IntroductionLong COVID (LC) poses a persistent challenge in clinical practice due to limited understanding of its etiology. LC is hypothesized to stem from aberrant immune responses in COVID-19. Vaccinations, which boost immune cells to restore function, could help ease LC symptoms.MethodsTo exclude the impact of vaccination, we examined the immune cell profiles of recovering COVID-19 patients before vaccines were available. White blood cell differentials were monitored in ninety-twohealthy unvaccinated controls. Seventy-six unvaccinated COVID-19 patients were monitored upon admission and on the 50th day post-symptom onset (DPSO50). Peripheral lymphocyte subsets were analyzed using flow cytometry.ResultsMild cases showed no significant changes in lymphocyte counts or subsets from admission to DPSO50. By DPSO50, severe and critical cases showed almost complete recovery from lymphopenia, with critical cases having CD19+ B-cell counts approximately 45% lower than the mild group. Severe and critical cases exhibited reduced B-cell frequencies, with critical cases displaying around 48% higher natural killer (NK) cell counts. In mild cases, NK cell counts negatively correlated with B-cell counts (r=-0.528, p=0.02). Additionally, critical cases showed positive correlations between NK cell counts and CD4+ T-cell counts (r=0.83, p<0.01), and between NK cell counts and CD8+ T-cell counts (r=0.74, p<0.01). Severe cases demonstrated decreased counts of CD4+CD25+CD127lowFoxP3+ regulatory T-cells (Tregs), which positively correlated with B-cell counts (r=0.37, p<0.05).DiscussionOur findings indicate that aberrant immune cell profiles in COVID-19 patients change dynamically during recovery, depending on disease severity. This study suggests that convalescent patients from critical COVID-19 may experience long-lasting B-cell lymphopenia.

病理学是医学领域中经典而古老的学科,起源于早期尸体解剖器官形态观察,随着显微镜发明和细胞学说建立,逐渐从宏观走向组织细胞的微观形态.近一个多世纪来,病理学因揭示疾病发生发展及病变机制的核心内容,而成为医学教育基础与临床的"桥梁"学科,也因病理改变是疾病诊断"金标准"而进入临床实践.近数十年来由于生物学技术和生命科学快速进步、个体化医学和靶向治疗时代开启、生物医学与工程门类渗透融合,促进病理学朝向器官系统亚专科方向、细胞大分子分析的分子病理,以及数字和人工智慧化方向发展,成为精准医学的重要支撑学科[1-2].

Purpose Our previously study showed that recombinant human endostatin (Endostar) combined with chemotherapy had significant activity to increase the mPFS in patients with advanced sarcomas with tolerable side effects. However, the small cohort size and short follow-up time made it difficult to screen sensitive sarcoma subtypes and determine whether there is an overall survival benefit. With the largest sarcoma cohort to our knowledge, we try to confirm the efficacy and safety of chemotherapy combined with Endostar in stage IV sarcomas, with the specific purpose of finding out the sensitive sarcoma types for this combined treatment. Methods After the exclusion of ineligible patients, 156 patients with stage IV bone and soft tissue sarcomas were included in this study according to the inclusion criteria. Results By the end of follow-up, the ORR was 10.7% (9/84) vs 1.4% (1/72) (p=0.041), the DCR was 26.2% (22/84) vs 5.6% (4/72) (p=0.001) in the combined group and chemotherapy group, respectively. The mPFS of combined group was significantly longer than the chemotherapy group (10.42 vs 6.87 months, p=0.003). The mOS were 26.84 months and 23.56 months, without significant difference (p= 0.481). In osteogenic sarcoma, there was no statistically significant difference in the mPFS between the two groups (p=0.59), while in the soft tissue sarcoma, the mPFS in the combined group was significantly higher than that of the chemotherapy group (11.27 vs 8.05 months, p=0.004). Specifically, undifferentiated polymorphic sarcoma (UPS) was the possible sarcoma subtypes that benefited from the combined therapy. For the 38 UPS patients (28 patients in the combined group and 10 patients in the chemotherapy group), the mPFS in the combined group was up to 14.88 months, while it was only 7.1 months in the chemotherapy group, with a significant difference (p=0.006). The most common adverse events in the combined group were myelosuppression, gastrointestinal reactions and abnormal liver function, without significant difference in two groups. Conclusion Chemotherapy plus Endostar could prolong mPFS and improve ORR and DCR in patients with stage IV soft tissue sarcoma, suggesting that the combined therapy could improve the patient prognosis in soft tissue sarcomas, especially the UPS patients.

