Liquid biopsy of circulating free DNA has become an auxiliary method to evaluate cancer.Because of the advantages of minimally invasive and simple, it plays an increasingly important role in the screening, diagnosis, monitoring treatment, prognosis and drug resistance detection of breast cancer.This article reviews the clinical application and related research progress of circulating free DNA (cfdna) in breast cancer.

Objective:To investigate the expressions and significance of cyclin D2 and bcl-2 in diffuse large B-cell lymphoma (DLBCL) and their clinical significances.Methods:The tissues of 87 DLBCL patients undergoing resection and 23 patients with lymphoid tissue reactive hyperplasia (RLH) in the First Central Hospital of Baoding from January 2015 to March 2020 were collected. Immunohistochemistry method was used to detect the expressions of cyclin D2 and bcl-2 in tissues of DLBCL and RLH, and the relationship between the expressions of cyclin D2 and bcl-2 as well as the association with the clinicopathological features of DLBCL patients.Results:The positive rates of cyclin D2 protein in DLBCL and RLH were 33.3% (29/87) and 2.0% (1/23), the positive rates of bcl-2 protein in DLBCL and RLH were 60.9% (54/87) and 7.0% (3/23); the positive rates of cyclin D2 and bcl-2 in DLBCL were higher than those in RLH, and the differences were statistically significant( χ2=7.566, P=0.006; χ2=17.512, P < 0.01). The expressions of cyclin D2 and bcl-2 proteins were related to the Ann Arbor staging and immunophenotype of DLBCL patients (all P < 0.05), while not related to age, gender, cancer location, tissue type (all P > 0.05). There was a positive correlation between the expressions of cyclin D2 and bcl-2 protein in DLBCL ( r=1.000, P < 0.01). Conclusions:cyclin D2 and bcl-2 may be related to the development and progression of DLBCL, and both may have some synergies.

Objective To investigate the expressions of Apaf-1 and Caspase-9 protein in ovarian serous carcinoma(OSC) and their clinicopathological significance.Methods The expressions of Apaf-1 and Caspase-9 protein in 45 cases of OSC,60 cases of ovarian serous cystadenomas and 32 cases of ovarian borderline serous cystadenomas were detected by immunohistochemical SP method.The relationship between the expressions of these two proteins and the clinicopathological features of OSC and the correlation between Apaf-1 and Caspase-9 expressions in OSC were analyzed.Results The positive rates of Apaf-1 in OSC,ovarian serous cystadenoma and ovarian borderline serous cystadenoma were 24.4%(11/45),75.0%(45/60) and 46.9%(15/32),the difference was statistically significant (χ2=26.734,P<0.01).Apaf-1 expression was correlated with pathological grade,clinical stage and lymph nodes metastasis (χ2=6.318,7.565,5.554,P<0.05).The expression rates of Caspase-9 in OSC,ovarian serous cystadenoma and ovarian borderline serous cystadenoma were 28.9%(13/45),83.3%(50/60) and 56.3%(18/32),the difference was statistically significant (χ2=31.682,P<0.01).The expression of Caspase-9 was correlated with pathological grade,clinical stage and lymph nodes metastasis (χ2=5.750,4.391,5.466,P<0.05).In OSC,the expressions of Apaf-1 and Capase-9 were positively correlated (k=0.433,P=0.003).Conclusion The low expressions of Apaf-1 and Caspase-9 in OSC may be related to the occurrence and development of OSC.

Objective To study the expressions of hypoxia inducible factor-1 α (HIF-1 α) and carbonic anhydrase 9 (CA9) in different molecular subtypes of breast carcinoma and their relationships with basallike subtype.Methods The expressions of HIF-1α and CA9 were detected by immunohistochemistry in five subtypes of breast carcinoma (n =803).The relationships of the expressions of HIF-1α and CA9 with the clinicopathologic parameters of basal-like subtype and with the prognosis of patients were analyzed.Results The positive expression rates of HIF-1α and CA9 protein in breast carcinoma were much higher than those in paratumor breast tissues (43.96％ vs.3.86％,x2 =354.858,P ＜ 0.001;19.80％ vs.4.61％,x2 =86.495,P ＜ 0.001).The positive expression rates of HIF-1α in Luminal A,Luminal B,epidermal growth factor receptor-2 (HER-2) over-expression,basal-like and normal breast-like subtypes were 31.97％,25.00％,56.06％,77.89％ and 55.76％ respectively,and the positive expression rates of CA9 were 7.48％,5.56％,60.61％,40.00％ and 27.88％ respectively,with significant differences (x2 =88.628,P ＜ 0.001;x2 =147.128,P ＜0.001).The positive expression rates of HIF-1α and CA9 in estrogen receptor (ER)-negative breast carcinomas(HER-2 over-expression,basal-like and normal breast-like) were significantly higher than those in ER-positive subtypes (Luminal A and Luminal B,all P ＜ 0.005).There was a significant positive correlation between HIF-1 α and CA9 (C =0.315,P =0.001),and both were correlated with the lymph node metastasis (x2 =4.542,P =0.033;x2 =7.025,P =0.008),pTNM stage (x2 =5.390,P =0.020;x2 =10.179,P =0.001) and poor prognosis (x2 =4.353,P =0.037;x2 =6.056,P =0.014) of basal-like subtype.Conclusion Over-expressions of HIF-1α and CA9 may be related to the ER-negative breast cancer more closely,and both may play important roles in the occurrence and development of basal-like subtype.Overexpressions of HIF-1α and CA9 may suggest the poorer prognosis of patients with basal-like breast carcinoma.

