The template-independent teminal transferase activity of Taq DNA polymerase results in an overhanging dA at the 3′end of its PCR products. The pGEMX vector constructed in this experiment forms a single overhanging dT at its 3′end as the result of cleavage with Xcm I restriction enzyme. This vector is very efficient for direct cloning of PCR product obtained by using Taq DNA polymerase.Recombinant colonies can be selected by Blue/white screening. Moreover,insertion fragment can be easily released from the vector simply with either BamH I or Hind III digestion.

Objective To explore the expression and significance of Kiss-1, Ki-67 and VEGF-C in papillary thyroid carcinoma(PTC) and thyroid follicular adenoma (FA).Methods Forty-four cases of PTC and twelve cases of FA paraffin-embedded tissues were used .Immunohistochemical staining and microscopic image analysis technique were used to analyze the expression of Kiss-1, Ki-67 and VEGF-C.Results The integrated optical density (IOD) of Kiss-1, and VEGF-C in the PTC groups was 475.56 ±126.02 and 805.29 ±226.05,respectively.The proliferation index of Ki-67 protein was (3.36 ±1.11) %and the difference between the PTC and FA groups was statistically significant (P<0.05).The IOD of the above two indices was 408.12 ±124.05 and 912.63 ±108.12 in the PTC with lymph node metastasis group , respectively, while the proliferation index of Ki -67 protein was (3.93 ±0.92) % and the difference vs the group without lymph node metastasis was significant ( P <0.05 ) .In the PTC with capsular infiltration group the IOD of above two was 425.58 ±87.38 and 891.37 ±149.36, the proliferation index of Ki -67 protein was (3.79 ±1.09) %and the difference with PTC group without capsular infiltrtion was statistically significant (P<0.05).Linear correlation analysis showed that Ki-67 and VEGF-C were with positively correlated in PTC and FA tissues (P<0.05),while Kiss-1 and Ki-67, VEGF-C were with negatively correlated in PTC and FA tissues (P<0.05).Conclusion Kiss-1, Ki-67 and VEGF-C can facilitate the differential diagnosis of PTC and FA , serving as prognostic indicators in patients with PTC .

Objective To investigate the correlation between serum HCY (Homocysteine) and carotid artery stenosis,plaque stability in patients with ischemic cerebrovascular disease.Methods 154 patients with ischemic cerebrovascular disease in Tangdu Hospital were enrolled in the study from June to December 2016.The serum levels of HCY were detected.CT angiography (CTA) was uesd for patients with neck vascular scanning.According to the difference of serum HCY level,patients were divided into 80 cases of high HCY group (observation group) and 74 cases of normal HCY group (control group).The degree of carotid artery stenosis,number and stability of plaque were compared between the two groups and the correlation between serum HCY level and degree of carotid artery stenosis and plaque stability were analyzed.Results The total stenosis rate in the observation group was significantly higher than that in the control group,the moderate stenosis rate and severe stenosis rate in the observation group were significantly higher than those in the control group,with the statistically significant differences (x2 =5.594～ 22.506,all P＜0.05).The levels of serum HCY in mild,moderate and severe stenosis group were 13.16 ± 6.73,15.19± 5.93 and 26.13 ±11.18 μmol/L respectively.The levels of H CY in moderate stenosis group and severe stenosis group were significantly higher than that in mild stenosis group,and the levels of HCY in severe stenosis group was significantly higher than that in moderate stenosis group,with the statistically significant differences (t=2.684～ 5.270,all P＜0.01).The rate of carotid plaque in the observation group was significantly higher than that in the control group,and the differences statistically significant (x2 =25.053,P＜0.01).The rate of unstable plaque and mixed plaque in the observation group was significantly higher than that in the control group,and the rate of stable plaque was significantly lower than that in the control group (x2 =4.067～ 14.95,all P＜0.05).The levels of serum HCY in stable plaque group,mixed plaque group and unstable plaque group were 16.14±5.49,21.91 ± 6.32 and 26.74 ± 10.59 μmol/L respectively.The levels of HCY in mixed plaque group and unstable plaque group were significantly higher than that in stable plaque group,and the differences were statistically significant (t=4.370,4.628,all P＜0.01).The level of HCY in unstable plaque group was significantly higher than that in mixed plaque group,and the difference was statistically significant (t =2.249,P＜ 0.05).Conclusion Serum HCY levels were closely related to carotid artery stenosis and plaque stability.Hyperhomocysteinemia can increase the incidence and degree of carotid artery stenosis as well as the number of carotid plaques and unstable plaques.