BackgroundThe rarity and complexity of soft tissue sarcoma (STS) make it a challenge to determine the incidence, survival, and metastasis rates. In addition, the clinicopathological risk factors for lymph node metastasis have rarely been reported. MethodsData on patients diagnosed with STS in the SEER database from 2000 to 2018 were extracted by SEER*Stat 8.3.9.1, and the incidence trend was calculated by Joinpoint 4.9 software. The KM method was used to calculate the survival curve, and the log-rank method was used to compare differences in the survival curves. The clinicopathological risk factors for lymph node metastasis were screened by logistic regression. ResultsAmong the 35987 patients, 4299 patients (11.9%) had distant metastasis. The overall lymph node metastasis rate was 6.02%, which included patients suffering from both lymph node and distant metastasis. Considering that some lymph node metastases might be accompanying events of distant metastasis, the rate of only lymph node metastasis in STS patients decreased to 3.42% after excluding patients with distant metastasis. Patients with only lymph node metastases (N1/2M0) had a significantly worse prognosis than those without metastases (N0M0) but a better prognosis than those with only distant metastases (N0M1) (p<0.0001). In the multivariate logistic analysis, STS patients with larger tumors located in the head and neck, viscera, retroperitoneum, and certain specific pathological subtypes (compared with the liposarcoma), such as undifferentiated pleomorphic sarcoma, rhabdomyosarcoma, endometrial stromal sarcoma, gastrointestinal stromal tumor, synovial sarcoma, and angiosarcoma, had a higher risk of lymph node metastasis. ConclusionsLymph node metastasis is rare in STS, and the metastasis rate is significantly different among the different pathological types. Tumor size, location, and pathological subtype are significantly associated with the risk of lymph node metastasis. The overall survival of patients with lymph node metastasis is better than that of patients with distant metastasis, which suggests a more precise prognosis evaluation should be performed in these AJCC stage IV STS patients.

Hyalinising clear cell carcinoma (HCCC) of the lung is an extremely rare tumour that is just recently recognised as one of the salivary gland-type tumours (SGTT) in the latest WHO classification of thoracic tumours. Eleven cases have been reported in English literature since Joaquín et al. reported the first case. Given the very limited number of cases, the clinical and histological features of pulmonary HCCC are equivocal. Herein, we present two cases of pulmonary HCCC. The patients were a 66-year-old man and a 48-year-old woman. The mass was located on the right main bronchus and right middle lobar bronchus separately. One was 2 cm and the other was 3.3 cm in the greatest dimension. The tumours were comprised of small monomorphic cells with clear or eosinophilic cytoplasm and infiltrated in a hyalinising stroma arranged in nests, cords, sheets and trabeculae. Their morphology resembled their head and neck counterparts. Immunohistochemically, the tumour cells were positive for AE1/AE3, P63, while negative for TTF1, Calponin, S-100, HMB45 and PAX8. Ki-67 labeling ranges from 3% to 10%. Fluorescence in situ hybridisation (FISH) demonstrated EWSR1 rearrangement and Next-generation sequencing (NGS) demonstrated EWSR1- ATF1 (exon 11: exon 3) fusion in case one and EWSR1- ATF1 (exon 2: exon 12) fusion in case two. This is the first time to report the EWSR1-ATF1fusion point other than exon 11: exon 3 in pulmonary HCCC. Case one recurred two years after local resection but didn't metastasise during follow-up 36 months. Case two is alive without disease after lobectomy during follow-up 14 months.

