Correlation between RAS and BRAF V600E gene mutations and clinicopathological characteristics of colorectal cancer / 肿瘤研究与临床

Objective:To investigate the correlation between KRAS, NRAS and BRAF V600E gene mutations and the clinicopathological characteristics of patients with colorectal cancer.Methods:Specimens from 217 patients with colorectal cancer who underwent surgical resection and were pathologically confirmed in Shanxi Province Cancer Hospital from January 2020 to December 2021 were selected, and the clinical data of the patients were retrospectively analyzed. The mutation status of KRAS, NRAS and BRAF V600E genes were detected in the paraffin specimens of surgically-resected tissues by direct sequencing. The mutation rates of KRAS, NRAS and BRAF V600E were compared among patients with different clinicopathological characteristics.Results:The mutation rates of KRAS, NRAS and BRAF V600E in 217 patients with colorectal cancer were 48.4% (105/217), 4.1% (9/217) and 3.7% (8/217), of which 1 patient (0.5%) had both KRAS and NRAS mutations. NRAS gene mutation was not correlated with gender, age, tumor size, tumor location, pathological type, degree of differentiation, depth of invasion, lymph node metastasis, distant metastasis, TNM stage, hemangioma thrombus/nerve invasion (all P>0.05); KRAS mutation rate in patients ≥ 60 year old was higher than that in patients < 60 year old [55.3% (63/114) vs. 40.8% (42/103), χ2 = 4.55, P = 0.033),and there was no correlation between KRAS gene mutation and other clinicopathological features (all P > 0.05); the mutation rate of BRAF V600E gene in colorectal cancerpatients with distant metastasis was higher than that in patients without distant metastasis [16.7% (4/24) vs. 2.1% (4/193), P = 0.006], and there was no correlation between BRAF V600E gene mutation and other clinicopathological features (all P > 0.05). Conclusions:Older colorectal cancer patients may be prone to KRAS gene mutation, and the BRAF V600E gene mutation rate is higher in patients with distant metastasis, and there is no correlation between NRAS gene mutation and clinicopathological characteristics.

Correlation between 21-gene recurrence score and clinicopathological characteristics of patients with Lumina type breast cancer and its clinical significance / 肿瘤研究与临床

Objective:To analyze the correlation between 21-gene recurrence score (RS) and clinicopathological characteristics of patients with Lumina type breast cancer, and to explore its significance in individualized treatment.Methods:The clinicopathological data of 59 patients with surgical resection and pathological diagnosis of Lumina type breast cancer in Shanxi Provincial Cancer Hospital from May 2018 to May 2019 were retrospectively analyzed. Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of 21 gene and RS was calculated. According to the 21-gene RS, the patients were divided into low recurrence risk group (RS < 18 points), intermediate recurrence risk group (RS 18-31 points) and high recurrence risk group (RS > 31 points). Univariate and multivariate logistic regression analyses were made to evaluate the correlations between different recurrence risk and clinicopathological characteristics of patients and their influence on the choice of adjuvant chemotherapy.Results:Based on the 21-gene RS, 29 patients were in low recurrence risk group, 22 cases were in intermediate recurrence risk group, and 8 cases were in high recurrence risk group. Single-factor analysis showed that age ( P = 0.012), maximum mass diameter ( P = 0.031), histological grade ( P = 0.036), progesterone receptor (PR) level ( P = 0.015), Ki-67 positive index ( P = 0.049) and molecular typing ( P = 0.010) were influencing factors of 21-gene RS recurrence risk. Multivariate logistic regression analysis showed that the age and Ki-67 positive index were negatively correlated with 21-gene RS recurrence risk (both P < 0.05). After grouping according to the 21-gene RS, 17 patients in the intermediate recurrence risk group (according to the traditional postoperative recurrence risk grouping method for breast cancer) were classified as low recurrence risk group, and 4 patients in the low recurrence risk group were classified as intermediate recurrence risk group ( χ2 = 4.535, P = 0.033). After grouping based on 21-gene RS, the number of patients who needed chemotherapy in individualized treatment decreased. Of the 17 cases, 11 cases did not undergo postoperative chemotherapy, and the remaining patients received chemotherapy. The postoperative follow-up period was 11-22 months. As of March 2020, there was no recurrence or disease progress. Conclusion:The 21-gene RS can provide objective basis for the individualized precise treatment and prognosis prediction for patients with early-stage Lumina type breast cancer.

