Our knowledge of molecular events and interactions in cell biology and molecular biology are mostly based on findings in nonliving body condition while ignoring the whole process of the events,which inevitably involves false assumption that all molecules behave in the same environment and in the same way. With the emergence of new fluorescent molecular probes prepared through linking new fluorescent material such as quantum dots (QDs) to micromolecule antibody such as antibody simulacrum, the development of methods for transducing extrinsic fluorescent probes into live cells,the application of laser technology, the finer fluorescent micro-imaging system, and the fast and sensitive fluorescence collection system in the application of life sciences, it is now possible to image the trajectory, kinetic behavior, and interaction of biomolecules in live cells. Visualizing single molecule in live cells has opened a new chapter for cell biology and molecular biology. Recent research developments and foreground of single molecule visualizing are discussed in this article.

Serum Cholyglycine (CG)is a main component of human bile acid,one of the conjuga-ted bile acids formed by the combination of bile acid and glycine is synthesized in the liver cells.Glycochol-ic acid (CG)as a clinical indicator for detecting hepatobiliary disorders,and the traditional index of liver
function test compared has greater advantages,its detection for intrahepatic cholestasis,liver disease and biliary system diseases diagnosis,treatment and prognosis analysis of pregnancy to provide an important ba-sis.

Objective To evaluate the prognostic values of leukocyte count,hemoglobin,biochemical parameters,erythrocyte sedimentation rate and immunoglobulin on mortality in patients aged 80 years and over.Methods Totally 342 patients(aged 85.6±4.0 years)were followed up for (82.0±36.9) months,and the cause and time of death were recorded.Results During the period of follow up,198 patients suffered from death.Compared with the survival group (132 cases),the death group had older age [ (86.5±4.4)years vs.(84.5±3.2)years,t=-4.86,P＜0.01 ],higher white blood cell [ (6.2± 1.7) ＞ 109/L vs.(5.5±1.3) × 109/L,t=-3.93,P＜0.01 ],lower hemoglobin [(134.4±14.4)g/L vs.(140.0± 12.6)g/L,t= 3.65,P＜0.01 ],slightly faster erythrocyte sedimentation rate [ 11 mm/h(15 mm/h) vs.9 mm/h (10 mm/h),U=- 3.31,P＜0.01 ],lower immunoglobulin M [ (0.9±0.5)mg/L vs.(1.1±0.8)mg/L,t =2.55,P＜0.05 ],slightly higher urea nitrogen [ (7.5±2.6) mmol/L vs.(6.8±1.6) mmol/L,t=2.81,P＜0.01]and creatinine [(113.0±32.5) μmol/L vs.(100.5±15.8) μmol/L,t=-4.65,P＜0.01 ].Cox multivariate analysis revealed that older age (RR=1.083,95％CI:1.040 1.127,P＜0.01),white blood cell count (RR=1.134,95％CI:1.021-1.260,P＜0.05),creatinine (RR=1.011,95％CI=1.0021.020,P＜0.05),hemoglobin(RR=0.835,95％CI:0.714-0.975,P＜0.05)andimmunoglobulin M(RR=0.710,95％CI:0.521-0.966,P＜0.03),aorticaneurysm(RR=2.144,95％CI:1.163-3.951,P ＜ 0.05 ) were the independent risk factors for death.Conclusions Aging,increased WBC count,decreased hemoglobin and immunoglobulin M,elevated creatinine and aortic aneurysm are the independent risk factors for death,which are powerful parameters for the prognostic evaluation in the elderly aged 80 years and over.

Objective To analyze relevant factors of cervical intraepithelial neoplasia residual or recurrence after loop electri‐cal excision procedure(LEEP) .Methods We observed the patients who were histopathologically confirmed CIN and treated with LEEP in our hospital from January 2006 to September 2012 ,and investigated the postoperative residual and recurrence .Then we an‐alysed relevant factors of residual or recurrence .Results (1)After at least 2 years of follow‐up ,26 cases showed residual or recur‐rence(the residual rate was 0 .97% and the recurrence rate was 4 .08% ) .(2)Overall severity of residual or recurrence had been re‐duced ,but the severity and recurrence of preoperative severity was positively correlated(r=0 .354 0 ,P<0 .05) .(3)Age ,smoking , preoperative high‐risk HPV load ,post‐operative the same type of HPV continuous infection ,endocervical involvement ,lesions in‐vo1ving quadrants ,and post‐operative TCT results between the recurrence group and non‐recurrence group were statistically signifi‐cant different(P<0 .05) .Conclusion CIN residual or recurrence after LEEP might be affected by many factors ,and the treatment should be individualized .

