Kaposi Varicelliform Eruption With Contact Dermatitis in a Person Living with AIDS.

Kaposi varicelliform eruption (KVE) is a cutaneous dissemination of a viral infection, which is mostly caused by herpes simplex virus (HSV) in the setting of certain underlying skin diseases. KVE occurs mainly in infants and children, but very rarely in adults. Here, we report a case of KVE with contact dermatitis in a 36-year-old man with acquired immunodeficiency syndrome (AIDS), who was referred to our deparment with pruritic well-defined facial erythema and multiple vesicular lesions. A punch biopsy and immunohistochemical examination established the diagnosis of KVE with contact dermatitis. After treatment with valacyclovir and antihistamines, facial lesions achieved complete remission. With this case report, KVE has specific manifestation in clinic, histopathology and immunohistochemistry, which could guide the early diagnosis and improve prognosis.

Unilateral and localized subcorneal pustular dermatosis.

We present an unusual case of unilateral and localized subcorneal pustular dermatosis, which has not been described previously.

Adipophilin expression in skin lesions with clear cell histology.

AIMS: Clear cells formed due to depositions of glycogen or lipids in the cytoplasm commonly occur in various tissues. Adipophilin (ADP), a lipid regulatory protein, is closely related to lipid droplets. This study aims to examine adipophilin expression in clear cells of various skin lesions. METHODS: ADP expression was examined using immunohistochemistry in 108 sections from 15 skin lesion types with clear cell histology, namely, sebaceoma (n=16), sebaceous adenoma (n=3), sebaceous carcinoma (n=12), xanthomata cutis (n=10), xanthogranuloma (n=8), Paget's disease (n=10), Bowen disease (n=10), hidradenoma (n=9), atypical lipoma (n=5), superficial lipomatous nevus (n=5), metastatic renal cell carcinoma (n=5), squamous cell carcinoma (n=4), seborrheic keratosis (n=4), dermatofibroma (n=4) and clear cell sarcoma (n=3). RESULTS: ADP was not expressed in Bowen disease, hidradenoma or seborrheic keratosis. Four expression patterns, foamy, reticular, granular and punctate, were summarised based on their expression in clear cells. Different expression patterns were related to tissue origin and differentiation degree. Foamy expression was commonly observed in lesions with mature sebaceous glands and xanthomas; reticular expression in adipocytes; granular expression in xanthoma, xanthogranuloma and metastatic renal carcinoma and punctate expression in sebaceoma, sebaceous carcinoma, Paget's disease, squamous cell carcinoma and clear cell sarcoma. Furthermore, stronger staining with focal vesicular labelling was noted in sebaceoma than in sebaceous carcinoma. Characteristic labelling was noted, including the circular distribution in Touton giant cells of xanthogranulomas and focal distribution in the clear cells along the edge of necrotic tissue in clear cell sarcoma. CONCLUSIONS: ADP is useful in identifying intracytoplasmic lipids and can be used to diagnose skin lesions with clear cell histology, especially in some lesions with characteristic labelling.

Effect of RNA Interference Targeting STAT3 Gene Combined with Ultrasonic Irradiation and SonoVue Microbubbles on Proliferation and Apoptosis in Keratinocytes of Psoriatic Lesions.

