Objective: To investigate the regulation of respiratory syncytial virus CTL epitope fused with G protein antigen fragment G1 to the specific immunoresponses. Methods: The recombinant plasmid pET-DsbA-G1 or pET-DsbA-G1F/M2 was transferred into E.coli BL21(DE3) and the fusion protein DsbA-G1F/M2 or DsbA-G1 was expressed.The expressing product was induced and purified by affinity chromatography. The two proteins were used to immunized BALB/c mice i.p, respectively. Serum and spleen cells were collected regularly. RSV-specific CTL responses were measured by MTT, IgG and IgG1 and IgG2a antibodies by ELISA, neutralizing antibodies by plaque reduction assay. Results: The recombinant proteins were expressed successfully and the purity was over 90% after purified by affinity chromatography. The protein DsbA-G1F/M2 induced significant RSV-specific CTL responses, while DsbA-G1 without CTL epitope did not induce detctable CTL responses. Strong IgG antibody responses were elicited,indicated by both. IgG1 and IgG2a titers induced by DsbA-G1F/M2, while only IgG1 was induced by DsbA-G1.The ratio of IgG1/IgG2a was downregulated significantly. Both antigens induced high level of neutralizing antibodies, but the latter was a little lower. Conclusion: DsbA-G1F/M2, the fusion protein of a CTL epitope and G protein fragment G1 can induce both cellular immunity and humoral immunity. The activation of CTLs downregulates the ratio of IgG1/IgG2a.The more balanced immunoresponse is advantageous for improving safety of the candidate vaccine.

Objective To investigate the clinical outcomes of terrible triad of the elbow treated with single lateral incision versus combined lateral and medial incisions.Methods This retrospective cohort research involved 57 patients with terrible triad of the elbow operated between July 2010 and June 2013.According to the surgical approaches,the patients were assigned to single lateral incision (Group A,n =32) and combined lateral and medial incisions (Group B,n =25).Group A consisted of 21 males and 11 females,with a mean age of (40.2 ± 3.3) years.Group B consisted of 18 males and seven females,with a mean age of (39.1 ± 2.6) years.Operation time,blood loss,postoperative complications,elbow range of motion,Mayo elbow performance score (MEPS) and Hastings and Graham heterotopic ossification classification were compared between groups.Results There were no significant differences in operation time and blood loss between Group A and B (P ＞ 0.05).Incidence of ulnar nerve injury and strength decrease were higher in Group A than Group B (P＜0.05).Mter a mean follow-up of 25.9 months,elbow extension-flexion and forearm pronation-supination in Group A [(123.4 ± 6.5) ~,(167.5 ± 6.0) °] were better than those in group B [(84.9 ± 27.2) °,(136.5 ± 8.5)°] (P ＜ 0.01),and MEPS in Group A [(95.8 ± 1.2) points] was also better than that in Group B [(84.9 ± 3.1) points] (P ＜ 0.01).Heterotopic ossification was more serious in Group B than Group A (P ＜ 0.01).Conclusion Single lateral incision is associated with better function and lower rate of heterotopic ossification than combined lateral and medial incisions in treating terrible triad of the elbow.

Objective To investigate the isokinetic strength of knee extensors and flexors in patients with knee osteoarthritis (KOA), and to establish the correlation between the isokinetic strength and function in patients with KOA. Methods 23 patients with bilateral KOA and 14 matched normal controls finished the isokinetic test of knee extensors and flexors, the Five Times Sit-to-Stand Test (FTSST), Gait analysis, and Balance test. The KOA patients were evaluated with Visual Analog Scale (VAS) for pain and Western Ontario and McMaster University Osteoarthritis Index (WOMAC). Results It was less of the peak torque, average peak torque, average power, max rep total work, total work of knee extensors and flexors in the mainly involved limbs than the contralateral limbs (P<0.05), and more of the hamstring/quadriceps peak torque (H/Q) percentage (P<0.05) in the patients. It was less of all the observed indicators of knee extensors and average power of knee flexors in the patients than in the normal controls (P<0.05), and was more of the H/Q percentage (P<0.05). The peak torque of knee extensors correlated with the results of FTSST, walking speed, walking distance, falling index, VAS, and WOMAC-pain (P<0.05), but the peak torque of knee flexors did not (P>0.05). Conclusion It is weak of the isokinetic strength of knee extensors and flexors in patients with KOA in the mainly involved limb, as well as the isokinetic strength of knee extensors compared with the normal controls. The changes in the extensors and flexors are not equivalent. The peak torque of knee extensors significantly correlated with the pain and functions of the knee.

