Objective To establish methodology to detect telomerase activity based on real-time quantitative PCR technique combined with telomeric repeat amplification protocol (TRAP). Methods RQ-TRAP system was developed by combining real-time quantitative PCR technique with conventional TRAP method. Telomerase activity was assessed and compared by RQ-TRAP assay and TRAP connected with enzyme-linked immunosorbent assay (TRAP-ELISA) respectively in 12 kinds of cells. Results The RQ-TRAP method was both accurate and specified in measuring telomerase activity in a series dilution of protein extracts from 293T cells. The sensitivity of this method was 8 cells and the amplification efficiency was 98.92%. Telomerase activity was not detected in negative control group. Statistical analysis revealed a strong correlation between the two assays (r2=0.762 5). Conclusion The feasibility of RQ-TRAP was proved in this article. Compared with TRAP-ELISA, RQ-TRAP has many advantages. Apart from sample extraction and real-time PCR cycling, no other extra time-consuming steps are needed for telomerase quantification;RQ-TRAP is less costly and more rapid and reliable than TRAP-ELISA for quantification of telomerase activity and it also support high throughput.

Objective To explore the relationship of different body mass index, waist circumference and serum lipid levels with essential hypertension among residents of Ningbo. And to explore the predictive value of BMI and WC (Waist Cir-cumference) for hypertension. Methods Adopting cluster sampling, 2 303 people (1 081 cases of hypertension and 1 222 people with normal blood pressure) were collected in the four townships of Ningbo City. We conducted questionnaires, physi-cal examination and blood lipid testing for all of subjects. WC group:WC≥85 cm in male or WC≥80 cm in female is central obesity, and the others are normal. BMI groups:BMI≥24 kg/m2 is overweight, and BMI<24 kg/m2 is normal. The relationship of BMI, WC and serum lipid levels with hypertension was analyzed by multivariate unconditional logistic regression model. The predictive value of BMI and WC for hypertension was analyzed by receiver operating characteristic (ROC) curve. Results WC, BMI, total cholesterol (TC), triglyceride (TG) and low density lipoprotein cholesterol (LDL-C) in hypertension group were higher than those in control group (P<0.01). And WC, BMI, TG, LDL-C is the impact factor for hypertension. The overweight rate and central obesity rate in hypertension group were higher than control group (P<0.01). TG and LDL-C level abnormality, overweight and central obesity were the risk factors of hypertension. As a risk factor of hypertension, over-weight with central obesity group is higher than overweight with normal waistline group, simple central obesity group and nor-mal group. In the area under the ROC curve, BMI is higher than WC in both male and female population. Conclusion WC, BMI, TG and LDL-C abnormalities were risk factors for hypertension, and BMI was a better predictive factor for hyper-tension than WC.

After the feasibility to reform the blended teaching model for health information management and its im-plementation were described , the reform of blended teaching model for health information management and its prob-lems were analyzed in terms of its model establishment , process implementation and effect assessment according to the target of professional training in Bengbu Medical College .

Objective To evaluate the biomechanical characteristics and the clinical advantage of transoral atlantoaxial reduction plate(TARP)Ⅲ.Methods Design of TARP-Ⅲ was based on TARP-Ⅱ.The screw hole in the axis was moved 1-2 mm upwards and inwards in a plate which turned a vertebral screw into a pedicle screw or an articular process screw.A polyaxial self-lock ring and polyaxial guiding drill were added to the crew hole of the plate.Finally,the withdrawal resist ence force of the three axis screws was tested and TARP-Ⅲ was used in 44 patients with complicated irreducible atlantoaxial dislocation.The axis was fixed with the pedicle screw or the articular process screw.Results The maximum withdrawal resist ence force of the anterior pedicle screw,the articular process screw and the vertebral screw in the axis was(593.1 ± 97.8)N,(469.9 ± 73.3)N and(395.2 ± 75.1)N respectively,with statistical difference between groups among three fixation methods(P ＜ 0.05).All 44 patients were followed up for 5-38 months(average 18 months),which showed complete anatomic reduction in 36 patients and appropriate anatomic reduction in eight,with basic correction of the angles between the brain stem and the spinal cord and sufficient decompression of the spinal cord.The decompression rate of the cervical spinal cord was average 88.2% according to the Yin evaluating method of cervical cord decompression.The improvement rate of spinal cord function was average 76.6% according to Japanese Orthopaedic Association(JOA)score.Conclusion With the design of polyaxial self-lock mechanism,TARP-Ⅲ with the pedicle screw or the articular process screw surpasses TARP-Ⅱ with vertebral screw in aspect of biomechanics.

