PEG10 amplification at 7q21.3 potentiates large-cell transformation in cutaneous T-cell lymphoma.

Mycosis fungoides (MF), the most common form of cutaneous T-cell lymphoma, undergo large-cell transformation (LCT) in the late stage, manifesting aggressive behavior, resistance to treatments, and poor prognosis, but the mechanisms involved remain unclear. To identify the molecular driver of LCT, we collected tumor samples from 133 MF patients and performed whole-transcriptome sequencing on 49 advanced-stage MF patients, followed by integrated copy number inference and genomic hybridization. Tumors with LCT showed unique transcriptional programs and enriched expressions of genes at chr7q. Paternally expressed gene 10 (PEG10), an imprinted gene at 7q21.3, was ectopically expressed in malignant T cells from LCT, driven by 7q21.3 amplification. Mechanistically, aberrant PEG10 expression increased cell size, promoted cell proliferation, and conferred treatment resistance by a PEG10/KLF2/NF-κB axis in in vitro and in vivo models. Pharmacologically targeting PEG10 reversed the phenotypes of proliferation and treatment resistance in LCT. Our findings reveal new molecular mechanisms underlying LCT and suggest that PEG10 inhibition may serve as a promising therapeutic approach in late-stage aggressive T-cell lymphoma.

Mutations in SREBF1, Encoding Sterol Regulatory Element Binding Transcription Factor 1, Cause Autosomal-Dominant IFAP Syndrome.

IFAP syndrome is a rare genetic disorder characterized by ichthyosis follicularis, atrichia, and photophobia. Previous research found that mutations in MBTPS2, encoding site-2-protease (S2P), underlie X-linked IFAP syndrome. The present report describes the identification via whole-exome sequencing of three heterozygous mutations in SREBF1 in 11 unrelated, ethnically diverse individuals with autosomal-dominant IFAP syndrome. SREBF1 encodes sterol regulatory element-binding protein 1 (SREBP1), which promotes the transcription of lipogenes involved in the biosynthesis of fatty acids and cholesterols. This process requires cleavage of SREBP1 by site-1-protease (S1P) and S2P and subsequent translocation into the nucleus where it binds to sterol regulatory elements (SRE). The three detected SREBF1 mutations caused substitution or deletion of residues 527, 528, and 530, which are crucial for S1P cleavage. In vitro investigation of SREBP1 variants demonstrated impaired S1P cleavage, which prohibited nuclear translocation of the transcriptionally active form of SREBP1. As a result, SREBP1 variants exhibited significantly lower transcriptional activity compared to the wild-type, as demonstrated via luciferase reporter assay. RNA sequencing of the scalp skin from IFAP-affected individuals revealed a dramatic reduction in transcript levels of low-density lipoprotein receptor (LDLR) and of keratin genes known to be expressed in the outer root sheath of hair follicles. An increased rate of in situ keratinocyte apoptosis, which might contribute to skin hyperkeratosis and hypotrichosis, was also detected in scalp samples from affected individuals. Together with previous research, the present findings suggest that SREBP signaling plays an essential role in epidermal differentiation, skin barrier formation, hair growth, and eye function.

Targeted inhibition of YAP/TAZ alters the biological behaviours of keloid fibroblasts.

Abnormal activation of fibroblasts plays a crucial role in keloid development. However, the mechanism of fibroblast activation remains to be determined. YAP/TAZ are key molecules in the Hippo signalling pathway that promote cell proliferation and inhibit apoptosis. Here, we show that keloid fibroblasts have higher levels of YAP/TAZ mRNA and proteins on primary culture. Targeted knockdown of endogenous YAP or TAZ significantly inhibited cell proliferation, reduced cell migration, induced cell apoptosis and down-regulated collagen1a1 production by keloid fibroblasts. Moreover, we demonstrate that verteporfin, an inhibitor of YAP/TAZ, has similar but stronger inhibitory effects on fibroblasts compared to YAP/TAZ knockdown. Our study provides evidence that YAP/TAZ may be involved in the pathogenesis of keloids. Targeted inhibition of YAP/TAZ could change the biological behaviours of fibroblasts and can potentially be used as therapy for keloids.