Objective:A large single-center, premature acute myocardial infarction (AMI) age (≤45 years) cohort was established to investigate the clinical features and the factors affecting major adverse cardiac events (MACE).Methods:This is a prospective and observational study. 603 patients with a clear diagnosis of AMI admitted to the Tianjin Chest Hospital from March 2015 to December 2017 were continuously selected. All patients were aged ≤45 years old, and a single-center large-sample premature AMI cohort was established. The patient's clinical basic conditions, laboratory indicators, imaging data, coronary angiography and treatment were collected. All patients were followed up for 1 year. MACE events such as cardiac death, recurrent AMI, revascularization, severe heart failure requiring hospitalization and stroke were recorded. Kaplan Meier method was used to draw the survival curve. Cox regression analysis was used to analyze the influence of risk factors, clinical characteristics and intervention methods on the long-term prognosis of MACE events.Results:A total of 603 AMI patients were included, 575 males (95.36%), 28 females (4.64%), and median age 41 (37, 44) years old. There were 422 patients (69.98%) with acute ST segment elevation myocardial infarction (STEMI), 206 patients (48.82%) with anterior myocardial infarction, and 181 patients (30.02%) with non ST segment elevation myocardial infarction (NSTEMI). Smoking was the most common risk factor for premature AMI (77.45%), followed by hyperlipidemia (48.42%) and hypertension (48.09%); smoking was the most common risk factor for male patients (80.35%), and hyperlipidemia was the most common risk factor for female patients (35.71%). 302 (50.08%) patients with premature AMI were treated with symptom onset to first medical contact (SO-to-FMC) ≤12 h; 563 patients (93.37%) had coronary angiography; coronary angiography showed that no significant stenosis, single-vessel disease, double-vessel disease, three-vessel disease, and patients with left main disease were 15(2.66%), 212(37.66%), 153(25.37%), 167(29.66%), 16(2.84%) cases; 318(56.48%) patients with vascular occlusion; The proportion of male combined with left main lesions was lower than that of female group (2.41% vs 12.50%, P=0.026); A total of 45 patients (7.46%) were recorded MACE. The 1-year MACE incidence was lower in the male group than in the female group (6.96% vs 17.86%, P=0.032). Multivariate COX regression analysis: there were 5 indicators that entered the regression model and were statistically significant: female ( HR:4.184; 95% CI:1.583-11.064; P=0.004), SO-to-FMC≤12 h ( HR:0.447; 95% CI:0.224-0.889; P=0.022), left ventricular ejection fraction (LVEF)≤40% ( HR:3.727; 95% CI:1.876-7.405; P<0.001), low-density lipoprotein (LDL) ( HR:1.315; 95% CI:1.041-1.662; P=0.022), homocysteine (Hcy) ( HR:1.011; 95% CI:1.002-1.019; P=0.011) were independent predictor of MACE occurrence in patients with early-onset AMI within 1 year. Conclusions:Smoking is the most common risk factor for young men with AMI. The most common risk factors for young women's AMI is hyperlipidemia, and the proportion of patients with left main artery disease is higher than that of men, but the proportion of patients receiving emergency intervention is lower than that of men, and the long-term prognosis of young women is poor. Early detection and control of these risk factors is a key measure to prevent the onset of AMI.