Value of droplet digital polymerase chain reaction in detecting epidermal growth factor receptor mutations in peripheral blood circulating tumor DNA of patients with non-small cell lung cancer / 肿瘤研究与临床

Objective:To analyze the value of droplet digital polymerase chain reaction (ddPCR) in detecting epidermal growth factor receptor (EGFR) mutations in peripheral blood circulating tumor DNA (ctDNA) of patients with non-small cell lung cancer (NSCLC).Methods:Peripheral blood samples of 63 patients with NSCLC who were treated in Shanxi Provincial Cancer Hospital from August 2018 to March 2019 were collected, and EGFR sensitive mutations in peripheral blood of patients were detected by ddPCR, and the results were compared with the results of amplification refractory mutation system polymerase chain reaction (ARMS-PCR). Kappa test was used to analyze the consistency of the two methods.Results:The EGFR sensitive mutations were found in 31 cases (49.2%) by ddPCR in peripheral blood of 63 patients with NSCLC. Among them, 1 case (1.6%) had G719X, 12 cases (19.0%) had E19-Del, 11 cases had T790M (17.5%), 7 cases (11.1%) had L858R. Seven cases (22.6%) of the patients had double mutations. In comparison, the above 4 mutations were found in 26 cases (41.3%) by the ARMS-PCR method, with 0 case, 12 cases (19.0%), 6 cases (9.5%), and 8 cases (12.7%), respectively, including 5 cases (19.2%) with double mutations. L858R in one case was positive when detected by ARMS-PCR, while it was negative when detected by ddPCR. The consistency rate of the two methods was 90.3% (κ = 0.8, P < 0.05). The median abundance of EGFR mutations in peripheral blood ctDNA of 31 cases was 1.7% (range 0.04%-23.60%). The median abundance of E19-Del was 2.50% (0.35%-22.70%), that of T790M was 0.6% (0.04%-14.00%), and that of L858R was 2.3% (0.20%-23.60%). Ten cases with the abundance of EGFR mutations < 1% when detected by ddPCR, accounting for 32.6% (10/31) of total patients with mutations, but only 5 cases of them were detected by ARMS-PCR. The detection rate of T790M by ddPCR in patients who had received tyrosine kinase inhibitor (TKI) and had acquired drug resistance was 57.9% (11/19), while it was 0 in patients without TKI treatment. Among patients with T790M mutation, 1 case had a mutation abundance < 0.1%, 7 cases had a mutation abundance of 0.1%-2.0%, 3 cases had a mutation abundance > 2.0%.Conclusions:The ddPCR provides a non-invasive, highly sensitive and absolutely quantitative method for detecting EGFR mutations in peripheral blood ctDNA of NSCLC patients. It provides a new detection method for EGFR-TKI targeted therapy in NSCLC patients with difficult sampling or with acquired drug resistance who need to repeatedly sample. The approach provides an important basis for the individualized targeted therapy.

Consistency of ALK Ventana-D5F3 immunohistochemistry interpretation in lung adenocarcinoma among Chinese histopathologists / 中华病理学杂志

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.

Efficacy of reinforcement on duodenal stump using single purse-string suture during laparoscopic radical gastrectomy for gastric cancer / 中华胃肠外科杂志

Objective@#To evaluate the efficacy of reinforcement on duodenal stump using single purse-string suture during laparoscopic radical gastrectomy for gastric cancer in preventing duodenal stump leakage.@*Methods@#A descriptive cohort study was conducted to retrospectively collect clinical data of 211 patients with gastric adenocarcinoma who underwent laparoscopic radical gastrectomy with Roux-en-Y or Billroth Ⅱ reconstruction and reinforcement on duodenal stump using laparoscopic single purse-string suture in Zhongshan Hospital of Fudan University between January 2013 and December 2016. Of 211 patients, 136 were male and 75 were female with mean age of (57.5±11.1)(24 to 87) years. Tumors locating at gastric upper 1/3, middle 1/3 and low 1/3 were found in 62, 68 and 81 patients respectively. Eighty-three cases underwent total gastrectomy, 128 underwent distal subtotal gastrectomy, 107 underwent Roux-en-Y reconstruction and 104 underwent Billroth II reconstruction. The procedure of reinforcement on duodenal stump using single purse-string suture during laparoscopic radical gastrectomy was as follows: (1) after cutting the duodenal stump to about 2.0 cm in length, use a 3-0 single-strand absorbable suture to make a muscle layer purse at a distance of 1.0 to 1.5 cm from the duodenal stump; (2) use the purse line to make a slipknot; (3) push the duodenum stump into the purse with a needle holder or grasper; (4) tighten the knot of the purse string, and then make 4 to 5 knots for reinforcement. Postoperative complications were defined and graded according to the Clavien-Dindo grading criteria, and the incidence of early complications was recorded. Clinicopathologic features and postoperative outcomes were analyzed.@*Results@#All patients completed operations successfully. The mean time of laparoscopic single purse-string suture was (5.1±1.6) (3.6 to 10.2) minutes. Postoperative early complication occurred in 31 cases (14.7%), of whom 27 cases developed surgery-related complications (12.8%), including 7 cases (3.3%) of peritoneal infection, 6 (2.8%) of pancreatic leakage, 4 (1.9%) of wound infection, 4 (1.9%) of gastroplegia, 2 (0.9%) of peritoneal hemorrhage, 2 (0.9%) of intestinal obstruction, 2 (0.9%) of lymphatic leakage, and no duodenal stump leakage; while 4 cases (1.9%) developed internal non-surgical complication, including 3 cases (1.4%) of pulmonary infection and 1 (0.5%) of cardiovascular event. The patient with peritoneal hemorrhage was healed after re-operation and all other patients were discharged uneventfully after conservative treatment. Four cases (1.9%) developed complications beyond grade III a of Clavien-Dindo criteria.@*Conclusion@#Reinforcement on duodenal stump using laparoscopic single purse-string suture during laparoscopic radical gastrectomy with Roux-en-Y or Billroth II reconstruction is simple and effective, and can prevent the risk of development of duodenal stump leakage.

