Tanshinone IIA alleviates vitiligo by suppressing AKT mediated CD8+ T cells activation in a mouse model.

Tanshinone IIA has been reported to exhibit anti-inflammatory effects, while it is not clear whether Tanshinone IIA has protective role in vitiligo. Premelanosome (PMEL) CD8+ T cells were adoptive transferred into Krt14- Kitl* mice with Kit ligand (KITL) over-expressed, to construct the vitiligo model. Pdk1fl/fl and Stat3fl/fl mice were crossed with Cd8cre mice to establish Pdk1TKO and Stat3TKO mice. Tanshinone IIA (200 µg) was intravenous injected to treat vitiligo in mice every 3 days. The accumulation of macrophages and CD8+ T cells in the ear skin was assayed by flow cytometry. Bone marrow-derived macrophages (BMDMs) were induced and stimulated with lipopolysaccharides (LPS) and IL-4. It was found that Tanshinone IIA alleviated the development of vitiligo, impaired PMEL CD8+ T cells accumulation in the ear skin, and inhibited LPS-induced TNF-α, IL-6, and IL-1ß expression and secretion in BMDMs, which could also inhibit IL-4-induced Arg-1 and Mrc-1 expression in BMDMs. In addition, Tanshinone IIA could inhibit the proliferation and cytotoxic function of CD8+ T cells indicated by the expression of Perforin, Granzymeb, and IFN-γ. Furthermore, Tanshinone IIA treated Pdk1TKO mice, not Stat3TKO mice, showed impaired PMEL CD8+ T cells accumulation in the ear skin. In summary, Tanshinone IIA alleviates vitiligo development with impaired CD8+ T cells accumulation and activation of Pdk1-Akt pathway.

Clinical outcomes of a combined arthroscopic and mini-open Outerbridge-Kashiwagi procedure for elbow osteoarthritis.

BACKGROUND: To evaluate the short-term clinical outcomes of a modified Outerbridge-Kashiwagi (O-K) procedure in the treatment of elbow osteoarthritis. METHODS: Between January 2012 and December 2016, 27 patients with elbow osteoarthritis were treated with a modified O-K procedure combining mini-open and arthroscopic technique in our institution. All patients with primary osteoarthritis and post-traumatic degenerative osteoarthritis of the elbow were included in the study if they had undergone the modified O-K procedure. Clinical outcomes were assessed using the visual analog scale (VAS), degree of flexion, extension loss, arc of motion, Mayo Elbow Performance Score (MEPS), and radiographs. RESULTS: Twenty-five patients with a mean age of 47.2 years (range, 21-69 years) at surgery were followed up for a mean of 54.5 months (range, 27-86 months). The VAS improved from 8.0 ± 1.4 (range, 6-10) preoperatively to 1.3 ± 1.1 (range, 0-3) at the final follow-up (P < .001), degree of flexion from 115.2° ± 12.0° (range, 90°-135°) to 130.6° ± 6.3° (range, 120°-140°) (P < .001), extension loss from 31.2° ± 15.0° (range, 10°-60°) to 10.2° ± 7.7° (range, 0°-30°) (P < .001), arc of motion from 84.0° ± 18.8° (range, 55°-120°) to 120.4° ± 9.3° (range, 105°-135°) (P < .001), and MEPS from 55.8 ± 8.1 (range, 40-70) to 88.4 ± 7.2 (range, 70-100) (P < .001). Radiographs at the final follow-up showed that 9 patients (36%) had significant recurrence of bone formation within the fenestration of the olecranon fossa. One patient developed delayed-onset ulnar neuropathy, with only slight numbness in the ulnar nerve distribution 6 months after surgery. CONCLUSIONS: The modified O-K procedure is safe and effective in pain relief and function restoration in patients with elbow osteoarthritis.

Systemic contact dermatitis in a family: Case report.

