Objective:To investigate the clinical and pathologic features and diagnosis of follicular lymphoma (FL) with a predominantly diffuse growth pattern (DFL) with 1p36 deletion.Methods:Eight cases of DFL with 1p36 deletion diagnosed at Department of Pathology, Beijing Friendship Hospital, Capital Medical University ( n=5) and the Affiliated Cancer Hospital of Zhengzhou University, Henan Cancer Hospital ( n=3) from January 2017 to January 2023 were included. Their clinicopathologic features and follow-up data were analyzed. Immunohistochemistry and fluorescence in situ hybridization (FISH) were performed. Results:There were five males and three females, with a median age of 67 years, and inguinal lymphadenopathy was found as the main symptom. Histologically, similar morphologic features were sheared among all cases, with effaced nodal structure and characterized by proliferation of centrocytes in a diffuse pattern, with or without follicular components. The germinal center-related markers such as CD10 and/or bcl-6 were expressed in the tumor cells, and 1p36 deletion but not bcl-2 translocation was appreciable in these cases.Conclusions:DFL with 1p36 deletion is a rare subtype of FL, with some overlaps with other types of FL or indolent B-cell lymphomas in their pathologic features. An accurate diagnosis requires comprehensive considerations based on their clinical, pathologic, immunohistochemical, and molecular features.

Objective:To compare the consistency of lymphoma multigene detection panels based on next-generation sequencing (NGS) with FISH detection of B-cell lymphoma gene rearrangement.Methods:From January 2019 to May 2023, fusion genes detected by lymphoma-related 413 genes that targeted capture sequencing of 489 B-cell lymphoma tissues embedded in paraffin were collected from Henan Cancer Hospital, and the results were compared with simultaneous FISH detection of four break/fusion genes: BCL2, BCL6, MYC, and CCND1. Consistency was defined as both methods yielding positive or negative results for the same sample. The relationship between fusion mutation abundance in NGS and the positivity rate of cells in FISH was also analyzed.Results:Kappa consistency analysis revealed high consistency between NGS and FISH in detecting the four B-cell lymphoma-related gene rearrangement ( P<0.001 for all) ; however, the detection rates of positive individuals differed for the four genes. Compared with FISH, NGS demonstrated a higher detection rate for BCL2 rearrangement, a lower detection rate for BCL6 and MYC rearrangement, and a similar detection rate for CCND1 rearrangement. No correlation was found between fusion mutation abundance in NGS and the positivity rate of cells in FISH. Conclusions:NGS and FISH detection of B-cell lymphoma gene rearrangement demonstrate overall good consistency. NGS is superior to FISH in detecting BCL2 rearrangement, inferior in detecting MYC rearrangement, and comparable in detecting CCND1 rearrangement.

Objective:To evaluate the effect of gonadotropin-releasing hormone agonist(GnRH-a)protocol and gonadotropin-releasing hormone antagonist(GnRH-ant)protocol in POSEIDON group 1.Methods:From January 2019 to December 2020,399 patients in POSEIDON group 1 who underwent assisted reproductive technology in the Center of Reproductive Medicine,Affiliated Hospital of Nantong University were retrospectively analyzed.Cohorts with similar baseline characteristics were screened by 1 ∶1 propensity score matching(PSM),and base-line data,clinical and laboratory parameters,and clinical outcomes were compared between GnRH-a and Gn-RH-ant groups.Results:①100 patients from GnRH-a group and 100 patients from GnRH-ant group were matched.②The total dose and total use time of gonadotropin(Gn)in GnRH-a group were higher than those in GnRH-ant group(P<0.001,P =0.048),and the hormone levels of luteinizing hormone,estradiol and progester-one on the day of HCG injection were lower than those in GnRH-ant group(P<0.001,P =0.011,P<0.001).There was no statistical difference in the number of follicles between the two groups(P>0.05),the number of oocytes retrieved,mature oocytes,normal fertilized oocytes,high-quality embryos and available embryos were lower in GnRH-a group than those in GnRH-ant group(P<0.01).③In GnRH-a group,the time to live birth(TTLB)was higher than GnRH-ant group(P =0.005),and the cumulative live birth rate(CLBR)was lower than GnRH-ant group(P =0.048),and there was no statistical difference in the number of transplants(P =0.536)and cumulative pregnancy rates(P =0.084)between the two groups.④The economic cost to live birth of GnRH-a group was higher than that of GnRH-ant group(P =0.02).Conclusions:Compared with GnRH-a group,Gn-RH-ant group could improve the cumulative live birth rate of a single ovulation induction cycle in patients in PO-SEIDON group 1,shorten the time to live birth and reduce the treatment cost of patients,and was the preferred protocol for patients in POSEIDON group 1.

