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 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.

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.

Objective To detect the protein expressions of S100A7 and CD34 in basal-like breast carcinoma and analyze the clinical relationship between S100A7 and basal-like breast cancer and the connection with CD34 markers microvessel denisity (MVD).To study the function of S100A7 in basal-like breast cancer angiogenesis,invasion and metastasis.Methods Immunohistochemistry (IHC) was used to detect the expressions of S100A7 and CD34 in 60 cases of basal-like breast cancer,40 cases of non basal-like breast cancer and 20 cases of normal breast tissues.Results The expressions of S100A7 in basal-like breast cancer,non basallike breast cancer and normal breast tissues were 83.33％,62.50％ and 35.00％ respectively.There were significant differences among them (x2 =5.556,P ＜ 0.051; x2 =17.107,P ＜ 0.05; x2 =4.051,P ＜ 0.05).There was positive correlation between S100A7 and lymph node metastasis,TNM staging and MVD of basal-like breast cancer patients.But there was no significant correlation between S100A7 and patient's age and tumor sizes.Conclusion The up-regulation of S100A7 expressions may be associated with the occurrence and development,invasion and metastasis,angiogenesis of basal-like breast cancer.The effects of S100A7 promoting angiogenesis may be an important factor in the acceleration of the local invasion and distant metastasis in basallike breast cancer.

Objective To explore the application value of detecting HPV E6/E7 mRNA in cervical cancer screening and prognosis.Methods bDNA technology was used to detect the HPV E6/E7 mRNA levels in 301 cases of paraffin-embedded cervical samples that were grouped based on pathological diagnosis.Statistic differece between multiple groups were assessed by chi-square segmentation method.Results The positive detection rate of HPV E6/E7 mRNA is higher in cervical cancer group than that in CIN1-3 groups and cervicitis group,which were 94.29 ％ (66/70),30.30 ％ (20/66),50.00 ％ (19/38),59.46 ％ (44/74),5.66 ％ (3/50),respectively (all P ＜ 0.004 5).The HPV E6/E7-positive rates in all three CIN groups were higher than that in cervicitis group (P ＜ 0.004 5).The positive rates in CIN2 and CIN3 group were higher than that in CIN1 group (P ＜ 0.004 5).HPV E6/E7 mRNA levels in cervical cancer group were higher than those in CIN groups and cervicitis group (16 508.730±8 741.402,929:337±460.880,3 239.681±1 447.399,5 286.224±2 933.445,4.941±2.263,respectively,all P ＜ 0.05).CIN groups had higher copy numbers of HPV E6/E7 mRNA than cervicitis group (P ＜ 0.05).More copies of HPV E6/E7 mRNA were detected in CIN3 group than those in CIN1 group (P ＜ 0.05).Conclusions The positive rate and expression levels of HPV E6/ E7 mRNA was related to the severity of cervical diseases.HPV E6/E7 mRNA detection in paraffin-embedded samples have a high clinical application value in predicting the risk of cervical cancer.It' s an effective measure for cervical cancer screening and follow-up.

Objective To establish a simple but quick method to improve the high fidelity of the synthesis of DNA frag -ments.Methods High fidelity DNA unidirection synthesis method (HFUS) was presented and used that involved Phusion DNA polymerase, BsrDⅠrestriction enzymes and λexonuclease.Using the same system at different temperatures , HFUS method synthesized one positive single-strand DNA and several reverse single -stranded DNA one by one into the target DNA fragment.Results Two random sequences DNA fragments of 340 bp and 450 bp were synthesized using HFUS meth-od.Conclusion This article explores a new method for the synthesis of genes .Through the harvest of 450 bp DNA, HFUS may promise to be a new approach to the synthesis of DNA .

Objective To detect the protein expression of Cripto-1 and CD34 in basal-like breast carcinoma and study the functions of Cripto-1 in basal-like breast cancer angiogenesis,invasion and metastasis.Methods Immunohistochemistry (IHC) was used to detect the expression of Cripto-1 and CD34 in 60 cases of basal-like breast cancer,40 cases of non basal-like breast cancer and 20 cases of normal breast tissues.The clinical relationship between Cripto-1 and basal-like breast cancer were analyzed as well as its connection with CD34 markers MVD.Results The expression of Cripto-1 protein in basal-like breast cancer (42/60,70.00 ％) was significantly higher than that of nonbasal-like breast cancer (19/40,47.50 ％) and normal breast tissue (4/20,20.00 ％) (both P ＜ 0.05).The expression of Cripto-1 protein was positively correlated with the tumor size,clinical stage,lymph nodes metastasis and MVD value in patients of basal-like breast cancer (all P ＜ 0.05),but there was no corelation with the age of the patients (P ＞ 0.05).Conclusions The up-regulation of Cripto-1 expression may be associated with angiogenesis and development in basal-like breast cancer.The effect of Cripto-1 promoting angiogenesis may be an important factor in the acceleration of the local invasion and distant metastasis in basal-like breast cancer.