BACKGROUND: Systemic contact dermatitis (SCD) is an allergic inflammatory skin disease. We report that 3 family members developed SCD after exposing to laundry detergent containing benzalkonium chloride, which is rare. SCD caused by benzalkonium chloride has been reported. However, Similar symptoms in the whole family caused by it have not been reported yet. In our case, a 36-year-old man was diagnosed as SCD, and his symptoms had not controlled after 7 days treatment, until he stopped dressing the clothes washed by the laundry detergent containing benzalkonium chloride. It was interesting that both his wife and the daughter developed SCD successively, and they have not exposed to any haptens besides the benzalkonium chloride in the laundry detergent. METHODS: Dermoscopic examination showed bright-red background, focal branching vessels and white scales. HE staining from the lesion revealed hyperkeratosis and parakeratosis, focal subcorneal microabscess, ocal hyperkeratosis, koilocyte in the epidermis, and erythrocyte extravasation, fibroplasia, hyaline degeneration and scattered aggregates of lymphocytes in the dermis. Then path test was performed 1 month after recovery with benzalkonium chloride 0.05% and 0.1% in petrolatum. RESULTS: Stop the laundry detergent containing benzalkonium chloride. The symptoms had controlled after they stopped the laundry detergent containing benzalkonium chloride. CONCLUSION: The case highlights that benzalkonium chloride with very low concentration and repeated exposure may be an active agent of SCD. It is of the utmost importance to pay close attention to patients presenting with similar symptoms within the family. A thorough examination of the medical history is essential to determine the underlying cause.

Study on the Effect of Different Concentrations of SO2 on the Volatile Aroma Components of 'Beibinghong' Ice Wine.

SO2 plays an important role in wine fermentation, and its effects on wine aroma are complex and diverse. In order to investigate the effects of different SO2 additions on the fermentation process, quality, and flavor of 'Beibinghong' ice wine, we fermented 'Beibinghong' picked in 2019. We examined the fermentation rate, basic physicochemical properties, and volatile aroma compound concentrations of 'Beibinghong' ice wine under different SO2 additions and constructed a fingerprint of volatile compounds in ice wine. The results showed that 44 typical volatile compounds in 'Beibinghong' ice wine were identified and quantified. The OAV and VIP values were calculated using the threshold values of each volatile compound, and t the effect of SO2 on the volatile compounds of 'Beibinghong' ice wine might be related to five aroma compounds: ethyl butyrate, ethyl propionate, ethyl 3-methyl butyrate-M, ethyl 3-methyl butyrate-D, and 3-methyl butyraldehyde. Tasting of 'Beibinghong' ice wine at different SO2 additions revealed that the overall flavor of 'Beibinghong' ice wine was the highest at an SO2 addition level of 30 mg/L. An SO2 addition level of 30 mg/L was the optimal addition level. The results of this study are of great significance for understanding the effect of SO2 on the fermentation of 'Beibinghong' ice wine.

Analysis of Lignan Content and Rhizosphere Microbial Diversity of Schisandra chinensis (Turcz.) Baill. Resources.

Genetic and environmental factors influence the growth and quality of medicinal plants. In recent years, rhizosphere microorganisms have also emerged as significant factors affecting the quality of medicinal plants. This study aimed to identify Schisandra resources with high lignan content and analyze the microbial diversity of the rhizosphere soil. High-performance liquid chromatography was used to measure the lignan content in nine Schisandra fruits. High-throughput sequencing was used to analyze the 16S rDNA sequences of rhizosphere bacteria to identify bacterial species diversity. The total lignan content of the nine Schisandra resources ranged from 9.726 mg/g to 14.031 mg/g, with ZJ27 having the highest content and ZJ25 the lowest. Among the six lignan components, Schisandrol A had the highest content, ranging from 5.133 mg/g to 6.345 mg/g, with a significant difference between ZJ25, ZJ27, and other resources (p < 0.05). Schizandrin C had the lowest content, ranging from 0.062 mg/g to 0.419 mg/g, with more significant differences among the resources. A total of 903,933 sequences were obtained from the rhizosphere soil of the nine Schisandra resources, clustered into 10,437 OTUs at a 97% similarity level. The dominant bacterial phyla were Actinobacteriota, Acidobacteriota, Proteobacteria, Chloroflexi, Gemmatimonadota, and Verrucomicrobiota. The dominant bacterial genera were Candidatus_Udaeobacter, Candidatus_Solibacter, RB41, Bradyrhizobium, Gaiella, and Arthrobacter. ZJ27 is the Schisandra resource with the highest lignan content, and the rhizosphere bacteria of Schisandra are rich in diversity. Schisandra B is negatively correlated with Bryobacter, Candidatus_Solibacter, and unnamed genera of Gaiellales.

Diversity of Endophytes of Actinidia arguta in Different Seasons.

