[Studies on clinicopathological features of duodenal-type follicular lymphoma of 18 patients].

To investigate the clinical and pathological characteristics of duodenal-type follicular lymphoma (D-FL), and to deepen the understanding of Duodenal-type follicular lymphoma. The clinical symptoms, endoscopic features, pathologic features, immunophenotype, molecular pathological features and treatment follow-up of 18 D-FL patients diagnosed in Department of Pathology, Beijing Tiantan Hospital affiliated to Capital Medical University between January 2020 and July 2023 were summarized. A total of 18 patients with D-FL were included, including 10 males and 8 females. The median age was 49 (32-69) years respectively. Most of the patients were found during gastroenteroscopy or presented with the common gastrointestinal symptoms of stomach pain, acid reflux, vomiting and diarrhea. Most endoscopic findings were multiple small gray and white polyposis. In the pathological morphology, the mucous layer and submucous layer showed lymphoid follicular structures with full and strained follicles. The immunophenotype showed that the tumor cells strongly expressed CD20 and BCL2 and had low proliferation activity. Immunoglobulin clonal analysis of 1 case showed IgK monoclonal rearrangement (1/1). FISH showed 1 case of BCL2 gene rearrangement (1/3). All patients did not receive targeted chemotherapy and adopted a wait-and-see strategy. Median follow-up was 12 (2-34) months. This study shows that D-FL is an indolent lymphoma, which tends to occur in the duodenum and has a good prognosis.

[Mid-term efficacy of China Net Childhood Lymphoma-mature B-cell lymphoma 2017 regimen in the treatment of pediatric Burkitt lymphoma].

Objective: To analyze the clinical characteristics of children with Burkitt lymphoma (BL) and to summarize the mid-term efficacy of China Net Childhood Lymphoma-mature B-cell lymphoma 2017 (CNCL-B-NHL-2017) regimen. Methods: Clinical features of 436 BL patients who were ≤18 years old and treated with the CNCL-B-NHL-2017 regimen from May 2017 to April 2021 were analyzed retrospectively. Clinical characteristics of patients at disease onset were analyzed and the therapeutic effects of patients with different clinical stages and risk groups were compared. Survival analysis was performed by Kaplan-Meier method, and Cox regression was used to identify the prognostic factors. Results: Among 436 patients, there were 368 (84.4%) males and 68 (15.6%) females, the age of disease onset was 6.0 (4.0, 9.0) years old. According to the St. Jude staging system, there were 4 patients (0.9%) with stage Ⅰ, 30 patients (6.9%) with stage Ⅱ, 217 patients (49.8%) with stage Ⅲ, and 185 patients (42.4%) with stage Ⅳ. All patients were stratified into following risk groups: group A (n=1, 0.2%), group B1 (n=46, 10.6%), group B2 (n=19, 4.4%), group C1 (n=285, 65.4%), group C2 (n=85, 19.5%). Sixty-three patients (14.4%) were treated with chemotherapy only and 373 patients (85.6%) were treated with chemotherapy combined with rituximab. Twenty-one patients (4.8%) suffered from progressive disease, 3 patients (0.7%) relapsed, and 13 patients (3.0%) died of treatment-related complications. The follow-up time of all patients was 24.0 (13.0, 35.0) months, the 2-year event free survival (EFS) rate of all patients was (90.9±1.4) %. The 2-year EFS rates of group A, B1, B2, C1 and C2 were 100.0%, 100.0%, (94.7±5.1) %, (90.7±1.7) % and (85.9±4.0) %, respectively. The 2-year EFS rates was higher in group A, B1, and B2 than those in group C1 (χ2=4.16, P=0.041) and group C2 (χ2=7.21, P=0.007). The 2-year EFS rates of the patients treated with chemotherapy alone and those treated with chemotherapy combined with rituximab were (79.3±5.1)% and (92.9±1.4)% (χ2=14.23, P<0.001) respectively. Multivariate analysis showed that stage Ⅳ (including leukemia stage), serum lactate dehydrogenase (LDH)>4-fold normal value, and with residual tumor in the mid-term evaluation were risk factors for poor prognosis (HR=1.38,1.23,8.52,95%CI 1.05-1.82,1.05-1.43,3.96-18.30). Conclusions: The CNCL-B-NHL-2017 regimen show significant effect in the treatment of pediatric BL. The combination of rituximab improve the efficacy further.

[The outcomes of Tiantan first-aid protocol on critically ill patients with primary central nervous system lymphoma].

