The relationship between aquaporins and skin diseases.

Aquaporins (AQPs) are a family of transmembrane channel proteins that can rapidly transport water molecules. The main subtype expressed in the epidermis and dermis is AQP3. Studies have confirmed that AQPs exert certain physiological functions in the skin, such as the maintenance of normal shape, the regulation of body temperature, moisturization and hydration, anti-aging, damage repair and antigen presentation. The abnormal expression of AQPs in skin cells can lead to a variety of skin diseases. This review summarizes the relevance of AQPs in dermatophysiological and pathophysiological processes, highlighting their potential as new drug targets for the treatment of skin diseases.

Cold-Induced RNA-Binding Protein and RNA-Binding Motif Protein 3: Two RNA Molecular Chaperones Closely Related to Reproductive Development and Reproductive System Diseases.

Cold-induced RNA-binding protein (CIRP) and RNA-binding motif protein 3 (RBM3) have recently been reported to be involved in cold stress in mammals. These proteins are expressed at low levels in various normal cells, tissues, and organs but can be upregulated upon stimulation by multiple stressors. Studies have shown that CIRP and RBM3 are multifunctional RNA molecular chaperones with different biological functions in various physiological and pathophysiological processes, such as reproductive development, the inflammatory response, the immune response, nerve injury regulation, and tumorigenesis. This paper reviews recent studies on the structure, localization and correlation of CIRP and RBM3 with reproductive development and reproductive system diseases.

Engineering Escherichia coli to bind to cyanobacteria.

We engineered Escherichia coli cells to bind to cyanobacteria by heterologously producing and displaying lectins of the target cyanobacteria on their surface. To prove the efficacy of our approach, we tested this design on Microcystis aeruginosa with microvirin (Mvn), the lectin endogenously produced by this cyanobacterium. The coding sequence of Mvn was C-terminally fused to the ice nucleation protein NC (INPNC) gene and expressed in E. coli. Results showed that E. coli cells expressing the INPNC::Mvn fusion protein were able to bind to M. aeruginosa and the average number of E. coli cells bound to each cyanobacterial cell was enhanced 8-fold. Finally, a computational model was developed to simulate the binding reaction and help reconstruct the binding parameters. To our best knowledge, this is the first report on the binding of two organisms in liquid culture mediated by the surface display of lectins and it may serve as a novel approach to mediate microbial adhesion.

Promoting effect of licorice extract on spermatogonial proliferation and spermatocytes differentiation of neonatal mice in vitro.

Licorice (glycyrrhiza uralensis) is known as an herb with detoxication, and it has been widely used in clinical prescription of Oriental herbal medicine. Studies on the effects of licorice in the reproductive system were very rare, especially in spermatogenesis. In order to elucidate the effects of licorice on spermatogonial proliferation and spermatocyte differentiation during neonatal mice spermatogenesis, the organ culture model of testis tissue from neonatal C57BL/6N mice (born 6 d) was established. Then, in the presence of licorice extract (LE), the proliferation activity of spermatogonia was identified with the positive rate quantitative analysis of 5-bromo-2-deoxyuridine (BrdU) and anti-proliferating cell nuclear antigen (PCNA) antibody by immunohistochemical staining. The results showed that, compared to the control group, the percentage of positive cells by BrdU staining enhanced dramatically and that the expression of PCNA protein increased significantly in the spermatogonia from the LE group and showed a concentration-dependent manner (P < 0.05). This indicated that the LE can significantly promote the proliferation of spermatogonia in the spermatogenesis of neonatal mice. Furthermore, proteins related to spermatocyte differentiation, synaptonemal complex protein 3 (SCP3) and meiotic recombinant protein Spo11, were detected by immunohistochemical staining. The results showed that the differentiated spermatocyte in the LE group was significantly increased compared with that of the control group and showed a concentration-dependent manner (P < 0.05). The above results suggested that the LE can significantly accelerate the proliferation of spermatogonia and the differentiation of spermatocytes in the testicular tissue of the neonatal mice, which may be a potential drug for male infertility.

[Neuregulin promotes spermatogonia proliferation in mice].

