[Long-pulsed 1064 nm Nd: YAG laser combined with terbinafine against chromoblastomycosis caused by Fonsecaea nubica and the effect of laser therapy in a Wistar rat model].

We report a case of chromoblastomycosis caused by Fonsecaea nubica, which was successfully treated by long-pulsed 1064 nm Nd: YAG laser combined with terbinafine. A 60-year-old man was admitted for the presence of a 30 mm×40 mm erythematous plaque on the dorsum of his right hand for about 10 months without any subjective symptoms. Both microscopic examination and tissue biopsy of the lesion showed characteristic sclerotic bodies of chromoblastomycosis. Lesion tissue culture on SDA at 26 ℃ for 2 weeks resulted in a black colony, and slide culture identified the isolate as Fonsecaea species. ITS sequence analysis of the isolate showed a 99% homology with F. nubica strain KX078407. The in vitro susceptibility of the isolate to 9 antifungal agents was determined using the microdilution method according to the guidelines of CLSI M38-A2 protocol, and terbinafine showed the lowest MIC (0.125 µg/ml). We subsequently established a Wistar rat model of chromoblastomycosis using the clinical isolate F. nubica and treated the rats with long-pulsed 1064 nm Nd: YAG laser (pulse width of 3.0 ms, fluence of 24 J/cm2, spot size of 3 mm, frequency of 4 Hz, repeated 3 times at an interval of 30 s) twice a week for a total of 8 sessions. Although the laser treatment alone was not able to eliminate the fungi, histopathological examination showed the aggregation of numerous lymphocytes in the local affected tissue, indicating an immune response that consequently facilitate the regression of the lesion. The patient was successfully treated by long-pulsed 1064 nm Nd: YAG laser once a week combined with terbinafine (0.25 /bid) for 8 weeks, and follow-up for 20 months did not reveal any signs of recurrence.

[Effect of 0.9-ms 1064-nm Nd:YAG laser combined with itraconazole for treatment of toenail onychomycosis].

OBJECTIVE: To evaluate the efficacy and safety of 0.9-ms 1064-nm Nd:YAG laser alone or combined with itraconazole for treatment of toenail onychomycosis. METHODS: A total of 37 patients with onychomycosis (178 toenails) were randomly assigned to groups A and B, and each group was further divided into different subgroups according to the Scoring Clinical Index of Onychomycosis (SCIO) and Onychomycosis Severity Index (OSI) scoring. All the patients were treated with 0.9-ms Nd:YAG laser once a week for 8 times. The patients in group A were treated with laser alone, and those in group B were treated with laser combined with itraconazole. The clinical effect, clinical scores, appearance of the toenails and adverse reactions in the two groups were analyzed, and the patients' satisfaction rate was also investigated. RESULTS: At the 12th months of follow-up, the clinical response rate and mycological cure rate in group A were 31.33% and 30.00%, respectively, similar to the rates in group B (35.79% and 41.18%, respectively) (P>0.05). After the treatments, the SCIO and OSI scores showed no significant changes in group A (P>0.05) but both increased significantly in group B (P<0.05). The response rates did not differ significantly among the subgroups with SCIO<12 or with OSI<16 (P>0.05), but showed significant differences among the subgroups with SCIO≥12 or with OSI≥16 (P<0.05). Of the total of 178 toenails, 33.71%, 74.72% and 70.79% toenails showed improvements in terms of clear nail growth, shape and color, respectively. The overall patients' satisfaction rate was 62.16%, and no adverse reactions related with the therapy were recorded in these patients. CONCLUSION: For treatment of toenail onychomycosis, 0.9-ms 1064-nm Nd:YAG laser can effectively improve the aesthetic appearance of the toenails, and a combined treatment with Nd:YAG laser and itraconazole can be better option in severe cases of onychomycosis.

[Expression and characterization of porcine epidermal growth factor in Lactobacillus plantarum].

Epidermal growth factor (EGF) is an epithelial cell growth factor that can stimulate intestinal development, repair the damage of epidermal cells as well as reduce the incidence of pathogen infection and diarrhea. In order to produce a recombinant Lactobacillus plantarum (L. plantarum) expressing porcine epidermal growth factor (pEGF), we constructed a recombinant vector stably expressing pEGF in L. plantarum strains. First, L. plantarum strain Lp-1 was isolated from intestinal contents of piglets. Then the functional domain of pEGF, M6 precursor protein signal peptide (SP) and super strong constitutive promoter (SCP) were connected with the backbone plasmid pIAß8 to construct the recombinant vector that was transformed into Lp-1 by electroporation. Afterwards, pEGF was expressed in Lp-1 and detected by Tricine-SDS-PAGE and ELISA. After orally irrigated early-weaned BALB/c mice with the recombinant L. plantarum every morning and late afternoon for 10 consecutive days, body weight, villous height and crypt depth in the intestine were measured to examine the influence of the recombinant bacteria on the intestinal development of early-weaned mice in vivo. Finally, the results of our experiments demonstrated that pEGF was successfully expressed in Lp-1 and the molecular weight of pEGF was 6 kDa. In addition, the recombinant pEGF can enhanced the daily gain and exerted significance influence (P < 0.05) to the small intestinal morphology of early-weaned BALB/c mice. In conclusion, pEGF could be expressed in L. plantarum and the recombinant pEGF possesses good biological activity.

Production and immunogenicity of VP2 protein of porcine parvovirus expressed in Pichia pastoris.

Viral protein 2 (VP2) of porcine parvovirus (PPV) is the major viral structural protein and is responsible for eliciting neutralizing antibodies in immunized animals. In this study, we constructed and characterized a recombinant yeast vector encoding the VP2 protein, designated as pGAPZαA-VP2. The construct was confirmed by restriction enzyme digestion, PCR, and sequencing and then introduced into P. pastoris strain SMD1168 by electroporation. The expressed VP2 protein was analyzed by SDS-PAGE and western blot. Immunization of mice with the VP2 protein elicited a PPV-specific humoral immune response. Notably, a preparation of VP2 protein containing adjuvant induced a much better antibody response than VP2 alone. Clearly, the adjuvant strongly enhanced the immunogenicity of VP2. This study provides a foundation for the application of the VP2 protein in the clinical diagnosis of PPV and in vaccination against PPV in the future.