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 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 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 investigate the possible association of circulating components of GH-IGFs-IGFBPs system with the GHR-exon 3 genotype in idiopathic short stature (ISS) children. Methods Genomic DNA was extracted and isolat-ed from peripheral leukocytes in 108 ISS children. GHR-exon 3 polymorphism was analyzed with multiplex poly-merase chain reactions (PCR) assay. According to the results of genotype, ISS children were divided into GHRfl group and GHRd 3 group. The height and weight were recorded in two groups. The body mass index (BMI) and BMI standard deviation score (SDS) were measured. The serum levels of insulin-like growth factor (IGF)-1, IGF-binding protein (IGFBP)-3, IGF-1 SDS and IGFBP3 SDS were calculated. GH stimulation test was used to measure the serum GH peak value. Fifty-five ISS chil-dren were treated with recombine human GH [0.15 IU/(kg·d)] for three months to analyse the association of IGF-1 response of GH treatment and genotypes. Results There were 63 GHRfl and 45 GHRd3 in 108 ISS children. There were no signifi-cant differences in BMI, IGF-1, IGFBP3, GH peak, IGF-1 SDS and IGFBP3 SDS between two groups (P>0.05). Multiple stepwise regression analysis showed that age, IGFBP3, lg (BMI) and lg (GH peak) were influencing factors of lgIGF-1 (P<0.05). In 55 ISS children treated with rhGH, there were 34 cases of GHRd3. The differences of △IGF-1 and △IGF-1 SDS were higher in GHRd3 group than those of GHRfl group (n=21). Conclusion The GH sensitivity may be a risk factor in ISS children, which may not be related with GHR polymorphism.

Objective:To investigate the effect of the expression levels of CXCL12 and CXCR4 of carbon dioxide (CO2) pneu-moperitoneum on nude mice xenografts from human ovarian cancer SKOV3 cells in the same pressure at different times. Methods:A total of 40 animal models of human ovarian cancer SKOV3 cell-bearing nude mice xenografts were established. The animals were ran-domly divided into four groups (n=10), namely, the simple anesthesia group, 0.5 h laparotomy group, and 0.5 and 1 h CO2 pneumoperi-toneum groups. After the aforementioned treatments in the groups, the expression levels of CXCL12 and CXCR4 mRNAs of tumor tis-sues were detected by reverse transcription-polymerase chain reaction (RT-PCR), whereas those of the CXCL12 and CXCR4 proteins were detected by Western blot. Results:The 40 animal models of human ovarian cancer SKOV3 cell-bearing nude mice were success-fully established. After the treatment, RT-PCR results reveal that the CXCL12 mRNA expression was significantly higher in the 1 h CO2 pneumoperitoneum group than that in the other three groups. However, no differences were observed in the CXCR4 mRNA expression of each group. Western blot analysis also obtained the same results. Conclusion:The expression level of CXCL12 in SKOV3 cells of the 1 h CO2 pneumoperitoneum group increased, which possibly promoted the growth of peritoneal tumors, and had a slight effect on tu-mor metastasis.

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 explore the effects of deoxynivalenol (DON) on apoptosis of human gastric carcinoma cell line SNU in vitro. Methods SNU cells were treated with DON at different concentrations (50, 100, 1000, 2000 μg/L) for 12 hours, and then cells were harvested for cell apoptosis by flow cytometric (FCM) DNA analysis and the expression of Bax, Bcl-2 and Caspase-3 at protein level with FCM and Western blotting. Results FCM results showed that the apoptosis rates of SNU cells in DON treatment groups were all higher than that in control, especially in DON 1000 μg/L and 2000 μg/L groups (P<0.05). In the concentration range from 50 to 2000 μg/L, a significant concentration-depended response correlation could be found between apoptosis rate and DON concentration (r =0.940, P <0.01). FCM and Western blotting showed Bax and Caspase-3 expression in often SNU cells DON treatment for 12 hours were up-regulated while that of Bcl-2 was down-regulated. Conclusion DON can induce apoptosis of SNU cells in vitro in dose-dependent manner, and possible mechanisms of apoptosis induction effects may be up-regulation of the expression of Bax and down-regulation of that of Bcl-2 and activation of the key enzyme of apoptosis Caspase-3.

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 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 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.