Salt glands play a pivotal role in the salt resistance of four recretohalophyte Limonium Mill. species.

Limonium Mill. plants are typical recretohalophytes, as they withstand salt stress by secreting excess salt onto the leaf surface through salt glands. However, little is known on the salinity thresholds of these plants and the function of salt glands in salt tolerance. Here, we investigated the salinity thresholds of salt tolerance of the Limonium species L. aureum (Linn.) Hill, L. gmelinii (Willd.) Kuntze, L. otolepis (Schrenk) Kuntze and L. sinuatum (L.) Mill grown with various concentrations of NaCl. The salinity thresholds of L. otolepis, L. aureum, L. sinuatum and L. gmelinii were 300, 350, 400 and 420 mm NaCl, respectively. Correlation analysis indicated that total dry weight, chlorophyll content and intercellular CO2 concentration were highly positively correlated with the total fresh weights of all four Limonium species and could therefore be used as indicators of plant salt tolerance. Furthermore, as the salt gland density on the leaf surface increased, the rate of salt secretion per salt gland also increased, allowing more Na+ to be secreted from the plant. Redundancy discriminant analysis indicated that salt gland density, Na+ content and Na+ secretion rate per salt gland were positively correlated with salt concentration. These observations support the notion that salt glands play important roles in the adaptation of Limonium species to high salinity conditions.

Berberine binds RXRα to suppress ß-catenin signaling in colon cancer cells.

Berberine, an isoquinoline alkaloid, is a traditional oriental medicine used to treat diarrhea and gastroenteritis. Recently, we reported that it could inhibit the growth of intestinal polyp in animals and in patients with the familial adenomatous polyposis by downregulating ß-catenin signaling. However, the intracellular target mediating the effects of berberine remains elusive. Here, we provide evidence that berberine inhibits ß-catenin function via directly binding to a unique region comprising residues Gln275, Arg316 and Arg371 in nuclear receptor retinoid X receptor alpha (RXRα), where berberine concomitantly binding to and synergistically activating RXRα with 9-cis-retinoic acid (9-cis-RA), a natural ligand binding to the classical ligand-binding pocket of RXRα. Berberine binding promotes RXRα interaction with nuclear ß-catenin, leading to c-Cbl mediated degradation of ß-catenin, and consequently inhibits the proliferation of colon cancer cells. Furthermore, berberine suppresses the growth of human colon carcinoma xenograft in nude mice in an RXRα-dependent manner. Together, our study not only identifies RXRα as a direct protein target for berberine but also dissects their binding mode and validates that berberine indeed suppresses ß-catenin signaling and cell growth in colon cancer via binding RXRα, which provide new strategies for the design of new RXRα-based antitumor agents and drug combinations.

cRGD peptide-installed epirubicin-loaded polymeric micelles for effective targeted therapy against brain tumors.

Current therapeutic strategies against glioblastoma multiforme (GBM) are futile mainly because of the poor access of drugs into malignant tissues, which is hindered by the tight blood-brain tumor barrier in the GBM vasculature. Nanomedicines have shown potential for circumventing the vascular barriers of GBM, particularly by targeting markers on the luminal side of endothelial cells in the blood vessels of GBM for achieving effective and selective translocation into the tumor. Thus, as the αvß3 and αvß5 integrins overexpressed on the endothelial cells of GBM can be targeted by cyclic-Arg-Gly-Asp (cRGD) peptide, herein, we developed cRGD-installed micellar nanomedicines loading epirubicin, the potent antiglioblastoma agent, through a pH-sensitive hydrazone-bond for effective treatment of GBM. These cRGD-installed epirubicin-loaded polymeric micelles (cRGD-Epi/m) achieved faster and higher penetration into U87MG cell-derived 3D-spheroids than the micelles without cRGD, conceivably through a cRGD-integrin mediated pathway. In vivo, the cRGD-installed micelles effectively suppressed the growth of an orthotopic GBM model by delivering high levels of epirubicin throughout the tumor tissue. These results indicate significant prospects for cRGD-Epi/m as an effective and translationable treatment against GBM.

The CELF1 RNA-Binding Protein Regulates Decay of Signal Recognition Particle mRNAs and Limits Secretion in Mouse Myoblasts.