Expression of costimulatory molecules B7H3 and B7H4 in T lymphoblastic lymphoma/leukemia and clinical significance / 中华病理学杂志

Objective@#To investigate the expression of B7H3 and B7H4 in T lymphoblastic lymphoma/leukemia (T-LBL/ALL) in correlation with clinicopathological parameters and patient prognosis.@*Methods@#Immunohistochemistry (IHC) was used to detect the expression of B7H3 and B7H4 protein in 100 cases of T-LBL/ALL(test group) and 30 cases of lymph node reactive hyperplasia (LH) (control group), diagnosed at Shanxi Cancer Hospital from January 2001 to June 2017. Real-time RT-PCR was used to detect the mRNA expression of B7H3 and B7H4 in 50 cases of T-LBL/ALL and 30 cases of LH (control group).@*Results@#There were 79 males,21 females. Immunohistochemical results showed that the expression rates of B7H3 and B7H4 were 23%(23/100) and 54%(54/100), respectively. By real-time RT-PCR, the relative expression of B7H3 mRNA in the T-LBL/ALL group was 2.5 times of that of the LH group. The expression levels of B7H4 mRNA in T-LBL/ALL group and LH group were extremely low.Single factor analysis showed that B7H3 protein expression in T-LBL/ALL group was associated with B symptoms and primary nodal disease (P<0.05). B7H4 protein expression was associated with mediastinal broadening and bone marrow involvement (P<0.05). B7H3 protein, B7H3 mRNA, B7H4 protein expression and IPI score were associated with prognosis (P<0.05), and the combined expression of B7H3 and B7H4 was associated with T-LBL/ALL prognosis (P<0.05). Multivariate Cox regression analysis showed that overexpression of B7H3 mRNA was an independent risk factor for the prognosis of patients with T-LBL/ALL (P<0.05).@*Conclusion@#Expression of B7H3 and B7H4 is closely corelated with clinicopathological parameters and prognosis of patients with T-LBL/ALL, suggesting that B7H3 and B7H4 expression play an important role in the development of T-LBL/ALL.

Expressions of programmed death-ligand 1 and 2 and phosphorylated protein kinase B in diffuse large B-cell lymphoma and their clinical significances / 白血病·淋巴瘤