Objective: To examine the outcomes of Tiantan first-aid protocol on critically ill patients with primary central nervous system lymphoma (PCNSL). Methods: The clinical data of 18 patients with PCNSL who were treated according to Tiantan first-aid protocol at Department of Neurosurgery,Beijing Tiantan Hospital, Capital Medical University from November 2019 to December 2021 were retrospectively analyzed. There were 9 males and 9 females, aged (56.9±11.1)years (range: 29 to 77 years). The median Karnofsky performance status(KPS) score at admission was 40 (range: 20 to 60). Three patients were mild coma, 3 were lethargy and 12 were conscious. The mean midline shift was 0.7 cm (range: 0 to 1.8 cm). After admission, all patients were treated according to the plan of rapid biopsy, rapid routine pathology and rapid salvage chemotherapy. The treatment procedures, clinical and radiographic outcomes, KPS score and adverse reactions of patients after chemotherapy were collected. Results: All of the 18 patients completed the first-aid treatment. The median duration from admission to the biopsy was 1 day (range: 0 to 5 days), from biopsy to routine pathological diagnosis was 1 day (range: 1 to 4 days) and from routine pathology to salvage chemotherapy was 1 day (range: 0 to 4 days). All the patients were pathologically confirmed with diffuse large B cell lymphoma, 1 patient was double-hit lymphoma. Seventeen patients underwent clinical remission and 1 died of cardiac dysfunction. The successful salvage rate was 17/18. Radiologically, complete remission was observed in 1 case, partial remission in 16 cases, and stable disease in 1 case. The median KPS score at discharge was 60 (range: 30 to 80). The mild gastrointestinal, hematological and hepatic adverse effects were observed after chemotherapy. Conclusion: Tiantan first-aid protocol is effective for critically ill patients with PCNSL, which has the merit to be popularly used and improved.

[Investigation for pathological interpretation criteria and its prognostic value for P53 expression in Chinese diffuse large B-cell lymphoma].

Objective: To explore the feasibility of predicting TP53 mutation risk by immunohistochemical staining (IHC) pattern of P53 in Chinese diffuse large B-cell lymphoma (DLBCL) and its correlation with a prognostic difference. Methods: Between January 2021 and December 2021, 51 DLBCL cases at Beijing Boren Hospital were gathered. These cases had both IHC and next-generation sequencing (NGS) results. IHC classified the P53 protein expression pattern into a loss (<1% ) , diffuse (>80% ) , and heterogeneous (1% -80% ) . The sensitivity and specificity of the predicting TP53 mutation by IHC were assessed by comparing the results of the NGS, and the TP53 high mutation risk group included both loss and diffuse expression of P53. From June 2016 to September 2019, Peking University Cancer Hospital collected 131 DLBCL cases with thorough clinicopathological and follow-up data. From their tumor blocks, tissue microarray blocks were made for IHC evaluation of P53 expression pattern, and prognosis effect of P53 studies. Results: Among 51 cases with both IHC and NGS results, 23 cases were classified as TP53 high mutation risk (7 cases loss and 16 cases diffuse) , 22/23 cases were proved with mutated TP53 by NGS. Only 1 of the 28 cases classified as TP53 low mutation risk was proved with mutated TP53 by NGS. IHC had a sensitivity and specificity of 95.7% and 96.4% for predicting TP53 mutation. NGS identified a total of 26 TP53 mutations with a mutation frequency of 61.57% (13.41% -86.25% ) . In the diffuse group, 16 missense mutations and 2 splice mutations were detected; 6 truncating mutations and 1 splice mutation were detected in the loss group; 1 truncating mutation was detected in the heterogeneous group. Multivariate analysis demonstrated that TP53 cases with high mutation risk have impartial adverse significance for the 131 patients included in survival analysis (HR=2.612, 95% CI 1.145-5.956, P=0.022) . Conclusion: IHC of P53 exhibiting loss (<1% ) or diffuse (>80% ) pattern indicated TP53 high mutation risk, IHC can predict TP53 mutation with high specificity and sensitivity. TP53 high mutation risk is an independent predictor for adverse survival.

[The incidence of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements in diffuse large B-cell lymphoma].