Rare Concurrence of Alopecia Areata in the Setting of the Lipedematous Scalp.

ABSTRACT: The lipedematous scalp (LS) is characterized by a thickened scalp because of the increased thickness of the subcutaneous fat layer. When the soft and boggy scalp is associated with shortened hairs and hair loss, it is referred to as lipedematous alopecia (LA). We report a case of alopecia areata with LS, which may be misdiagnosed as LA. However, the histopathologic features showed a thickened subcutaneous fat layer and hair bulb inflammation. Hair regrowth was appreciable after treatment with topical steroids, minoxidil, oral compound glycyrrhizin, and vitamin D. This case report aims to show that LS and alopecia areata may coexist, and histopathologic examination is necessary for precise diagnosis.

Concurrent extramammary Paget's disease involving the genitalia and axilla: Case report and literature review.

Concurrent multiple primary extramammary Paget's disease (EMPD) is rare. Herein, we present two Chinese cases of concurrent primary EMPD involving both the genitalia and the axilla, and they also had a history of other malignancy. We also summarise the cases of multiple primary EMPD previously described in literature. Careful examination of all apocrine sweat gland-bearing sites and additional internal malignancies is recommended for patients with EMPD.

IQGAP1 promotes anoikis resistance and metastasis through Rac1-dependent ROS accumulation and activation of Src/FAK signalling in hepatocellular carcinoma.

BACKGROUND: Hepatitis B virus (HBV) has a crucial role in the progression of hepatocellular carcinoma (HCC). Tumour cells must develop anoikis resistance in order to survive before metastasis. This study aimed to investigate the mechanism of IQGAP1 in HBV-mediated anoikis evasion and metastasis in HCC cells. METHODS: IQGAP1 expression was detected by immunohistochemistry, real-time PCR and immunoblot analysis. Lentiviral-mediated stable upregulation or knockdown of IGAQP1, immunoprecipitation, etc. were used in function and mechanism study. RESULTS: IQGAP1 was markedly upregulated in HBV-positive compared with HBV-negative HCC cells and tissues. IQGAP1 was positively correlated to poor prognosis of HBV-associated HCC patients. IQGAP1 overexpression significantly enhanced the anchorage-independent growth and metastasis, whereas IQGAP1-deficient HCC cells are more sensitive to anoikis. Mechanistically, we found that HBV-induced ROS enhanced the association of IQGAP1 and Rac1 that activated Rac1, leading to phosphorylation of Src/FAK pathway. Antioxidants efficiently inhibited IQGAP1-mediated anoikis resistance and metastasis. CONCLUSIONS: Our study indicated an important mechanism by which upregulated IQGAP1 by HBV promoted anoikis resistance, migration and invasion of HCC cells through Rac1-dependent ROS accumulation and activation of Src/FAK signalling, suggesting IQGAP1 as a prognostic indicator and a novel therapeutic target in HCC patients with HBV infection.

A benzoxazole derivative PO-296 inhibits T lymphocyte proliferation by the JAK3/STAT5 signal pathway.