Objective To investigate the expressions of programmed death-ligand 1 (PD-L1) and PD-L2 and phosphorylated protein kinase B (p-AKT) in diffuse large B-cell lymphoma (DLBCL) patients and their correlations with clinicopathological features and prognosis. Methods A total of 68 paraffin-embedded specimens of DLBCL patients diagnosed in Shanxi Provincial Cancer Hospital with detailed follow-up record from January 2010 to December 2012 were included in the study. The expressions of PD-L1, PD-L2 and p-AKT proteins in DLBCL were detected by using immunohistochemistry (IHC). Results The positive rate of PD-L1 protein in DLBCL patients was 22.1％ (15/68), which was related to germinal center B-cell (GCB) subtype or not (χ2= 5.591, P= 0.018), clinical stage (χ2= 3.969, P= 0.046), international prognostic index (IPI) grades (χ2=4.178, P=0.041) and treatment remission rate (χ2=6.587, P=0.010). The positive rate of PD-L2 protein in DLBCL patients was 14.7％ (10/68), which was related to extranodal metastasis or not (χ2=6.772, P= 0.009). The positive rate of p-AKT for DLBCL patients was 61.8％ (42/68), which was correlated with age (≥60 years old) or not (χ2=6.227, P=0.013), Eastern Cooperative Oncology Group (ECOG) grades (χ2=4.005, P=0.045), B symptoms (χ2=10.187, P=0.001) and treatment remission rate (χ2=4.096, P=0.043). Univariate survival analysis showed that the overall survival (OS) rate and progression free survival (PFS) rate of PD-L1 protein positive expression group were lower than those of PD-L1 protein negative expression group (both P< 0.05). In the patients with non-GCB subtype, OS rate and PFS rate of PD-L1 protein positive expression group were lower than those of PD-L1 protein negative expression group (both P<0.05). p-AKT protein positive expression group had poorer OS rate and PFS rate compared to p-AKT negative expression group (both P< 0.05). Correlation analysis showed that PD-L1 protein expression was correlated with PD-L2 and p-AKT proteins expressions (r= 0.380, P= 0.001;r= 0.273, P= 0.025). The prognosis was worse when p-AKT and PD-L1 proteins was co-expressed (P< 0.05). Multivariate analysis suggested high expressions of PD-L1 and p-AKT proteins were independent prognosis risk factors in DLBCL (both P<0.05). Conclusions The expressions of PD-L1 and p-AKT proteins may be involved in the occurrence and development of DLBCL. Blocking PD-1 and PD-L1 access or combined blocking could provide a promising future for the clinical therapy.

Ultrasonography-guided wire-localization and nano-carbon staining for detection of sentinel lymph node in breast cancer / 肿瘤研究与临床

Objective@#To evaluate the usage of ultrasound guided wire-localization, nano-carbon staining and the combination of the above two methods in detecting sentinel lymph node (SLN) in breast cancer.@*Methods@#A total of 159 cases of breast cancer from May 2015 to December 2017 in Shanxi Provincial Cancer Hospital were selected, and they were treated with ultrasound guided wire-localization, nano-carbon staining and combination of the two methods separately to detect SLN before the operation. After the operation, SLN and axillary lymph node in each group were marked and made pathological diagnosis.@*Results@#There were 69 cases with pathological diagnosis of SLN metastasis and 90 cases without abnormal representation. With the patient as the unit, the sensitivity of ultrasound guided wire-localization was 100.0% (69/69), the sensitivity of nano-carbon staining was 98.6% (68/69), and the sensitivity of combination of the two methods was 97.1% (67/69). The specificity of ultrasound guided wire-localization was 3.3% (3/90), the specificity of nano-carbon staining was 2.2% (2/90), and the specificity of combination of the two methods was 5.6% (5/90). With the count of SLN as the unit, the combination of the two methods had the highest diagnostic efficiency in detecting SLN, and the difference was statistical significant (χ2 = 34.31, P < 0.001).@*Conclusions@#Ultrasound guided wire-localization and nano-carbon staining are safe and accessible methods for detecting SLN. It provides a precise treatment for early breast cancer, and it can protect medical staff from radiation, which is expected to be the best method for detection of SLN in breast cancer.

Relationship between expressions of OCT-4, CD117 and clinicopathological features and prognosis of patients with ovarian cancer / 肿瘤研究与临床

Objective@#To investigate the relationship between expressions of OCT-4, CD117 and clinicopathological features and prognosis of patients with ovarian cancer.@*Methods@#A total of 70 paraffin-embedded tissues of patients with ovarian cancer from January 2010 to February 2016 in Shanxi Provincial Cancer Hospital were collected. The expressions of OCT-4 and CD117 were detected by immunohistochemistry.@*Results@#OCT-4 was mainly expressed in cytoplasm, while CD117 was expressed in cell membrane and cytoplasm. The positive expression rate of OCT-4 was 74.3% (52/70), and the positive expression rate of CD117 was 68.6% (48/70). The positive expression rates of OCT-4 in ovarian cancer tissues with poorly differentiation and high CA125 levels (≥500 U/ml), no peritoneal effusion and sensitive to chemotherapy drugs were 92.1% (35/38), 87.5% (28/32), 88.9% (24/27), and 78.7% (48/61), respectively, which were higher than those in ovarian cancer tissues with well and moderately differentiation, low CA125 levels (<500 U/ml), peritoneal effusion and resistance to chemotherapy drugs, the differences were statistically significant (P values were 0.000, 0.020, 0.047, and 0.043). The positive expression rates of CD117 in ovarian cancer tissues with poorly differentiation and peritoneal effusion were 84.2% (32/38) and 79.1% (34/43), respectively, which were higher than those in ovarian cancer tissues with well and moderately differentiation and no peritoneal effusion, the differences were statistically significant (P values were 0.006 and 0.017). Patients with OCT-4-positive expression, peritoneal effusion, and poorly differentiation had a shorter overall survival time (all P < 0.05). The peritoneal effusion and differentiation were independent prognostic factors in patients with ovarian cancer (both P < 0.05).@*Conclusion@#OCT-4 can be used as an important reference for predicting drug sensitivity and evaluating prognosis in patients with ovarian cancer.