We previously identified several mRNAs encoding components of the secretory pathway, including signal recognition particle (SRP) subunit mRNAs, among transcripts associated with the RNA-binding protein CELF1. Through immunoprecipitation of RNAs crosslinked to CELF1 in myoblasts and in vitro binding assays using recombinant CELF1, we now provide evidence that CELF1 directly binds the mRNAs encoding each of the subunits of the SRP. Furthermore, we determined the half-lives of the Srp transcripts in control and CELF1 knockdown myoblasts. Our results indicate CELF1 is a destabilizer of at least five of the six Srp transcripts and that the relative abundance of the SRP proteins is out of balance when CELF1 is depleted. CELF1 knockdown myoblasts exhibit altered secretion of a luciferase reporter protein and are impaired in their ability to migrate and close a wound, consistent with a defect in the secreted extracellular matrix. Importantly, similar defects in wound healing are observed when SRP subunit imbalance is induced by over-expression of SRP68. Our studies support the existence of an RNA regulon containing Srp mRNAs that is controlled by CELF1. One implication is that altered function of CELF1 in myotonic dystrophy may contribute to changes in the extracellular matrix of affected muscle through defects in secretion.

Hydroxychloroquine-conjugated gold nanoparticles for improved siRNA activity.

Current technology of siRNA delivery relies on pharmaceutical dosage forms to route maximal doses of siRNA to the tumor. However, this rationale does not address intracellular bottlenecks governing silencing activity. Here, we tested the impact of hydroxychloroquine conjugation on the intracellular fate and silencing activity of siRNA conjugated PEGylated gold nanoparticles. Addition of hydroxychloroquine improved endosomal escape and increased siRNA guide strand distribution to the RNA induced silencing complex (RISC), both crucial obstacles to the potency of siRNA. This modification significantly improved gene downregulation in cellulo. Altogether, our data suggest the benefit of this modification for the design of improved siRNA delivery systems.

Polymeric micelles incorporating (1,2-diaminocyclohexane)platinum (II) suppress the growth of orthotopic scirrhous gastric tumors and their lymph node metastasis.

Nano-scaled drug carriers have great potential for the treatment of solid tumors. Nevertheless, hypovascularity and fibrosis in some types of solid tumors have been demonstrated to reduce the penetration and accumulation of nano-scaled drug carriers. Diffuse-type scirrhous gastric cancers present such characteristics as well as frequent metastasis to the lymph nodes; therefore, it remains a great challenge to eradicate scirrhous gastric cancers based on the drug targeting using nanocarriers. Herein, we demonstrated that polymeric micelles with 30-nm diameter incorporating (1,2-diaminocyclohexane)platinum(II) (DACHPt), the parent complex of the anticancer drug oxaliplatin, efficiently penetrated and accumulated in an orthotopic scirrhous gastric cancer model, leading to the inhibition of the tumor growth. Moreover, the elevated localization of systemically injected DACHPt-loaded micelles in metastastic lymph nodes reduced the metastatic tumor growth. These results suggest DACHPt-loaded micelles as a promising nanocarrier for the treatment of scirrhous gastric cancers and their lymphatic metastases.

[Carcinoma of kidney collecting duct: an analysis of 10 cases].

OBJECTIVE: To study the clinicopathologic characteristics of renal collecting duct carcinoma (CDC). METHODS: A retrospective study was done in 10 cases of CDC. RESULTS: Among 466 cases of renal cell carcinoma admitted in our Institute between January 1989 and June 30, 1999, 10(2.1%) cases of CDC were identified. Seven presented with gross hematuria and 3 with abdominal pain. Radical nephrectomy was done in 9 patients, enucleation of tumor in one. The primary tumor was located predominantly in the renal medulla. Histologic examination showed prominent tubular or tubulopapillary structures. Sarcomatoid carcinoma, cystadenocarcinoma, nests and cords of tumor cells in desmoplastic stroma were identified in some cases. High molecular weight cytokeratin 34 beta E12 was positive in 8 cases and peanut agglutinin in 7 cases. According to Fuhrman's nuclear grade, one was in G2, 4 were in G3 and 5 in G4. Six patients died of metastases within 3 to 23 months (mean 13.3 months), one died of heart disease with tumor free after 19 months, two survived with tumor free for 14 months and 39 months, respectively, one lost from follow-up. CONCLUSION: CDC is a distinctive renal cell carcinoma with prominent clinical appearance and progressive clinical course.

Local regulation of immune responses: corneal endothelial cells alter t cell activation and cytokine production.