Objective To investigate the correlation between HCY (Homocysteine),folate,vitamin B12 and head and neck vascular stenosis in patients with ischemic cerebrovascular disease.Methods 225 patients with ischemic cerebrovascular disease in Tangdu Hospital of the Fourth Military Medical University were enrolled in the study from April 2016 to October 2016.The serum levels of HCY,folate and vitamin B12 were detected.CT angiography (CTA) was uesd for patients with head and neck vascular scanning.According to whether the presence of vascular stenosis,patients were classified as no vessel stenosis group and vascular stenosis group.According to the degree of stenosis,patients were classified as no vascular stenosis group,mild moderate stenosis group and severe stenosis group.Results The HCY levels in the vascular stenosis group were significantly higher than no vessel stenosis group,while the levels of folate and vitamin B12 were significantly lower than no vessel stenosis group,the differences were statistically significant (t=9.656,7.140 and 8.350,all P＜0,01).The HCY levels in mild moderate stenosis group and severe stenosis group were significantly higher than no vessel stenosis group,and the HCY levels in severe stenosis group were significantly higher than mild moderate stenosis group,the differences were statistically significant (t=6.108,9.401 and 5.273,all P＜0.01).The folate levels in mild moderate stenosis group and severe stenosis group were significantly lower than no stenosis group,the differences were statistically significant (t=5.574 and 5.988,all P＜0.01).The vitamin B12 levels in mild moderate stenosis group and severe stenosis group were significantly lower than no stenosis group,the differences were statistically significant (t=4.548 and 7.816,all P＜0.01).The degree of head and neck vascular stenosis and serum levels of HCY were positively correlated (r=0.331,P＜0.01).The degree of head and neck vascular stenosis and levels of vitamin B12 were negatively correlated (r=-0.279,P＜0.05).Conclusion The levels of HCY,folate and vitamin tB12 were closely related to the degree of head and neck vascular stenosis.HCY,folate,vitamin B12 and head and neck CTA play important roles in patients with ischemic cerebrovascular disease clinically.

BACKGROUND:Previous studies have demonstrated that amblyopic patients have a variety of dysopsia compared with normal people.Engineered visual system could find amblyopia through detection of human visual system.OBJECTIVE:To screen sensitive indexes for amblyopia through visual function examination to establish a intemet-based amblyopia screening method.DESIGN,TIME AND SETTING:Randomized,blinded,and controlled experiment.The study was performed at Department of Optometry,People's Hospital of Guangxi Zhuang Autonomous Region from September to November 2008.PARTICIPANTS:A total of 80 volunteers undergoing visual examination in Department of Optometry,People's Hospital of Guangxi Zhuang Autonomous Region were selected,aged 4-19 years.All people had no organic diseases of eyes.They were divided into normal group (n=40) and ambiyopic group (n=40) according to their corrected visual acuity (whether ＞ 4.9).METHODS:Using vision system,the various visual function indexes of each participant were examined.The data from single eye and both eyes were collected and analyzed using Fisher.The sensitive indexes were selected to establish identification function for amblyopia screening.MAIN OUTCOME MEASURES:Age,visual noise,orientation identification and contour integration of participants were examined.RESULTS:Results showed that age,visual noise,orientation identification and contour integration were clinically significant indexes,and cross validation suggested that the identification correct rate was 92.5%.The discriminant function of amblyopia was Y1=1.175X1+0.786X2+0.928X3+1.061X4-0.225X5+2.547X6+1.313X7-18.651;the discdminant function of normal vision was Y2=1.369X1+ 1.728X2+1.779X3+1.549X4-1.912X5+2.665X6+0.387X7 26.640.CONCLUSION:Visual noise,orientation identification and contour integration in vision detection system could be used to screen amblyopia in children,in particular with assistance of internat.

Objective To investigate the relationship between serum homocysteine (HCY)levels and oxidative stress in pa-tients with coronary heart disease (CHD).Methods According to the diagnostic criteria of CHD in 2010 (WS 319-2010)is-sued by the Ministry of Health,79 patients with CHD in Xi’an Central Hospital from June 2014 to December 2014 were se-lected as the experimental group,and 5 5 cases of healthy physical examination for the same period were selected as the nor-mal control group.The levels of oxidative stress marker malondialdehyde (MDA)and the activity of antioxidant enzyme su-peroxide dismutase (SOD)in serum were detected to analyze the state of oxidative stress.The levels of serum HCY were de-tected,and the correlation between HCY levels and MDA levels and SOD activity was analyzed.Results The levels of MDA in the CHD group were significantly higher than that in the control group,while SOD activity was significantly lower than that in the control group,the differences were statistically significant (t=3.112,2.684,all P<0.05).The levels of HCY in the CHD group were significantly higher than that in the control group,the difference was statistically significant (t=3.268,P<0.05).In the CHD group,the levels of HCY were positively correlated with the levels of MDA (r=0.236,P<0.05),and negatively correlated with SOD activity (r=-0.221,P<0.05).Conclusion Serum HCY levels were increased and oxidative stress reaction was enhanced in patients with CHD.Oxidative stress was associated with elevated serum HCY levels.Serum HCY levels and oxidative stress played an important role in the occurrence and development of CHD.

