Objective:To investigate the difference of lymphocyte subsets between elderly patients with rheumatoid arthritis and non-elderly patients and its clinical significance.Methods:A total of 124 patients with rheumatoid arthritis in Affiliated Hospital of Nantong University from January, 2017 to December, 2019 were enrolled.The patients were divided into elderly group(≥60 years old, 34 cases)and non-elderly group(<60 years old, 90 cases). Rheumatoid arthritis activity(DAS-28)scoring was performed for each patient.Peripheral blood mononuclear cells(PBMCs)were extracted by Ficoll density centrifugation.Lymphocytes were labeled and detected by 18-color flowcytometry with more than 30 fluorescent antibodies.Results:DAS-28 scoring showed that the disease activity score of the elderly group(4.56±1.89)was higher than that of the non-elderly group(3.37±1.49)( t=3.633, P<0.001). Flow cytometry showed that MAIL%T(mucus-associated lymphoid tissue T cell subset)( Z=-2.798, P=0.005), Tn%CD8 T cells(initial CD8 T cells)( Z=-2.179, P=0.029), VD2% T(Vδ2+ T, γδT cell subtype)( Z=-2.806, P=0.005), PD1-CD28-%Th( Z=-2.050, P=0.040)and IGM+ D-%B( Z=-2.376, P=0.017)were lower in the elderly group than in the non-elderly group.While, CD45+ CD27+ %CD8 T cells( Z=-3.069, P=0.002), abT%T cell(αβT cells)( Z=-2.103, P=0.035), CD27-CD28+ %T cells( Z=-2.341, P=0.019), ASC%PBMC( Z=-2.341, P=0.019)and ASC%CD19+ ( Z=-2.000, P=0.046)subgroup expression were higher in the elderly group than in the non-elderly group. Conclusions:The disease activity of elderly patients with rheumatoid arthritis is significantly higher than that of younger patients.The expressions of abT%T and CD4% abT in effector T cells of elderly patients with rheumatoid arthritis are higher than those of younger patients, while the expression of VD2% T is lower.The expression level of CD45RA+ CD27+ %CD8 T with cytotoxic effect is higher; However, the expression level of Tn%CD8 T in naive cells is lower.

Objective To prepare specific mouse monoclonal antibody (mAb) against human adenovirus type 55 Hexon protein (HAdV55 Hexon). Methods The Hexon genes of HAdV55, 3, 4, 7, 16 and 21 were chemically synthesized as templates for PCR amplification. The prokaryotic expression plasmids pET28a-HAdV55 Hexon and eukaryotic expression plasmids pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon were constructed respectively. The pET28a-HAdV55 Hexon plasmid was transformed into E. coli competent cell BL21 (DE3) and was induced by IPTG. After the purified inclusion body was denatured and renatured, Hexon55 protein was purified by tangential flow filtration system. pCAGGS-HAdV55 Hexon was used to immunize BALB/c mice by cupping, and HAdV55 Hexon protein was used to booster immunization. The anti-HAdV55 Hexon mAb was prepared by hybridoma technique and the titer and subclass were determined. The specificity of antibody was identified by Western blot using HEK293T cells transfected with pCAGGS-HAdV55 Hexon and by immunofluorescence assay (IFA) using BHK cells transfected with pCAGGS-HAdV55 Hexon. Both clones with high titer were selected, and the cross-reactivity of pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon transfected cells were analyzed by Western blot analysis and IFA. Results PET28a-HAdV55 Hexon and pCAGGS-HAdV55 Hexon, 3, 4, 7, 16 and 21 expression plasmids were successfully constructed. BL21 transformed with pET28a-HAdV55 Hexon was induced by IPTG. The HAdV55 Hexon protein was mainly expressed in the form of inclusion body. After denaturation and renaturation, the purified HAdV55 Hexon protein was obtained by ultrafiltration. Six hybridoma cell lines secreting HAdV55 Hexon mAb were obtained. The antibody subclass analysis showed that 2 strains were IgG2a subtypes and 4 strains were IgG2b. Two specific HAdV55 Hexon antibodies with high titer were obtained, and there was no cross-reactivity with HAdV3, 4, 7, 16, 21 Hexon. Conclusion The specific mice mAb against HAdV55 Hexon provides an experimental basis for establishing its antigen detection method.
Animals ; Mice ; Humans ; Adenoviruses, Human/genetics* ; Escherichia coli/genetics* ; HEK293 Cells ; Isopropyl Thiogalactoside ; Blotting, Western ; Immunoglobulin G ; Antibodies, Monoclonal ; Antibody Specificity ; Mice, Inbred BALB C