Objective: To investigate the incidence of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangement in Chinese diffuse large B-cell lymphoma (DLBCL) . Methods: From January 2013 to August 2020, 922 DLBCL cases were collected. C-MYC and BCL2 protein expression levels were analyzed by immunohistochemistry staining. Fluorescence in situ hybridization was used to detect the structural abnormalities of MYC, BCL2, and BCL6, including gene breaks and copy number changes. Results: MYC and BCL2 and/or BCL6 gene breaks were found in 29 out of 922 DLBCL cases (3.15%) , including 25 cases of double-hit lymphoma (DHL; 14 cases involving MYC and BCL2 rearrangements and 11 cases involving MYC and BCL6 rearrangements) and four cases involving MYC, BCL2, and BCL6 rearrangements, referring to triple-hit lymphoma. According to the threshold of C-MYC ≥40% and BCL2 ≥50%, 541 cases (58.68%) overexpressed C-MYC and BCL2 proteins, including 22 DHL cases. Moreover, according to the threshold of C-MYC ≥70% and BCL2 ≥50%, 52 cases (5.64%) overexpressed C-MYC and BCL2 proteins, including nine DHL cases. The P53 protein expression was detected by immunohistochemistry staining. The mutant P53 expression pattern was shown in 101 out of 709 cases (14.25%) , whereas 13 cases (1.83%) were negative, likely indicating P53 gene fragment deletion. Conclusion: The incidence of high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 rearrangements was low in DLBCLs, and no significant correlation between gene abnormality and protein overexpression was shown. The correct diagnosis of DHL depends on molecular genetic detection.

[The pathological diagnosis and differential diagnosis of hematopoietic cell tumors of ambiguous lineage].

Objective: To explore the key points of the pathological and differential diagnoses of extra-medullary masses of hematopoietic cell tumors of ambiguous lineage, and to discuss the possible solutions. Methods: Five hematopoietic cell tumors of ambiguous lineage cases were collected, including myeloid sarcoma, mixed phenotype acute leukemia, B/myeloid, T-lymphoblastic lymphoma combined with acute myeloid leukemia, acute undifferentiated leukemia with cutaneous MPDCP and early T-precursor cell acute lymphoblastic leukemia. The data including morphology, immunostaining, and flow cytometry analysis were collected, and we explored the problems and differential diagnosis in the diagnosis of hematopoietic cell tumors of ambiguous lineage. Results: The five cases showed that the accurate pathological diagnosis and classification of hematopoietic cell tumors of ambiguous lineage should be based on lineage-specific antigens. Moreover, tumor cells have the potential of multi-directional differentiation. In different sites or different periods, the differentiation of tumor cells may be different. Biopsy and detection of all related markers should be performed for the initial diagnosis, and the detection should be repeated when the condition of the patient changes. Combined application of multi-techniques, including morphology and flow cytometry analysis, is recommend for the diagnosis of hematopoietic cell tumors of ambiguous lineage, since the conventional morphology and immunophenotyping methods are limited. Conclusion: Hematopoietic cell tumors of ambiguous lineage are derived from hematopoietic stem cells with a potential of multi-differentiation. The differentiation of tumor cells is variable. We need to integrate cell morphology, flow cytometry, pathology, clinical data, and molecular genetics to make a comprehensive diagnosis.

Suppression of αvß6 downregulates P-glycoprotein and sensitizes multidrug-resistant breast cancer cells to anticancer drugs.

Multidrug resistance (MDR) in breast cancer treatment is the major cause leading to the failure of chemotherapy. P-glycoprotein (P-gp), the product of the human MDR1 gene, plays a key role in resistance to chemotherapy and confers cross-resistance to many structurally unrelated anticancer drugs. We have previously reported that integrin αvß6 plays a critical role in breast cancer invasion and metastasis. However, whether and how αvß6 is associated with P-gp and regulated by potential genetic mechanisms in breast cancer remains unclear. In the present study, we further investigated the reversal effect and underlying mechanisms of MDR in breast cancer. Two small interfering RNA constructs (pSUPER-ß6shRNAs) targeting two different regions of the ß6 gene have been designed to inhibit αvß6 expression by transfecting them into adriamycin-resistant MCF-7/ADR cell lines. Suppression of αvß6 dramatically downregulated the levels of MDR1 gene mRNA and P-gp. In particular, ß6shRNA-mediated silencing of αvß6 gene increased significantly the cellular accumulation of Rhodamine 123 and markedly decreased drug efflux ability, suggesting that ß6shRNAs indeed inhibit P-gp mediated drug efflux and effectively overcome drug resistance. In addition, inhibition of integrin αvß6 suppressed the expression of ERK1/2. Interestingly, our data demonstrate that suppression of integrin αvß6 caused significant downregulation of Bcl-2, Bcl-xL and upregulation of caspase 3, Bad, accompanied by increasing activity of cytochrome C. A possible connection between αvß6 and P-gp in drug resistance biology is suggested. Taken together, ß6shRNA could efficiently inhibit αvß6 and MDR1 expression in vitro and these findings may offer specifically useful means to reverse MDR in breast cancer therapy.