BACKGROUND:An accurate genotyping analysis is one of the critical prerequisites for patients with colorectal cancer receiving matched therapies. Conventional genotyping analysis is currently used to detect either gene mutations or MSI status, delaying the detection of critical tumor biomarkers and thus the optimal time for treatment. An assay that analyzes both biomarkers in a streamlined process is eagerly needed. METHODS:We developed an assay combining Multiplex PCR Amplification, Single-base Extension and capillary electrophoresis (CE) analysis (MASE-CE) for synchronous detection of KRAS/NRAS/BRAF mutations and MSI status. In a 190 colorectal cancer cohort, we identified seven somatic mutations in KRAS, NRAS and BRAF as well as five MSI loci (D2S123/D5S346/D17S250/BAT-25/BAT-26) simultaneously. KRAS/NRAS/BRAF mutations were detected by NGS and MASE-CE, and MSI status were detected by PCR-CE and MASE-CE methods. RESULTS:The MASE-CE method showed high consistency with NGS for mutation detection (Kappa value ≥0.8) and PCR-CE (Kappa value = 0.79). In addition, the limits of detection (LOD) of MASE-CE assay for MSI and somatic mutation were 5% and 2%, respectively. CONCLUSIONS:In somatic mutation detection and MSI detection, the LOD of MASE-CE assay was superior to that of qPCR and NGS. MASE-CE assay is a highly sensitive, time-saving and specimen-saving method, which can greatly avoid the cumbersome testing process and provide clinical decision for doctors in time.

目的 探讨智能化抗酸杆菌检测在结核病理诊断中的应用.方法 收集150例肉芽肿性改变的石蜡包埋组织,同时进行手工抗酸染色和自动抗酸染色,分别进行人工阅片病理诊断.使用自主开发的结核杆菌识别人工智能(tuber-culosis bacillus finder,TB-Finder)作为初筛,另请病理医师复核150张自动抗酸染色切片并作出病理诊断,分析两种染色模式的效率及准确率.结果 150例智能化平台染色的阳性率为29.3％(44/150),传统模式染色的阳性率为6.6％(10/150).染色加诊断全流程智能化平台每张切片耗时12 min,传统模式每张切片耗时57 min,智能化平台节约时间成本约78％.结论 智能化抗酸杆菌检测平台的应用可提高病理医师的工作效率和诊断准确率,有助于规范质量控制.

目的 探讨p16表达在胃癌前病变、早期胃癌及癌旁非肿瘤性胃黏膜中的表达及临床意义.方法 使用内镜黏膜下剥离术(ESD)获得的早期胃癌标本30例(除黏膜内癌,还包括低级别异型增生21例、高级别异型增生23例、黏膜下浸润癌9例),采用免疫组织化学法检测p16、Ki-67、MUC5AC、MUC6、MUC2表达,分析标志物之间,以及与癌分化、临床特征的关系.结果 p16在非肿瘤性胃黏膜组织中呈阴性或个别细胞核表达(≤1+),而低级别异型增生、高级别异型增生、黏膜内癌及黏膜下浸润癌细胞中其过表达(≥2+)阳性率分别为4.8％、39.1％、80.0％、88.9％,差异均具有统计学意义(x2=34.515,P＜0.001).p16表达方式分析显示8例低级别异型增生中均为核阳性,而15例高级别异型增生阳性中,2例为核/质阳性(2/15,13.3％),28例黏膜内癌阳性中,22例为核/质阳性(22/28,78.6％),9例黏膜下浸润癌8例为核/质阳性(8/9,88.9％).p16核/质阳性在黏膜内癌及黏膜下浸润癌与高级别异型增生间具有显著性差异(x2=16.856,P＜0.001;x2=10.286,P=0.001).p16表达与Ki-67指数在低级别异型增生、高级别异型增生、黏膜内癌相关,但浸润癌中p16过表达高而Ki-67指数较低(22.2％).p16过表达与癌分化程度、细胞类型,以及其他临床特征无关.但同一样本的不同区域p16表达在高级别异型增生及早癌中有一定的异质性,在检测中需要注意.结论 p16过表达存在于胃癌前病变及早期胃癌,与良性增生有鉴别意义;尤其p16核/质过表达对于高级别异型增生、黏膜内癌、浸润癌具有提示意义;同时,在胃癌前病变及早期胃癌p16表达与Ki-67增殖指数具有一致性.