Objective:To investigate the clinicopathological and molecular characteristics of pulmonary enteric adenocarcinoma (PEAC).Methods:The clinical and pathological data of 19 cases of PEAC in the Affiliated Cancer Hospital of Zhengzhou University were retrospectively collected from 2015 to 2019. Immunohistochemistry (IHC) was used to detect the relevant immunophenotypes, amplification refractory mutation system (ARMS) and fluorescence in situ hybridization (FISH) were used to detect the expression of EGFR, KRAS and ALK genes. The patients were followed up, and the relevant literature was reviewed and analyzed.Results:There were 19 cases, including 10 males and 9 females, with a mean age of 58 years (range 33-71 years). Microscopically, the tumors showed moderately to highly differentiated adenoid and/or papillary growth patterns. The tumor cells were highly columnar and sometimes showed pseudostratification. Inflammatory necrosis and scattered nuclear fragmentation were seen in some glandular lumens. IHC showed variable expression of CK7 (19/19), TTF1 (8/19), Napsin A (6/19), villin (17/19), CK20 (16/19) and CDX2 (10/19). Molecular testing showed KRAS mutation in nine cases (9/19), EGFR mutation in one case (1/19), and positive ALK split signal in one case (1/19). In the literature, the reported mutation rate of KRAS in PEAC was much higher than that of EGFR and ALK. All 19 cases underwent surgical resection and 11 cases were subjected to chemotherapy or radiotherapy.Conclusions:PEAC is a rare variant of invasive pulmonary adenocarcinoma, and has similar histological and cytological features to that of colorectal adenocarcinoma. However, detailed medical history, histologic heterogeneity, an IHC combination of CK7 +/villin + and high KRAS mutation rate are the key points of diagnosis. The prognosis needs long-term follow-up and big data statistics.

Objective: To understand the consistency of ALK Ventana-D5F3 immunohistochemistry (IHC) interpretation in Chinese lung adenocarcinoma among histopathologists from different hospitals, and to recommend solution for the problems found during the interpretation of ALK IHC in real world, with the aim of the precise selection of patients who can benefit from ALK targeted therapy. Methods: This was a multicenter and retrospective study. A total of 109 lung adenocarcinoma cases with ALK Ventana-D5F3 IHC staining were collected from 31 lung cancer centers in RATICAL research group from January to June in 2018. All cases were scanned into digital imaging with Ventana iSCANcoreo Digital Slide Scanning System and scored by 31 histopathologists from different centers according to ALK binary (positive or negative) interpretation based on its manufacturer′s protocol. The cases with high inconsistency rate were further analyzed using FISH/RT-PCR/NGS. Results: There were 49 ALK positive cases and 60 ALK negative cases, confirmed by re-evaluation by the specialist panel. Two cases (No. 2302 and No.2701) scored as positive by local hospitals were rescored as negative, and were confirmed to be negative by RT-PCR/FISH/NGS. The false interpretation rate of these two cases was 58.1% (18/31) and 48.4% (15/31), respectively. Six out of 31 (19.4%) pathologists got 100% accuracy. The minimum consistency between every two pathologists was 75.8%.At least one pathologist gave negative judgement (false negative) or positive judgement (false positive) in the 49 positive or 60 negative cases, accounted for 26.5% (13/49), 41.7% (25/60), respectively, with at least one uncertainty interpretation accounted for 31.2% (34/109). Conclusion There are certain heterogeneities and misclassifications in the real world interpretation of ALK-D5F3 IHC test, which need to be guided by the oncoming expert consensus based on the real world data.