[Value of PET/CT in the prognosis of extranodal NK/T cell lymphoma].

Objective: To explore the value of Positron-Emission Tomography/Computed Tomography (PET/CT) in the prognosis of extranodal NK/T cell lymphoma. Methods: The patients of NK/T cell lymphoma diagnosed from January 2007 to July 2016 in Department of Pathology of Beijing Tongren Hospital were enrolled in this study. Seventy-two in-hospital patients were examined on the invasion of adjecent tissue or organ by PET/CT. The PET/CT results were analyzed retrospectively. Kaplan-Meier method was used to analyze the prognostic value of the positive results by PET/CT on overall survival (OS). Results: There were 54 males and 18 females with median age of 44.5 years (13-75 years). According to Ann Arbor staging system, there were 16 cases (22.2%) in stage Ⅰ, 29 cases (40.3%) in stage Ⅱ, 6 cases (8.3%) in stage Ⅲ and 21 cases (29.2%) in stage Ⅳ. According to the IPI scoring system, there were 34 cases (47.2%) in the low risk group (0-1 point), 21 cases (29.2%) in the low-middle risk group (2 points), 16 cases (22.2%) in the middle-high risk group (3 points), and 1 case (1.4%) in the high risk group (4-5 points) . The median follow-up time was 29.2 months (1-118 months). The disease occured in unilateral nasal cavity in 26 cases (36.1%), bilateral nasal cavities in 36 cases (50.0%), nasopharynx, oropharynx and pharynx in 10 cases (13.9%). The tumors of 51 cases involved the surrounding tissue, including nasal wings in 29 cases (40.3%), nasal sinus in 29 cases (40.3%), maxillofacial soft tissue in 18 cases (25.0%), orbital in 12 cases (16.7%), maxilla and skull base in 8 cases (11.1%), eyelid in 6 cases (8.3%), brain tissue in 3 cases (4.2%), eyeball in 2 cases (2.8%). In addition, cervical and inguinal lymphadenopathy were found in 54 cases (75.0%) . Splenomegaly and hepatomegaly were found in 10 cases (13.9%) and 2 cases (2.8%), respectively. Survival analysis showed that the clinical stage and IPI were significantly associated the clinical prognosis (P=0.000, 0.001, respectively). Involvement of the maxillofacial soft tissue, eyelid, orbital, maxilla and skull base and brain tissue were significantly related to reduced the overall survival time (P=0.006, 0.000, 0.024, 0.001 and 0.015, respectively). Involvement of palate or tonsil, the nosewingand nasal sinus did not show significant shorter overall survival (P=0.091, 0.063, and 0.139, respectively). Cox regression multivariate analysis showed maxilla and skull base involvement was independent adverse prognostic factor (P=0.047). Conclusions: The PET/CT examination can accurately detect the extent of adjacent and distant tissues of tumor involvement of NK/T cell lymphoma by showing the tumor structure and metabolic status, thus has important value in the clinical staging and prognosis predication.

[Bortezomib inhibits hypoxia-induced increase of Orai-1 expression in pulmonary arterial smooth muscle cell].

Objective: In this study, a primary culture system for the rat distal pulmonary arterial smooth muscle cell (PASMC) was established to observe the effect of Bortezomib a treatment on the basal intracellular calcium concentration ([Ca(2+) ](i)), store operated calcium entry (SOCE) and Orai-1 expression in rat PASMC. Methods: We employed the primary culture method for the rat distal PASMC including the enzymatically dissociation of PASMC from the freshly isolated distal pulmonary artery and the culture of PASMC. The In Cyte system was used to measure the basal [Ca(2+) ](i) and SOCE after substantial treatment.Orai-1 protein expression in rat pulmonary artery smooth muscle was detected by Western blot. Results: Compared with Hypoxia group, the basal [Ca(2+) ](i) were significantly reduced in Hypoxia+ BTZ group(P<0.01). The basal [Ca(2+) ](i) A340/A380 ratio of Normoxia group was(1.07±0.02). The basal [Ca(2+) ](i) of Hypoxia group was(1.49±0.03); The Hypoxia+ BTZ group was(1.17±0.03). Compared with Hypoxia group, the store operated calcium entry were significantly reduced in Hypoxia+ BTZ group(P<0.01). The SOCE A340/A380 ratio of Normoxia group was(0.56±0.02). The SOCE of Hypoxia group was(0.84±0.02); The Hypoxia+ BTZ group was(0.66±0.02). The level of Orail-1 protein in pulmonary artery smooth muscle of Hypoxia group was (181.5±12.7)% higher than control group which was(100±0)%, (P<0.05). In the Hypoxia+ BTZ group Orai-1 protein expression was recovered(146.7±15.1)%, (P<0.05). Conclusion: Bortezomib inhibit chronically hypoxic enhancement of Orail-1 protein expression, basal [Ca(2+) ](i) and SOCE in rat distal pulmonary arterial smooth muscle cells.

