[Angioimmunoblastic T-cell lymphoma: histopathological grading and prognosis].

Objective: To investigate the histological features and prognostic factors of angioimmunoblastic T-cell lymphoma (AITL). Methods: The pathological data of 62 patients with AITL with complete follow-up information were retrospectively collected and analyzed from Changhai Hospital during September 2012 and September 2017. Histological and immunohistochemical (IHC) examination, in situ hybridization (ISH), and single nucleotide polymorphisms (SNP) gene mutation analysis were done. Subgroup evaluation with histology, IHC, ISH, SNP gene mutation, and association with clinical progression were performed. Results: The cohort included 62 cases of AITL, including 46 males and 16 females patients, with a median age of 64 years. Follicular dendritic cells (FDC) area showed significantly expansion (≥30%) in 40 cases; increased plasma cells (≥10%) was seen in 37 cases; B cells were distributed around blood vessels in 37 cases; and increased p53 mutation positive cells (≥40%) were seen in 39 cases; high Ki-67 index (≥40%) was seen in 39 cases; RHOA mutation was seen in 19 cases; TET2 mutation was seen in 9 cases. Overall survival analysis showed these factors were significantly correlated with tumor prognosis (P<0.05). Multivariate analysis showed that CD38 positive cells<10%, Ki-67≥40%, RHOA and TET2 mutations were risk factors associated with overall survival. Conclusions: AITL could be divided into two different prognostic groups, low-grade and high-grade, with statistically significance outcome, based on the FDC area expansion, degree of plasma cell proliferation, B cells distribution pattern combined with gene mutations and clinical progression. Low-grade malignant group progresses slowly, and high-grade malignant group is highly invasive.

[Comparison of epidermal growth factor receptor gene mutation in lung adenocarcinoma using biopsied tissue, pleural effusion and blood samples].

Objective: To compare different specimen types of lung adenocarcinoma in the detection of epidermal growth factor receptor (EGFR) gene and to correlate EGFR mutations with patient clinical features. Methods: One hundred lung adenocarcinoma cases were collected from June to December in 2015, at the First Affiliated Hospital of Xinjiang Medical University.Of the 100 lung adenocarcinoma samples, 43 were male and 57 were female. The age was from 40 to 88 years old, and the average age was 66 years. One hundred lung adenocarcinoma cases were divided equally into two groups. Mutation analysis of EGFR gene by real-time PCR was performed using biopsied tissue and paired blood samples in one group (n=50) and using pleural effusion and paired blood samples in the other group (n=50). Results: The mutation rate of EGFR gene in biopsy samples was 54% (27/50) , higher than that of blood samples (46%, 23/50), but without statistical differences (χ(2)=0.640, P=0.424). In contrast, mutation rate of EGFR gene in pleural effusion samples (42%, 21/50) was higher than that of blood samples (34%, 17/50), but without statistical differences(χ(2)=0.679, P=0.409). Two patients had EGFR mutation detected in paired blood samples but not in the corresponding biopsy samples, and four patients had EGFR mutation detected in pleural effusion samples but not in their paired blood samples. The mean progression-free survival of patients with detectable EGFR mutation were 9.5 months (tissue samples), 8.6 months (pleural effusion) and 8.5 months (blood). However, there was no statistical difference. Conclusions: Blood samples may be used to assess EGFR mutations for patients with lung adenocarcinoma. However, further studies are needed to improve the sensitivity and accuracy in the detection of EGFR mutations using blood samples.

[Impact of PRDM1 gene inactivation on C-MYC regulation in diffuse large B-cell lymphoma].