To the Editor: Raynaud phenomenon (RP) is a transient, vasospastic phenomenon that is prominent in systemic sclerosis (SSc). Most patients with SSc develop RP and can result in recurrent digital ulcers (DU) and critical ischemic events. General management of RP, including lifestyle changes, pharmacological and surgical intervention, can hardly lead to optimal remission of RP symptoms and ischemic complications quickly. Botulinum toxin (BTX) injection has been investigated as a treatment option in RP. In 2004, two RP patients were successfully treated with BTX-A for the first time. In recent years, clinical trials have been carried out to assess the therapeutic efficacy of local injections with BTX in improving primary and secondary RP.[1] However, the efficacy, best injection protocol, and side effects of BTX in treating RP secondary to SSc (SSc-RP) need to be systemically reviewed. Here, we analyzed the published clinical studies relevant to the therapeutic effect of BTX in treating SSc-RP, aiming to renew our recognition of the treatment. This systemic review has been registered on PROSPERO (International Prospective Register of Systematic Reviews, ID: CRD42020158574). We utilized a string made up of relevant keywords ("scleroderma" or "systemic sclerosis" or "SSc") AND ("botulinum toxin" or "clostridium botulinum toxins" or "botulin") [Supplementary Table 1, https://links.lww.com/CM9/A860]. A literature search was conducted using PubMed, Cochrane Library, Embase, and Web of Science in January 2021 for all articles published from 1995 to 2021. The initial search yielded 251 articles, which were narrowed down to five according to the inclusion and exclusion criteria [Supplementary Table 2, https://links.lww.com/CM9/A860]. The study selection process is shown in [Supplementary Figure 1, https://links.lww.com/CM9/A860]. We had gotten three case series studies[2–4] and two randomized controlled trials (RCTs)[5,6] finally. The risk of bias of the included studies is illustrated in [Supplementary Figure 2, https://links.lww.com/CM9/A860]. A total of five articles involving 155 patients were assessed in this systemic review. 10.3% (16/155) of the enrolled patients were males, and 89.7% (139/155) were females. Patient characteristics are listed in [Supplementary Table 3, https://links.lww.com/CM9/A860]. All patients included in this study suffered from severe symptoms of SSc-RP and had failed conventional medical treatment or surgical therapy. A variety of protocols for BTX injection were used. The studies differed with respect to doses and sites of BTX injection as well as the type of BTX [Supplementary Table 3, https://links.lww.com/CM9/A860]. Four studies used BTX-A[2–5] while one RCT done by Motegi et al[2] used BTX-B for injection.[6] The dose of BTX-A varied from 10 U to 100 U,[3–5] and the dose of BTX-B included 250 U, 1000 U, and 2000 U.[6] In three studies, BTX was injected into the palmar aspect of the hand, targeting the neurovascular bundles[2,3,6] and in two studies, BTX was injected into the dorsal surface near the proximal phalanx base.[4,5] The researches support the efficacy of local BTX injection for the treatment of SSc-RP. Most of the studies (four studies including 115 patients) showed that BTX injection was effective in the treatment of RP and related DUs in patients with SSc.[2–4,6] It could improve the symptoms of SSc-RP, reduce the pain, and heal intractable DUs. Motegi et al[2] measured severity of RP, Raynaud score including frequency, pain, color, and duration, as the primary endpoint, and change in temperature after cold-water challenge, number of DUs, and pain Visual Analogue Scale (VAS) as the secondary endpoint in 45 and 10 patients, respectively (no overlap between two groups).[6] They found that Raynaud score and pain VAS score in the BTX group was significantly lower than that in the control group (P < 0.05), and skin temperature recovery after cold-water stimulation was significantly improved after BTX injection. The numbers of DUs in the BTX group were also significantly lower in 4 to 16 weeks after injection. Uppal et al[3] assessed hand function, joint movement, ulcer healing, and subjective feeling including pain, color change, and cold intolerance in 20 patients. They found that hand function and the ranges of joint movement of the fingers were statistically significantly improved. Besides, 75% of the patients who had ulceration in fingers showed complete healing and 80% of the patients showed improvement in symptoms of the injected hands. However, Bello et al[5] reported a negative result showing that although the enrolled 40 patients had slightly better outcomes after injecting BTX-A into affected hands, the difference was not statistically significant. They performed a subgroup analysis according to SSc subtype, RP disease duration, RP severity, and baseline treatment, and found that patients with longer disease duration since RP onset (>15.56 years) and diffuse scleroderma subtype might respond worse to BTX injection.[5] We further focused on the influence of BTX type (BTX-A and BTX-B) on the efficacy of SSc-RP. In the included studies, four studies including 110 patients used BTX-A,[2,3,5] one study including 45 patients used BTX-B.[6] Among the four studies using BTX-A, three case series studies showed great efficacy,[2–4] and one RCT showed the differences were not statistically significant.[5] Both BTX-A and BTX-B can produce good efficacy although the optimal doses were different. BTX-B was effective at approximately 20 to 40 times the dose of BTX-A in the treatment of SSc-RP according to Motegi et al.[6] The doses of as low as 10 U to 100 U BTX-A have been reported to show favorable results.[2–4] We did not find the difference in treatment effects among different dosages. As for the injection dose of BTX-B, Motegi et al[6] found that the effective dose might be ≥1000 U, and both 1000 U and 2000 U had similar treatment effects while the 2000 U group might improve skin surface temperature better.[6] The studies reported different onset times of pain reduction and times of ulcer healing. The onset time of pain reduction varied from as soon as 2 weeks to within 16 weeks after injection. The healing time of DUs was 12 weeks. The duration of efficacy was between 4 and 6 months. The injection sites were described differently, but most of the studies injected BTX around the neurovascular bundles proximal to the A1 pulley, the space next to the metacarpophalangeal joint. Motegi et al[6] and Uppal et al[3] injected BTX into the palmar aspect of the hand, while Bello et al[5] who got the negative result injected BTX into the dorsal surface of the hand. It seems that the palmar aspect might be a better injection site. The major finding of this systemic review is that researches support the efficacy and safety of local BTX injection for the treatment of SSc-RP. The injection site getting close to the sclerosis lesions, near the digital neurovascular, might be suitable. The dose of as low as 10 U and up to 100 U of BTX-A, and 1000 U or 2000 U BTX-B could all produce great efficacy, while the higher dose may lead to more side effects. Larger double-blind, prospective, randomized, placebo-controlled studies are needed to confirm the beneficial efficacy of BTX on the treatment of SSc-RP, determine injection protocol, and explore different subgroups of patients responding to BTX injection, hence providing high-quality evidence to support future clinical decision-making. Funding This work was supported by a grant from the National Natural Science Foundation of China (No. 81771706). Conflicts of interest None.