OBJECTIVE: To explore the role of neuregulin (neural regulation of protein, NRG) in the process of mouse spermatogonia proliferation. METHODS: Mouse testis fragments were cultured in the medium DMEM containing purified NRG1beta or NRG3 at the concentration of 50, 100 and 200 ng/ml, respectively, followed by BrdU immunohistochemical staining and determination of the proliferation rate of spermatogonia. RESULTS: Compared with the control group, neuregulin significantly promoted the proliferation of spermatogonia (P < 0.05). The proliferation rates of spermatogonia cultured in the medium with 50, 100 and 200 ng/ml of NRG13 were 1.69, 1.55 and 1.86 times, and those with 50, 100 and 200 ng/ml of NRG3 were 1.35, 1.54 and 2.11 times that of the control. CONCLUSION: NRG1beta and NRG3 can promote the proliferation of mouse spermatogonia, and NRG is expected to be applied in the treatment of male infertility.

Phosphorylated SAP155, the spliceosomal component, is localized to chromatin in postnatal mouse testes.

SAP155 is an essential component of the spliceosome and its phosphorylation is required for splicing catalysis, but little is known concerning its expression and regulation during spermatogenesis in postnatal mouse testes. We report that SAP155 is ubiquitously expressed in nuclei of germ and Sertoli cells within the seminiferous tubules of 6- and 35-day postpartum (dpp) testes. Analyses by fractionation of testes revealed that (1) phosphorylated SAP155 was found in the fraction containing nuclear structures at 6 dpp in amounts much larger than that at other ages; (2) non-phosphorylated SAP155 was detected in the fraction containing nucleoplasm; and (3) phosphorylated SAP155 was preferentially associated with chromatin. Our findings suggest that the active spliceosome, containing phosphorylated SAP155, performs pre-mRNA splicing on chromatin concomitant with transcription during testicular development.

[Effect of the chemically assisted enucleation on the enucleation of sheep oocytes and the development of their reconstructed embryos].

In order to enhance the efficiency of sheep somatic cell nuclear transfer, we used a chemically assisted enucleation with colchicine to study the effects of the concentration of colchicine, the incubation time of oocytes in colchicine and the maturation time of oocytes on the enucleation rates and the development of reconstructed embryos. The results showed that 1) there were no significant differences in the rates of cytoplast protrusion and enucleation between oocytes that were incubated in colchicine (0.4 microg/mL) for 0.5 h and oocytes that were incubated in colchicine (0.4 microg/mL) for 1 h, and the rate of cytoplast protrusion can be 85.4% while the rate of cytoplast enucleation is 100%. 2) There was no significant difference in oocyte enucleation between oocytes treated with medium containing 0.2 microg/mL colchicine for 0.5 h and oocytes treated with medium containing 0.4 microg/mL colchicine for 0.5 h. 3) A maturation time of 18-23 h did not affect the rates of cytoplast protrusion and enucleation by chemically assisted enucleation, whereas the rate of enucleation of oocytes by blind enucleation was found to decrease with a prolonged incubation time. 4) The development rates of reconstructed embryos could not be influenced by these two enucleation methods, increased from oocytes matured for 21-23 h. These results demonstrate that sheep oocytes can be enucleated fast and effectively by optimized colcholine chemically assisted enucleation, which can enhance the enucleation rate of sheep oocytes and the early development of reconstructed embryos in vitro.

[Development and clinical application of male human papillomavirus genotyping by membrane DNA chip].

OBJECTIVE: To develop a new method for the detection of male human papillomavirus (HPV) genotypes and to investigate its clinical application value. METHODS: With computer assistance and based on the classical common primers MY09/11, modified PGMY09/11 with 23 HPV subtypes for PCR and Genbank data on HPV, we designed probes for the simultaneous detection of 18 high-risk subtypes (HPV-16, 18, 31, 33, 35, 39, 45, 51, 52, 53, 56, 58, 59, 66, 68, 73, 83 and MM4) and 5 low-risk subtypes (HPV-6, 11, 42, 43 and 44) and fixed them to the special membrane to make a DNA chip. A total of 112 male urethral samples were collected with swabs and studied for the clinical value. Meanwhile the single subtypes of HPV positive were sequenced and the standard samples detected for their sensitivity. RESULTS: Of the total number, 25 samples were found to be HPV positive, 13 single HPV infection and 12 multiple infection. Nine HPV gene subtypes were noted in the samples: 6, 11, 16, 18, 33, 35, 43, 56 and 73, with sensitivity up to 10 copies of HPV DNA. CONCLUSION: Human papillomavirus genotyping by the membrane DNA chip is applicable to the diagnosis of male HPV infection as well as to the related epidemic and etiological investigation.

Size-selective junctional barrier and Ca(2+)-independent cell adhesion in the testis of Cynops pyrrhogaster: expression and function of occludin.