Objective To investigate the relationship between the plasma level of interleukin 15 (IL-15) and the quantitative myasthenia gravis (QMG) score in late-onset myasthenia gravis (MG).Methods The blood samples of 86 patients (over 50 years old) who admitted to Henan Provincial People's Hospital between 2010 and 2016 were collected at two different stages:pre-treatment and six months post-treatment.The diagnosis was verified by characterizing clinical manifestation,neostigmine testing,electromyographic recording,thymic imaging and plasma anti-acetylcholine receptor (AchR).All the patients were divided further by modified Osserman classification and treated with cholinesterase inhibitor and glucocorticoid.The blood samples of 42 healthy controls were also collected from their physical examination at the end of 2016.Enzyme linked immunosorbent assay was used to evaluate the levels of IL-15 in plasma.Thymectomy was performed on patients with MG accompanied by thymoma.The possible correlation between the expression of IL-15 and the QMG scores,different types were analyzed.Results Before treatment,the levels of IL-15 in the plasma were much higher in the late onset MG patients (both ocular and generalized) than in the healthy controls ((5.75± 1.57) pg/ml vs (4.40±0.50) pg/ml,t=2.925,P＜0.01).And the late onset MG patients with thymoma showed higher level of IL-15 compared to the healthy controls ((7.39±0.84) pg/ml vs (4.40±0.50) pg/ml,t=3.925,P＜0.01).Further analysis showed that the IL-15 levels in all the MG patients with different pathological types of thymoma were higher than in the healthy controls.In the mild type of late-onset MG patients without thymoma,the IL-15 level was not increased ((4.49±0.74)pg/ml vs (4.40±0.50) pg/ml,t=1.752,P＞0.05) and in the severe type of late-onset MG patients without thymoma,the IL-15 level was mildly increased ((4.76±0.75) pg/ml vs (4.40±0.50) pg/ml,t=2.462,P＜0.05) compared to the healthy controls.Furthermore,the IL-15 level decreased dramatically upon the therapy in all the MG patients,especially in the late-onset MG patients with thymoma.Moreover,IL-15 was positively correlated with QMG scores before treatment (r=0.375,P＜0.01),especially in the late-onset MG patients with thymoma (r=0.823,P＜0.01),but not in the late-onset MG patients without thymoma (r=0.039,P＞0.05).IL-15 and QMG scores returned to normal six months after treatment.Conclusions IL-15 is increased in the plasma of late-onset MG patients,and is positively correlated with the QMG scores,especially in the late-onset MG patients with thymoma.In addition,IL-15 is decreased upon the therapy in the MG patients.

Objective: To explore the expression of IQGAP1 (Ras GTPase-activating-like protein containing IQ domain) in esophageal squamous cell carcinoma (ESCC) tissues and cell lines and its effect on the proliferation and invasion of TE-2 cell. Methods: Totally 125 pairs of cancer tissues and para-cancerous tissues from ESCC patients, who underwent surgical resection inAffiliated Tumor Hospital of Zhengzhou University from January 2015 to December 2016, were included in this study; in addition, ESCC cell lines (TE-2, TE3, ECA109) and normal esophageal epithelial cell line Het-1A were also collected. The expression of IQGAP1 was detected by immunohistochemical staining and its relationship with cliniopathological features was also analyzed. IQGAP1 mRNA and protein expressions in ESCC cell lines were detected by Real-time quantitative PCR (qPCR) and Western blotting, respectively. TE-2 cells were transfected with si-IQGAP1 (positive transfection group) and si-CTRL (negative control group) plasmids, and the effects of IQGAP1 silencing on the proliferation and invasion of TE-2 cells were detected by MTT and Transwell assay. The expressions of E-cadherin and Ncadherin were detected by Western blotting. Results: The positive expression rate of IQGAP1 in ESCC tissues was significantly higher than that in para-cancerous tissues (P<0.05), which was closely related to tumor stage and histologic grade (all P<0.05). The mRNAand protein expressions of IQGAP1 in TE-2, TE-3 and ECA109 cells were significantly higher than those in Het-1Acells (all P<0.05).After IQGAP1 was silenced, compared with the negative control group and the blank group, the expression of IQGAP1 mRNAand protein in the positive transfection group significantly decreased (all P<0.05); the proliferation and invasiveness of TE-2 cells significantly decreased (all P<0.05); E-cardherin was up-regulated while N-cardherin was down-regulated (all P<0.05) in the positive interference group. Conclusion: IQGAP1 is highly expressed in ESCC tissues, and si-IQGAP1 can inhibit the proliferation and invasion of TE-2 cells, which plays an important role in the occurrence and development of ESCC.