BACKGROUND: Signal transducer and activator of transcription 3 (STAT3) was strongly expressed and activated in psoriatic keratinocytes (KCs) and correlated with the severity of psoriasis. The study aimed to investigate the effects of STAT3 small interfering RNA (siRNA) combined with ultrasonic irradiation and SonoVue microbubbles on the proliferation and apoptosis in KCs of psoriatic lesions and the relative mechanisms. METHODS: Psoriatic KCs were transfected under four experimental conditions: (1) STAT3 siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group); (2) STAT3 siRNA only carried by Lipofectamine 3000 (L group); (3) the negative control of siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (siRNA-NC); (4) not treated as Blank. Cell Counting Kit-8 assay was used to evaluate the cell proliferation. Cell cycle analysis was detected with cycle test Plus DNA reagent kit associated with flow cytometer. FITC Annexin V apoptosis detection kit associated with flow cytometer was applied for apoptosis analysis. Fluo calcium indicator associated with flow cytometer was used to analyze intracellular free calcium concentration ([Ca2+]i). The expressions of cyclin D1 and Bcl-xL were detected both at the mRNA level by real-time reverse transcription-polymerase chain reaction (RT-PCR) and at the protein level by Western blotting. The obtained data were statistically evaluated by two-way analysis of variance. RESULTS: STAT3 siRNA inhibited the growth of KCs in a time-dependent manner showing the highest proliferation inhibition in LUS group with proliferation ratio of 45.38% ± 5.85% at 72h (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced an altered cell cycle distribution of KCs showing the highest increases in G2/M-phase population up to 18.06% ± 0.36% in LUS group (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced late apoptosis of KCs with the highest late apoptosis percentage of 22.87% ± 1.28% in LUS group (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced the elevation of [Ca2+]iof KCs with the highest calcium fluorescence intensity mean of 1213.67 ± 60.51 in LUS group (P < 0.05 vs. L group, siRNA-NC, or Blank). STAT3 siRNA induced the downregulation of cyclin D1 and Bcl-xL expressions of KCs at mRNA and protein levels with the lowest expressions in LUS group with cyclin D1 expression of 51.81% ± 9.58% and 70.17% ± 4.22% at mRNA level and at protein level, respectively, and with Bcl-xL expression of 37.58% ± 4.92% and 64.06% ± 7.78% at mRNA level and at protein level, respectively (P < 0.05 vs. L group, siRNA-NC, or Blank). CONCLUSIONS: STAT3 siRNA inhibited the growth and induced the apoptosis in psoriatic KCs likely partly through altering cell cycle distribution, elevating [Ca2+]i, and downregulating cyclin D1 and Bcl-xL expressions. Silencing the target gene STAT3 in psoriatic KCs with siRNA combined with ultrasonic irradiation and microbubbles would contribute to a significant innovation as a new clinical therapy for psoriasis.

[Expressions of survivin, PI3K and AKT in keratinocytes in skin lesions and their pathogenic role in psoriasis vulgaris].

OBJECTIVE: To explore the role of survivin and PI3K/AKT pathway in the pathogenesis of psoriasis vulgaris (PV). METHODS: Plaque-like lesions collected from 22 patients with PV in progressive stage and 18 normal control skin specimens were examined using immunohistochemical staining, Western blotting and real-time quantitative PCR for expressions of survivin, PI3K and AKT in the keratinocytes, and their correlation was analyzed. A small interfering RNA (siRNA) was used to knock down AKT in cultured HaCaT cells, and Western blotting was used to detect the changes in the expression of survivin. RESULTS Compared with normal skin, PV lesions showed obviously up-regulated expressions of survivin, PI3K and AKT in the keratinocytes. Survivin expression was positively correlated with PI3K (r=0.4510, P=0.0351) and AKT (r=0.4423, P=0.0393) in the keratinocytes in PV lesions. In cultured HaCaT cells, siRNA-mediated knockdown of AKT caused down-regulation of survivin expression. CONCLUSION: Survivin and PI3K/AKT signaling pathway may participate in the occurrence and progression of PV.

Effects of RNA interference combined with ultrasonic irradiation and SonoVue microbubbles on expression of STAT3 gene in keratinocytes of psoriatic lesions.

