Effects of CD4+ T lymphocytes from ovariectomized mice on bone marrow mesenchymal stem cell proliferation and osteogenic differentiation.

The present study was designed to investigate the effects of T cells on the proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMMSCs). BMMSCs were co-cultured with CD4+ T cells that had been pretreated with anti-TNF-α or controls and were derived from ovariectomized (OVX) mice or sham control mice. MTT was used to assess the proliferative ability of BMMSCs and flow cytometry was used to analyze the BMMSC cell cycle. Following the induction of osteogenic differentiation in BMMSCs, calcium nodules were observed using alizarin red staining and alkaline phosphatase (ALP) staining. The expression levels of the osteogenesis-associated genes, runt related transcription factor 2 (Runx2) and osteocalcin (OCN) in BMMSCs were quantified using reverse transcription-quantitative PCR and western blotting. Osteogenesis-related signaling pathways, including ERK, JNK and p38 MAPK were also examined by western blotting. BMMSCs co-cultured with CD4+ T cells from OVX mice exhibited reduced proliferative ability compared with sham mice and the cell cycle was arrested at the G2/M phase. Additionally, BMMSCs co-cultured with CD4+ T cells from OVX mice presented with reduced levels of osteogenic differentiation and lower ALP activity, less calcium deposition and reduced expression of Runx2 and OCN compared with sham mice. The reduced levels of proliferation and osteogenic differentiation of BMMSCs induced by CD4+ T cells were not seen when the T cells were had been pretreated with anti-TNF-α. The results indicated that CD4+ T cells from OVX mice inhibited the proliferation and osteogenic differentiation of BMMSCs by producing high levels of TNF-α and may provide a novel insight into the dysfunction of BMMSCs caused by estrogen deficiency.

Two Nanometer-Sized High-Nuclearity Homometallic Bromide Clusters (M26Br38)12- (M = Cu, Ag): Syntheses, Crystal Structures, and Efficient Adsorption Properties.

The spontaneous formation of discrete spherical nanosized molecules is ubiquitous in nature; however, the actual structural imitation of such high-symmetry polyhedra from the edge sharing of regular polygons has still proved to be elusive. Herein, two high-nuclearity metal clusters, namely (TTB)4·M26Br38 (M = Cu (1), Ag (2), TTB·Br3 = 1, 3, 5-tris(triethylammoniomethyl)benzene tribromide), have been rationally and solvothermally synthesized and structurally characterized. Single-crystal X-ray analysis confirmed that 1 has I4̅3m symmetry with a (Cu25Br34)12- anion shell enwrapping a CuBr4 tetrahedron and 2 has I4̅3m symmetry with a [Ag26Br34]12- anion shell enwrapping a Br4 pyramid. The diffuse-reflectance UV-vis measurements showed that both compounds possess proper semiconductor behaviors with tunable band gaps of 1.87 eV for 1 and 1.90 eV for 2. Interestingly, all the samples feature distinct adsorption speed and compound 1 shows good adsorption activity for methyl orange (MO) under the same reaction conditions. The effects of pH, temperature, and cyclicity on dye adsorption, together with the thermal stabilities and luminescence properties of the compounds, were also studied. From the cyclic adsorption of compound 1 to the anionic dye MO, it was found that the adsorption of MO was good over three cycles, and the third adsorption rate was still 93.90%.

Spontaneous Remission of Subcutaneous Scedosporiosis Caused by Scedosporium dehoogii in a Psoriatic Patient.

To date, only one case of post-traumatic endophthalmitis caused by Scedosporium dehoogii has been reported, but its contamination or colonization might not be precluded due to the absence of pathogenic isolation and/or pathological examination. We report the first case to our knowledge of S. dehoogii-induced subcutaneous scedosporiosis in a psoriatic patient. A 58-year-old man with 5-year history of psoriasis vulgaris and immunosuppressant therapy developed pyrexia and multiple subcutaneous abscesses on both knees. Direct microscopy of the yellowish pus showed masses of bright green short spores. Skin biopsy revealed some branched septate hyphae within the granuloma. Two aspirated pus specimens collected at a 1-week interval produced white cottony colonies on Sabouraud dextrose agar. Bacterial cultures of one blood and two purulent samples were negative, and fungal culture of blood sample was not performed. The isolate was identified as S. dehoogii using ß-tubulin phylogeny and species-specific PCR with primer MSDE1/MSA2. Without addition of antifungal treatment, subcutaneous lesions disappeared spontaneously after immunosuppressant withdrawal and no relapse occurred during 64-month follow-up. The spontaneous recovery may result from immune reconstitution following immunosuppressant discontinuation.