The seasonal changes in environmental conditions can alter the growth states of host plants, thereby affecting the living environment of endophytes and forming different endophytic communities. This study employs Illumina MiSeq next-generation sequencing to analyze the 16SrRNA and ITS rDNA of endophytes in 24 samples of Actinidia arguta stem tissues across different seasons. The results revealed a high richness and diversity of endophytes in Actinidia arguta, with significant seasonal variations in microbial community richness. This study identified 897 genera across 36 phyla for bacteria and 251 genera across 8 phyla for fungi. Notably, 69 bacterial genera and 19 fungal genera significantly contributed to the differences in community structure across seasons. A distinctive feature of coexistence in the endophytic community, both specific and conservative across different seasons, was observed. The bacterial community in winter demonstrated significantly higher richness and diversity compared to the other seasons. Environmental factors likely influence the optimal timing for endophyte colonization. Solar radiation, temperature, precipitation, and relative humidity significantly impact the diversity of endophytic bacteria and fungi. In addition, seasonal variations show significant differences in the nutritional modes of fungal endophytes and the degradation, ligninolysis, and ureolysis functions of bacterial endophytes. This study elucidates the potential role of endophytes in assisting Actinidia arguta in adapting to seasonal changes and provides a theoretical basis for further exploration of functional microbial strains.

Stabilizing Mechanisms in Patients Treated Using Hill-Sachs Remplissage With Bankart Repair in Abduction-External Rotation Position.

BACKGROUND: Hill-Sachs lesion (HSL) remplissage with Bankart repair (RMBR) provides a minimally invasive solution for treating HSLs and glenoid bone defects of <25%. The infraspinatus tendon is inserted into the HSL during the remplissage process, causing the infraspinatus to shift medially, leading to an unknown effect on glenohumeral alignment during the resting abduction-external rotation (ABER) and muscle-active states. PURPOSE/HYPOTHESIS: The purpose of this study was to evaluate the possible check-rein effect and muscle-active control in stabilizing the glenohumeral joint after RMBR in vivo. We hypothesized that the check-rein effect and active control would stabilize the glenohumeral joint in the ABER position in patients after RMBR. STUDY DESIGN: Controlled laboratory study. METHODS: We included 42 participants-22 patients in group A who met the inclusion criteria after RMBR and 20 healthy participants in group B without shoulder laxity. Three-dimensional magnetic resonance imaging was performed to analyze the alignment relationship of the glenohumeral joint with and without muscular activity. Ultrasonic shear wave elastography was used to evaluate the elastic properties of the anterior capsule covered with the anterior bands of the inferior glenohumeral ligament. RESULTS: Patients who underwent RMBR demonstrated more posterior (-1.81 ± 1.19 mm vs -0.76 ± 1.25 mm; P = .008) and inferior (-1.05 ± 0.62 mm vs -0.45 ± 0.48 mm; P = .001) shifts of the humeral head rotation center and less anterior capsular elasticity (70.07 ± 22.60 kPa vs 84.01 ± 14.08 kPa; P = .023) than healthy participants in the resting ABER state. More posterior (-3.17 ± 0.84 mm vs -1.81 ± 1.19 mm; P < .001) and less-inferior (-0.34 ± 0.56 mm vs -1.05 ± 0.62 mm; P < .001) shifts of the humeral head rotation center and less anterior capsular elasticity (36.57 ± 13.89 kPa vs 70.07 ± 22.60 kPa; P < .001) were observed in the operative shoulder during muscle-active ABER than in resting ABER states. CONCLUSION: The check-rein effect and muscle-active control act as stabilizing mechanisms in RMBR during the ABER position. CLINICAL RELEVANCE: Stabilizing mechanisms in RMBR during the ABER position include the check-rein effect and muscle-active control.

[Study on the evaluation of glenoid bone defects by MRI three-dimensional reconstruction].