The most effective sequence of small interfering RNA (siRNA) silencing STAT3 of psoriatic keratinocytes (KCs) was screened out, and the effects of the most effective siRNA combined with ultrasonic irradiation and SonoVue microbubbles on the expression of STAT3 of KCs and the dose- and time-response were investigated. Three chemically-synthetic siRNAs targeting STAT3 carried by Lipofectamine 3000 were transfected into KCs, and the effects on STAT3 expression were detected, then the most effective siRNA was selected for the subsequent experiments. The negative controls of siRNA (siRNA-NC) labeled with Cy3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles were transfected into KCs, then the optimal parameters of ultrasonic irradiation were determined. The most effective siRNA carried by Li-pofectamine 3000 combined with ultrasonic irradiation at the optimal parameters and SonoVue microbubbles was transfected into KCs, and the dose- and time-response of RNA interference was determined. The effect of RNA interference by the most effective siRNA at the optimal time and dose carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group) was compared with that only carried by Li-pofectamine 3000 (L group). The results showed that siRNA-3 achieved the highest silencing efficacy. 0.5 W/cm2 and 30 s were selected as the parameters of ultrasonic irradiation. The siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3 expression at mRNA and protein levels in dose- and time-dependent manners determined at 100 nmol/L with maximum downregulation on mRNA at 48 h, and on protein at 72 h after transfection. The LUS group achieved the highest silencing efficacy. It was concluded that siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3 expression in psoriatic KCs, and the optimized transfection condition and the sequence of siRNA-3 could serve for further research on gene therapy of psoriasis.

Effects of RNA Interference Combined with Ultrasonic Irradiation and SonoVue Microbubbles on Expression of STAT3 Gene in Keratinocytes of Psoriatic Lesions / 华中科技大学学报（医学）（英德文版）

The most effective sequence of small interfering RNA (siRNA) silencing STAT3 of psoriatic keratinocytes (KCs) was screened out,and the effects of the most effective siRNA combined with ultrasonic irradiation and SonoVue microbubbles on the expression of STAT3 of KCs and the dose-and time-response were investigated.Three chemically-synthetic siRNAs targeting STAT3 carried by Lipofectamine 3000 were transfected into KCs,and the effects on STAT3 expression were detected,then the most effective siRNA was selected for the subsequent experiments.The negative controls of siRNA (siRNA-NC) labeled with Cy3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles were transfected into KCs,then the optimal parameters of ultrasonic irradiation were determined.The most effective siRNA carried by Lipofectamine 3000 combined with ultrasonic irradiation at the optimal parameters and SonoVue microbubbles was transfected into KCs,and the dose-and time-response of RNA interference was determined.The effect of RNA interference by the most effective siRNA at the optimal time and dose carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles (LUS group) was compared with that only carried by Lipofectamine 3000 (L group).The results showed that siRNA-3 achieved the highest silencing efficacy.0.5 W/cm2 and 30 s were selected as the parameters of ultrasonic irradiation.The siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue mierobubbles could effectively knock down the STAT3 expression at mRNA and protein levels in dose-and time-dependent manners determined at 100 nmol/L with maximum downregulation on mRNA at 48 h,and on protein at 72 h after transfection.The LUS group achieved the highest silencing efficacy.It was concluded that siRNA-3 carried by Lipofectamine 3000 combined with ultrasonic irradiation and SonoVue microbubbles could effectively knock down the STAT3 expression in psoriatic KCs,and the optimized transfection condition and the sequence of siRNA-3 could serve for further research on gene therapy of psoriasis.

[Treatment of Guishen Zhiyang Recipe for Senile Pruritus Patients with Blood Deficiency and Hy- peractivity of Gan-yang Syndrome].

Objective To observe the clinical efficacy of Guishen Zhiyang Recipe (GZR) in trea- ting senile pruritus patients with blood deficiency and hyperactivity of Gan-yang syndrome (BDHGYS) and to study the mechanism in nervous-endocrine-immune systems. Methods Ninety senile pruritus pa- tients were assigned to the treatment group and the control group by complete randomized lot, 45 in each group. Totally 41 patients in the treatment group and 38 patients in the control group completed the trial. Patients in the treatment group took GZR, while those in the control group took Fuyang Granule (FG). Ex- ternal application of Binghuang Fule Soft Ointment was performed to all patients. The therapeutic course for all was 8 weeks. Symptoms and efficacy changes in skin lesion were observed in the two groups. Lev- els of stem cell factor (SCF) , dynorphin (DYN) , testosterone (T) , estradiol (E2) , and IL-4 were detec- ted in the two groups using double antibody sandwich ELISA. Results Compared with before treatment, pruritus degree, scratching area, scaling, lichenoid lesion, irritability and restlessness, dysphoria, dry pharynx, dizziness, and tinnitus all decreased in the two groups after 8 weeks of treatment (P <0. 05). Scores of Chinese medical syndromes all decreased after 4 and 8 weeks of treatment (P <0. 05). Levels of SCF and DYN obviously decreased in the two groups after 4 and 8 weeks of treatment (P <0. 05,P < 0. 01). Compared with the control group, obvious improvement was seen in pruritus degree, scratching number, scaling, irritability and restlessness , dysphoria, dizziness , and tinnitus(P <0. 05) ; total scores of Chinese medical syndromes decreased (P <0. 05) ; SCF also decreased (P <0. 05) in the treatment group after 8 weeks of treatment. Conclusions Based on external application of Binghuang Fule Soft Ointment, GZR showed better efficacy than that of the control. It had certain roles in regulating related mediator levels that might affect nervous-endocrine-immune systems.