目的:探讨PD-L1在Ⅰ~Ⅱ期肺腺癌的表达情况及其与临床病理参数和常见基因突变的相关性。方法:选取2019年4—11月北京大学第一医院胸外科手术切除Ⅰ~Ⅱ期的肺腺癌病例共253例。依据临床病理特征进行分组，采用免疫组织化学方法检测标本中PD-L1的表达，计算肿瘤比例评分（tumor proportion score，TPS）。TPS<1%为无表达，TPS≥1%为有表达，TPS≥50%为高表达。采用二代测序技术对其中85例样本的13个肺癌常见基因突变进行全外显子检测。比较PD-L1的表达在各临床病理分组间的差异以及与基因突变状态的相关性。结果:PD-L1表达阳性率为9.9%（25/253），高表达率为2.0%（5/253）。ⅡB期病例PD-L1表达率高于Ⅰ~ⅡA期，浸润性癌PD-L1表达率高于微小浸润性癌，PD-L1表达率随着浸润性癌病理学分级的升高而升高。PD-L1的表达显示出与表皮生长因子受体（EGFR）基因突变具有负相关趋势。结论:PD-L1在Ⅰ~Ⅱ期肺腺癌的表达明显低于Ⅲ~Ⅳ期；在Ⅰ~Ⅱ期病例内部，其阳性率随着肿瘤分期和分级的升高而升高。PD-L1的表达与EGFR基因突变具有负相关趋势，可能是EGFR突变型肺腺癌预后好于野生型的原因之一。