In urodeles which has testicular structure different from that in mammals, blood-testis barrier was reported to exist like in mammals. However, molecular and functional analyses of the components of the blood-testis barrier in urodeles have not been reported yet. Toward elucidation of the barrier functions and their molecular components in newt testis, we aimed to isolate occludin cDNAs and obtained two kinds of occludin partial cDNAs (occludin 1 and 2) encoding the putative second extracellular loop. Immunoblot and immunofluorescence studies using antibodies against peptides each corresponding to a part of the second extracellular loop of occludin 1 and 2, and those against beta-catenin and zonula occludens-1 (ZO-1) showed that occludin, as well as beta-catenin and ZO-1, was expressed not only in Sertoli cells but also in germ cells throughout all the stages from spermatogonia to elongate spermatids. Tracer experiments revealed a size-selective barrier which allows small molecules ( approximately 500 Da) to get into cysts through Sertoli cells' barrier, but not larger ones (>1.9 kDa) in the stages from spermatogonia to almost mature sperm. No occludin peptides corresponding to a part of the second extracellular loop destroyed the junctional barrier, while both the peptides and antibodies significantly inhibited reaggregation of the dissociated testicular cells which was to a large extent Ca(2+)-independent. These results indicate that the second extracellular loop of occludin is involved in cell adhesion rather than in size-selective barrier in newt testis, though the possibility cannot be excluded that the peptides were not long enough to inhibit the barrier function.

Delivery of 125I-cobrotoxin after intranasal administration to the brain: a microdialysis study in freely moving rats.

In order to determine the contribution of intranasal (i.n.) administration to the uptake of large molecular weight (MW) substances into central nervous system (CNS), concentration in brain of the centrally acting polypeptide cobrotoxin (NT-I) versus time profiles were studied using dual-probe microdialysis in awake free-moving rats. NT-I, radiolabeled with sodium (125)I-Iodide ((125)I-NT-I), was administered at the dose of 105 microg/kg intravenously and intranasally in the same set of rat (n=15). The (125)I-NT-Inasal preparations were formulated with borneol/menthol eutectic mixture (+BMEM) as an absorption enhancer and without (-BMEM). After application, the dialysates sampled simultaneously from olfactory bulb and cerebellar nuclei were measured in a gamma-counter for radioactivity. The real concentrations of NT-I were recalculated by in vivo recoveries of microdialysis probes. The results showed that the area under the curve (AUC) value in cerebellar nuclei (2283.51+/-34.54 min ng/ml) following i.n. administration (+BMEM) was significantly larger than those (AUC(olfactory)=1141.92+/-26.42 min ng/ml; AUC(cerebellar)=1364.62+/-19.35 min ng/ml) after intravenous (i.v.) bolus, respectively. A prolonged time values to peak concentrations after i.n. application (+BMEM) were observed compared with those following i.v. administration. Also, following i.n. application (+BMEM) the measured time value to peak concentration in cerebellar nuclei (85 min) was statistically longer than that in olfactory bulb (75 min), which could be plausibly an indication for NT-I delivery into brain via nose-brain pathway in the presence of absorption enhancer. i.n. administration (-BMEM) had little or no ability of NT-I delivering into brain. In conclusion, i.n. administration (+BMEM) significantly enhanced brain transport of NT-I with uneven distribution in discrete regions of brain compared with i.v. administration. Additionally, multi-probe microdialysis technique should be considerably valuable in brain delivery studies.

HSP90beta is involved in signaling prolactin-induced apoptosis in newt testis.

We have shown in vivo and in vitro that prolactin induces apoptosis in the 7th generation of spermatogonia during newt spermatogenesis, but the underlying molecular mechanism remained unknown. To determine the role of heat shock protein (HSP) 90beta, a molecular chaperone for client proteins in signal transduction and transcriptional regulation, in prolactin-induced apoptosis, we cloned HSP90beta cDNA from newt testis. HSP90beta was detected highly at spermatogonial stage and in both the membrane and cytosol fractions only in germ cell-enriched fraction, while prolactin receptor was expressed in the membrane fraction of only germ cells. Co-immunoprecipitation demonstrated that HSP90beta associated constitutively with prolactin receptor on the plasma membrane of germ cells, suggesting that prolactin receptor is also one of the client proteins for HSP90beta. Inhibition of HSP90beta function by geldanamycin was shown to promote spermatogonial apoptosis. Taken together, these results suggest that HSP90beta is involved in signaling prolactin-induced apoptosis through the receptor.