Objective: To investigate the role of PRDM1 gene inactivaion in the regulation of C-MYC in diffuse large B-cell lymphoma (DLBCL), and to explore the correlation of its immunophenotype and prognosis. Methods: 100 cases paraffin-embedded DLBCL tissues were collected from January 2009 to December 2015 at the First Affiliated Hospital of Xinjiang Medical University along with 20 cases of reactive proliferative lymph nodes as control. Immunohistochemical methods were used to detect the expression of CD20, CD10, MUM1, Ki-67, bcl-6, PRDM1/Blimp1, C-MYC and PAX5 protein. The tumors were classified into two subtypes according to Hans classification.The expression of PRDM1 and C-MYC gene in tumor group and control group was detected by reverse transcription PCR (RT-PCR) and the relationship between PRDM1 and C-MYC gene was analyzed.OCI-LY1 (GCB subtype) and OCI-LY3 (non-GCB subtype) cell lines were transfected with small interfering RNA by cationic liposome reagent transfection, and the expression of C-MYC in the transfected cell lines was detected by RT-PCR and Western blot. The Kaplan-Meier method was used to analyze the prognostic significance of PRDM1/Blimp1 and C-MYC at protein and mRNA levels. Results: There were 27 cases of GCB subtype and 73 cases of non-GCB subtype according to Hans classification. The positive expression of Blimp1 in DLBCL group and proliferative lymph nodes in control group was seen in 26(26.0%) and 20 cases(100%), respectively. There were 58 cases with high expression of PRDM1 at mRNA level, including 22 cases of GCB subtype and 36 cases non-GCB subtype, and the difference was statistically significant (P=0.004). There were differences in PRDM1 gene expression between the two immunological subtypes, serum lactate dehydrogenase (serum LDH) level, presence of B symptoms, tumor primary sites and other clinical pathological parameters, while C-MYC expression was different in gender, IPI score, and serum LDH levels. Upon PRDM1/Blimp1 gene silencing in the two cell lines, C-MYC protein and gene expression were up-regulated in the transfection group, compared with the blank control group and negative control group by reverse transcription PCR and Western blot analyses. Moreover, PRDM1 expression was significantly associated with C-MYC(χ(2)=7.648, P=0.006) at mRNA level. Conclusion: The up-regulation of C-MYC gene expression induced by PRDM1 inactivation in DLBCL may play an important role for the development of DLBCL.PRDM1 protein and mRNA are associated with immunophenotyping and PRDM1 mRNA is a marker of poor prognosis.

[Clinicopathologic characteristics of epidermal growth factor receptor mutations in non-small cell lung cancer in Xinjiang region].

Objective: To investigate the point mutation of epidermal growth factor receptor (EGFR) gene and clinicopathologic characteristics in patients with non-small cell lung cancers(NSCLC)of Xinjiang region. Methods: Five-hundred and eighty-two cases of paraffin-embedded tissue in patients with NSCLC were collected between January 2013 and December 2015 in the First Affiliated Hospital of Xinjiang Medical University. The DNA was extracted from these tissues by Qiagen kit, to test thirty-two mutations in EGFR exons 18, 19, 20 and 21 using fluorescent quantitative qRT-PCR technology by TaqMan probe; the clinicopathologic features of patients were analyzed according to the mutation status of EGFR. Results: There were 173 cases with EGFR gene mutation in 582 cases of paraffin-embedded tissue in patients with NSCLC, and the mutation rate was 29.7%(173/582). There were statistical difference in female patients (50.5%, 98/194), no history of smoking(47.3%, 96/203), high differentiation(6/9), adenosquamous carcinoma(6/11), peripheral location (34.9%, 88/252), and surgical specimens(38.2%, 83/217), respectively (P<0.05). Multiple factors Logistic analysis showed that gender, degree of differentiation, and pathologic types had statistical differences to EGFR when α=0.05. There were no statistical differences between other variants. Conclusions: There are higher rate EGFR gene mutation in women patients, non-smokers, and well-differentiated, adenocarcinoma. Gender, degree of differentiation and pathological patterns are independent influencing factors on EGFR mutation status.

[BRCA1/2 gene mutation and clinicopathologic features of triple negative breast cancer].