Abstract. Background. Methylene blue is the most commonly used tracer for sentinel lymph node (SLN) biopsy (SLNB) in China. This study aimed to investigate the feasibility of clinical application of SLNB using methylene blue dye (MBD) for early breast cancer and the prognosis of patients with different SLN and non-SLN statuses. Methods. We retrospectively analyzed the clinicopathological data of patients with early breast cancer treated at the Peking University First Hospital between 2013 and 2018. We calculated the SLN identification rate (IR) in SLNB with MBD and the false-negative rate (FNR), and analyzed the prognosis of patients with different SLN and non-SLN statuses using Kaplan-Meier curves. Results. Between January 2013 and December 2018, 1603 patients with early breast cancer underwent SLNB with MBD. The SLN IR was 95.8% (1536/1603). Two SLNs (median) were detected per patient. There were significant differences in FNR between patients with SLN micrometastasis and macrometastasis (19.0% vs. 4.5%, χ2 = 12.771, P < 0.001). Chi-square test showed that there were significant differences in SLN successful detection rates among patients with different vascular tumor embolism status (96.3% vs. 90.8%, χ2 = 9.013, P = 0.003) and tumor (T) stages (96.6% vs. 94.1%, χ2 = 5.189, P = 0.023). Multivariate analysis showed that vascular tumor embolism was the only independent factor for SLN successful detection (odds ratio: 0.440, 95% confidence interval: 0.224−0.862, P = 0.017). Survival analysis showed a significant difference in disease-free survival (DFS) between patients with non-SLN metastasis and patients without non-SLN metastasis (P = 0.006). Conclusion. Our single-center data show that, as a commonly used tracer in SLNB in China, MBD has an acceptable SLN IR and a low FNR in frozen sections. This finding is consistent with reports of dual tracer-guided SLNB. Positive SLNs with non-SLN metastasis are associated with DFS.

Paraneoplastic autoimmune multiorgan syndrome is a complex and deadly disease. We retrospectively reviewed the clinical features and risk factors for paraneoplastic autoimmune multiorgan syndrome in 145 Chinese patients. The most common neoplasm was Castleman disease (56%), and patients with Castle-man disease tended to be younger (<= 42 years old: 83% vs. 29%) and to have a greater proportions of lichen planus-like lesions (47% vs. 27%) and bronchiolitis obliterans (49% vs. 29%), compared to other neoplasm-associated patients. Among all 145 patients in the study, the survival rates were 84% at 1 year, 65% at 3 years, and 54% at 5 years. Kaplan-Meier curve analysis revealed that mortality was associated with older age (> 42 years), neoplasm type, labial lesions, and larger skin lesion area (> 17.5% of the body surface area). However, only older age and larger skin lesion area were independent factors associated with mortality in multivariate analysis. We suggest that patients with Castleman disease and paraneoplastic autoimmune multiorgan syndrome have many unique characteristics and the underlying risk factors for death require further exploration.

OBJECTIVES:We aimed to describe the clinical and histopathologic features of Castleman disease (CD), particularly emphasizing its associations with paraneoplastic pemphigus (PNP) and prognosis.METHODS:We retrospectively enrolled 123 CD patients at our center. Clinical, pathologic, and laboratory data were reviewed.RESULTS:Fifty percent of the patients had PNP. Compared with those without PNP, patients with PNP-associated CD had more hyaline vascular (HV) variants (83.9% vs 57.4%), fewer mixed cellular variants (16.1% vs 24.6%), and no plasmacytic variants (0% vs 18.0%). Thirty-eight of 87 patients with the HV variant of CD (HV-CD) had stroma-rich (SR) features, and the incidence rate was higher in those with PNP-associated CD than in those without PNP (48.4% vs 13.1%, P < .001). The SR variant was associated with higher PNP-associated IgG titers than SR absence before surgery (median 1:160 vs 1:80, P = .019) or after surgery (median 1:160 vs 1:40, P = .013). The SR variant was also an unfavorable prognostic factor for CD survival in univariate analysis. The 3-year survival rates were 47.5% among those with PNP and 87.7% among those without PNP (P < .001).CONCLUSIONS:PNP is associated with specific subtypes of CD and affects survival. The SR variant of HV-CD positively correlates with the incidence of PNP.

