Objective To study the condition and method of hydrodynamics-based transgene(HDT) in rat fatty liver. Methods Inject different dosages and concentrations of green fluorescent protein plasmid pEGFP-C1 at different speeds, then collect 4 rats' liver leaves at different time points after injection and prepare their frozen section, finally observe and quantify the GFP expression with fluo rescence microscope at 488 nm excitation wavelength. Results Plasmid pEGFP-C1 concentration 33mg/L, injection speed 2ml/s, injection volume 8.5%of rat body weight, injected plasmid. After 6 hours of injection, GFP-positive cells rate of pedicel leaf is about 18%, left leaf about 14%, middle leaf about 12.5%;R3ight leaf about 10% and tail leaf about 8%. GFP begin to gradually reduce since 24 hours, until 72 hours almost no GFP-positive cells were checked in all liver leaves. Conclusion Hydrodynamics-based transgene can be applied to rat fatty liver, the appropriate conditions of this method are 33mg/L plasmid concentration, 8.5% rat avoirdupois, 2ml/s injection speed, and the suitable time to observe the proportion of GFP-positive cells is 6-24 hours after gene injection.

Objective To discuss the clinical value of serum tumor specific growth factor (TSGF) and vascular endothelial growth factor (VEGF) in early diagnosis of thyroid cancer.Methods Eighty thyroid cancer patients (thyroid cancer group),120 nodular goiter patients (nodular goiter group),and 80 healthy adult (blank control group) were enrolled in this study.The level of TSGF and VEGF in two groups were detected by ELISA and compared.Results The level of TSGF and VEGF in thyroid cancer group and nodular goiter group were significantly higher than those in blank control group (P ＜ 0.05).Conclusion Serum TSGF and VEGF have some clinical value in early diagnosis of thyroid cancer.

Objective To study the causes, significance of alteration of PtiO 2, PtiCO 2 and pHti in liver tissue during liver ischemia reperfusion (I R). Methods After rabbits were anesthetized, liver ischemia was induced by complete occlusion of the hepatoduodenal ligment for 45 min, then the portal and arterial flow were released, and observed for 120 min for measuring the PtiO 2, PtiCO 2 and pHti in liver tissue and the pathology of the liver during ischemia reperfusion. Results After 15 min of hepatic vascular occlusion, PtiO 2 decreased to 4 mmHg, PtiCO 2 increased fast to (149.63?9.80) mmHg (P

Objective To determine the association between homocysteine and severity of large cerebral artery atherosclerosis.Methods Total 152 ischemic stroke patients were evaluated using transcranial Doppler(TCD),magnetic resonance angiography(MRA) or digital substraction angiography(DSA).Severity of cerebral artery atherosclerosis was scaled by number of stenosed or occluded artery.Results There were 83 patients in low level homocysteine group with mean Hcy level(15.31?3.08)?mol/L,while 69 patients in high level homocysteine group with mean Hcy(34.47?21.38)?mol/L.Number of abnormal intra-and extra-artery was 1.86?1.51 and 1.52?1.46 respectively.No significant difference was demonstrated in the number of abnormal artery between two groups.No correlation was noted between Hcy and number of abnormal artery.Conclusion There is no relationship between high homocysteine and severity of large cerebral artery atherosclerosis in ischemic stroke patients.Homocysteine might be the marker of the disease.

ObjectiveTo develop an antibody-array system for multiple detection of antibodies against Japanese B encephalitis virus (JEV),Tick-borne encephalitis virus (TBV),Dengue virus ( DENV ),West Nile virus (WNV),Western equine encephalitis virus (WEEV) and East Equine encephalitis virus (EEEV).MethodsRecombined antigens were spotted on array as capture antigens.Specific antibodies were detected by using a sandwich ELISA format.Rabbit antiserum was employed to select and confirm the specificity of antigens and to optimize the conditions of the assay.The detection efficiency of the system was validated by 40 clinical suspected serum samples and compared with the relative ELISA assays.ResultsEleven recombined antigens were selected as diagnostic antigens with high specificity.Better detection could be achieved when scale of antigen concentrations were within 0.125-0.900 mg/ml and the serum dilutions were 1:100-1:1000.When detecting the 26 clinical suspected TBE serum samples,20 were IgG positive (76.9％),and 17 were IgM positive (65.3％) which was 96.1％ and 84.6％ consistent with the relevant ELLSA tests,the 8 clinical suspected JEV serum samples,4 were IgG positive (50.0％),and 5 were IgM positive (62.0％),which was 86.3％ and 90.1％ consistent with the relevant ELLSA tests.As for the 22 DEN serum samples,13 were IgG positive (60％) and 15 were IgM positive (68％) which was 85％ and 93％ consistent with ELISA.The specificity of the assay was 100％ and the sensitivity was higher than the relative ELISAs.ConclusionThe developed antibody-array is highly specific and reliable,which could be used for the detection of antibodies against the 6 arboviruses.

Objective To evaluate the Flavivirus specific monoclonal antibody(McAb) 2A10 as detective antibody for simultaneously identify tick borne encephalitis virus( TBEV), Japanese encephalitis virus( JEV), dengue ( DEN )-2, DEN-4 and yellow fever virus ( YFV ) by antibody microarray technique.Methods The antibody microarray was developed by spotting TBEV, JEV, DEN-2, DEN-4 and YFV specific McAb on chip as capture antibodies. After incubating with cultured viral supernatants of the above viruses, CY3 labeled detective antibody 2A10 was added to the chips. After reaction, the antibody microarray was scanned and the results were analyzed. By comparing the signal intensities of different spots on chips,the detecting titre and sensitivity of 2A10 for Flavivirus were determined, and the value of 2A10 in detection of Flavivirus was evaluated. Results The hybridization results demonstrated that the titre of 2A10 for Flavi2A10 was specific for Flavivirus and could be used as universal detective antibody for Flavivirus on antibody microarray.