Objective:To investigate the occurrence of carotid artery abnormalities in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients and the related risk factors affecting the occurrence of carotid artery abnormalities.Methods:A total of 169 regular follow-up outpatients with HIV/AIDS from April 2003 to December 2019 in Peking Union Medical College Hospital, whose carotid artery ultrasound examination were performed from July 2015 to December 2019 were included. The patients were divided into young (≤44 years old), middle-aged and elderly (≥45 years old) according to their ages, and the physical examination data of the included patients were collected.The statistical methods were logistic regression analysis and single sample t test. Results:Among the 169 HIV/AIDS patients, 40(23.7%) had abnormal carotid artery and 129(76.3%) had no abnormal carotid artery. Middle-aged and elderly people (odds ratio ( OR)=3.85, 95%confidence interval (95% CI) 1.54-9.65, P<0.01), hypertension ( OR=6.24, 95% CI 1.95-20.00, P<0.01), hyperlipidemia ( OR=2.44, 95% CI 1.00-5.93, P<0.05), and elevated human leucocyte antigen (HLA)-DR + CD8 + /CD8 + ( OR=1.03, 95% CI 1.01-1.06, P<0.05) were the risk factors for carotid artery abnormality. The common carotid artery inner medium film thickness (IMT) of patients in HIV/AIDS group Ⅰ (20 to 30 years old), group Ⅱ (31 to 40 years old), group Ⅲ (41 to 50 years old) were (0.061 0±0.001 2), (0.062 9±0.001 4) and (0.065 6±0.002 6) cm, respectively, which were thicker than the control groups ((0.051±0.003), (0.056±0.004) and (0.063±0.002) cm, respectively). The differences were all statistically significant ( t=5.119, 4.775 and 1.739, respectively, all P<0.05). The common carotid artery IMT of patients in HIV/AIDS group A (30 to 44 years old) and group B (45 to 59 years old) were (0.062 6±0.001 1) and (0.072 3±0.003 4) cm, respectively, which were thicker than the control groups ((0.052±0.011) and (0.064±0.015) cm, respectively), the differences were both statistically significant ( t=9.520 and 3.012, respectively, both P<0.01). Conclusion:Younger HIV-positive people have a higher probability of abnormal carotid arteries and may be at greater risk of cardiovascular disease than HIV-negative people of the same age, suggesting that HIV-positive people, especially young people, should be examined early with ultrasound of the neck arteries to detect abnormalities and intervene as soon as possible.

Objective:To explore the curative efficacy of pelvic reconstruction plate and T-plate elastic fixation in the treatment of acetabular fractures with comminuted posterior wall.Methods:A retrospective analysis was conducted of the 21 patients who had been treated at Department of Orthopaedics, Affiliated Hospital to the Second Mongolia Medical University for acetabular fractures with comminuted posterior wall from January 2017 to June 2019. They were 15 males and 6 females, aged from 29 to 55 years (average, 41.5 years). According to the Letournel-Judet classification, there were 15 cases of simple posterior wall fracture with dislocation of the femoral head, 4 cases of posterior wall and posterior column fracture, and 2 cases of double-column and posterior wall fracture. The time from injury to surgery averaged 3 days (from 2 to 5 days). The posterior acetabulum was treated by pelvic reconstruction plate combined with T-plate elastic fixation through the posterior acetabular Kocher-Langenbeck approach. Postoperative fracture reduction, fracture union time, function of the affected hip and complications at the last follow-up were evaluated.Results:This group of 21 patients were followed up for 6 to 24 months (average, 15 months). By the Matta imaging scoring, the postoperative reduction of the posterior wall fracture was evaluated as excellent in 18 cases and as good in 3, giving an excellent to good rate of 100%. The fracture union time averaged 10 weeks (from 8 to 12 weeks) for this group. By the improved Merle d'Aubigné & Postel evaluation at the last follow-up, the affected hips scored from 12 to 18 points (average, 16 points), yielding 18 excellent, 2 good and one poor cases, giving an excellent to good rate of 95.2%. There was no major hemorrhage, nerve injury or deep vein thrombosis intraoperatively. During the follow-up, mild ectopic ossification occurred in one case, and subluxation of the femoral head and traumatic arthritis were observed in another, but no patient had other complications like avascular necrosis of the femoral head.Conclusion:In the treatment of acetabular fracture with comminuted posterior wall, pelvic reconstruction plate and T-plate elastic fixation through the posterior acetabular Kocher-Langenbeck approach can lead to fine short-term outcomes.

