RD mice at a later stage. The therapy of rAd-gfp-bcl-X(L) for retinal degenerative diseases still needs further study.

[Effect of bcl-2 antisense oligonucleotides on multidrug resistance of cultured uveal melanoma cells].

OBJECTIVE: To evaluate the chemotherapeutic sensitivity of uveal melanoma in vitro and reverse its drug resistance by the delivery of bcl-2 antisense oligodeoxynucleotide (AS-ODN). METHODS: The drug sensitivity tests of primary cultured uveal melanoma cells toward 5-flurouracil (5-FU), thiophosphamide (TSPA), cisplatin (DDP), adriamycin (ADM), vinblastine (VLB), dacarbazine (DTIC) were performed with 3, -4, 5 dimethyliazol-2,5 diphenyl tetrazolium bromide (MTT). AS-ODN bcl-2 were delivered with cationic lipid to down-regulate bcl-2 expression. Immunohistochemistry and Western-blot methods were used to detect bcl-2 expression. According to the principle of multi-drug mutual action, the influence of anti-apoptosis gene bcl-2 on tumor cell drug sensitivity was measured. RESULTS: Uveal melanoma was resistant to different chemotherapeutic drugs in different degrees. AS-ODN bcl-2 could down-regulate bcl-2 expression, was synergetic with all tested drugs and partially reversed the multi-drug resistance. CONCLUSIONS: Clinically, with ordinary dosages the above 6 drugs have little cytotoxicity to uveal melanoma cells, bcl-2 over-expression is associated with multi-drug resistance and AS-ODN bcl-2 can partially reverse the multi-drug resistance.

[Clinical analysis of tumors of the iris and ciliary body].

OBJECTIVE: To evaluate the clinical features, different diagnosis, pathological features, management, and prognosis of tumors of the iris and ciliary body. METHODS: Medical records, photographs, pathological findings and the results of follow-up of 37 cases with tumors of the iris and ciliary body were reviewed as a retrospective study. RESULTS: Of the 37 cases with tumors of the iris and ciliary body, 26 were male and 11 were female. The mean age at diagnosis was 38 years, ranged from 5 - 64 years. According to the histopathological examination, melanoma was the most common tumor in the iris and ciliary body (15 cases, 40.5%), followed by metastatic tumors (8 cases, 21.6%), teratoid medulloepitheliomas (3 cases, 8.1%) and iris nevus (2 cases, 5.4%). MANAGEMENT: The tumors were excised in 14 cases. Enucleation was performed in 21 cases. Two cases were observed without any surgical treatment. Thirty-four cases were followed-up for 2 months to 15 years, averaged 31 months. Most melanomas of the iris and ciliary body are round or semi-spherical dark brown vascularized mass, with engorged episcleral sentinel vessels in some cases. The tumor showed a shadow during transillumination. Ultrasound biomicroscopy revealed a low to medium echoic solid lesion, with echoic changes in adjacent infiltrated tissues. Melanoma showed positive immunoreactivity for melanoma-specific antigen, and had a good prognosis. Metastatic tumors of the iris and ciliary body were flat or near round, dirty, single or multiple neoplasms, growth rapidly, with abundant neovascularization, and had a poor prognosis. Primary carcinomas could be found in other parts of the body. CONCLUSIONS: Melanoma of the iris and ciliary body has typical features that may help to distinguish them from other tumors. Metastatic tumor has characteristic features, but the diagnosis can be made only with supplementary examination and immunocytochemical studies. Medulloepitheliomas should be differentiated from retinoblastoma.

[Subretinal transplantation of retinal cells by outer approach].

OBJECTIVE: To investigate the approaches and effect of pure retinal photoreceptor cells and mixed retinal cells subretinal transplantation on retinal degenerative diseases. METHODS: 16 Kunming mice were divided into two groups: group A and group B (n = 8). Through sclera and choroid, mixed retinal cells were injected into the subretinal space of mice eyes of group A and pure retinal photoreceptors were injected into the subretinal space of mice eyes of group B by using a specific micro-syringe under operative microscopy. 30, 90 and 180 d after transplantation, experimental eyes were enucleated, stained with HE and analyzed by light microscopy. RESULTS: Most transplants (13/15) were exactly in host subretinal space without the infiltration of inflammatory cells and interfere of retina. The infiltration of inflammatory cells and damage of retina were only found in few eyes (2/15). The transplanted retinal cells survived well at 180 days after transplantation. The mixed retinal cells all formed rosette, but the pure retinal photoreceptors formed an orderly layer of photoreceptors. CONCLUSIONS: This specific technique of subretinal microinjection is an easy, safe and optimal technique to meet the need of subretinal drug administration and retinal cells transplantation for retinal degenerative diseases. It is easier for the transplanted pure retinal photoreceptor cells to establish functional connection with host cells through subretinal microinjection.