OBJECTIVE:To compare the differences in the contents of water-soluble total protein,total phenolic acid and total polysaccharides among the water decoctions of crude Cibotium barometz and processed products and to illuminate the effect of pro-cessing on 3 kinds of components of C. barometz. METHODS:UV-visible spectrophotometry was adopted to determine the con-tents of water-soluble total protein,total phenolic acid and total polysaccharides in the water decoction of crude C. barometz and 4 processed products,namely sand-scorch C. barometz,yellow wine C. barometz,salt C. barometz and steamed C. barometz,at wavelengths of 590,760 and 489 nm respectively. RESULTS:The contents of water-soluble total protein in 5 samples were 4.03%,3.32%,3.13%,3.33% and 3.49%,those of total phenolic acid therein were 0.25%,1.34%,1.38%,2.34% and 1.41%,and those of total polysaccharides therein were 28.56%,36.06%,45.21%,49.60% and 49.01%,respectively. CONCLU-SIONS:All above processing methods have an effect to some degree on the contents of the 3 kinds of components of C. barometz, where the contents of water-soluble total protein are lower after processing,while those of total phenolic acid and total polysaccha-rides are higher thereafter.

Objective To analyze of internal strength in patients with chronic disease,make clear the concept development,property,prerequisites,outcome and evaluation index of internal force.Methods To retrieve the relevant research literature in the knowledge service platform of Pubmed,PsycINFO and Chinese CNKI,Wanfang data,using the Rodgers analysis method to analyze the evolution of the concept of internal force.Results The molding process of internal strength of patients with chronic diseases included 4 defining characteristics:self concept,support,development and transcendence.Conclusions Rodgers evolution concept analysis method can help medical workers make clear concept essence of internal strength,deepening their cognition and understanding,so as to provide the basis for the implementation of the related research and clinical interventions.

Objective To study the expression and clinical significance of CD34 and smooth muscle actin (SMA) in stroma of breast tissues and lesions. Methods Seventy cases of breast tissues and lesions, including 20 fibroadenomas, 10 sclerosing adnoses, 30 invasive ductal carcinomas and 10 invasive lobular carcinomas were investigated, and 10 normal breast tissues were served as controls. Immunohistochemical staining was applied to compare the distribution of CD34+ fibrocytes and SMA-reactive myofibroblasts. Results The stroma of normal breast tissue contained CD34+ fibrocytes, whereas SMA-reactive myofibroblasts were absent (100% for both). All benign breast lesions exhibited astromal CD34+ fibrobytes, and fibroadenomas showed SMA-reactive myofibroblasts as well. In invasive ductal and lobular carcinoma the stroma was devoid of CD34+ fibrocytes, but a varying number of stromal SMA-reactive myofibroblasts were detectable (100%). Conclusion In breast carcer, immunohistochemical staining used in detecting expressions of CD34 and SMA is helpful in distinguishing benign lesions from malignancies.

Objective To evaluate the relationship between sternum protection and bone marrow suppression in postoperative radiotherapy for esophageal carcinoma. Methods Total of 98 postoperative patients with thoracic esophageal carcinoma were randomly divided into experimental group (52 cases) and control group (46 cases). All patients were given intensity-modulated radiotherapy (IMRT), with the dose of 50-50.4 Gy. The patients in experimental group were irradiated by 6 fields (4-fields in front, 2-fields behind) which were crossed to avoid direct exposure to the sternum. The patients in control group were irradiated by 5 fields (3-fields in front, 2-fields behind) with front-middle of the field passing through the sternum. Concurrently all patients received 2 cycles of cisplatin chemotherapy. Results Dmean, V20 and V30 of the sternum in the experimental group were (20.21 ±3.60) Gy, (40.78 ±7.19) % and (33.78 ±9.44) %, which were lower than those in the control group [(30.91±5.21) Gy, (81.01±4.81) %, (51.60±6.84) %], respectively (P<0.05). However, the volume and dose distribution of lung, spinal cord and heart were similar between the two groups (P> 0.05). Both the incidence rates of bone marrow suppression at 14th day and 35th day after radiotherapy were significantly higher in the control group (52.2%, 73.9%) than those in the experimental group (28.8 %, 50.0 %) (P< 0.05), and the incidence rate of bone marrow suppression at 7th day after radiotherapy was similar between the two groups. Conclusion Protecting and sketching for sternum in postoperative radiotherapy for esophageal carcinoma can reduce the incidence of bone marrow suppression effectively, which would not increase the radiation dose in the lung, heart and spinal cord.