Kerion and Tinea Corporis Caused by Rabbit-Derived Trichophyton interdigitale in Three Siblings and One Consulting Doctor Using ß-Tubulin Gene to Identify the Pathogen.

Trichophyton interdigitale is generally deemed as an anamorph of Arthroderma vanbreuseghemii based on internal transcribed spacer (ITS) sequencing, but recently their anamorph/teleomorph connection should be cautioned based on ß-tubulin phylogeny. We report three siblings and one consulting doctor who developed kerion and tinea corporis after contact with domestic rabbits. Seven same strains were isolated from four patients and three regions of a sick rabbit. The ITS and D1/D2 sequences of our isolate were 99 % homologous to A. Vanbreuseghemii, while ß-tubulin sequence was 100 % identical to T. interdigitale. Our isolate was identified as T. interdigitale based on maximum likelihood analysis of ß-tubulin. Random amplified polymorphic DNA revealed that the band patterns of five isolated strains and another rabbit-derived strain WCH023 were identical for OPF-03 and OPF-12. Skin lesions of all patients resolved completely for 2- to 6-week therapy of oral terbinafine and topical 1 % bifonazole or 1 % terbinafine cream. This study demonstrates that T. interdigitale of rabbit origin can cause various types of human dermatophytosis by mild scratch. Terbinafine may be the first choice for dermatophytosis caused by T. interdigitale.

Cutaneous Chromoblastomycosis Caused by Veronaea botryosa in a Patient with Pemphigus Vulgaris and Review of Published Reports.

Chromoblastomycosis and phaeohyphomycosis represent two poles of a disease spectrum caused by melanized fungi. Veronaea botryosa belongs to a small genus of saprobic fungi that occasionally cause human infections. To date, 11 cases of V. botryosa-induced cutaneous phaeohyphomycosis have been actually reported since 1990 after exclusion of 2 duplicated cases. We report the first case to our knowledge of cutaneous chromoblastomycosis caused by V. botryosa in a patient with pemphigus vulgaris. A 61-year-old man with 5-year history of pemphigus vulgaris and long-term treatment of corticosteroids and immunosuppressive agents developed multiple nodules on the dorsum of right wrist and hand after wrist trauma. Skin biopsy showed numerous brown muriform cells and a few septate hyphae in the tissue. Veronaea botryosa was isolated from the biopsy samples and then identified based on morphologic observation and DNA sequencing. The patient underwent immediate withdrawal of cyclophosphamide and gradual decrease in prednisone. Skin lesions healed after 5-month itraconazole therapy with an interval of 1-month terbinafine and one cycle of liquid nitrogen cryotherapy. Our results demonstrate that V. botryosa could induce both chromoblastomycosis and phaeohyphomycosis. Combined use of itraconazole and cryotherapy may be preferable to treat this infection.

Comparison of modified Chicago sky blue stain and potassium hydroxide mount for the diagnosis of dermatomycoses and onychomycoses.

The diagnostic value of modified Chicago sky blue (CSB) stain and potassium hydroxide (KOH) mount for superficial mycoses was compared using fungal culture as gold standard. The sensitivity and screening time of the CSB stain were superior to the KOH mount. The CBS stain is simple, quick and reliable for diagnosing superficial mycoses.

Molecular identification and antifungal susceptibility of 186 Candida isolates from vulvovaginal candidiasis in southern China.