Expression of death receptor 5 upreguleted by chloroquine enhances the sensitivity of Huh7 cells to tumor necrosis factor related apoptosis-inducing ligand / 肿瘤研究与临床

Objective To explore the effect of chloroquine on death receptor 5 (DR5) expression of hepatocellular carcinoma Huh7 cells and cell proliferation and apoptosis induced by tumor necrosis factor related apoptosis-inducing ligand (TRAIL).Methods Huh7 cells were divided into four groups:the control group (1∶1 000 dimethyl sulfoxide),TRAIL group (50 μg/L),chloroquine group (10 μmol/L) and TRAIL +chloroquine group (TRAIL 50 μg/L + chloroquine 10 μmol/L).Thiazolyl blue tetrazolium bromide (MTT) assay was used to determine the proliferation activity of cells,immunofluorescence was used to detect the expression of DR5,4',6-diamidino-2-phenylindole (DAPI) staining was used to observe cell apoptosis and Western blot was used to detect the expression of cleaved poly ADP-ribose polymerase (PARP).Results TRAIL treatment could decrease Huh7 cells proliferation activity;when compared with the cell viability in the control group,the cell proliferation inhibition rate of chloroquine group,TRAIL group and TRAIL+ chloroquine group was (89±8) ％,(53±10) ％ and (27±7) ％,respectively;compared with TRAIL group alone,cell proliferation activity was decreased in TRAIL+ chloroquine group (t =3.922,P =0.017).The expression of DR5 was upregulated in chloroquine group,and the cell apoptosis signaling was activated in TRAIL + chloroquine group.The cell apoptosis rate of TRAIL group and TRAIL + chloroquine group was (10.0±2.3) ％ and (20.4±4.0) ％,respectively,and there was a statistical difference (t =3.894,P =0.018).Conclusion Chloroquine can enhance the cell chemosensitivity to TRAIL treatment by upregulating the expression of DR5 in Huh7 cells.

Correlation of BRAF V600E mutation with the clinicopathological features of papillary thyroid carcinoma / 肿瘤研究与临床

Objective To discuss the BRAF V600E mutation rate in papillary thyroid carcinoma (PTC) and its relationship with the clinicopathological features. Methods Two hundred and sixty-five PTC patients(including 226 cases of classical type,29 cases of follicular type, 3 cases of high cell type, 2 cases of diffuse sclerosis type, 2 cases of eosinophilic type, 3 cases of cystic type) from August 2014 to October in Shanxi Provincial Cancer Hospital, were collected with completely clinical and pathological information. The BRAF V600E mutation was detected by real-time polymerase chain reaction (RT-PCR) method. Pearson χ 2 test and the exact probability method were used to analysis the relationship between gene mutations and clinicopathological data. Results BRAF V600E mutation rate in PTC patients was 73.21 %(194/265). There was no significant difference in the mutation rate of BRAF V600E among patients with different age, gender, tumor location,tumor number and extravaginal invasion(all P>0.05),but the mutation rates of BRAF V600E gene in patients with different tumor size, histopathological subtypes, lymph node metastasis and clinical stage were significantly different(all P<0.05).Conclusion The PTC patients with positive BRAF V600E mutation have poor clinicopathological features,and BRAF V600E mutation may be a predictor of advanced PTC.

Promoter methylation of MEG3 gene in hepatocellular carcinoma and its clinical significance / 肿瘤研究与临床

Objective To investigate promoter methylation state of MEG3 gene in hepatocellular carcinoma and its clinical significance. Methods Methylation-specific polymerase chain reaction (MSP) was performed to detect methylation of MEG3 promoter in 58 cases of hepatocellular carcinoma tissues and 20 cases of normal control from December 2014 to December 2016 in Shanxi Provincial Cancer Hospital, and to analyze the relationship between methylation and the clinicopathological features of the patients. Results The methylation incidence rate of MEG3 promoter in hepatocellular carcinoma tissues (55.2 %, 32/58) was higher than that in normal liver tissue (25.0 %, 5/20), and there was a significant statistical difference (χ2=6.72,P =0.02).There was no significant difference in the incidence of MEG3 methylation of the patients with different age, gender, α-fetoprotein levels, tumor number and differentiation (all P> 0.05). There was a significant difference in the incidence of MEG3 methylation of the patients with different tumors diameter, liver cirrhosis, hepatitis B surface antigen (HBsAg), TNM staging and distant metastasis (all P< 0.05). Conclusion Aberrant promoter methylation in MEG3 genes may be associated with the occurrence of hepatocellular carcinoma,tumor diameter,liver cirrhosis,HBsAg,TNM staging and distant metastasis.

