Pemphigus is an autoimmune mucocutaneous blistering disease caused by autoantibodies against desmogleins. Rituximab effectively treats pemphigus by inducing remission and rapidly reducing corticosteroid dosage. In Korea, the high cost of rituximab had been a burden until the National Health Insurance began to cover 90% of rituximab costs via reimbursement for severe pemphigus patients. We analyzed 214 patients with pemphigus who were treated with their first round of rituximab. The time to initiate rituximab and the time to partial remission under minimal therapy (PRMT) were both significantly shorter after the rituximab reimbursement policy. The total steroid intake for PRMT and complete remission (CR) was less in patients who were diagnosed after the reimbursement. The interrupted time series (ITS) model, a novel analysis method to evaluate the effects of an intervention, showed a decrease in total systemic corticosteroid intake until PRMT after reimbursement began. In peripheral blood mononuclear cells from patients with pemphigus vulgaris, the relative frequencies of desmoglein 3-specific CD11c+CD27−IgD− atypical memory B cells positively correlated with the periods from disease onset to rituximab treatment and to PRMT and the total systemic corticosteroid intake until PRMT. We found that early rituximab therapy, induced by the reimbursement policy, shortened the disease course and reduced the total corticosteroid use by pemphigus patients. The decreased frequency of circulating desmoglein-specific atypical memory B cells can be used as a surrogate marker for a good prognosis after rituximab.

Purpose: This study was designed to determine whether a novel anti-melanogenic agent, PF3758309, has the potential to cause acute cutaneous irritation using a human skin equivalent model (HSEM). Methods: Human skin equivalent was constructed through incubation of normal human derived epidermal keratinocytes (HEKs) on collagen matrix insert with proliferation media. Using constructed human skin equivalent, the irritation potential of PF3758309 were investigated whether the viability of this agent-treated HESM is under 50% (irritant) or not (non-irritant) using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay after applying the agent to the epidermal surface of the HSEM. Also, the PF3758309mediated anti-pigmentation effects were analyzed using Fontana-Masson staining in the HSEM. Results: The integrity of the constructed HSEM was confirmed using immunohistochemical staining with differentiation markers of epidermis, and observed that Keratin 5, Keratin 1 and Involucrin were stained in basal, supra-basal and granular layers, respectively. In vitro irritation assay showed that the mean viabilities of the PF3758309 was 83.6%, 78.8% and 77.8% at the treatment doses of 0.2, 0.5 and 1 mg, respectively; however, the mean viability of the positive control (5% sodium dodecyl sulfate)-treated HESM was 2.8%. Also, in vitro corrosion assay showed that the mean viabilities of the PF3758309 was 95.3%, 95.0% and 94.2% at the treatment doses of 0.2, 0.5 and 1 mg, respectively. Furthermore, using a Fontana-Masson staining assay, the melanin levels in the PF3758309-treated HSEM was significantly decreased compared with the levels in control HSEM. Conclusion: The anti-melanogenic agent, PF3758309, has no skin irritation potential under the conditions used in this study.

Rehmannia glutinosa (Gaertn.) DC., belonging to the family Scrophulariaceae, is an important medicinal herb cultivated in East Asia. Traditionally, R. glutinosa is steam processed to increase its efficacy in treating various ailments such as diabetes, hematinic deficiencies and adrenal disorder. However, standardization of processed R. glutinosa is highly needed to increase its quality to fulfill global market demand that is safe and possess high level of efficacy. Therefore, this study aimed to optimize the R. glutinosa steam processing methods by evaluating some key parameters such as steaming temperature, number of steaming times, steaming duration, and additive supplementation. R. glutinosa samples were steam processed at different temperatures (100 °C, 110 °C, and 120 °C), various steaming times (1 to 5 times), several steaming duration (1 to 4 h), and additives supplementation (rice wine, 5% EtOH, 10% EtOH, 20% EtOH, 30% EtOH, and 40% EtOH). As the result, 2 h, 3 replications, and supplementation with 20% EtOH at 120 °C were identified as the optimal conditions for R. glutinosa steam processing. Optimized processed R. glutinosa (SPRR 20%EtOH) resulted in significantly higher content of 5-HMF (7648.60 ± 150.08 µg/g) and iso-verbacoside (203.80 ± 10.72 µg/g) compared with unprocessed R. glutinosa (UPR). Compared to those of other samples, SPRR 20% EtOH samples had higher total flavonoid (55.36 ± 1.68 mg/g) and phenolic (69.24 ± 4.56 mg/g) contents and stronger DPPH antioxidant activity (56%). Furthermore, SPRR 20% EtOH had excellent anti-inflammatory activity, as evidenced by the suppression of inducible nitric oxide synthase (iNOS) caused by activation of nuclear factor-κB (NF-κB) through p-p65 pathway in LPS-stimulated RAW 264.7 cells. These findings will provide a basis towards industrialization of R. glutinosa processing technology that will be very helpful for oriental medication field.

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