There is limited information regarding the molecular epidemiology and antifungal susceptibilities of Candida isolates using the Neo-Sensitabs method in patients with vulvovaginal candidiasis (VVC). From August 2012 to March 2013, 301 non-pregnant patients aged 18-50 years with suspected VVC were prospectively screened at a teaching hospital in southern China. The vaginal isolates were identified by DNA sequencing of internal transcribed spacer and the D1/D2 domain. Antifungal susceptibility testing of seven antifungal agents was performed using the Neo-Sensitabs tablet diffusion method. Candida species were isolated from 186 cases (61.79 %). The most common pathogen was Candida albicans (91.4 %), followed by Candida glabrata (4.3 %), Candida tropicalis (3.2 %) and Candida parapsilosis (1.1 %). The susceptibility rates to C. albicans were higher for caspofungin, voriconazole and fluconazole than those for itraconazole, miconazole, ketoconazole and terbinafine (P<0.01). The resistance rates to C. albicans were 4.7, 6.5, 7.1, 7.6, 12.3, 27.7 and 74.7 % for caspofungin, miconazole, itraconazole, voriconazole, fluconazole, ketoconazole and terbinafine, respectively. No drugs tested apart from fluconazole exhibited differences in resistance between C. albicans and non-albicans Candida isolates. The results demonstrate that, using DNA sequencing, C. albicans is the most common isolate from Chinese patients with VVC. Caspofungin, voriconazole and fluconazole may be preferable to other azoles and terbinafine in the treatment of VVC.

PKG and NHR-49 signalling co-ordinately regulate short-term fasting-induced lysosomal lipid accumulation in C. elegans.

Lysosomes act as terminal degradation organelles to hydrolyse macromolecules derived from both the extracellular space and the cytoplasm. In Caenorhabditis elegans fasting induces the lysosomal compartment to expand. However, the molecular and cellular mechanisms for this stress response remain largely unclear. In the present study, we find that short-term fasting leads to increased accumulation of polar lipids in lysosomes. The fasting response is co-ordinately regulated by EGL-4, the C. elegans PKG (protein kinase G) orthologue, and nuclear hormone receptor NHR-49. Further results demonstrate that EGL-4 acts in sensory neurons to enhance lysosomal lipid accumulation through inhibiting the DAF-3/SMAD pathway, whereas NHR-49 acts in intestine to inhibit lipids accumulation via activation of IPLA-2 (intracellular membrane-associated calcium-independent phospholipase A2) in cytoplasm and other hydrolases in lysosomes. Remarkably, the lysosomal lipid accumulation is independent of autophagy and RAB-7-mediated endocytosis. Taken together, our results reveal a new mechanism for lysosomal lipid metabolism during the stress response, which may provide new clues for investigations of lysosome function in energy homoeostasis.

[Construction of a red fluorescent shuttle vector controlled by recA operon promoter of Streptococcus mutans].

OBJECTIVE: To construct a red fluorescent shuttle vector controlled by recA operon promoter to transform Streptococcus mutans. METHODS: The promoter of recA was amplified from Streptococcus mutans UA159, and connected to plasmid pDsRed2-N1 to construct pRred with a red fluorescent coding gene, which was then inserted into the shuttle vector pDL276 to construct pLRred. RESULTS: pLRred was successfully constructed, and Escherichia coli transformed with the pLRred plasmid could express reporter gene DsRed. CONCLUSIONS: The recombination plasmid pLRred can be used in the further research of the expression of cariogenic virulence factor gene by Streptococcus mutans in biofilm.

Brain abscess caused by Cladophialophora bantiana in China.

A case of a 38-year-old male farmer with a brain abscess caused by Cladophialophora bantiana is described. He had a 2 year history of non-insulin-dependent diabetes and myelodysplastic syndrome. A cranial computed tomography scan demonstrated a hypodense ring lesion with peripheral oedema and a midline shift in the left frontal lobe. A darkly pigmented mould was isolated from the brain abscess. The isolate was identified as C. bantiana based on its morphological features and DNA sequence analysis. The patient was unresponsive to burr hole aspiration and irrigation, as well as liposomal amphotericin B infusion, and died after discharge from the hospital. This is believed to be the first case of a cerebral abscess due to C. bantiana in China.

Experimental pathogenicity of a clinical isolate of Trichosporon dermatis in a murine model.