Immunosuppressants have shown striking achievements in treating autoimmune diseases in recent years. It is urgent to develop more immunosuppressants to provide more options for patients. PO-296 [2-(6-chlorobenzo[d]oxazol-2-yl)-4,5,6,7-tetrahydro-2H-indazol-3-ol] was identified as a novel benzoxazole derivative. We observed that it exhibits an obvious immunosuppressive activity to T lymphocytes. PO-296 significantly inhibited the proliferation of activated human T lymphocyte without cytotoxicity. Moreover, PO-296 did not affect the expression of cluster of differentiation (CD)-25 or CD69 but induced T lymphocyte cycle arrest in the G0/G1 phase. Furthermore, PO-296 inhibited interleukin (IL)-6, IL-17, and interferon gamma expression but had no effect on IL-2, IL-4, or IL-10. Yet, importantly, PO-296 inhibited the phosphorylation of signal transducer and activator of transcription 5 (STAT5), increased the phosphorylation of p70S6K, but did not affect the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/mitogen-activated protein kinase pathway. In conclusion, these findings indicate that PO-296 inhibits human activated T-lymphocyte proliferation by affecting the janus kinase 3 (JAK3)/STAT5 pathway. PO-296 possesses a potential lead compound for the design and development of new immunosuppressants for the treatment of autoimmune diseases.

K313, a novel benzoxazole derivative, exhibits anti-inflammatory properties via inhibiting GSK3ß activity in LPS-induced RAW264.7 macrophages.

Benzoxazole and its derivatives have been widely studied in recent years due to their various biological properties. A previous study has demonstrated that K313 (1H-indole-2,3-dione 3-(1,3-benzoxazol-2-ylhydrazone)), a novel benzoxazole derivative, inhibits T cell proliferation to yield immunosuppressive effects. However, there are no related reports about its anti-inflammatory effects. In the present study, we investigated the anti-inflammatory properties and the underlying molecular mechanism of K313 in lipopolysaccharide (LPS)-induced RAW264.7 macrophages. K313 dose-dependently (5, 10, and 20 µM) inhibited LPS-stimulated nitric oxide (NO), interleukin (IL)-6, tumor necrosis factor (TNF)-α, and 3-nitrotyrosine (3-NT) production and significantly decreased the gene transcription levels of inducible nitric oxide (iNOS), IL-6, and TNF-α. In addition, the results showed that the inflammatory cytokines suppressed by K313 were not regulated by p65 NF-κB, ERK1/2, AKT, or p38 MAPK. Instead, K313 increased phosphorylation of glycogen synthase kinase-3 beta (GSK-3ß) (Ser9) resulting in GSK-3ß deactivation. Moreover, in LPS-stimulated RAW264.7 macrophages, K313 and lithium chloride (LiCl) had a synergistic effect on the anti-inflammatory response. These results indicated that K313 exhibited anti-inflammatory properties and revealed the potential mechanism. K313 can increase GSK-3ß (Ser9) phosphorylation to decrease GSK-3ß activation in LPS-induced RAW264.7 macrophages.

Tolerogenic dendritic cells induced by BD750 ameliorate proinflammatory T cell responses and experimental autoimmune encephalitis in mice.

BD750, a novel JAK3/STAT5 inhibitor, can inhibit T cell proliferation. This study aims to evaluate whether BD750 can induce tolerogenic dendritic cells (tolDC) and their function in experimental autoimmune encephalitis (EAE) in mice. Following BD750 treatment, LPS-induced maturation of DC, allogeneic T cell proliferation, Th1 and Th17 cell functional differentiation, the STAT5 and AKT activation were determined. The effect of tolDC loaded with antigen peptide on the development and severity of EAE and their splenic Th1 and Th17 cell responses were determined. In comparison with LPS-induced mature DC (mDC), BD750 treatment induced tolDC with lower expression levels of costimulatory molecules and pro-inflammatory cytokines and lower levels of STAT5 phosphorylation. TolDC inhibited allogeneic T cell proliferation and reduced Th1 and Th17 responses. Adoptive transfer of tolDC loaded with MOG35-55 inhibited the development and severity of EAE in mice, accompanied by reduced numbers of inflammatory infiltrates and decreased levels of demyelination in the spinal cord tissues of mice. In addition, treatment with tolDC loaded with antigen peptide also significantly reduced the frequency of splenic Th1 and Th17 cells in EAE mice. The effects of tolDC were similar to that of the JAK/STAT inhibitor, CP690550-treated DC. In conclusion, treatment with BD750 induced tolDC that inhibited pro-inflammatory T cell immunity in vitro and in vivo. BD750 and tolDC may be valuable for development of new therapies for EAE and other autoimmune diseases.