Objective To observe the effect of nitroglycerin(NTG) on pulse wave velocity(PWV) in patients with hypertension(HTN).Methods36 volunteers,mean age 48.1±10.2 years,were divided into HTN group and non-hypertension(NHTN) group according to whether he or she had hypertension or not.The baseline PWV and PWV at 5th minute and 10th minute after sublingual NTG were detected.PWVs of the two groups were compared.ResultsThe basal PWV in HTN group was higher than that in NHTN group.PWV was reduced significantly after NTG were given sublingually 5 minutes or 10 minutes compared with baseline condition(both P<0.01).PWV 10 minutes after sublingual NTG raised a little compared with that after 5 minutes(P<0.01).PWVs 5 minutes or 10 minutes after sublingual NTG in HTN group were higher than those in NHTN group(both P<0.05).ConclusionNTG can reduce PWV in patients with HTN.

Objective To investigate the prevalence rate of heart valve calcification and its relativity with pathological changes and clinical pathogenic factors in elder patients at autopsy Methods Pathology data at autopsy of 1047 patients with age from 60 to 106 years in Beijing Hospital from November 1954 to March 2016 were collected.Cases of heart valve calcification verified at autopsy were retrospectively reviewed.The prevalence of heart valve calcification and its relativity with age,clinical disease and coronary atherosclerosis was investigated.Results Among 1047 autopsies,aortic valve calcification(AVC)was found in 15.2 ％ (n=159),mitral valve calcification(MVC)in 9.6 ％ (n=101),both AVC and MVC calcification in 5.4 ％ (n =57)and heart valve calcification in 19.4 ％ (n =203).The prevalence of heart valve calcification was 6.4％ (15/234)at age of 60-69,12.8％(37/289)at age of 70-79,22.5％(70/311)at age of 80-89 and 38.0％ (81/213) at age of 90-106 years,respectively(tendency x2 =82.523,P＜0.01).Calcification prevalence was significantly increased when complicated with coronary artery stenosis,hypertension,coronary artery disease (CAD),diabetes and chronic kidney disease (CKD).Multivariate regression analysis showed that age and CAD were independently risk factors for heart valve calcification(OR=1.066,95％ CI:1.048-1.086,P＜ 0.01;OR =2.238,95％ CI:1.396-3.589,P＜0.01,respectively),while hypertension,diabetes and CKD were not independent risk factors(OR =1.223,95％ CI:0.859-1.741,P＞ 0.05;OR =1.053,95％ CI:0.700-1.586,P ＞0.05;OR =0.924,95％ CI:0.610-1.399,P＞ 0.05,respectively).As compared with patients without heart valves calcification,patients with heart valve calcification had more increased risk for coronary atherosclerosis(OR =2.983,95a％CI:1.868-4.765,P＜0.01).Conclusions Prevalence of heart valve calcification is increased in elder patients with increasing age.Prevalence of heart valve calcification is higher in CAD patients than in non-CAD patients.And heart valve calcification is sigmficantly associated with coronary atherosclerosis.

Objective To explore the effect of electroporation mediated gene therapy on angiogene-sis of the distraction area during early mandibular distraction osteogenesis (DO). Methods Thirty-two New-Zeland rabbits were randomly divided into 4 groups: group A: recombinant plasmid pIRES-VEGF165-EGFP and electroporation; group B: recombinant plasmid pIRES-VEGF165-EGFP; group C: normal saline (NS) and electroporation and group D: control group. The rabbits were sacrificed at 1d, 3d, 7d and 14d after injection, respectively. The distraction area tissue was removed for histological examination and electron microscopy, and immunohistochemical stain for CD34 was performed to detect the microvessel density. Results Generation of vascular endothelial cells (VEC) in the group A and group B were active, and majority of VEC in groups C and D took on early change of cataplasia and apoptosis. The immunohistochemical stain for CD34 showed that it expressed weakly at the first day after transfection, and at 3,7,14 days after transfection, CD34 of VECs in the distraction area expressed positively. CD34 expression was the strongest in group A (P<0. 05), and there was significant difference among three groups and different time, respectively.Compared to each other, CD34 of VECs expressed positively with a tendency to rise in the groups A and B. But it fluctuated at the level of the expression at the first day in the groups C and D. Conclusion Electroporation-mediated transfecting recombinant plasmid could promote angiogenesis during early stage of mandibular DO. It could promote local vascular proliferation and penetration, increase the blood flow of broken ends in fractured bone. It indicates that electroporation-mediated transfecting recombinant plasmid play an important role in regulating and promoting growth and reparative process of the bone.