Objective: To investigate the feasibility of MRI three-dimensional (3D) reconstruction model in quantifying glenoid bone defect by comparing with CT 3D reconstruction model measurement. Methods: Forty patients with shoulder anterior dislocation who met the selection criteria between December 2021 and December 2022 were admitted as study participants. There were 34 males and 6 females with an average age of 24.8 years (range, 19-32 years). The injury caused by sports injury in 29 cases and collision injury in 6 cases, and 5 cases had no obvious inducement. The time from injury to admission ranged from 4 to 72 months (mean, 28.5 months). CT and MRI were performed on the patients' shoulder joints, and a semi-automatic segmentation of the images was done with 3D slicer software to construct a glenoid model. The length of the glenoid bone defect was measured on the models by 2 physicians. The intra-group correlation coefficient ( ICC) was used to evaluate the consistency between the 2 physicians, and Bland-Altman plots were constructed to evaluate the consistency between the 2 methods. Results: The length of the glenoid bone defects measured on MRI 3D reconstruction model was (3.83±1.36) mm/4.00 (0.58, 6.13) mm for physician 1 and (3.91±1.20) mm/3.86 (1.39, 5.96) mm for physician 2. The length of the glenoid bone defects measured on CT 3D reconstruction model was (3.81±1.38) mm/3.80 (0.60, 6.02) mm for physician 1 and (3.99±1.19) mm/4.00 (1.68, 6.38) mm for physician 2. ICC and Bland-Altman plot analysis showed good consistency. The ICC between the 2 physicians based on MRI and CT 3D reconstruction model measurements were 0.73 [95% CI (0.54, 0.85)] and 0.80 [95% CI (0.65, 0.89)], respectively. The 95% CI of the difference between the two measurements of physicians 1 and 2 were (-0.46, 0.49) and (-0.68, 0.53), respectively. Conclusion: The measurement of glenoid bone defect based on MRI 3D reconstruction model is consistent with that based on CT 3D reconstruction model. MRI can be used instead of CT to measure glenoid bone defects in clinic, and the soft tissue of shoulder joint can be observed comprehensively while reducing radiation.

Discovery of resident memory T cells in inflammatory vitiligo: A case report.

RATIONALE: The purpose of this report was to describe resident memory cluster of differentiation 8 (CD8) + T cells may contribute to the progression of inflammatory vitiligo. PATIENT CONCERNS: A 32-year-old male has a stable vitiligo for 1 year, then some patches present inflammatory erythema. Two years later, the inflammatory patches enlarged and joined together, and the remaining 2 common patches shows repigmentation and no change respectively. Both CD69 + CD8 + T cells and CD103 + CD8 + T cells showed marked increase in inflammatory vitiligo than common vitiligo. DIAGNOSIS: Histological findings show that the numbers of lymphocytes are increased in inflammatory vitiligo than common vitiligo. Immunofluorescence staining show that the numbers of CD69 + CD8 + T cells demonstrated a marked increase in inflammatory vitiligo than common vitiligo. INTERVENTIONS: Without any intervention. OUTCOMES: The previous upper 2 patches on the abdomen with erythematous rim were enlarged and joined together. However the lowest lesion with uninflamed common rim on the abdomen remained static, the one on the right groin showed spot-like repigmentation. LESSONS: This case report demonstrates that resident memory CD8 + T cells may contribute to the progression of inflammatory vitiligo.

Polymeric nanoparticles containing rapamycin and autoantigen induce antigen-specific immunological tolerance for preventing vitiligo in mice.

Vitiligo is an autoimmune disease in which pigment is lost in patches of the skin. CD4+ T cells are implicated in vitiligo while regulatory T cells (Tregs) could ameliorate vitiligo. Rapamycin together with autoantigen have been shown to induce immunological tolerance and promote Tregs in multiple autoimmune diseases. In the current study, we synthesized nanoparticles containing rapamycin and autoantigen HEL46-61 (NPHEL46-61/Rapa) and investigated their effects on vitiligo. We treated bone marrow-derived dendritic cells (BMDCs) from TrpHEL mice with NPHEL46-61/Rapa and monitored the phenotype of BMDCs. We investigated the effects of NPHEL46-61/Rapa-treated BMDCs on CD4+ T cell proliferation and differentiation. We administrated NPHEL46-61/Rapa to TCR-TrpHEL mice and investigated the effects on vitiligo. We found that BMDCs can uptake the NPHEL46-61/Rapa, which resulted in decreased expression of costimulation molecules CD80 and CD86 in BMDCs. BMDCs treated with NPHEL46-61/Rapa suppressed antigen-specific CD4+ T cell proliferation while promoted the differentiation of these CD4+ T cell to Tregs in vitro. Administration of NPHEL46-61/Rapa to TCR-TrpHEL mice ameliorated vitiligo, promoted Treg production, and suppressed IFN-γ and IL-6 production, while induced IL-10 production. Therefore, our study provides experimental evidence that nanoparticles containing rapamycin and autoantigen could induce antigen-specific immunological tolerance and prevent vitiligo.