Objective: To explore the changes in ribosomal DNA copy number in peripheral blood among patients with pneumoconiosis and its influencing factors, so as to provide insights into prevention and treatment of pneumoconiosis. Methods: Eighty-eight patients with pneumoconiosis who visited a designated hospital and 71 community residents with no history of pneumoconiosis or dust exposure were selected as the pneumoconiosis group and control group, and age, smoking history, drinking history and cumulative years of exposure to dust were collected through questionnaire surveys. The copy number of 45S rDNA and 5S rDNA was detected using real-time fluorescence quantitative PCR, and the differences between the two groups were compared. Factors affecting the copy number of 45S rDNA and 5S rDNA were identified by a multiple linear regression model. Results: The pneumoconiosis group had a median age of 56.00 (interquartile range, 15.25) and a mean cumulative dust exposure duration of (12.40±8.08) years, with 56.82% smoking and 62.50% drinking. The control group had a median age of 64.00 (interquartile range, 37.00) years, with 32.39% smoking and 26.76% drinking. The median copy number of 45S rDNA in the pneumoconiosis group was 1.29 (interquartile range, 0.59), which was lower than 2.10 (interquartile range, 1.88) in the control group; the median copy number of 5S rDNA in the pneumoconiosis group was 5.33 (interquartile range, 0.85), which was higher than 4.66 (1.34) in the control group (both P<0.05). Multiple linear regression analysis identified age (β=-0.034) and pneumoconiosis (β=-1.595) as factors affecting 45S rDNA copy number, age (β=-0.013) as a factor affecting 5S rDNA copy number, and age (β=0.018) as a factor affecting 5S rDNA copy number in the pneumoconiosis group (all P<0.05). Conclusions Compared with community residents with no history of pneumoconiosis or dust exposure, the copy number of 45S rDNA in peripheral blood among patients with pneumoconiosis is reduced and the copy number of 5S rDNA is increased.

In this study, the swabs were collected among patients with an influenza-like illness (ILI) admitted to 2 sentinel surveillance hospitals of Yantai from April 2014 to August 2017. All specimen were cultured and identified by hemagglutination inhibition assay. Complete sequences of Hemagglutinin (HA) of influenza A were amplified, sequenced and analyzed using molecular and phylogenetic methods. The potential vaccine efficacy were calculated using Pepitope model. The results showed that the antigenicity of A (H3N2) had changed greatly. 8 strains of influenza A (H1N1) pdm09 belonged to subclade 6B.1 and 14 strains clustered in 6B.2. 12 strains of influenza A (H3N2) fell into subgroup 3C.3a and 33 strains clustered in 3C.2a. Several residues at antigen sites and potential glycosylation sites had changed in influenza A strains. Vaccine efficacy of influenza A (H1N1) pdm09 in 2015/2016 and 2016/2017 seasons were 77.29% and 79.11% of that of a perfect match with vaccine strain, meanwhile vaccine efficacy of influenza A (H3N2) in 2014/2015, 2015/2016 and 2016/2017 were-5.18%, 16.97% and 42.05% separately. In conclusion, the influenza A virus circulated in Yantai from 2014 to 2017 presented continual genetic variation. The recommended vaccine strains still afforded protection against influenza A (H1N1) pdm09 strains and provided suboptimal protection against influenza A (H3N2) strains.

Objective:To evaluate the performance of the combinative index-a product of pressure (P) and respiratory rate (RR), to predict the outcome of weaning.Methods:Single-center, case-control study method was used to prospectively collect the data of 76 patients who had been mechanically ventilated for at least 24 h in ICU of the Affiliated Changshu Hospital of Xuzhou Medical University from November 2017 to April 2019, excluding the patients with central respiration repression and primary neuromuscular disease. The spontaneous breath test (SBT) was performed for 1 h after the patient was ready to be weaned. All patients were divided into two groups according to whether they were successfully weaned from the mechanical ventilation: 50 cases in the success group and 26 cases in the failure group. Clinical data and values of peak inspiratory pressure (P peak), driving pressure (DP) and RR of two groups were collected. The independent sample t test and Mann-Whitney U test were used for the comparison between the two groups. The differences between enumeration data were assessed by Chi-square test. Finally, receiver operating characteristic (ROC) curve analysis was used to evaluate the predictive value of each index. Results:Of the 76 patients studied, 26 patients failed extubation. The values of P peak×RR and DP×RR in the success group were significantly less than those in the failure group; P peak×RR (cmH 2O·breaths/min): 291.8±76.5 vs 519.8±108.8 ( P<0.01), DP×RR (cmH 2O·breaths/min): 131.6±34.2 vs 227.0±47.5 ( P<0.01). The AUC of P peak×RR, DP×RR was 0.960 and 0.941, respectively. A decrease in P peak×RR index less than 362 cmH 2O·breaths/min had a sensitivity of 96.2%, a specificity of 88.0%, a positive predictive value (PPV) of 97.8% and a negative predictive value (NPV) of 80.6%, to predict successful weaning. A decrease in DP×RR index less than 170 cmH 2O·breaths/min had a sensitivity of 88.5%, a specificity of 92.0%, a PPV of 94.0% and a NPV of 88.5%, to predict successful weaning. Conclusion:The combination of pressure and respiratory rate can accurately predict whether the patients can be successfully weaned from mechanical ventilation.