Objective To evaluate the prognostic value of ~(99m)Tc-MIBI myocardial perfusion single-photon emission computed tomography (MPS) with adenosine triphosphate in patients aged 80 years or older. Methods A total of 265 patients [ mean age (84. 2 ±3.6) years old ] who underwent adenosine triphosphate and rest ~(99m)Tc-MIBI myocardial SPECT imaging were followed-up for (36. 7 ± 22. 8 ) months. Results During the period of follow-up, 57 patients (20. 4% ) suffered from cardiac events, including 20 major events: 5 cardiac death and 15 acule non-fatal myocardial infarction, 14 unstable angina pectoris,7 heart failure and 16 cases undergoing PCI. The cardiac event rate in patients with fixed or mixed perfusion defects (n = 54) was 50%, which was significantly higher lhan that in patients with reversible perfusion defects(n=67, 31. 3% ,P<0. 05) and normal perfusion imaging( n = 144, 6. 2% ,P <0. 01 ) . The major cardiac event rate in patients with fixed or mixed perfusion defects was 27. 8% , which was significantly higher than that in those with reversible perfusion defects ( 6. 0% , P < 0. 05) and normal perfusion imaging (0.7%, P < 0. 01 ) . Cox multivariate analysis revealed that an abnormal MPS was the most important independent predictor of major or total cardiac events. Conclusions ~(99m) Tc-MIBI MPS with ATP is demonstrated to be a powerful tool for the prognostic evaluation in octogenarian population. Octogenarians with a normal MPS have a low risk of major or total cardiac events, but when an abnormal MPS is present, the risk is significantly higher, being highest in patients with fixed or mixed perfusion defects.

Objective To explore in vitro cytotoxic activities of DCIKs against hepatocarcinoma cells by co-culturing cytokine-induced killer cells (CIKs) with dendritic cells (DCs) derived from peripheral blood of patients with hepatocellular carcinoma (HCC).Methods Peripheral blood mononuclear cells (PBMCs) were isolated from 23 patients with HCC and cultured with cytokines to induce DCs and CIKs.DCIKs were induced by co-culturing CIKs with DCs.After 14 days of co-culture,the phenotypes of DCIKs and CIKs were analyzed by flow cytometry,and their in vitro cytotoxic activities against SMCC-7721 and HepG2 hepatocarcinoma cells were measured by MTT assay.Levels of IL-12,IL-4 and IFN-γ in the supernatants of cell culture were detected by enzyme-linked immunosorbent assay (ELISA).Results High expressions of CD3+CD8+ and CD3+CD56+ were observed on DCIKs.The percentages of effector cells,cytotoxic activity and cytokine secretion were all significantly increased with DCIKs as compared with those CIKs without DC co-culture (P＜0.05).Conclusion Co-culture of CIKs with DCs can enhance the differentiation of effector cells and the cytolytic activities of CIKs against hepatocarcinoma cells in vitro.Immunotherapy with DCIKs may be a promising strategy for the treatment of patients with HCC.

Objective To investigate the accuracy of transthoracic echocardiography (TTE) for diagnosing left cardiac valve calcification in elderly patients.Methods In this retrospective study,410 elderly patients died in Beijing Hospital from 1982 to 2015 were collected.They all had full pathological cardiac valve examination data in the autopsy examination and full mortem TTE data.They were classified into three groups according to the year of death:group 1982-1995,group 1996-2005 and group 2006-2015,as well as into three groups according to the age of death:group 60-75,group76-90 and group 91-106.Results The sensitivity,specificity,diagnostic coincidence rate,positive likelihood ratio (PLR),negative likelihood ratio (NLR),positive predictive value (PPV) and negative predictive value(NPV) of TTE versus autopsy pathological findings as golden standard for diagnosing left cardiac valve calcification were 88.6％,28.2％,43.7％,1.234,0.405,0.298 and 0.878.Compared with group 1982-1995,the group 1996-2005 and group 2006-2015 showed the sensitivity and PPV of TTE versus golden standard autopsy findings for diagnosing left cardiac valve calcification in the elderly were increased while the specificity,PLR,NLR and NPV were decreased.Compared with group 60-75,the group 76-90 and group 91-106 showed that the sensitivity and PPV of TTE versus golden standard autopsy findings for diagnosing left cardiac valve calcification were increased,while the specificity and diagnostic coincidence rate were decreased.The sensitivity,specificity,diagnostic coincidence rate,PLR,NLR,PPV,NPV of TTE versus golden standard autopsy findings for the diagnosis of aortic valve calcification and mitral valve calcification in the elderly were 88.2％ and 44.0％,30.8％ and 75.3％,42.7％ and 71.5％,1.275 and 1.780,0.382 and 0.744,0.250 and 0.198,0.909 and 0.906,respectively.Conclusions TTE provides high sensitivity for diagnosing left cardiac valve calcification in the elderly,especially for diagnosing the aortic valve calcification,but the specificity is not satisfactory.