Objective To investigate histo-pathological distribution and clinico-pathological significance in a large Chinese triple-negative breast cancer (TNBC) patients serials based on the latest understanding of its clinico-pathological diversity, and to provide more information to clinicians to improve precision of individualized treatment of TNBC. Methods A retrospective analysis was performed on patients with TNBC at Breast Disease Center, Peking University First Hospital between January 2010 and December 2019. Histo- and clinico-pathological characteristics were analyzed by Chi-square test and Student’s t-test, and prognoses were calculated using Kaplan-Meier method and a Cox proportionate hazards model. Bonferroni correction was used to correct for multiple comparison. Results Conventional type of TNBC (cTNBC) were identified in 73.7% of 582 TNBC, while special type of TNBC (sTNBC) were 26.3%, including 71 apocrine carcinoma, 20 medullary carcinoma, 31 metaplastic carcinoma, 18 invasive lobular carcinoma, 7 invasive micropapillary carcinoma, 5 adenoid cystic carcinoma and 1 acinic cell carcinoma. Compared to sTNBC, cTNBC was associated with high histologic grade (P<0.001) and lower androgen receptor (AR) expression (P<0.001). TNM stage of low-grade cTNBC was significantly lower than that of high-grade cTNBC (P=0.002). Although no significant difference, there was a trend that the rate of 5-year disease-free survival (DFS) and 5-year overall survival (OS) were longer in high-grade cTNBC than in high-grade sTNBC (P=0.091 and 0.518), and were longer in low-grade sTNBC than in high-grade sTNBC (P=0.051 and 0.350). Metaplastic carcinomas showed larger tumor size (P=0.008) and higher proliferative Ki67 index (P=0.004) than cTNBCs. Conclusions Results from our cohort imply that sub-categorization or subtyping and histological grading could be meaningful in pathological evaluation of TNBC, and need to be clarified in more large collections of TNBC.

Krüppel-like factor 4 (KLF4) is a transcription factor and plays a vital role in cancer initiation and development. However, the role of Krüppel-like factor 4 in the metastasis of non-small cell lung cancer (NSCLC) is not clear. Here, we demonstrated that the expression of Krüppel-like factor 4 was significantly decreased in human non-small cell lung cancer tissues compared with that in normal tissues using Western blot. We performed immunohistochemical staining and observed the decreased expression of Krüppel-like factor 4 in human lung cancer tissues, and metastatic tumor tissues located in the trachea and main bronchus. We also found that the E-cadherin expression was decreased, while vimentin expression was increased in human NSCLC tissues and metastatic tumor tissues located in the trachea and main bronchus. Additionally, enforced expression of Krüppel-like factor 4 in mouse lungs significantly inhibited the metastasis of circulating Lewis lung carcinoma cells to the lungs by attenuating mesenchymal-epithelial transition (MET). Furthermore, cell scratch assays and Matrigel invasion assays revealed that overexpression of Krüppel-like factor 4 inhibited the migration and invasion of non-small cell lung cancer cell lines A549, H1299, H226, and H1650 cells. Moreover, overexpression of Krüppel-like factor 4 attenuated TGF-β1-induced epithelial-mesenchymal transition (EMT) in A549, and inhibited the phosphorylation of c-Jun-NH2-terminal kinase (JNK), an important pathway in metastasis in non-small cell lung cancer. Our in vivo and in vitro findings illustrate that Krüppel-like factor 4 inhibited metastasis and migration of non-small cell lung cancer, and indicate that Krüppel-like factor 4 could be a potential therapeutic target for the treatment of non-small cell lung cancer.