[Deoxycholate induces apoptosis in cultured normal human esophageal mucosal epithelial cells].

OBJECTIVE: To study the effect of deoxycholate in inducing apoptosis of human normal esophageal mucosal epithelial cells in vitro and investigate the molecular mechanism. METHODS: Cultured normal human esophageal mucosal epithelial cells were treated with deoxycholate, and the cell apoptosis were evaluated with TUNEL, DNA ladder, flow cytometry with PI-staining, Annexin V-FITC conjugated with PI staining, and Western blotting. RESULTS: Flow cytometry, TUNEL and DNA ladder demonstrated that deoxycholate could induce apoptosis in normal human esophageal mucosal epithelial cells in a dose- and time-dependent manner. A percentage of 21.3% of the cell population treated with deoxycholate at 500 micromol/L for 30 min exhibited detectable caspase-3 activity shown by flow cytometry, which was significantly higher than the control level (1.5%, P<0.01). Western blotting suggested that deoxycholate down-regulated Bcl-2 protein expression and up-regulated Bax expression, but Fas was negative in the cells before and after deoxycholate treatment. CONCLUSIONS: Deoxycholate could induce apoptosis in cultured human esophageal mucosal epithelial cells. Aaspase-3 activation, Bcl-2 protein down-regulation and Bax up-regulation are involved in deoxycholate-induced apoptosis, which does not involve Fas-L/Fas.

[Effects of histamine on proliferation, apoptosis and differentiation of human keratinocytes].

OBJECTIVE: To investigate the effects of histamine on the proliferation, apoptosis and differentiation of human keratinocytes (HKC) and the mechanisms. METHODS: The effect of histamine on the growth of HKC in vitro was examined by MTT assay and trypan blue exclusion assay. Cell cycle analysis and early apoptosis analysis by double staining with Annexin V-FITC and PI were carried out by flow cytometry. DNA ladder assay was performed for the detection of cell apoptosis. HKC free calcium concentration ([Ca(2+)](i)) was measured by laser scanning confocal microscopy in combination with calcium fluorescence probe Fluo-3/AM. HKC differentiation markers keratin 10 (K10) and involucrin was detected by streptavidinbiotin complex immunocytochemical assay. RESULTS: Histamine at high concentration inhibited the proliferation of HKC with cell viability ratio of 65.6% at 1 x 10(-4) mol/L, while histamine at low concentrations promoted proliferation of HKC with the cell viability ratio of 130.7% at 1 x 10(-8) mol/L. Histamine at 1 x 10(-4) mol/L altered cell cycle distribution of HKC with an increase in G0/G1-phase cell population to 30.97%, a decrease in S-phase population to 73.81% and inhibition of G1/S switching. Histamine at 1 x 10(-4) mol/L induced obvious apoptosis of HKC with early apoptosis ratio of 18.64% as compared with the control (5.60%, P<0.05). Histamine 1 x 10(-4) mol/L induced an increase of HKC [Ca(2+)](i) up to 58.9% and cimetidine (an H(2) receptor antagonist) decreased HKC [Ca(2+)](i) down to 25.4%. Histamine at this concentration down-regulated the expressions of K10 and involucrin of HKC but these changes were not significantly different from those of the control (P>0.05). CONCLUSIONS: Histamine at high concentrations inhibits cell cycle progress of HKC, mediates cell apoptosis and induces the increase of [Ca(2+)](i), which might be a partial explanation for growth arrest of HKC elicited by histamine. Histamine may regulate epidermal tissue turnover under physiological conditions, whereas under pathological circumstances of the skin as in trauma and inflammation, histamine at high concentrations may inhibit the regeneration of epidermis and the differentiation of HKC.