OBJECTIVE: To investigate the incidence of BRCA1/2 gene mutation among triple negative breast cancer (TNBC) patients. METHODS: Polymerase chain reaction and DNA sequencing were used to detect mutations of BRCA1 (exons 2, 11 and 20) and BRCA2 (exon 11) genes using paraffin-embedded tissue samples of 33 TNBC patients (21 Uyghur patients and 12 Han patients ) in Xinjiang Uygur Autonomous Region. RESULTS: Among 33 cases of TNBC, 5 cases (5/33, 15.2%) were found to have BRCA1 gene mutation. The exon 11 of BRCA1 gene mutation proportion was seen in 4/5 and exon 20 of BRCA1 gene mutation proportion was seen in 1/5. No germline mutation was found in exon 2 of BRCA1 gene and exon 11 of BRCA2 gene. The onset age of patients with BRCA1 mutations were younger than 50 years. Four BRCA1 mutation patients were premenopausal. The proportions of BRCA1 gene mutation of Uygur TNBC patients and Han TNBC patients were 2/12 and 3/21, respectively, without significant difference (P=0.854). BRCA mutations were mainly found in stageⅠtoⅡ (4/5) and only 1 case (1/5) was found with stage Ⅲ disease. No significant difference was found in fertility status, menstrual status and age at menarche between mutated and non-mutated patients (P>0.05) . CONCLUSIONS: Germline mutation of BRCA1 may be more often associated with TNBC than BRCA2 for its higher mutation rate. Compared with non-mutated TNBC cases, several clinicopathologic features could be identified among mutated cases.

[Potential mechanism and prognostic value of promoter methylation of PRDM1 gene in diffuse large B cell lymphoma].

Objective: To investigate PRDM1 gene methylation status, immune classification and their prognostic significance in diffuse large B cell lymphoma (DLBCL). Methods: Immunohistochemical (IHC) staining for CD20, CD10, bcl-2, bcl-6, PRDM1/Blimp-1 and MUM1 was carried out in 100 cases of DLBCL specimens and 20 reactive lymphoid proliferation samples. All patients were classified into germinal center B cell-like (GCB) and activated B cell-like (ABC) subtype according to Hans' algorithmin. PRDM1 gene methylation was detected by methylation-specific PCR (MSP) and its relationship with clinicopathologic parameters was analyzed. OCI-Ly1 (GCB type) and OCI-Ly3 (ABC type) cell lines were transfected by Small interfering RNA(siRNA) with cationic lipid reagent transfection mediated, and the PRDM1/Blimp-1 expression in before and after transfected cell lines were detected with reverse transcription-PCR and Western blot methods. The relationship between PRDM1 gene methylation, clinicopathologic parameter and survival was analyzed using one-way analysis of variance. Results: One hundred patients were classified into 73 (73%) cases of GCB subtypes and 27 (27%) cases of ABC. PRDM1/Blimp-1 was expressed in 21 DLBCL and highly expressed in 20 reactive lymphoid proliferation. PRDM1 gene methylation was detected in 23% (23/100) of DLBCL, while no methylation was detected in all 20 reactive lymphoid proliferation. The difference of the PRDM1 methylation status between DLBCL and the control samples was statistically significant (P=0.004). However, there was no significant correlation between the PRDM1 gene methylation and clinicopathologic parameters (P>0.05). Reverse transcription-PCR and Western blot showed that PRDM1 gene expression was reduced in siRNA-induced group compared with blank control group and negative control group. One-way analysis of variance revealed that aged ≥60 years, performance status score above 3, and the presence of general symptoms were associated with significantly lower overall survival rate. Conclusions: PRDM1 gene silencing with aberrant CpG methylation is probably one of the critical events in the oncogenesis of DLBCL. This may have important implications as a candidate marker for diagnosis and targeted gene therapy. Meanwhile in vitro siRNA transfected OCI-Ly1 and OCI-Ly3 cell lines confirm that PRDM1 gene is a suppressor gene in DLBCL and may represent a novel therapeutic target.

[Primary central nervous system diffuse large B cell lymphoma: a clinicopathologic and molecular study].