Lung cancer is one of the malignant tumors with the highest incidence and mortality rate worldwide and has a poor prognosis. With the popularization of thin layer CT, more and more early lung cancers are being detected. In recent years, thoracoscopic technology and robotic surgery technology have been innovated, and multiple adjuvant therapies such as chemotherapy, targeted therapy and immunotherapy have progressed rapidly, providing lung cancer patients with multiple treatment options. However, the prognosis of lung cancer patients has not been significantly improved, which is related to the overall poor prognosis of lung cancer patients on the one hand, and the urgent need to find effective predictive markers for patients' prognosis, to accurately screen patients, and to carry out targeted treatment in order to improve patients' prognosis on the other hand. In this paper, we review the progress of tertiary lymphoid structures (TLSs) in lung cancer and discuss the prognostic predictive value of TLSs in lung cancer and their feasibility as biomarkers for adjuvant therapy.

Objective To investigate the serum levels of brain derived neurotrophic factor (BDNF) and heat shock protein 70 (HSP70),and evaluate their clinical significance in brain injury of children with complicated febrile convulsions (CFC).Methods Forty-three patients with CFC,41 children with simple febrile convulsions and 36 healthy controls were recruited in this study; their serum levels of BDNF and HSP70 were detected by enzyme linked immunosorbent assay (ELISA); correlation between levels of BDNF and HSP70 and both time of twitch duration and twitch frequency were analyzed.Results The mean serum concentrations of BDNF (4.96±1.21 ng/mL) and HSP70 (2.86± 0.93 ng/mL) in patients of CFC group were significantly higher than those in patients of simple febrile convulsions and healthy controls (P＜0.05); while no significant difference was noted between patients of simple febrile convulsions and healthy controls (P＞0.05).These changes were closely related to the duration time and frequency of convulsions(r=0.553,P=0.015; r=0.587,P=0.012; r=0.526,P=0.024; r=0.562,P=0.018).Conclusions It is sure that some brain injury exists in children with CFC; BDNF and HSP-70 might be one of the molecular mechanisms for the brain injury and protective effect.

Objective To explore the effect of let?7 miRNA silencing on Kaposi′s sarcoma? associated herpesvirus ( KSHV) lytic replication and the underling mechanism. Methods The pEGFP?C2?let?7 sponge vector was transfected into BCBL?1 and 293T cells with Lipofectamine 2000 to silence the expression of let?7 miRNAs. Quantitative real?time PCR ( qRT?PCR) was used to quantify the expression of let?7 miRNAs, the transcriptional levels of mitogen?activated protein kinase kinase kinase kinase 4 ( MAP4K4) , cyclooxygenase?2 ( COX?2) and matrix metalloproteinase 13 ( MMP?13) , and the DNA copy numbers of KSHV open reading frame 50 ( ORF50) and open reading frame 72 ( ORF72) . Western blot was used to detect the total and phosphorylated protein levels of MAP4K4, COX?2, extracellular regulated protein kinases (ERK1/2), c?Jun N?terminal kinase (JNK) and p38 MAPK. Results The expression of let?7 miRNAs was dramatically decreased in the let?7 sponge transfected BCBL?1 and 293T cells compared with that in the vector?transfected cells ( P<0.05 for all) . The gene copy number and mRNA transcriptional level of KSHV ORF50 were significantly increased in the let?7 sponge transfected BCBL?1 cells compared with that in the vector?transfected cells ( 1. 00 ± 0. 10 vs. 2. 33 ± 0. 18 and 1. 08 ± 0. 48 vs 3. 22 ± 0. 27, respectively, P<0.001 for both) . The gene copy number and mRNA transcriptional level of KSHV ORF72 were also significantly increased in let?7 sponge transfected BCBL?1 cells compared with those in the vector?transfected cells(1.07±0.49 vs 1.67±0.45 and 1.01±0.19 vs 1.54±0.11, respectively, P<0.05 for both). Furthermore, the mRNA transcriptional levels of MAP4K4, COX?2 and MMP?13 were significantly increased in the let?7 sponge transfected BCBL?1 cells compared with those in the vector?transfected cells (1.00±0.05 vs 5.73±0.96, 1.00±0.05 vs 2.68±0.19, 1.00±0.02 vs 2.69±0.25, respectively, P<0.001 for all). Let?7 miRNAs silencing also increased the protein expression levels of MAP4K4, COX?2 and phospho?ERK1/2, while the phospho?JNK and phospho?p38 were not changed in the BCBL?1 and 293T cells. Conclusions Let?7 silencing may activate the replication of KSHV, possibly through up?regulating MAP4K4 and its downstream molecules COX?2, MMP?13, and phosphorylation of ERK1/2, finally results in the progression of Kaposi sarcoma.

