清营汤加减对急性湿疹大鼠的干预及对大鼠皮肤TNF-α、IL-4表达的影响

目的探究清营汤加减对急性湿疹大鼠背部皮肤TNF-α、IL-4表达的影响及作用机制。方法 SD大鼠60只随机分为6组:氯雷他定组,清营汤高、中、低剂量组,模型组和正常组,每组10只。除正常组外,剩余各组对大鼠皮肤右侧背部,使用2,4-二硝基氯苯(DNCB)试剂造模,给予清营汤加减药液、氯雷他定药液、0.9%NaCl溶液对造模成功大鼠进行灌胃治疗。治疗后对实验组大鼠湿疹严重程度进行评分评估;运用称重法比较治疗后皮肤的重量差异;并采用免疫组化法(SP法)检测大鼠右侧背部皮肤组织中TNF-α、IL-4含量。结果①与模型组比较,清营汤灌胃中、低剂量组实验大鼠湿疹程度评分均明显降低(P0.05),高剂量组降低尤为显著(P0.01),说明清营汤加减可明显改善DNCB所致实验大鼠皮肤糜烂、水肿、红斑症状。②与模型组比较,氯雷他定组、清营汤灌胃组实验大鼠皮肤肿胀程度显著减轻(P0.01)。与清营汤低剂量组比较,清营汤中、高剂量组肿胀度明显减轻(P0.05)。与清营汤中剂量组比较,清营汤高剂量组肿胀度明显减轻(P0.05)。与氯雷他定组比较,清营汤灌胃高剂量组大鼠皮肤肿胀程度明显减轻(P0.05)。③与模型组比较,氯雷他定组、清营汤灌胃组大鼠病变皮肤组织中TNF-α、IL-4含量明显下降(P0.01);与氯雷他定组比较,清营汤灌胃高剂量组实验大鼠病变皮肤组织中TNF-α、IL-4含量显著降低(P0.05)。结论清营汤加减治疗急性湿疹可明显改善渗出、糜烂、红斑、肿胀等症状,其治疗机制可能为通过调控TNF-α、IL-4的水平来发挥治疗作用。     

The external validation of the 2019 ACR/EULAR classification criteria for IgG4-related disease in a large cohort from China

ObjectiveTo externally validate the performance of the 2019 American College of Rheumatology (ACR)/European League Against Rheumatism (EULAR) classification criteria for IgG4-related disease (IgG4-RD) within a cohort from China and to compare the criteria with the 2020 revised comprehensive diagnostic (RCD) criteria for IgG4-RD.MethodsThis study included 875 IgG4-RD and 302 non-IgG4-RD cases (213 mimickers and 89 patients with other diseases). Using expert clinical judgment as the gold standard for diagnosis of IgG4-RD, the performance (sensitivity, specificity, area under the curve (AUC) of the 2019 ACR/EULAR criteria for IgG4-RD was evaluated. We also compared it with the 2020 RCD criteria.ResultsThe 2019 ACR/EULAR classification criteria had a sensitivity of 76.6% (95% CI: 73.8% to 79.4%) and a specificity of 98.0% (96.0%-99.4%), an AUC of 0.873 (0.857–0.889) in the overall cohort. Those false negative cases under the 2019 ACR/EULAR classification criteria had significantly lower levels of serum IgG4, and fewer had pathological information, with a higher frequency in the involvement of those uncommon organs compared with the true positive cases. The cases judged as negative by the 2019 ACR/EULAR classification criteria yet judged as “definite” by the 2020 RCD criteria had more involvement of uncommon organs.ConclusionsThe 2019 ACR/EULAR classification criteria for IgG4-RD show outstanding specificity and good sensitivity in real-world clinical practice. The 2020 RCD criteria are helpful for the diagnosis of IgG4-RD in clinical scenarios where IgG4-RD presents as involving an isolated organ, especially the unusual sites.     

The hsp27kD heat shock protein and p38-MAPK signaling are required for regular epidermal differentiation

Background

In human epidermal keratinocytes the expression of hsp27 is closely related to differentiation in vitro and in situ.

Objective

We aimed to gain further insight into the role of hsp27 in epidermal differentiation by specific inhibition through siRNA and inhibition of p38-MAPK, the key enzyme of hsp27 phosphorylation.