The effects of 308-nm excimer laser on the infiltration of CD4+, CD8+ T-cells, and regulatory T cells in the lesional skin of patients at active and stable stages of nonsegmental vitiligo.

BACKGROUND: To explore the associations between this treatment and CD4+, CD8+ T-cells, and regulatory T cells (Treg). METHODS: We collected the skin biopsies from 295 patients with nonsegmental vitiligo treated with or without 308-nm excimer laser. We revealed that patients at stable stage showed a better response to 308-nm excimer laser than those at active stage. RESULTS: In comparison with untreated patients, CD4+ and CD8+ T cells were both reduced while Foxp3+ Treg cells, TGF-ß, and IL-10 were elevated in the lesional sites of patients who showed effective reaction to the treatment. In the treated patients at active stage, the infiltration of CD4+ and CD8+ T cells was significantly declined but Foxp3+ Treg cells, TGF-ß, and IL-10 were increased compared to those in untreated patients at active stage. These changes of cell infiltration were more obvious in the treated patients at stable stage, explaining why there were more patients at stable stage response better than patients at active stage. CONCLUSIONS: 308-nm excimer laser is effective to reduce the infiltration of CD4+ and CD8+ T cells but promote the infiltration Treg cells and secretion of TGF-ß and IL-10 in the lesion skin of vitiligo patients, especially at stable stage.

Redirection Using Double Pulley Technique for Snapping Triceps Tendon: A Case Report and Technique Note.

BACKGROUND: Snapping triceps tendon is an increasingly recognized clinical entity, which is associated with a variety of pathologic factors. The causative factors include inherited structural or developmental variations, post-traumatic malalignment, and other reasons. The main complaint of patients with snapping lateral triceps are the snapping sensation, mild muscle weakness of elbow extension, with or without tenderness in distal triceps tendon. Many treatment options have been reported previously, including tendon resection and redirection. CASE PRESENTATION: We present the case of a 19-year-old boy with post-traumatic distal lateral head of triceps tendon dislocation who complained of extension weakness and snapping sensation in his left elbow. Then, we used two-strand-overhand locking (TSOL) knot combined with double pulley technique to redirect the snapping triceps tendon. CONCLUSION: The patient recovered well after the operation without complaining of discomfort. This fixing and redirection tendon technique, described previously for repairing rotator cuff tears, may be applied in a similar fashion for the snapping triceps tendon with promising clinical results.

Apigenin protects human melanocytes against oxidative damage by activation of the Nrf2 pathway.

Vitiligo is a chronic, autoimmune destruction of melanocytes, resulting in progressively expanding depigmented skin patches. Severity of the disorder, which affects approximately 1% of humans, may be mitigated using topical corticosteroids combined with phototherapy; along with other clinical strategies; however, no definitive cures are currently available. Here, the capacity of apigenin, a plant-derived aglycone, to inhibit oxidative stress-mediated melanocyte depletion in vitro using a PIG3V vitiligo perilesional melanocyte cell model is evaluated. PIG3V cells, treated with selected doses of apigenin, were challenged with H2O2, then assessed for viability and the oxidative stress-related parameters: superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA) by enzyme-linked immunoabsorbent assay (ELISA). Additionally, expression of nuclear factor erythroid 2p45 (NF-E2)-related factor 2 (Nrf2) and downstream targets was detected using Western blotting. Outcomes demonstrated that compared with negative control cultures, apigenin-treated cells exhibited enhanced viability. Likewise, apigenin enhanced expression of the cellular anti-oxidants SOD, CAT, and GSH-Px, but inhibited production of MDA, an oxidative stress biomarker. Interestingly, the expression and nuclear localization of the Nrf2 transcription factor, an important regulator oxidative stress and its downstream target genes, was significantly increased by apigenin treatment. Apigenin influence on Nrf2 was further validated by experiments demonstrating that Nrf2 knockdown cells failed to exhibit significant apigenin-mediated effects on cell viability and oxidative stress. Apigenin's non-toxicity and ability to affect multiple oxidative stress-related parameters through its effects on Nrf2 signaling in melanocytes suggests that it may prove to be a valuable therapeutic tool in long-term management of vitiligo.

Yes-associated protein regulates the growth of human non-small cell lung cancer in response to matrix stiffness.