Monoterpenoid Indole Alkaloids from Kopsia officinalis and the Immunosuppressive Activity of Rhazinilam.

Six new monoterpenoid indole alkaloids, kopsinidines C-E (1-3), 11,12-methylenedioxychanofruticosinic acid (4), 12-methoxychanofruticosinic acid (5), and N(4)-methylkopsininate (7), as well as chanofruticosinic acid (6, as a natural product) and 23 known alkaloids, were obtained from the twigs and leaves of Kopsia officinalis. Their structures were characterized by physical data analysis. All isolated compounds were evaluated for their immunosuppressive activity on human T cell proliferation. Rhazinilam (29) significantly inhibited human T cell proliferation activated by anti-CD3/anti-CD28 antibodies (IC50 = 1.0 µM) and alloantigen stimulation (IC50 = 1.1 µM) without obvious cytotoxicity for naïve human T cells and peripheral blood mononuclear cells (0-320 µM). Although it did not affect T cell activation, it induced T cell cycle arrest in the G2/M phase and inhibited proinflammatory cytokine production in activated T cells.

Hemorrhagic Fever with Renal Syndrome, Zibo City, China, 2006-2014.

Analysis of hemorrhagic fever with renal syndrome cases in Zibo City, China, during 2006-2014 showed that it occurred year-round. Peaks in spring and fall/winter were caused by Hantaan and Seoul viruses, respectively. Rodent hosts were the striped field mouse for Hantaan virus and the brown rat and house mouse for Seoul virus.

[Modified Mohs micrographic surgery in the treatment of cutaneous melanoma].

OBJECTIVE: To study the significance of modified Mohs micrographic surgery in the treatment of cutaneous melanoma with the most appropriate margins. METHODS: A total of 20 cases with cutaneous melanoma treated by the technique of modified Mohs micrographic surgery from July 2013 to July 2014 were reviewed. Modified Mohs micrographic surgery set safe margin according to Guideline of Chinese Melanoma Treatment (2011) first, and then decide whether to adopt wider margins according to the pathology of the resected margin. Data concerning the numbers of additional resection and the final total width of margins were analyzed and the latter compared with the initial margins. RESULTS: Among 20 cases, 7 cases (35%) needed more than two resections to obtain a clear margin. The final widths of margin in 7 cases (35%) are different from the guideline. CONCLUSIONS: Modified Mohs micrographic surgery could ensure the thorough removal of primary tumor, while avoiding residual tumor as well as over-excision, which reflect the trend toward individualized treatment and minimally traumatic surgery.

[Clinicopathological features of extramammary Paget's disease: a report of 75 cases].

OBJECTIVE: To evaluate the clinicopathological features of extramammary Paget's disease. METHODS: Statistical analyses were performed for the clinicopathological features for 75 cases of extramammary Paget's disease from 2004 to 2013. RESULTS: Extramammary Paget's disease exhibited a male predominance (5.25:1). The mean age of onset was (64.23 ± 12.02) years. Single lesion accounted for 92.0% (69/75) and most of them were located in genital area. Pruritus occurred in 61.6% (45/73) patients. Invasive extramammary Paget's disease accounted for 22.7% (17/75). The concordance between clinical and pathological diagnoses was 85.3% (64/75). Among 11 misdiagnosed cases, 7 cases were misdiagnosed as eczema. Comparing Paget cells located in epithelium of adnexa or dermis and those confined to epidermis, significant differences existed in age of onset, course, recurrence rate (all P<0.05), but not in lesion size. Among 67.1% (47/70) of patients undergoing Mohs surgery, 38.3% (18/47) had a complete one-time resection while 61.7% (29/47) required more than twice for thorough resection. The 5-year recurrence rate was 25.0%. The postoperative 5-year recurrence rate of Mohs micrographic surgery (MMS) was 18.0% and the rate of non-MMS 36.5%. From 2011 to 2013, 3 recurrent cases had no radiotherapy. However another 15 cases on radiotherapy were non-recurrent. CONCLUSIONS: With a slow progression, extramammary Paget's disease is more common among elder males. Recurrence rate is associated with the degree of invasion. Thus an early diagnosis is essential. Due to obvious differences between gross and pathological boundaries, Mohs surgery may help to determine the margin. Meanwhile, adding radiotherapy reduces the recurrence rate.