Objective: To investigate the level of interleukin-6 (IL-6) and the effect of valproic acid(VPA) administration on IL-6 promoter methylation, further to explore the epigenetic mechanism in febrile seizures. Methods: Sprague-Dawley (SD) rats (21 day) were randomly divided into control group (n=12) and febrile seizure (FS) group (n=12), and seizures were generated using hot bath methods and evaluated by racine score and electroencephalogram (EEG). The IL-6 protein and mRNA levels were detected using ELISA and quantitative reverse transcription polymerase chain reaction (qRT-PCR), respectively.Rat C6 cell line was cultured and randomly divided into control, VPA(0.2 mol/L), hyperthermia(43.5 ℃, 1 h), hyperthermia (43.5 ℃, 1 h)+VPA (0.2 mol/L) groups.The methylation status of IL-6 promoter in FS rats and the effect of VPA on IL-6 methylation in C6 cell line were examined by bisulfite sequencing polymerase chain reaction (BSP) to determine the epigenetic regulation of IL-6 level in FS. Results: Racine score and EEG results showed that the frequency and amplitude of neuronal discharge increased after hot water bath in rats, and the latency of convulsion was shorter.The expression levels of IL-6 mRNA and protein were significantly increased in Fs group compared with those in control group(mRNA: Control: 0.98±0.34; FS: 2.85±0.39, P<0.05; Protein: Control: (2 824.33±169.20)pg/ml; FS: (3 514.58±86.4)pg/ml, P<0.01). The methylation of IL-6 promoter in FS group (84% (21/25)) was lower than that in control group (100% (25/25)) (P<0.05). A significant increase in methylation of IL-6 promoter was observed in C6 cell line from VPA combined with hyperthermia (96% (24/25)) but no change was showed in that from VPA alone (100% (25/25)) compared with that from control(100% (25/25)). Conclusion IL-6 level is up-regulated in FS rats, and the hypomethylation or demethylation of the IL-6 promoter region might increase the IL-6 level.VPA administration can elevate its methylation level, which provides a strong experimental basis for the future study of the epigenetic mechanism in FS.

Objective:To investigate the expression and clinical significance of PD-L1 in colorectal cancer (CRC). Methods:A total of 210 CRC patients who accepted radical surgery in our hospital from January 2015 to January 2016 were divided into three groups, namely, high-frequency microsatellite instability (MSI-H), low-frequency microsatellite instability (MSI-L), and microsatellite stable (MSS). The expression of PD-L1 was detected by immunohistochemistry, and the expression characteristics of PD-L1 in different types of CRC were analyzed. Results:CRC cases with low differentiation had a higher expression of PD-L1 than CRC patients with high differ-entiation (P<0.05). PD-L1 had a positive rate of 75.8%in the MSI-H group and a rate of 9.3%in the MSI-L and MSS groups, wherein the difference between the two groups was statistically significant (P<0.05). Conclusion:PD-L1 was positively expressed in some CRC tu-mor tissues, and its positive rate was significantly higher in MSI-H than in MSI-L and MSS. The therapeutic effect of a PD-L1 blocker for patients with MSI-H CRC might be preferable.

Objective To study PTEN protein expression and clinical significance in patients with diffuse large B cell lymphoma. Methods Immunohistochemical staining was used to determine the PTEN protein expression in 40 cases of primary diffuse large B lymphoma tissuse. The results were analyzed by Kaplan-Merie survival analysis, Log-Rank test and Logistic regression analysis. Results PTEN protein was positive in 16 cases and negative in 24 cases. There was no significant difference between two groups in twoyear overall survival rate(62.5 % vs 66.7 %, P >0.05). Survival analysis showed that patient' s survival time gradually were reduced with extended time between PTEN protein-positive group and negative group, lower in PTEN-positive group than the negative group, but there was no significant difference in survival curve (P >0.05) in the two groups. We compared characteristics of patients between PTEN protein positive and negative groups,including molecular type, patient' s age, stage, LDH, physical score and extranodular invasion, there was no significant difference among them. PTEN protein was not correlated with prognosis, while International Prognosis Index(IPI) was still a risk factor (OR >1). Conclusion PTEN protein expression may not predict the outcome in diffuse large B cell lymphoma, but IPI still is a predictor.