Corneal endothelial cells (CE cells) inhibit antigen- and mitogen-activated lymphocyte proliferation assays, although interleukin 2 receptor (IL-2R) expression and responsiveness to exogenous IL-2 are unaffected. To examine this activity further, co-cultures of CE cells and T cell clones were studied. CE cells inhibited IL-2 and IL-4 production by T cells stimulated with Ag and APC, but not IL-5 or IL-6 production. CE cells also inhibited NFAT-driven lacZ reporter gene production following Ag stimulation of transfected KZO T hybridoma cells. Conversely, stimulation of IL-2 production by ionomycin, with or without PMA, was unaffected by the CE cells. Preincubation of KZO hybridoma or Jurkat cells with CE cells, or CE cell-conditioned culture supernatant, inhibited the intracellular calcium ([Ca(2+)](i)) increase induced by TCR ligation, but not the [Ca(2+)](i)increase induced by ionomycin or thapsigargin. The inhibitory effect was independent of APC and did not act by blocking costimulation, since IL-2 production stimulated by immobilized anti-CD3 alone was also inhibited by CE cells. The supernatant factor was heat labile. This novel activity is unlike other immunoregulatory molecules, including transforming growth factor beta (TGF-beta) and may contribute to local immune privilege.

[Expression of VEGF and tumor angiogenesis in bladder cancer].

OBJECTIVE: To evaluate expression of vascular endothelial growth factor (VEGF) and its relation to angiogenesis in bladder cancer. METHODS: The expression of VEGF by immunohistochemical staining was examined in 68 cases of primary transitional carcinoma (TCC) and 7 subjects with normal urothelium. Microvessel density (MVD) in 40 cases of invasive bladder cancer was determined by Factor VIII immunohistochemical staining. RESULTS: The expression of VEGF was negative or low level in all normal bladder tissue but high in bladder cancer. VEGF and MVD were closely related to tumor cell nuclear grade and clinical stage. The average MVD was higher in the cases with the high level expression of VEGF, than in those with the low level expression of VEGF. CONCLUSIONS: This study indicates that VEGF is positively correlated with the occurrence and progression of bladder cancer. VEGF is one of the primary angiogenic factors for bladder cancer, and is able to induce tumor angiogenesis and accelerate tumor growth. Quantification of VEGF may provide a valuable marker for a poorer prognosis of patients.

[The effect of combined endocrine therapy on prostate and testis].

OBJECTIVE: To study the pathological changes of the effect of combined endocrine therapy on normal prostate, benign prostatic hyperplasia, prostatic adenocarcinoma and testis. METHODS: 11 radical prostatectomy specimens, 3 core-needle biopsies of prostate and 3 testes obtained after at least 3 months of enatone-flutamide inhibition therapy were studied. Step-section was performed on radical specimens and average 16 sections per case were reviewed. PSA, PSAP and AE1/AE3 were immunostained on 14 cases of prostate cancer and a comparative study pre- and post-treatment was made. RESULTS: No residual tumor was recognized in 2 cases. Such characteristic changes were found in 9 cases as prominent acinar atrophy, decreased ratio of acini to stroma, stromal fibrosis, squamous metaplasia of carcinoma, cytoplasmic vaculation, nuclear shrinkage, and nucleolar shrinkage. No apparent change was discovered in 3 cases. Secretory epithelial atrophy and basal cell hyperplasia were the popular change in BPH and normal prostate. The expressions of both PSA and PSAP were markedly reduced in prostate carcinoma and nonneoplastic glands. Pathological downstaging of the tumor was not found statistically. Epithelial atrophy also existed in seminal vesicles and Leydig cells of the testes after hormone therapy. CONCLUSIONS: Combined endocrine therapy results in histologically distinctive changes that can be found in both nonneoplastic and neoplastic prostate tissue. However, the drugs can not eradicate prostate cancer completely. Testis atrophy is the direct action of the therapy.

[Expression and significance of Fas/APO-1 and bcl-2 protein in renal cell carcinoma].

OBJECTIVE: To investigate the role of Fas/APO-1 and bcl-2 in occurrence and progression of renal cell carcinoma. METHOD: Immunohistochemistry method was used to detect the expression of Fas/APO-1 and bcl-2 protein in 35 cases of renal cell carcinoma tissues and 26 cases of normal renal tissues. RESULT: The expression rate of Fas/APO-1 protein in renal cell carcinoma tissues was 57.14%, significantly lower than that in normal renal tissues (84.62%) (P < 0.05), and the intensity of expression was also lower. The expression rate of bcl-2 protein reached 80.00%, much higher than that in normal renal tissues (53.85%) (P < 0.05). CONCLUSION: Both Fas/APO-1 and bcl-2 take part in the occurrence and progression of renal cell carcinoma.