Predictive values of clinicopathological features and expression of thyroid transcription factor 1 for epidermal growth factor receptor mutations in non-small cell lung cancer / 肿瘤研究与临床

Objective To study the values of clinicopathological features and expression of thyroid transcription factor 1 (TTF-1) in predicting the mutation status of epidermal growth factor receptor (EGFR) gene in patients with non-small cell lung cancer (NSCLC). Methods Mutation status of exons 18, 19, 20 and 21 in EGFR, and expression of TTF-1 protein in 283 cases of NSCLC diagnosed in Shanxi Provincial Cancer Hospital from January 2013 to December 2014 were analyzed by using amplification refractory mutation system (ARMS) and immunohistochemical method. The correlation of EGFR mutations with the clinicopathological features and TTF-1 expression were studied to explore the values of them in the prediction of EGFR mutations. Results Among 283 cases of NSCLC, the rate of EGFR gene mutation was 30.0 ％(85/283), including 3 cases with double mutations(exon 18 and exon 20 double mutations in one case, exon 19 and exon 21 double mutations in one case, exon 20 and exon 21 double mutations in one case). The EGFR gene mutations were associated with gender, histological type, history of smoking, and expression of TTF-1 (all P<0.001), but not related to age and tumor location (P= 0.785, P= 0.138). The combination of factors with high mutation rates (women, adenocarcinoma, no smoking, and TTF-1 positive) made the positive predictive value of EGFR mutations up to 57.6 ％. And the combination of factors with low mutation rates (male, nonadenocarcinoma, smoking history, TTF-1 negative) made the EGFR negative predictive value up to 90.3％. Conclusion The combination of clinicopathological features and TTF-1 expression status in patients with NSCLC has a great predictive value for EGFR mutations, which can provide a useful reference for clinical treatment decision-making.

Correlation between expression of C-met and epidermal growth factor receptor-tyrosine kinase inhibitor resistance in lung adenocarcinoma / 肿瘤研究与临床

Objective To detect C-met protein expression and gene amplification in lung adenocarcinoma, and to analyze their relationship with epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) resistance and prognosis. Methods A total of 120 cases of lung adenocarcinoma diagnosed in Shanxi Provincial Cancer Hospital from January 2011 to May 2013 were selected. The expressions of C-met protein and C-met gene amplification were conducted by immunohistochemistry (IHC) method and fluorescence in situ hybridization (FISH), and all patients were followed up. The relationship between the expression of C-met protein and gene amplification with clinicopathological features and EGFR-TKI resistance and prognosis were analyzed. Results The high expression of C-met protein and gene amplification in 120 tissues were 17.5 ％ (21/120), 10.83 ％ (13/120). Of the 80 patients treated with EGFR-TKI, the incidence of C-met protein high expression was 30.43 ％ (14/46) in patients with drug resistance, which was significantly higher than that in patients without drug resistance (11.76 ％, 4/34), the difference was statistically significant (χ2= 3.908, P= 0.048). The rate of C-met gene amplification was 19.57 ％ (9/46) in patients with drug resistance,which was significantly higher than that in patients without drug resistance (2.94 ％, 1/34) the difference was statistically significant (P= 0.038). The expression of C-met protein in 46 patients with drug resistance was positively correlated with gene amplification (r= 0.388, P= 0.008), but in 40 patients without TKI, the expression of C-met protein was not correlated with gene amplification (r=0.279, P=0.081). The high expression of C-met protein was correlated with age, pathological grade and clinical stage (all P<0.05), while C-met gene amplification was related to clinical stage (P=0.036). Cox regression analysis suggested that C-met gene amplification was an independent prognostic factor (P= 0.034). Conclusions C-met protein expression and gene amplification are risk factors for EGFR-TKI resistance. C-met gene amplification suggests poor prognosis, and can be used as an independent factor for prognostic evaluation.