Objective To study the expressions and clinical significances of activating transcription factor 3 (ATF3) and Runt-related transcription factor 2 (Runx2) in breast carcinoma tissues.Methods The expressions of ATF3 and Runx2 were detected in 105 cases of primary breast invasive ductal carcinoma (IDC) and their matched para-tumor breast tissues by immunohistochemistry.The relationships between the two transcription factors and the clinical pathological features of IDC patients were analyzed.Results The positive expression rates of ATF3 and Runx2 in IDC group were 80.95％ (85/105) and 69.52％ (73/105) respectively,much higher than those in para-tumor breast tissues 11.43 ％ (12/105) and 8.57％ (9/105),with statistically significant differences (x2 =102.097,P =0.000;x2 =11.595,P =0.001).ATF3 and Runx2 expressions showed significant relationships with histological grade (x2 =14.623,P =0.001;x2 =24.891,P =0.000),lymph node metastasis (x2 =7.059,P =0.008;x2 =6.358,P =0.012) and pTNM stage of IDC (x2 =5.807,P =0.016;x2 =4.902,P =0.027),while both were not correlated with patients' age (x2 =0.274,P =0.601;x2 =1.554,P =0.213) and tumor size (x2 =2.476,P =0.290;x2 =5.261,P =0.072).There was a significant positive relationship between ATF3 and Runx2 in breast carcinoma (C =0.498,P =0.000).Conclusion The over-expressions of ATF3 and Runx2 may participate in the tumorigenesis,invasion,metastasis and clinical stage of breast carcinoma,which suggests that they may be key factors to evaluate malignant degree and prognosis of breast carcinoma.

Objective To investigate the expression and clinical significance of apoptotic protease activating factor 1 (Apaf-1) and bax in prostate cancer (PCa) and benign prostatic hyperplasia (BPH). Methods Immunohistochemistry was used to detect the expression of Apaf-1 and bax in the tissues from 45 PCa patients and 60 BPH patients. Results The positive rates of Apaf-1 and bax in PCa tissues were 22.22%(10/45) and 20.00 % (9/45), respectively, while those in BPH tissues were 48.33 % (29/60) and 46.67 % (28/60). There was a statistically significant difference in the expressions of Apaf-1 and bax between two groups (P< 0.05). The expressions of Apaf-1 and bax were not correlated with the age of patients and distant metastasis (P>0.05), but they were correlated with the pathological grade and clinical stage of PCa (P< 0.05). The expressions of Apaf-1 and bax in PCa tissues were lower than those in BPH tissues. There was a positive correlation between the expression of Apaf-1 and bax (r=0.535, P<0.01). Conclusion Apaf-1 and bax might be correlated with the carcinogenesis and development of PCa.

Objective To investigate the expressions and significances of Bcl-2 and Bax in basal-like breast carcinoma (BLBC).Methods The expressions of Bcl-2 and Bax were detected in 43 cases of BLBC, 57 cases of non-BLBC and 60 cases of normal breast tissues by immunohistochemistry,and their relationships with physiological and pathological characteristics of patients were analysized.Results The positive rate of Bcl-2 in BLBC was 69.77%,higher than 43.86% in non-BLBC (χ2 =6.647,P =0.01 0)and 21 .67% in normal breast tissues (χ2 =23.831 ,P =0.001 ).The positive rate of Bax in BLBC was 20.93%,lower than 45.61 % in non-BLBC (χ2 =6.564,P =0.01 0)and 76.67% in normal breast tissues (χ2 =31 .270,P =0.001 ).The expressions of Bcl-2 and Bax were correlated with lymphnode metastasis (χ2 =6.927,P =0.008;χ2 =6.203,P =0.01 3)and pTNMstaging of BLBC (χ2 =6.331 ,P =0.01 2;χ2 =5.972,P =0.01 5).There was negative correlation between the expression of Bcl-2 and Bax in BLBC (r = -0.408,P <0.01 0). Conclusion High expression of Bcl-2 and low expression of Bax interact with each other leading to unbalance of cell deferation and apoptosis,resulting in promoting genesis and progress of BLBC.