[Expression of CD137 in tumor cells of Hodgkin lymphoma from Northern China and its application in pathological differential diagnosis].

OBJECTIVE: To investigate the expression of CD137 in HRS cells of classical Hodgkin Lymphoma (cHL), and its application in the pathological differential diagnosis. METHODS: 54 cases of cHL with "HRS" cells, and 55 cases of non-cHL with "HRS-like" cells as control group were collected. "HRS" cells and "HRS-like" cells rich areas in slides were selected from relevant groups to produce two tissue microarrays. This study focused solely on "HRS" cells and "HRS-like" cells, immunohistochemical staining for antibodies including CD30, CD15, CD20, CD3, and PAX5 were performed on CHL cases, CD137 (clone BBK-2) immunostaining and EBER in situ hybridization were detected in both groups. RESULTS: All cHL cases aged 22.0-68.0 (median 45.5) years with the male to female ratio as about 1.7∶1 primarily involved lymph nodes; while in the control group, 54 cases aged 12.0-81.0 (median 50.0) years with male∶female=1.9∶1 primarily located in nodes with only 1 case in skin. In the cHL group, CD30, CD15, CD20 and CD3 were positive in 100.0%, 70.4%, 18.5% and 0 cases in order, and PAX5 showed weak to moderate positive in 70.4% cases; the positive rate of EBER was 25.9% in cHL group, and 21.8% in the control group; The CD137 positive rates were 57.4% in cHL and 14.5% in the control groups with a significant difference (P<0.001). There were no significant differences when CD137 expressions were further compared according to the differences in age [elder group (aged 60) /non elderly group], gender (male/female), EBV infection (yes/no), histological subtype and the expression of those major diagnostic markers (positive/negative) in either cHL or control groups (all P valued>0.05). The positive rates of CD137 expression in cHL were significantly different when sub-grouped according to accession year before or after 2013 (39.4% vs 85.7%, P=0.001); However, no difference was seen in the control group (12.5% vs 16.1% , P=0.705). For those cases after 2013, the CD137 positive ratio of cHL group was more significantly different with the control one (85.7% vs 16.1%, P<0.001). CONCLUSION: Frequent expression of CD137 in "HRS" cells of cHL cases from Northern China could be detected by IHC method, while the CD137 expression was much lower in "HRS-like" cells of the control group. The positive rate of CD137 expression by immunohistochemical staining significantly increased in this cohort stored less than 3 years, which might be more reliable. CD137 might be potentially applied on pathological differential diagnosis of cHL.

[TNFAIP3 deletion status in classical Hodgkin lymphoma and its relation to Epstein-Barr virus].

Objective: To investigate the TNFAIP3/A20 abnormalities and its association with Epstein-Barr virus (EBV) in classical Hodgkin lymphoma (CHL). Methods: Formalin-fixed, paraffinembedded tissue blocks of 54 CHL patients were collected and subjected to the construction of tissue microarray (TMA) for further analyses. EBV status was evaluated by in situ hybridization (ISH) for EBER1/2 and immunohistochemistry (IHC) with anti-LMP-1 antibody. Fluorescence in situ hybridization (FISH) and IHC were performed to determine the copy number alterations of TNFAIP3 and A20 protein expression respectively. Results: The concordance rate of IHC for LMP-1 and ISH for EBER1/2 was100%, and 25.9% (14/54) cases were identified with EBV infection. Immunohistochemistry analysis demonstrated 27.8% (15/54) cases with A20 expression deficiency. Of the 54 cases tested for A20 expression, 49 cases were simultaneously analyzed by FISH, which showed 10 (20.4% ) cases harboring TNFAIP3 deletion. However, discrepancy was observed between the results of A20 by IHC and TNFAIP3 deletion by FISH. Only 1 case with TNFAIP3 deletion demonstrated complete loss of A20 immunoreactivity. In addition, comparison of the frequency of either A20 expression loss or TNFAIP3 deletion between EBV-positive and-negative cases did not reveal any significance (P>0.05). Conclusion: TNFAIP3 deletion could be observed in both EBV-positive and - negative CHL cases. A20 expression by IHC could not confirm TNFAIP3 deletion by FISH, which might be related to the technical issues.