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Competing Interests: All the authors are Novartis employees. The authors report no other conflict of interest in this work.

Background: Gentiana siphonantha Maxim. is a traditional medicine for the treatment of rheumarthritis, icterepatitis, pain and hypertension; it is rich in gentiopicroside with anti-inflammatory, antibacterial, and free radical-scavenging activities. This study was to evaluate the antidermatophyte activity of G. siphonantha on a guinea pig model in vitro and in vivo. Material andMethods: The antidermatophyte activities of 10 plants were tested by the broth microdilution method. Fractions and an extract of G. siphonantha were tested against Trichophyton mentagrophytes by the disc diffusion method. The morphological changes of T. mentagrophytes were observed. Component analysis of the n-butanol (n-BuOH) fraction was made by HPLC. Finally, the antifungal activity in an in vivo guinea pig model of dermatophytosis was examined., Results: G. siphonantha had strong antidermatophyte activity with MIC50 values of 32-64 μg/mL. The n-BuOH fraction of G. siphonantha showed the most potent activity compared to the other fractions. After being exposed to the n-BuOH fraction at 80 and 160 μg/mL, the hyphae were distorted and collapsed. Gentiopicroside is the main active ingredient in the n-BuOH fraction of G. siphonantha. The lesion scores of the guinea pig model of dermatophytosis significantly declined in the 10% and 30% extract and positive control groups in comparison with the untreated control group. Periodic acid-Schiff and hematoxylin/eosin staining displayed similar results., Conclusion: The n-BuOH fraction of G. siphonantha demonstrated antidermatophyte efficacy in experimental dermatophytosis., (© 2018 S. Karger AG, Basel.)

Objective: To study the pharmacokinetics and selectivity of 5-aminolevulinic acid (5-ALA) from 5-ALA thermolabile gel formulation in cervical intraepithelial neoplasia (CIN). The novel thermogel Pluronic F127 is liquid at cold temperatures and turns into a gel-like consistency at body temperature, thereby improving adhesion of 5-ALA to the cervix uteri., Methods: 27 female patients with CIN 1-3 were included in this study. Thirty minutes to 12 h before conisation, 10 mL of thermo-gel containing either 4, 10, or 20% of 5-ALA were topically applied to the cervix. Biopsies were taken from the lesions as well as normal surrounding epithelial tissue for histological examination, fluorescence microscopy and spectrometry., Results: The thermogel Pluronic F127 was easy to handle and proved reliable as vehicle carrier. 5-ALA induced porphyrin fluorescence was maximal after application of 10% 5-ALA thermogel. We observed the higher porphyrin fluorescence intensity within the CIN lesions (1,116 +/- 241 AU), as compared to normal adjacent epithelium (704 +/- 166 AU). This difference was statistically significant (p < 0.05). Four to six hours after application, porphyrin fluorescence in dysplastic epithelium reached maximal intensity, and tumor selectivity was the highest in CIN 3 achieving a tumor-to-normal ratio of 3.5. Compared to other studies employing 5-ALA, the thermogel preparation of 5-ALA leads to twofold increase in the porphyrin selectivity., Conclusions: The thermogel Pluronic F127 seems to improve the local cervical drug application. Based on these results, we recommend the application of a 10% 5-ALA thermogel formulation 4-6 h prior to photodynamic therapy in CIN.

Purpose: Gap junctions are intercellular contacts important for the synchronization of muscle cell activity through electrical coupling. Since the role of gap junctions for the function of smooth bladder muscle is still a matter of debate, we investigated the occurrence of gap junctions and the gap junctional protein connexin (Cx) 45 in the detrusor of the nonobstructed stable human bladder., Materials and Methods: Detrusor biopsies from 6 patients aged 64 (55-72) years with stable nonobstructed bladders were investigated for the occurrence of gap junctions by electron microscopy, molecular biological techniques and immunohistochemistry., Results: Transmission electron microscopy and freeze fracture showed the presence of gap junction at plasma membranes of detrusor smooth muscle cells. By reverse transcriptase (RT) polymerase chain reaction (PCR) and in situ hybridization, we found an expression of Cx45 in the detrusor. These data were confirmed by immunolocalization of Cx45 on smooth muscle cells., Conclusions: This study provides morphological as well as molecular biological and immunohistochemical evidence that bladder smooth muscle cells are electrically coupled.

In order to determine the concurrent expression of DR and CD antigen of T lymphocyte surface markers, the fluorescence-rosette double-labelling technique was developed by employing monoclonal antibody against HLA-DR and sensitized sheep erythrocytes with OKT series of monoclonal antibodies against cell surface antigen. Its reliability was evaluated by comparison with two-step isolating technique for determining DR antigen of activated T lymphocyte subsets in 30 healthy volunteers. The result showed there was no statistic difference between results determined with two methods. The double-labelling technique was characterized by its samplity, no injury for cellular activity and ability for analysing the expression of the single antigen and double antigens on cell surface concurrently.