Relationship between mismatch repair protein expression and clinicopathological features of endometrial cancer / 肿瘤研究与临床

Objective To screen out probable lynch syndrome (LS) associated endometrial cancer (EC) by investigating the expression of mismatch repair (MMR) protein in EC, and to analyze the disease traits combined with clinicopathologic characteristics. Methods The expressions of MSH2, MSH6, MLH1 and PMS2 were detected by using immunohistochemistry (IHC) in 443 EC patients. Results In 443 EC patients, 328 cases (74%) with all MMR proteins expression were classified as sporadic EC, and 115 cases (26%) cases with loss expression of at least one MMR protein were regarded as probable LS. MMR-deficient cases mostly showed a loss of MLH1/PMS2 expression (42%), followed by the absence of MSH2/MSH6 (23%), MSH6 (17%), PMS2 (17%) and MSH6/MLH1/PMS2 (3%). Compared with the sporadic EC group, obesity was not found in probable LS group (body mass index<28 kg/m2) (P=0.040), and high tumor grade was common (P=0.012); There was no significant difference between the two groups in age, the incidence of diabetes or hypertension, family history of cancer or histological type, tumor location, the International Federation of Gynecology and Obstetrics (FIGO) stage, myometrial invasion, lymph node metastasis, lymphatic or vascular invasion (all P> 0.05). A higher tumor grade was more common in the MSH6 and PMS2 deficient groups. Conclusions Compared with sporadic EC, the absence of obesity, a high grade tumor are more common in probable LS cases.

Expression of programmed death 1 and programmed death-ligand 1 in T-lymphoblastic lymphoma/leukemia and their clinical significances / 白血病·淋巴瘤

Objective To study the expression of programmed death 1 (PD-1) and its ligand PD-L1 in T lymphoblastic lymphoma/leukemia (T-LBL/ALL), and to explore the relationship between their expression and clinical pathological factors and prognosis of the disease. Methods PD-L1 and PD-1 were detected in 56 patients with T-LBL/ALL by immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR). Twenty patients with reactive hyperplasia were selected as the control group. Results Immunohistochemical results showed that PD-1 was not expressed in T-LBL/ALL tumor cells and 17.9 %(10/56) in tumor infiltrating immune cells,which was statistically significant compared with control group(18/20,90 %) (P= 0.000). The positive rates of PD-L1 protein were 37.5 % (21/56) and 10.0 % (2/20) in the experimental group and the control group, respectively and accordingly the difference between the two groups was statistically significant (P =0.044). Results of qRT-PCR showed that the relative expression levels of PD-L1 and PD-1 mRNA in 56 cases of T-LBL/ALL were significantly higher than those in control group (12.255 vs. 2.575, 37.990 vs. 3.615), and the differences were both statistically significant (both P < 0.05). Univariate analysis showed that age, PD-L1 protein and mRNA expression were closely correlated with prognosis(all P <0.05). Multivariate Cox regression analysis showed that overexpression of PD-L1 protein and age (≤25 years old)were independent prognostic risk factors(both P <0.05).Conclusion In T-LBL/ALL,PD-1/PD-L1 may be involved in the immune escape of the tumor,which is expected to be a new target for the treatment of diseases.

Relationship between mismatch repair protein expression and clinicopathological features of endometrial cancer / 肿瘤研究与临床

Objective To screen out probable lynch syndrome (LS) associated endometrial cancer (EC) by investigating the expression of mismatch repair (MMR) protein in EC, and to analyze the disease traits combined with clinicopathologic characteristics. Methods The expressions of MSH2, MSH6, MLH1 and PMS2 were detected by using immunohistochemistry (IHC) in 443 EC patients. Results In 443 EC patients, 328 cases (74%) with all MMR proteins expression were classified as sporadic EC, and 115 cases (26%) cases with loss expression of at least one MMR protein were regarded as probable LS. MMR-deficient cases mostly showed a loss of MLH1/PMS2 expression (42%), followed by the absence of MSH2/MSH6 (23%), MSH6 (17%), PMS2 (17%) and MSH6/MLH1/PMS2 (3%). Compared with the sporadic EC group, obesity was not found in probable LS group (body mass index<28 kg/m2) (P=0.040), and high tumor grade was common (P=0.012); There was no significant difference between the two groups in age, the incidence of diabetes or hypertension, family history of cancer or histological type, tumor location, the International Federation of Gynecology and Obstetrics (FIGO) stage, myometrial invasion, lymph node metastasis, lymphatic or vascular invasion (all P> 0.05). A higher tumor grade was more common in the MSH6 and PMS2 deficient groups. Conclusions Compared with sporadic EC, the absence of obesity, a high grade tumor are more common in probable LS cases.