[Retrospective analysis for clinical and histopathological characteristics of basal cell carcinoma in 418 patients].

OBJECTIVE: To investigate the clinical and histopathological characteristics of basal cell carcinoma (BCC) in Chinese patients. METHODS: Clinical and pathological data of BCC confirmed by histology from 2010 to 2012 in Peking University First Hospital were retrospectively analyzed. RESULTS: Among 418 patients enrolled, the male/female ratio was 0.77:1. The average age was (65.39±13.51) years. Among the patients younger than 60 years who occupied 29% of all the cases, the male/female ratio was 1.16:1. In terms of the histology subtypes of the BCCs, 81.8% were nodular, followed by superficial (9.8%), and the others were in very small proportion. The head and face were the most common sites of BCC (86.6%). All morpheaform subtypes, and the majority of the nodular subtypes were located on the head and face, whereas the trunk and extremities were the most common locations for the others. Clinically, 86.6% of the BCC were pigmented and 80.4% were not ulcerated. The diagnostic accordance rates of BCC on the head and face (84.7%) and on the trunk (79.1%) were higher than those on the extremities (46.2%, P<0.05). CONCLUSION: The most clinical and histopathological characteristics of our cases were similar to those of Caucasian. This study displays some unique characteristics. The young and middle aged patients occupied relative higher proportion, and their gender ration was different from that of the aged group. Tumor with hyperpigmentation was popular and few cases were ulcerated. In this study, multiple BCC cases were seldom, and the BCC patients with nevus sebaceous were older than those in other reports. The research of the diagnostic accordance rates of BCC revealed that both doctors and patients should pay more attention to BCC.

Toosendanin inhibits T-cell proliferation through the P38 MAPK signalling pathway.

In recent years, immunosuppressants have shown significant success in the treatment of autoimmune diseases. Therefore, there is an urgent need to develop additional immunosuppressants that offer more options for patients. Toosendanin has been shown to have immunosuppressive activity in vitro as well as effects on autoimmune hepatitis (AIH) in vivo. Toosendanin did not induce apoptosis in activated T-cells and affect the survival rate of naive T-cells. Toosendanin did not affect the expression of CD25 or secretion of IL-2 by activated T-cells, and not affect the expression of IL-4 and INF-γ. Toosendanin did not affect the phosphorylation of STAT5, ERK, AKT, P70S6K. However, toosendanin inhibited proliferation of anti-CD3/anti-CD28 mAbs-activated T-cells with IC50 of (10 ± 2.02) nM. Toosendanin arrested the cell cycle in the G0/G1 phase, significantly inhibited IL-6 and IL-17A secretion, promoted IL-10 expression, and inhibited the P38 MAPK pathway. Finally, toosendanin significantly alleviated ConA-induced AIH in mice. In Summary, toosendanin exhibited immunosuppressive activity in vivo and in vitro. Toosendanin inhibits the proliferation of activated T-cells through the P38 MAPK signalling pathway, significantly suppresses the expression of inflammatory factors, enhances the expression of anti-inflammatory factors, and effectively alleviates ConA-induced AIH in mice, suggesting that toosendanin may be a lead compound for the development of novel immunomodulatory agents with improved efficacy and reduced toxicity.