The Yes­associated protein (YAP) transcriptional coactivator is recognized as a crucial regulator of human cancer. However, its involvement in human non­small cell lung cancer (NSCLC) in response to physical cues remains unclear. In this study, substrates with different rigidity were generated in order to evaluate the role of YAP, and its upstream regulators in the Hippo pathway, in the regulation of growth of an NSCLC cell line within particular environments. It was shown that the expression of the YAP protein in SPCA-1 NSCLC cells was significantly increased when cultured on a stiff substrate compared to a soft substrate. However, the expression of phospho­YAP protein and large tumor suppressor kinase 1 (LATS1) were markedly decreased after culturing on the stiff substrate. Phosphorylation of YAP by LATS1 leads to cytoplasmic retention of YAP, which inhibits its function as a nuclear transcription coactivator. The study also found that the stiff substrate promoted the growth of NSCLC cells in vitro, and an increase in the transcription levels of Survivin, connective tissue growth factor, amphiregulin and Ki67, as well as a decrease in the expression level of YAP in the cytoplasm, and adecrease in p-YAP. In conclusion, the findings showed that the stiffness of the subcellular matrix altered the behavior of NSCLC cells, and that YAP regulated the growth of NSCLC cells in response to matrix stiffness, thereby suggesting a role for the Hippo­YAP pathway in the response of NSCLC cell growth to specific microenvironments.

Attenuation of peripheral regulatory T-cell suppression of skin-homing CD8⁺T cells in atopic dermatitis.

PURPOSE: Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8⁺T cells have been known to play an important role in the pathogenesis of atopic dermatitis (AD). However, the mechanisms underlying the loss of self-tolerance remain unclear. Regulatory T cells (Tregs) play a key role in the development of homeostasis in the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8⁺CLA⁺T cells might be underlying mechanism in AD. MATERIALS AND METHODS: CD8⁺CLA⁺T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequencies of CD8⁺CLA⁺T cells were evaluated. The proliferative responses of CD8⁺CLA⁺T cells were assessed by flow cytometry, and the levels of transforming growth factor-ß1 (TGF-ß1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay. RESULTS: Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheral CD8⁺CLA⁺T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8⁺CLA⁺T cells in AD. Meanwhile, the levels of TGF-ß1 produced by Tregs were significantly lower in AD, and anti-TGF-ß1 abolished such suppression. CONCLUSION: The attenuated inhibitory ability of Tregs on hyper-activated autologous CD8⁺CLA⁺T cells, mediated by TGF-ß1, plays an important role in the pathogenesis of AD.

Ginsenosides Rb1 and Rg1 Stimulate Melanogenesis in Human Epidermal Melanocytes via PKA/CREB/MITF Signaling.

Reduced or defective melanin skin pigmentation may cause many hypopigmentation disorders and increase the risk of damage to the skin triggered by UV irradiation. Ginsenosides Rb1 and Rg1 have many molecular targets including the cAMP-response element-binding protein (CREB), which is involved in melanogenesis. This study aimed to investigate the effects of ginsenosides Rb1 and Rg1 on melanogenesis in human melanocytes and their related mechanisms. The effects of Rb1 and Rg1 on cell viability, tyrosinase activity, cellular melanin content and protein levels of tyrosinase, microphthalmia-associated transcription factor (MITF), and activation of CREB in melanocytes were assessed. Results showed that Rb1 or Rg1 significantly increased cellular melanin content and tyrosinase activity in a dose-dependent manner. By contrast, the cell viability of melanocytes remained unchanged. After exposure to Rb1 or Rg1, the protein levels of tyrosinase, MITF, and phosphorylated CREB were significantly increased. Furthermore, pretreatment with the selective PKA inhibitor H-89 significantly blocked the Rb1- or Rg1-induced increase of melanin content. These findings indicated that Rb1 and Rg1 increased melanogenesis and tyrosinase activity in human melanocytes, which was associated with activation of PKA/CREB/MITF signaling. The effects and mechanisms of Rb1 or Rg1 on skin pigmentation deserve further study.

Regulatory T cells from active non-segmental vitiligo exhibit lower suppressive ability on CD8+CLA+ T cells.