Background Non-Langerhans cell histiocytosis (non-LCH) is a collective term that encompasses a long list of rare "histiocytosis" that do not meet the criteria for Langerhans cell histiocytosis (LCH). Among cutaneous non-LCH, the xanthogranuloma (XG) family represents a distinct group of disorders derived from dermal dendritic cells (DDCs) at different stages of differentiation. Objective To investigate the clinicopathological characteristics of the XG family in adults and review the relevant literature. Materials and Methods We performed a retrospective clinicopathological study of five adult cases with a previous diagnosis of non-LCH. Clinicopathological features, immunophenotypes, genetic alterations and ultrastructural characteristics were analysed. Results Skin biopsies revealed that all five cases were characterized by diffuse infiltration of polymorphic cells, which were immunoreactive to factor XIIIa but negative for Langerin, CD1a, and S100. None of the cases harboured the BRAF V600E mutation. Electron microscopy of two cases exhibited abundant cytoplasmic processes with numerous lysosome-like dense bodies and electron-lucent vesicles in the cytoplasm and extracellular matrix. The overall features suggested that DDCs are the cellular origin, and these cases fulfilled the criteria for the XG family. Conclusion The XG family represents a spectrum of rare diseases with different clinical presentations, a wide range of morphological appearances, and a shared common origin (DDCs). This group of disorders has been proposed as a unique entity with diagnostic challenges that should not be underestimated.

BACKGROUND:Some drugs that target molecular pathways are available for the targeted treatment of lung cancer. Multiple tests are needed to detect the status of the known molecular targets to determine whether the patients can respond to the drugs. An integrated platform for various gene alteration detection including both mutations and rearrangements is necessary for patients, especially those without enough tissue.METHODS:In our study, detections of EGFR mutations, ALK rearrangement, ROS1 rearrangement, and alterations of other nine important lung cancer-related genes were integrated into a single next-generation sequencing (NGS) platform. The NGS analysis was performed in 107 cases of non-small cell lung cancer (NSCLC). Meanwhile, hot spots such as EGFR L858R, EGFR E746-A750Del mutations and gene rearrangement of ALK and ROS1 were detected by immunohistochemical (IHC) staining.RESULTS:NGS could explore various gene mutations and gene rearrangements with a reduced experiment time and lower amounts of tumor tissues than multiple IHC staining experiments. NGS results were more informative and reliable than IHC staining for EGFR gene alterations, especially for the exon 19 region. NGS could also increase the positive rate of ALK rearrangement and decrease the false positive results of ROS1 rearrangements detected by IHC staining.CONCLUSIONS:NGS is effective for confirmation the status of various important lung cancer-related gene alterations. Furthermore, NGS is necessary for the confirmation of the IHC results of ALK and ROS1 rearrangements.

The CD38 molecule (CD38) catalyzes biogenesis of the calcium-mobilizing messenger cyclic ADP-ribose (cADPR). CD38 has dual membrane orientations, and type III CD38, with its catalytic domain facing the cytosol, has low abundance but is efficient in cyclizing cytosolic NAD to produce cADPR. The role of cell surface type II CD38 in cellular cADPR production is unknown. Here we modulated type II CD38 expression and assessed the effects of this modulation on cADPR levels. We developed a photoactivatable cross-linking probe based on a CD38 nanobody, and, combining it with MS analysis, we discovered that cell surface CD38 interacts with CD71. CD71 knockdown increased CD38 levels, and CD38 knockout reciprocally increased CD71, and both could be cocapped and coimmunoprecipitated. We constructed a chimera comprising the N-terminal segment of CD71 and a CD38 nanobody to mimic CD71's ligand property. Overexpression of this chimera induced a dramatically large decrease in CD38 via lysosomes. Remarkably, cellular cADPR levels did not decrease correspondingly. Bafilomycin-mediated blockade of lysosomal degradation greatly elevated active type II CD38 by trapping it in the lysosomes but also did not increase cADPR levels. Retention of type II CD38 in the endoplasmic reticulum (ER) by expressing an ER construct that prevented its transport to the cell surface likewise did not change cADPR levels. These results provide first and direct evidence that cADPR biogenesis occurs in the cytosol and is catalyzed mainly by type III CD38 and that type II CD38, compartmentalized in the ER or lysosomes or on the cell surface, contributes only minimally to cADPR biogenesis.