Objective To observe the effect of traditional Chinese treating pressure ulcer compound on microcirculation of rats in-vivo.Methods Rats were divided into four groups according to the random number table,includeing high-dose group,low-dose group,positive control group and negative control group.Model rats of microcirculation disturbance were created by dropping epinephrine on partial of mesentery,and then the following indexes of four groups were detected: arteriole diameter,arteriole flow rate,junction of capillary network counts,etc.Results The rats mesentery arteriole diameter variation of positive control group in different time were(74.73±4.23)min,(73.46±4.02)min,(81.60±3.23)min,(93.37±3.84)min,(98.38±3.25) min respectively,and the variation of high-dose group in different time were (72.26± 2.90)min,(72.16 ± 3.20) min,(82.45±2.85)min,(94.64 ± 3.37)min respectively.There was no significant difference between the two groups (P＞0.05).The rats mesentery arteriole flow rate variation of negative control group in different time were (5.06±1.12)min,(3.47±0.97)min,(8.51±1.15)min,(43.55±3.24)min,(56.47±3.54)min respectively,the variation of positive control group were (19.37± 1.81)min,(21.54±2.69) min,(39.86±3.53)min,(86.41 ±4.27) min,(94.35 ± 4.23) min respectively,the variation of high-dose group were (13.73 ± 2.02) min,(22.67 ± 2.48) min,(46.32 ± 4.27) min,(89.18 ± 5.13) min,(95.07 ± 4.67) min respectively,and the variation of low-dose group were (6.22±1.48)min,(10.34±2.09)min,(25.41±2.49)min,(56.87±3.75)min,(74.30±3.18)min respectively.The rats mesentery arteriole flow rate has recovered to the level of before dropping epinephrine by 9 rmin in positive control group and high-dose group,and there were no significant difference between the two groups (P＞0.05),but there were statistical significance when high-dose group and positive control group were compared with low-dose group and negative control group (P＜0.05).The junction of capillary network counts variation in different time were: negative control group 0,16.92± 1.17,39.19±2.65,positive control group 20.66±1.89,36.78±3.04,76.04±4.29,high-dose group 14.33±1.46,43.75±3.74,81.23±3.34,low-dose group 9.50± 1.13,23.19 ± 2.44,55.62 ± 3.46,respectively,the junction of capillary network counts of high-dose group and low-dose group were more than negative control group (P＜ 0.05),and the high-dose group was more than the positive control group (P＜0.05).The resuming time of local microcirculation flow of positive control group (8.23 ±2.61)rmin was relatively equivalent to high-dose group (8.56 ±1.87) min,(P＞0.05).Conclusion Traditional Chinese compound can obviously modify microcirculation disturbance of rats mesentery caused by epinephrine.Its effect showed a dose-effect proportional relationship.

Objective To study the expression and clinical significance of FoxM1 and Cep55 protein in basal-like breast carcinoma (BLBC).Methods The expression of FoxM1 and Cep55 protein in 66 cases of BLBC,70 cases of non-BLBC and 66 cases of normal adjacent breast tissue were detected by immunohistochemistry.Their relationship with the clinical pathological factors was analyzed.Results The positive rate of FoxM1 protein in BLBC,non-BLBC and normal adjacent breast tissue was 77.3％ (51/66),60.0％(42/70) and 13.6％(9/66),Cep55 protein was 74.2％(49/66),57.1％(40/70) and 16.7％(11/66),respectively.BLBC and non-BLBC tissue were higher than normal adjacent breast tissue,and BLBC tissue was higher than non-BLBC,there was significant difference (P ＜0.05).In BLBC tissue,the positive expression of FoxM1 and Cep55 protein correlated with lymph nodes metastasis and TNM staging (P ＜0.05),while did not correlate with age,menopause,tumor size (P ＞ 0.05).There was direct correlation between the positive expression of FoxM1 and Cep55 protein (r =0.259,P ＜ 0.05).Conclusion FoxM1 and Cep55 protein may play an important role in the carcinogenesis and development of BLBC,and both of them have the synergistic effect.