Methods

Normal human keratinocytes (KC) and organotypic skin cultures (SE-skin equivalents) were used. Expression and phosphorylation of hsp27 was inhibited in these models by siRNA and SB203580, a specific inhibitor of p38-MAPK, respectively. Modification of morphology and expression of hsp27 and other differentiation associated proteins was investigated by immunofluorescence, western blot, and RT-PCR.

Results

Inhibition of p38-MAPK resulted in a downregulation of hsp27 in KC and SE. Additionally, in the presence of SB203580 Ca²⁺ induced expression of pro-filaggrin and loricrin was inhibited at the protein level and expression of filaggrin, keratin 10, and transglutaminase 1 at the mRNA level. Addition of SB203580 to SE, as well as hsp27 knockdown in this model resulted in identical patterns of irregular differentiation, disturbance of epidermal layers, and delayed expression of K10.

Conclusion

These results provide evidence that the expression of hsp27 and its phosphorylation by p38-MAPK are required for keratinocyte differentiation and for the formation of a regularly stratified epidermis.     

Efficacy and safety of pegylated liposomal doxorubicin in primary cutaneous B‐cell lymphomas and comparison with the commonly used therapies

Objectives: The therapy of advanced, relapsed or refractory primary cutaneous lymphomas is often unsatisfactory. Recent data indicate a favourable pharmacokynetic, pharmacodynamic and toxicity profile of pegylated liposomal doxorubicin (Peg‐Doxo) in primary cutaneous T‐cell lymphomas, while in primary cutaneous B‐cell lymphomas (PCBCLs), the drug efficacy has never been assessed so far. Methods: We performed a prospective phase II pilot clinical trial of Peg‐Doxo monotherapy (20 mg/m²) in PCBCLs. One patient had a marginal zone B‐cell lymphoma and four were affected by diffuse large B‐cell lymphoma‐leg type, all with widespread nodular lesions. Results: All the patients achieved a complete response (CR = 100%) in a short period of time (median 3 months), even when pretreated with radio‐chemotherapy. Two experienced a relapse. At follow‐up, one patient died for progressive disease; four are in CR after 5, 52, 63 and 69 months. As concerning the toxicity profile, the treatment was well‐tolerated, no one decreased or delayed the dose. The haematological toxicity was mild with only one case of grade III neutropenia; a patient showed a grade I neurotoxicity. Dermatological toxicity, in particular the palmar–plantar erythrodysesthesia, did not occurred, probably because of both the low dosages of Peg‐Doxo monotherapy and the oral prophylaxis with pyridoxine. Conclusions: In spite of the small number of patients, it emerges that monochemotherapy with Peg‐Doxo has a significantly high clinical activity and a good safety profile in PCBCLs, even in aggressive forms, compared with other therapeutic regimens, which are completely reviewed. It suggests the need of further investigations in this field.     

Identification of prognostic genes and construction of a novel gene signature in the skin melanoma based on the tumor microenvironment

Skin melanoma remains a highly prevalent and yet deadly form of cancer, with the exact degree of melanoma-associated mortality being strongly dependent upon the local tumor microenvironment. The exact composition of stromal and immune cells within this microenvironmental region has the potential to profoundly impact melanoma progression and prognosis. As such, the present study was designed with the goal of clarifying the predictive relevance of stromal and immune cell-related genes in melanoma patients through comprehensive bioinformatics analyses. We therefore analyzed melanoma sample gene expression within The Cancer Genome Atlas database and employed the ESTIMATE algorithm as a means of calculating both stromal and immune scores that were in turn used for identifying differentially expressed genes (DEGs). Subsequently, univariate analyses were used to detect DEGs associated with melanoma patient survival, and through additional functional enrichment analyses, we determined that these survival-related DEGs are largely related to inflammatory and immune responses. A prognostic signature comprised of 10 genes (IL15, CCL8, CLIC2, SAMD9L, TLR2, HLA.DQB1, IGHV1–18, RARRES3, GBP4, APOBEC3G) was generated. This 10-gene signature effectively separated melanoma patients into low- and high-risk groups based upon their survival. These low- and high-risk groups also exhibited distinct immune statuses and differing degrees of immune cell infiltration. In conclusion, our results offer novel insights into a number of microenvironment-associated genes that impact survival outcomes in melanoma patients, potentially highlighting these genes as viable therapeutic targets.