Objective To identify the infection and the replication of Tick-borne encephalitis virus(TBEV) in human neuroblastoma cells.Methods After being inffected with TBEV,the cell culture supernatant of human neuroblastoma cell line SK-N-SH was collected and assayed at different time points.Byusing real-time RT-PCR and plaque assay to measure the titer of virus in the supernatant,the replication andproliferation of TBEV in human neuroblastoma cell was identified.Meanwhile,the morphological change of SK-N-SH after TBEV infection was also visualized by observation under microscope and immunmquorescenceassay.Results Real-time RT-PCR and plaque assay both demonstrated that TBEV could replicate effectively in SK-N-SH cells,the peak titer could reach 2.92× 107 PFU/ml on 3 days post-inoculation.And significant morphological change occured on infected SK-N-SH cells after 2 days post inoculation.By immunofluorescence assay,the virus particles could be detected and visualized.Conclusion TBEV can replicate andproliferate effcctively and cause significant cell morphological changes in human neuroblastoma cell SK-N-SH,which demonstrated that SK-N-SH could be a suitable cell model for TBEV culture.

Objective: To test the effectiveness of psychological flexibility training on career adaptability among middle school students who undertook psychological courses based on acceptance commitment therapy and the adolescent mental flexibility model(DNA-V), and to provide a reference plan to improve the mental health of middle school students. Methods: This study recruited 110 junior high school students (60 boys and 50 girls) from a middle-school in Beijing. The students were randomly divided by class into a DNA-V face-to-face course group(offline group n=33), a DNA-V online course group(online group n=40), and a regular school psychology course group(control group n=37). Louise Hayes DNA-V intervention program was condensed into a six-hour middle-school DNA-V psychology curriculum. Using the Avoidance and Fusion Questionnaire for Youth and the Career Adaptability Scale, changes in psychological flexibility and career adaptability were measured before(T1), one week after(T2), and two months after (T3) the intervention. Results: Linear mixed models were used for the analysis, while controlling for demographic variables. Psychological flexibility and career adaptability in the offline group were higher at T2 and T3 than at T1(psychological flexibility t=4.22, 3.11; career adaptablity t=3.05, 4.16, P<0.01), while the difference between T2 and T3 was not statistically significant. The psychological flexibility and career adaptability of the online group were not statistically significant at T1, T2, and T3. The psychological flexibility and career adaptability of the control group increased from T1 to T2(t=4.64, 2.47, P<0.05), but T3 decreased back to a level close to T1. Conclusion In terms of both psychological flexibility and career adaptability, the DNA-V face-to-face psychology course resulted in a retention period of at least two months.

Objective:To analyze the correlation between chronic obstructive pulmonary disease (COPD) and cognitive dysfunction.Methods:This is a case-control study. From February 2022 to October 2022, 32 COPD patients (inpatient and outpatient) from the Department of Respiratory and Critical Care Medicine and Rehabilitation Medical Center of the First Affiliated Hospital of Nanjing Medical University and 32 healthy subjects were recruited. All participants underwent a thorough evaluation, which included Montreal Assessment of Cognitive Function (MoCA), visuospatial n-back task included accuracy (ACC) and mean response time (RT), the pulmonary functions including forced vital capacity (FVC), forced expiratory volume in the first second (FEV 1), one-second rate (FEV 1/FVC) and maximum volume per minute (MVV), Health Survey Short Form (SF-36), and St. George′s Respiratory Questionnaire (SGRQ). The correlation between cognitive dysfunction and lung function, SF-36 and SGRQ in COPD patients were analyzed. Results:The prevalence of smoking, hypertension and cardiovascular disease in the two groups were significantly different (all P<0.05). MoCA score, 1-back ACC and 2-back ACC in COPD group were significantly lower than those in healthy control group [(23.86±4.50) vs (27.55±1.29) points, (76.82%±16.60%) vs (90.61%±7.40%), (67.93%±10.10%) vs (78.74%±10.38%), all P<0.001]; 2-back RT was significantly higher than that of healthy group [(316.43±108.17) vs (254.09±101.62) ms, P<0.05]; and the Physiological function (PF), physiological function (RP), emotional function (RE), energy (VT), social function (SF), physical pain (BP) in SF-36 were significantly worse than the healthy control group (all P<0.05). The MoCA score of COPD group was positively correlated with FEV 1/FVC ( r=0.501, P=0.018). The 1-back ACC was positively correlated with FEV 1 and FEV 1/FVC ( r=0.568, 0.634; both P<0.05). The 1-back RT was negatively correlated with FEV 1/FVC and MVV ( r=-0.452, -0.534; both P<0.05). The 2-back ACC was positively correlated with FEV 1/FVC ( r=0.426, P=0.048). The 2-back RT was negatively correlated with MVV ( r=-0.571, P=0.006). In COPD group, MoCA score was negatively correlated with activity, influence and total score in SGRQ ( r=-0.533, -0.466, -0.521; all P<0.05). The 1-back ACC was negatively correlated with activity, influence and total score ( r=-0.552, -0.517, -0.584; all P<0.05). The 1-back RT was positively correlated with activity, influence and total score ( r=0.430, 0.379, 0.417; all P<0.05). The 2-back ACC was negatively correlated with impact and total score ( r=-0.398, -0.412; both P<0.05). Conclusion:COPD patients have impaired cognitive function, which is mainly manifested by the decline of working memory and executive function, and is correlated with the lung function, general health condition and quality of life.