Correlation of KRAS and PIK3CA gene status between primary tumors and paired metastases in colorectal cancer / 临床与实验病理学杂志

Purpose To investigate the mutation status of KRAS and PIK3CA gene in colorectal cancer (CRC) primary lesions and corresponding liver metastasis and its clinical significance.Methods The gene mutations of KRAS and PIK3CA were detected in 58 cases of primary lesions of CRC and corresponding liver metastasis tissue by real-time PCR.Results The mutation rates of KRAS were 31.03％ (18/58) and 25.86％ (15/58) in primary lesions of CRC and corresponding liver metastasis tissue,respectively,in which G12D was most commonly detected.The mutation rates of PIK3CA were 8.62％ (5/58) and 10.34％ (6/58) respectively,in which the most common mutation site was E545K.Only one case carried simultaneously both mutations of KRAS (G12D) and PIK3CA (E545K).The mutation of KRAS and PIK3CA had a good consistency between primary lesions and liver metastasis.Univariate analysis showed that the mutation of KRAS was related to the primary lesion of tumor location,the quantity of metastasis and the types of tumor (P ＜ O.05),PIK3 CA mutation was associated with the synchronous/metachronous liver metastasis and the quantity of metastasis (P ＜ 0.05).Multivariate Cox regression analysis showed that synchronous/metachronous liver metastasis and the mutation of KRAS were influencing factors for prognosis of CRC.The overall survival of patients with CRC who had simultaneous liver metastases was longer than those with heterotopic liver metastases;the overall survival of KRAS wild-type mutant patients was longer than those of mutant patients (P ＜ 0.05).Conclusion The G12D site of KRAS gene has the highest mutation frequency in CRC,KRAS/PIK3CA mutation has a good consistency of the primary lesions of CRC and corresponding liver metastasis.Primary lesions can be as the source of molecular detection.To achieve individualized treatment,we need to reassess the genetic status of metastasis based on the choice of targeted therapy for precision medicine.

Clinical features and treatment of primary nasal B-cell lymphoma / 白血病·淋巴瘤

Objective To study the clinical characteristics and treatment of primary nasal B-cell lymphoma (PNBCL). Methods A retrospective analysis was performed based on the clinical records of 18 PNBCL cases who were treated from January 2009 to June 2015. The clinical manifestations, imaging features, diagnosis approaches and treatment of them were analyzed. Results The main symptoms were nasal obstruction and rhinorrhea. Of all patients, 15 cases were in Ann Arbor stageⅠE-ⅡE, and 3 cases were in Ann Arbor stageⅢE-Ⅳ. The median age was 51 years (12-76 years). The ratio of men to women was 11:7. Only 1 patient had B symptoms. Elevated LDH levels were observed in 4 patients. 13 patients were diffuse large B-cell lymphoma(DLBCL), 3 patients were mantle cell lymphoma, and 2 patients were Burkitt lymphoma. CT examination showed the abnormal nasal soft tissue shadow, with unilateral location and light to moderate enhancement. 14 patients received combination chemotherapy only, and 3 patients received chemotherapy and radiotherapy. Total effective rate was 82.3 % (14/17). At the time of last follow-up, 5 patients died, and the 3-year OS rate was 54.5%(6/11). Conclusions Most PNBCL patients are in Ann Arbor stageⅠE-ⅡE and B symptoms are rare, and the most common pathological types is DLBCL. The treatment for PNBCL is chemotherapy, radiotherapy can assist, but the prognosis is poor, and innovative chemotherapy regimens are necessary.

Expression of programmed death 1 and programmed death-ligand 1 in T-lymphoblastic lymphoma/leukemia and their clinical significances / 白血病·淋巴瘤

Objective To study the expression of programmed death 1 (PD-1) and its ligand PD-L1 in T lymphoblastic lymphoma/leukemia (T-LBL/ALL), and to explore the relationship between their expression and clinical pathological factors and prognosis of the disease. Methods PD-L1 and PD-1 were detected in 56 patients with T-LBL/ALL by immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR). Twenty patients with reactive hyperplasia were selected as the control group. Results Immunohistochemical results showed that PD-1 was not expressed in T-LBL/ALL tumor cells and 17.9 %(10/56) in tumor infiltrating immune cells,which was statistically significant compared with control group(18/20,90 %) (P= 0.000). The positive rates of PD-L1 protein were 37.5 % (21/56) and 10.0 % (2/20) in the experimental group and the control group, respectively and accordingly the difference between the two groups was statistically significant (P =0.044). Results of qRT-PCR showed that the relative expression levels of PD-L1 and PD-1 mRNA in 56 cases of T-LBL/ALL were significantly higher than those in control group (12.255 vs. 2.575, 37.990 vs. 3.615), and the differences were both statistically significant (both P < 0.05). Univariate analysis showed that age, PD-L1 protein and mRNA expression were closely correlated with prognosis(all P <0.05). Multivariate Cox regression analysis showed that overexpression of PD-L1 protein and age (≤25 years old)were independent prognostic risk factors(both P <0.05).Conclusion In T-LBL/ALL,PD-1/PD-L1 may be involved in the immune escape of the tumor,which is expected to be a new target for the treatment of diseases.