The pathogenicity of Trichosporon dermatis isolated from skin lesions of a patient has been examined in mice. Balb/c mice were treated with two intraperitoneal injections of 100 mg/kg cyclophosphamide on days 4 and 1 and one subcutaneous injection of 10 mg/kg dexamethasone on day 1 pre-inoculation, and then challenged with 0.2 ml T. dermatis inoculum (1 × 10(8) CFU/ml) by topical application on an abrasive wound in the dermabrasive group and by hypodermic injection in the subcutaneous group. In the intravenous group, 0.2 ml of high (1 × 10(8) CFU/ml) or low (1 × 10(7 )CFU/ml) inoculum was injected into the tail vein. Histopathology and inverse fungal culture were performed on the skin lesion and viscera, and renal fungal burden was also determined. Inoculated sites developed localized infections after dermabrasive and subcutaneous challenge in all mice, but the maximum area of skin lesions, and number of positive cultures from the lesions, were higher for immunocompromised mice. In the intravenous group, all immunocompetent animals survived during the four-week period, whereas 100 and 70% of immunocompromised animals died by 3 and 5 days in the high and low-inoculum groups, respectively. The incidence of disseminated infection and the renal fungal burden of immunocompromised mice were higher than those of immunocompetent mice. Our results demonstrate that subcutaneous and intravenous injection of T. dermatis can successfully establish cutaneous and systemic infection models in immunocompromised mice, with the kidney and lung being most susceptible.

Cutaneous phaeohyphomycosis of foot caused by Curvularia clavata.

A case of cutaneous lesions on the left foot caused by Curvularia clavata in a 64-year-old immunocompetent man is described. Fungal elements were found in the exudate and biopsy specimen. The isolate was identified as C. clavata based on its colonial and microscopic morphology in pure culture. The skin lesions healed after a 12-week regimen with oral fluconazole. This is the second case of cutaneous phaeohyphomycosis caused by C. clavata.

Biased genotype distributions of Candida albicans strains associated with vulvovaginal candidosis and candidal balanoposthitis in China.

BACKGROUND: Vulvovaginal candidosis (VVC), which is most frequently caused by Candida albicans, is one of the most common vaginal infections and is a common problem worldwide. Despite the fact that extensive epidemiological studies have been performed, what triggers VVC, especially recurrence of the infection, is still uncertain. METHODS: Genotypes of C. albicans strains associated with VVC and balanoposthitis and of strains isolated from samples from vaginas of asymptomatic women and from various extragenital sites were determined with use of C. albicans microsatellite locus I polymorphism analysis. Genetic similarity of representative strains with the same and different C. albicans microsatellite locus I genotypes were examined by sequence analysis of housekeeping genes CaADP1, CaSYA1, and CaVPS13. RESULTS: The C. albicans microsatellite locus I genotypes of independent C. albicans strains isolated from samples from extragenital sites were mostly of individual specificity. In contrast, strains associated with VVC were mainly concentrated to a few genotypes, with genotypes 30-45 and 32-46 being the most common. The overall frequencies of the 2 genotypes among C. albicans strains from vaginal samples from patients with VVC and from asymptomatic women were 59.1% and 24.0%, respectively (P = .002); the frequencies among patients with complicated VVC and among patients with uncomplicated VVC were 69.2% and 35.7%, respectively (P = .003). A similar genotype distribution pattern of C. albicans strains associated with balanoposthitis was also revealed. The genetic similarity of strains with the dominant genotypes associated with both VVC and balanoposthitis was confirmed by sequence analysis of the 3 genes. CONCLUSIONS: The results suggest the existence of vaginopathic C. albicans strains with enhanced virulence and tropism for the vagina and the high possibility of sexual transmission of genital C. albicans infection. Identification of specific genotypes that correlate with severity of VVC is also of diagnostic and therapeutic significance.

Granulomatous skin infection caused by Malassezia pachydermatis in a dog owner.

BACKGROUND: Malassezia pachydermatis is part of the normal cutaneous microflora of dogs and many other mammals. M pachydermatis has not yet been reported as an agent that causes skin infection in humans, although it has been found to cause fungemia and other nosocomial infections in preterm newborns and immunocompromised adults. OBSERVATIONS: Malassezia pachydermatis was isolated from the facial granuloma of a healthy woman and her dog's skin scrapings and cerumen. The yeast identity was established by standard methods and scanning electron microscopy. A skin biopsy specimen showed chronic inflammatory granuloma, numerous purple-red round or ovoid spores in the superficial necrotic tissue, and sparse red spores in the dermis. The skin lesions healed after oral fluconazole and cryotherapy. CONCLUSIONS: Definite diagnosis of M pachydermatis-induced skin infection principally depends on the results of fungal culture and histologic examination, and the combination of oral fluconazole and adjunctive cryotherapy seems to be an effective therapeutic regimen.