Objective: To investigate clinicopathologic characteristics, immunophenotype and EB virus-related molecular genetic alterations in primary central nervous system diffuse large B cell lymphoma (DLBCL) along with correlation with clinical prognosis. Methods: A total of 30 cases of primary central nervous system DLBCL were retrospectively studied by retrieving clinical data, histological evaluation and immunophenotyping by EnVision two steps methods. The expression of EBER mRNA was detected by in situ hybridization and bcl-2, bcl-6 and C-MYC gene abnormalities were analyzed by interphase fluorescence in situ hybridization. Results: The cases included 18 males and 12 females (sex ratio of 1.5∶1.0) with an age ranging from 24 to 78 years (average age of 52 years, the median age of 53 years). The single primary clinical presentation was focal neurologic deficits. Tumor locations were supratentorial (21 cases), subtentorial (7 cases), involving both locations in 2 cases. Diffuse growth pattern was observed with large lymphoid cells mostly resembling centroblasts with abundant basophilic cytoplasm with oval to round, vesicular nuclei containing fine chromatin. An angiocentric and angiodestructive growth pattern was also present. Other features included perivascular space invasion. Immunohistochemical staining using a panel of CD10, bcl-6 and MUM1, six cases were germinal center-like (GCB) and 24 cases were non-germinal central-like (non-GCB). The positive rates of bcl-2, bcl-6 and C-MYC were 53.3% (16/30), 80.0% (24/30) and 20.0% (6/30), respectively. Genetic alterations were detected by FISH and the gene arrangement rates of bcl-2, bcl-6 and C-MYC were 3.3% (1/30), 16.7% (5/30) and 3.3% (1/30), respectively. There were 19 cases in stage 0-1 disease and 11 cases had stage 2-3 disease. Postoperative follow-up for average 13.6 months showed the median survival of 10 months, one-year survival of 46.7% and 16 patients died within a year. Conclusions: The clinical prognosis of primary central nerve system DLBCL depends on age, clinical performence status score, IPI score, immune classification and treatment. Patients typically progress rapidly with the high mortality within one year of diagnosis. Surgical resection combined with high-dose methotrexate or cytarabine chemotherapy offer the best treatment option.

Homogeneous high attenuation renal cysts and solid masses--differentiation with single phase dual energy computed tomography.

AIM: To assess the feasibility of using single-phase dual-energy computed tomography (DECT) to differentiate between homogeneous high attenuation renal cysts and solid renal masses. MATERIALS AND METHODS: Twenty-nine pathologically proven solid renal masses in 29 patients and 14 high attenuation renal cysts from 11 patients were evaluated retrospectively. Two readers independently measured CT values from each lesion using both unenhanced single-energy phase and nephrographic dual-energy phase scans. Enhancement was defined as the attenuation difference between average-weighted 120 kV images and unenhanced images. Diagnostic sensitivity, specificity and accuracy based on enhancement, D-value (CT: 80 kV-140 kV) and DE-ratio (CT: 80/140 kV) were compared by the receiver operator characteristic (ROC) curves. RESULTS: Using 17.6 HU as the cutoff value for enhancement, resulted in a sensitivity, specificity and accuracy of 96.6%, 100% and 97.7%, respectively. Corresponding values were 100%, 92.9% and 97.7% using a D-value cutoff of 15.6 HU, and 100%, 85.7% and 95.3% using a DE-ratio cutoff of 1.3. There were no significant differences in the AUCs obtained from the ROC curves for enhancement, D-value or DE-ratio. The mean effective radiation dose was 6.04 mSv with biphasic scanning compared with 2.91 mSv for single dual-energy nephrographic phase scanning. CONCLUSION: Single-phase dual-energy CT is able to differentiate high attenuation renal cysts and solid renal masses with high sensitivity, specificity and accuracy, based on either D-value or DE-ratio. Omitting unenhanced scanning reduces the radiation dose by more than 50%.