Objective To explore the effect of let?7 miRNA silencing on Kaposi′s sarcoma? associated herpesvirus ( KSHV) lytic replication and the underling mechanism. Methods The pEGFP?C2?let?7 sponge vector was transfected into BCBL?1 and 293T cells with Lipofectamine 2000 to silence the expression of let?7 miRNAs. Quantitative real?time PCR ( qRT?PCR) was used to quantify the expression of let?7 miRNAs, the transcriptional levels of mitogen?activated protein kinase kinase kinase kinase 4 ( MAP4K4) , cyclooxygenase?2 ( COX?2) and matrix metalloproteinase 13 ( MMP?13) , and the DNA copy numbers of KSHV open reading frame 50 ( ORF50) and open reading frame 72 ( ORF72) . Western blot was used to detect the total and phosphorylated protein levels of MAP4K4, COX?2, extracellular regulated protein kinases (ERK1/2), c?Jun N?terminal kinase (JNK) and p38 MAPK. Results The expression of let?7 miRNAs was dramatically decreased in the let?7 sponge transfected BCBL?1 and 293T cells compared with that in the vector?transfected cells ( P<0.05 for all) . The gene copy number and mRNA transcriptional level of KSHV ORF50 were significantly increased in the let?7 sponge transfected BCBL?1 cells compared with that in the vector?transfected cells ( 1. 00 ± 0. 10 vs. 2. 33 ± 0. 18 and 1. 08 ± 0. 48 vs 3. 22 ± 0. 27, respectively, P<0.001 for both) . The gene copy number and mRNA transcriptional level of KSHV ORF72 were also significantly increased in let?7 sponge transfected BCBL?1 cells compared with those in the vector?transfected cells(1.07±0.49 vs 1.67±0.45 and 1.01±0.19 vs 1.54±0.11, respectively, P<0.05 for both). Furthermore, the mRNA transcriptional levels of MAP4K4, COX?2 and MMP?13 were significantly increased in the let?7 sponge transfected BCBL?1 cells compared with those in the vector?transfected cells (1.00±0.05 vs 5.73±0.96, 1.00±0.05 vs 2.68±0.19, 1.00±0.02 vs 2.69±0.25, respectively, P<0.001 for all). Let?7 miRNAs silencing also increased the protein expression levels of MAP4K4, COX?2 and phospho?ERK1/2, while the phospho?JNK and phospho?p38 were not changed in the BCBL?1 and 293T cells. Conclusions Let?7 silencing may activate the replication of KSHV, possibly through up?regulating MAP4K4 and its downstream molecules COX?2, MMP?13, and phosphorylation of ERK1/2, finally results in the progression of Kaposi sarcoma.

BACKGROUND: Both aldehyde dehydrogenase 2 (ALDH2) rs671 polymorphism and lifestyle behaviors are involved in coronary artery disease (CAD), while the interaction between them is currently unknown. METHODS: A nested case-control study was conducted in 161 patients with CAD and 495 controls in dyslipidemia population in Yinzhou District, Ningbo, Zhejiang Province, China, in August 2013. Anthropometric data and blood samples were collected, demographic characteristics and lifestyle behaviors information were obtained by a face-to-face interview, dietary intake was assessed by a food frequency questionnaire, and genomic DNA was genotyped. RESULTS: Carriers with increasing number of A alleles had an elevated CAD risk compared with G allele carriers (adjusted OR = 1.483, 95% CI = 1.114-1.974). Carriers of rs671 A/G and A/A genotypes had a higher CAD risk than carriers of G/G genotype (adjusted OR = 1.492, 95% CI = 1.036-2.148). Similarly, individuals with rs671 A/A genotype had a higher CAD risk than individuals with A/G and G/G genotypes (adjusted OR = 2.161, 95% CI = 1.139-4.101). We found a borderline additive interaction between regular fried food intake and A/A and A/G genotypes, and a significantly additive interaction between sedentary/light physical activity and A/A and A/G genotypes. CONCLUSIONS Individuals with A/A or A/G genotypes of rs671 have a higher CAD risk, if they lack physical activity and take fried food regularly, than individuals with G/G genotypes. These findings can help to provide a guide to targeted heart health management.
Adult ; Aged ; Aged, 80 and over ; Aldehyde Dehydrogenase, Mitochondrial ; genetics ; Alleles ; Case-Control Studies ; China ; Coronary Artery Disease ; blood ; genetics ; Dyslipidemias ; blood ; genetics ; Female ; Genetic Association Studies ; Genetic Predisposition to Disease ; Genotype ; Humans ; Life Style ; Male ; Middle Aged ; Polymorphism, Single Nucleotide ; Risk Factors