Objective: To establish a convenient non-invasive tracheal perfusion method for constructing a mouse model of silicosis-induced pulmonary fibrosis. Methods: The specific pathogen-free C57BL/6 mice were randomly divided into control group and model group, with 15 mice in each group. After anesthesia, a 22G arteriovenous indwelling needle was used to inset into the trachea through the mice's mouth. The model group mice were perfused with 0.1 mL of silica suspension with a mass concentration of 25 g/L, and the mice in the control group were perfused with an equal volume of 0.9% sodium chloride solution. On the 7th, 14th, and 30th day after modeling, the body weight of the mice was measured, and the lung tissue morphology and pathological changes were observed. The expression of α-smooth muscle actin (α-SMA) and collagen type 1 alpha 1 (COL1A1) protein in lung tissue of mice was detected by immunofluorescence on the 30th day after modeling. Results: There was no death of mice in the two groups during the experiment. There was no significant difference in body weight between the two groups (P>0.05). The lung tissues of the mice in the model group were pinkish-gray and uneven in color on the 7th and 14th days after dust exposure. On the 30th day after dust exposure, the lung tissue of the mice in the model group was gray and hard, and unevenly distributed silicon nodules were visible by the naked eyes. The histopathology results of lung tissue showed that compared with the mice in control group, the model group mice exhibited persistent aggravation of pulmonary inflammation, thickening of alveolar septum, infiltration of inflammatory cells gradually clustering into clumps, and an increasing number of fibrous foci.On the 30th day after dust exposure, the relative expression of α-SMA and COL1A1 proteins in the lung of the model group was higher than those in the control group (median： 72.59 vs 5.91, 35.62 vs 10.07, both P<0.05). Conclusion: The method of tracheal perfusion silica suspension of mice using 22G arteriovenous indwelling needle can successfully construct an animal model of silicosis fibrosis. This method is convenient, safe and effective, and is worth promoting.

We developed a three-dimensional mini-type permanent magnetic resonance imaging (MRI) device in our lab. The purposes of this study were (1) for further development of MRI technologies, (2) for support of broadening practices of animal test modeling in medical research, and (3) for training more specialists from colleges or universities in the field of MRI. This paper describes the research and development at our lab(s), especially stressing on the design of the main magnet, the gradient coil and the radio frequency coil. In addition, the specific methodologies used in our lab(s) and the related data are emphasized. The 3D MRI technologies have met the needs of using small animals, super thin sections of live animal body and high imaging resolutions. MRI images of mice head and abdominal have been obtained successfully by using the imager that we developed. The imaging results and analyses have also been discussed.
Animals ; Equipment Design ; Imaging, Three-Dimensional ; instrumentation ; Magnetic Resonance Imaging ; instrumentation ; methods ; Mice

Objective:To investigate the influencing factors of significant liver fibrosis in patients with chronic hepatitis B (CHB) concurrent with non-alcoholic fatty liver disease (NAFLD).Methods:Those who underwent liver pathological examination and confirmed diagnosis of CHB and NAFLD in Tianjin Second People′s Hospital from August 2014 to September 2017 were enrolled. Data regarding their demographic information, laboratory tests results, and liver pathology results were analyzed. The latter results were used to categorize the patients either in non-significant liver fibrosis group (Metavir stage