Purpose To study the expression of HIF-1αand Glut-1 in the molecular subtypes of breast carcinoma and their correlation with basal-like breast carcinoma. Methods 803 cases of invasive breast carcinoma from our database were identified. The clinicopath-ologic findings and the biologic markers including estrogen receptor ( ER) , progesterone receptor ( PR) , and human epidermal growth factor receptor-2 (HER-2) status were reviewed. Immunohistochemical MaxVision stains for cytokeratin 5/6 (CK5/6) and epidermal growth factor receptor ( EGFR) were performed. All breast carcinomas were subclassified into Luminal A, Lumincal B, HER-2 over-expression, normal-like, and basal-like subtypes according to Nielsen criteria. Immunohistochemical stain was also used to detect the expression of HIF-1αand Glut-1. Results Positive expression rates of HIF-1αprotein in basal-like, HER-2 over-expression, normal-like, Luminal A and Luminal B substypes were 77. 89% (74/95), 56. 06% (37/66), 55. 76% (92/165), 31. 97% (141/441), 25. 00% (9/36), respectively. The positive expression rates of Glut-1 protein were 80. 00% (76/95), 57. 58% (38/66), 58. 18%(96/165), 34. 01% (150/441), 25. 00% (9/36), respectively. The positive expression rates of HIF-1α and Glut-1 in the basal-like, HER-2 over-expressing and normal-like subtypes were remarkably higher than that in Luminal A and Luminal B subtypes ( P<0. 004 5) and the expression of HIF-1a and Glut-1 was negatively correlated with the expression of ER (P<0. 01). In the ER-negative breast cancers, the positive expression rates of HIF-1a and Glut-1 in basal-like substype were much higher than that in the other sub-types (P<0. 004 5), and the expression of HIF-1α was positively correlated with expression of Glut-1 in basal-like breast carcinoma (P<0. 01). Conclusion The overexpression of HIF-1αand Glut-1 may be closely related to the ER-negative breast cancer and HIF-1α and Glut-1 might play an important role in the development of basal-like breast carcinoma.

Purpose To investigate the expression of Apaf-1 and Caspase-9 in prostate cancer ( PCa) and benign prostatic hyperplasia ( BPH) , and to analyze their correlation with clinicopathological parameters of PCa. Methods Immunohistochemistry was used to de-tect Apaf-1 and Caspase-9 protein in 45 cases of PCa and 60 cases of BPH. Results The positive rate of Apaf-1 and Caspase-9 in PCa tissues was significantly lower than that in BPH (P<0. 05). The expression of Apaf-1 and Caspase-9 was not correlated with the age of patients and distant metastasis (P>0. 05), but it was correlated with the pathological grade and clinical stage of PCa (P<0. 05). Conclusion Apaf-1 and Caspase-9 are lowly expressed in PCa. There is positive correlation between the expression of Apaf-1 and Caspase-9 (rs =0. 645, P<0. 01). Apaf-1 and Caspase-9 might be correlated with the carcinogenesis and development of PCa.

In this study, a fast carbonyl reductases colorimetric screening method for discovering stereoselective carbonyl reductases was established by combining the reverse alcohol oxidation with the azoreductase-catalyzed reduction of azo dye. When azo dye (Orange I , 4-(4-hydroxy-1-naphthylazo) benzenesulfonic acid) and azoreductase (AzoB) were added into the reaction system of alcohol oxidation catalyzed by carbonyl reductase, the produced NAD(P)H served as electron donor for the azoreductase to reduce the azo dye, resulting the color fade. Hence, the carbonyl reductases can be screened by the obvious color change. When chiral alcohol was used as the substrate, the activity and stereoselectivity of carbonyl reductases can be screened at the same time.
Alcohol Oxidoreductases ; chemistry ; Alcohols ; chemistry ; Azo Compounds ; chemistry ; Coloring Agents ; chemistry ; High-Throughput Screening Assays ; NADH, NADPH Oxidoreductases ; chemistry ; NADP ; chemistry ; Oxidation-Reduction ; Stereoisomerism