BACKGROUND: Recent studies have shown that vitiligo is a T-cell mediated autoimmune disease. Skin-homing cytotoxic T lymphocytes expressing cutaneous lymphocyte-associated antigen (CLA) have been suggested to be responsible for the destruction of melanocytes in vitiligo. An aberration in the suppressive function of regulatory T cells (Tregs) has been reported in vitiligo patients. However, whether the weakened suppressive ability of the Tregs contributes to hyper-activated skin homing CD8(+)CLA(+) T cells remains to be determined. OBJECTIVES: To investigate the inhibition of circulating Tregs on the proliferation of autologous CD8(+)CLA(+) T cells in non-segmental vitiligo patients. METHODS: CD8(+)CLA(+) T cells and Tregs were obtained from the peripheral blood of 13 non-segmental vitiligo patients and 7 controls. The proliferative responses of CD8(+)CLA(+) T cells were assessed in the absence or presence of autologous Tregs, and the levels of Transforming Growth Factor ß1(TGF-ß1) and IL-10 in culture supernatants were detected by enzyme-linked immunosorbent assay. RESULTS: The proliferative responses of circulating CD8(+)CLA(+) T cells in the presence of Tregs were significantly higher in the active vitiligo than in the stable vitiligo and control groups. Tregs from active vitiligo patients exhibited a lower inhibitory effect on proliferation of CD8(+)CLA(+) T cells. The levels of TGF-ß1 produced by Tregs were significantly lower in active vitiligo than other groups and anti-TGF-ß1 antibodies could abrogate the suppressive function of Tregs. CONCLUSIONS: The functional activity of Tregs is compromised in active vitiligo patients. TGF-ß1 plays an important role in the autoimmune mechanism of the disease.

Characterization of circulating CD8+T cells expressing skin homing and cytotoxic molecules in active non-segmental vitiligo.

BACKGROUND: Vitiligo is caused by melanocyte depletion. Studies have suggested that skin-homing cytotoxic T lymphocytes that express cutaneous lymphocyte-associated antigen (CLA) are responsible for melanocyte depletion. The characteristics of these skin-homing cytotoxic T cells have not been well established yet. OBJECTIVES: To investigate the frequency of skin-homing CD8(+)T cells (CD8(+)CLA(+)T cells) and their expression of cytotoxic molecules, as well as migration-related molecules in CD8(+)T cell in non-segmental vitiligo patients. MATERIALS & METHODS: The frequency of CD8(+)CLA(+)T cells and their expression of cytotoxic molecules (perforin, granzyme-B and FasL) in peripheral blood of patients with non-segmental vitiligo were assessed using flow cytometry. Levels of chemokine receptors (CCR4, CCR10) on CD8(+)T cells were evaluated. RESULTS: Our results revealed a higher frequency and increased expression of perforin and granzyme-B in circulating CD8(+)CLA(+)T cells from patients with active vitiligo. The expression levels of CCR4 increased in CD8(+)T cells in active vitiligo patients. CONCLUSION: Patients with active non-segmental vitiligo have a higher frequency of CD8(+)CLA(+)T cells and hyper-activated cytotoxic functions, which may be involved in the pathogenesis of non-segmental vitiligo.

Quantitative assessment of groundwater vulnerability using index system and transport simulation, Huangshuihe catchment, China.

Groundwater vulnerability assessment has been an increasingly important environment management tool. The existing vulnerability assessment approaches are mostly index systems which have significant disadvantages. There need to be some quantitative studies on vulnerability indicators based on objective physical process study. In this study, we tried to do vulnerability assessment in Huangshuihe catchment in Shandong province of China using both contaminant transport simulations and index system approach. Transit time of 75% of hypothetical injected contaminant concentration was considered as the vulnerability indicator. First, we collected the field data of the Huangshuihe catchment and the catchment was divided into 34 sub areas that can each be treated as a transport sub model. Next, we constructed a Hydrus1D transport model of Huangshuihe catchment. Different sub areas had different input values. Thirdly, we used Monte-Carlo simulation to improve the collected data and did vulnerability assessment using the statistics of the contaminant transit time as a vulnerability indicator. Finally, to compare with the assessment result by transport simulation, we applied two index systems to Huangshuihe catchment. The first was DRASTIC system, and the other was a system we tentatively constructed examining the relationships between the transit time and the input parameters by simply changing the input values. The result of comparisons between the two index systems and transport simulation approach suggested partial validation to DRASTIC, and the construction of the new tentative index system was an attempt of building up index approaches based on physical process simulation.