[Effects of bradykinin on the proliferation, apoptosis and differentiation of human keratinocytes].

OBJECTIVE: To investigate the effects of bradykinin (BK) on the proliferation, apoptosis and differentiation of human keratinocyte (HKC) and the underlying mechanisms. METHODS: HKCs were cultured together with 1 x 10(-4) - 1 x 10(-9) mol/L of BK. With methyl thiotetrazole (MTT) and trypan blue staining it was shown that the BK in dose of 1 x 10(-4) mol/L possessed most powerful inhibitory effect, and the survival rate of HKC was 69.3%. Therefore, BK was employed in the dose of 1 x 10(-4) mol/L in the following studies. When the growth of HKCs reached the logarithmic phase, BK in the concentration of 1 x 10(-4) mol/L was added, and it was categorized as the test group (E). HKCs without BK served as the control group (C). The cell cycle and apoptosis were detected by flow cytometry after being cultured for 24 and 48 hours. The change in intracellular calcium [Ca(2+)](i) was determined by means of laser scanning confocal microscopy with calcium fluorescence probe Fluo-3/AM technique. The expression of HKC differentiation labeling protein keratin10 (K10) and involucrin were detected with Strept Avidin-Biotin Complex (SABC) immunocytochemical assay. RESULTS: The cell ratio in G0/G1 phase in E group increased by 34.57% while in S phase decreased by 58.91% in reference to that in C group. The G1/S phase switching of HKCs was obviously inhibited by BK, and apoptosis was stimulated (apoptotic rate of 15.34% in E group vs 5.60% in C group, P < 0.05). The [Ca(2+)](i) increased transiently in HKCs by 163.0% in E group after 3 minutes of BK activation and decreased thereafter in reference to that in C group. The K10 expression in HKC was down-regulated in E group with positive cell rate of 2.20%, which was lower than that of C group (6.89%, P < 0.05). CONCLUSION: The cell cycle process of HKC could be inhibited by high concentration of BK with increased apoptosis and an increase in [Ca(2+)](i), which might be the mechanism of inhibition of growth of HKC in vitro. Furthermore, the epithelial regeneration and HKC differentiation can also be inhibited by BK.

[Effects of all-trans-retinoic acid, acitretin and tazarotene on apoptosis and Bax/Bcl-2 expressions of human melanoma cells A375 and the significance].

OBJECTIVE: To investigate the effects of all-trans retinoic acid (ATRA), acitretin and tazarotene on apoptosis and Bax/Bcl-2 protein expressions of human melanoma A375 cells. METHODS: The effects of retinoids on apoptosis and Bax/Bcl-2 protein expressions of A375 cells in vitro were examined. Apoptosis analysis with double staining with annexin V-FITC and PI was performed using flow cytometer. SABC immunocytochemistry was employed for detection of Bax/Bcl-2 protein expressions. RESULTS: At the concentration of 1 x 10(-5) mol/L, ATRA, acitretin and tazarotene all induced apoptosis of A375 cells with apoptosis ratio of 5.03% (P<0.05), 13.42% (P<0.05) and 2.88% (P>0.05), respectively, and acitretin induced more significant apoptosis than the other two agents (P<0.05). In addition, all the three agents significantly increased the number of cells positive for Bax expression and decreased the number of cells expressing Bcl-2 (P<0.05), among which acitretin had the strongest effects. CONCLUSIONS: ATRA, acitretin and tazarotene mediate apoptosis of A375 cells possibly through, at least partially, the mitochondrial pathway. Acitretin may be utilized as a valuable alternative for treating melanoma.