Modular scientific research teaching is an open teaching mode for undergraduates in medical colleges and universities. Under the premise of students' own choice, the lab-based training was performed for 15 teaching weeks, with three modular scientific research courses. Meanwhile, with the help of a series of scientific research lectures, literature reports, and discussions of experimental results, the teachers guided the students to carry out modular scientific research training and taught related theoretical knowledge during experimental teaching. The modular scientific research teaching for undergraduates in medical colleges and universities helps to enhance their knowledge of life science and basic medical knowledge, cultivate basic experimental abilities, and make them fully prepared for clinical research or postgraduate study.

Background and purpose:16α-[18F]lfuoroestradiol (18F-FES) is an in vivo speciifc imaging agent for estrogen receptor (ER). We investigated the concordance between tumor ER status as determined by FES-PET and in vitro immunohistochemical assays. Methods: 18F-FES was prepared by ourselves. Twenty-six patients were enrolled (17 primary and 9 metastatic/recurrent). Patients underwent both 18F-FES and 18F-FDG PET/CT. Results:We found good overall agreement (96.15%) between in vitro ER assays and FES-PET. The ER status diagnosis sensitivity of 18F-FES was 93.33%and the speciifcity was 100%when using cut-off value of SUVmax≥1.5. There was a positive correlation between in vitro ER, PR assays and the SUVmax of 18F-FES while in vitro HER-2/neu assays correlatived negatively with 18F-FES SUVmax. Conclusion:These results suggested 18F-FES may be useful for studying the ER expression of all malignant lesions in patients with breast cancer and guiding individual therapy.

Objective To investigate the factors related with final test for continued learning of rehabilitation. Methods The records of 511 continued learning students for physical therapy were analysed with linear regression, with the results of theoretical and operating test as independent variable, while sex, age, titles, profession, education, works, levels of hospital and evaluations as independent variable. Results The results of both theoretical and operating test related with profession, education, works levels and evaluation. Conclusion The models estimate the results of continued learning for rehabilitation has been established.

Objective To characterize the clinical course of biliary complications after right lobe living donor liver transplantation (RL-LDLT) and to identify the independent risk factors for biliary strictures.Methods 105 consecutive RL-LDLT recipients operated from April 2007 to April 2010 were followed up. The clinical and operative data were reviewed. The biliary complications and independent risk factors of biliary stricture were studied.Results The median follow-up duration was 49.5 months ranging from 562 to 1675 days.A total of 40 patients (38.1 ％) experienced 11 bile leak episodes (10.4％ ) and 37 (35.2％) biliary stricture episodes after transplantation.Bile leaks occurred at a median time of 9 days ranging from 4 to 54 days after transplantation.For biliary strictures,the occurring time was delayed and scattered wide with a median of 7.6 months ranging from 12 to 790 days after transplantation. Moreover, the biliary stricture incidence in the first year after transplantation was significantly higher than later.The independent risk factors for biliary strictures were CMV infection,bile leaks and bile duct size (≤3 mm).Conclusion The independent risk factors for biliary strictures after RL-LDLT were CMV infection,bile leaks and bile duct size (≤3mm).In order to avoid biliary complications,careful preoperative evaluations are necessary. The dissection of bile ducts should be meticulous to protect its blood supply.CMV infection should be prevented after transplantation.Close surveillance of biliary complications should be given to RL-LDLT recipients during the first year after transplantation.

Objective To observe the outcomes of living donor liver transplantation (LDLT) for children with biliary atresia (BA) and to summarize the clinical experiences. Methods Forty-four BA patients (26 boys and 18 girls) underwent LDLT between October 2006 and December 2010. Mean (SD) and median (range) age at operation was (12.1 ± 9.0) months and 9 (6-60) months,respectively. The 44 donors were lineal relatives to the consorted recipients. Their mean (SD) and median (range) age at operation was (32. 7 ± 8. 0) months and 31 (20～54) years, respectively. All donor graft types were the left lateral segments with compatible ABO blood groups. Clinical data,including pre-operative evaluations, surgical technique, postoperative management and outcomes in all donors and recipients were retrospectively analyzed. Results All donors were followed up for (17. 5 ± 13. 3) months. No donor mortality was encountered, with a minimal morbidity and no long-term sequelae. Nine out of 44 recipients died. Three patients died of portal vein thrombosis (PVT), one of hepatic artery thrombosis (HAT), two of biliary complications, one of surgical site infections, one of abdominal bleeding and one of pulmonary infection. The overall 1-year and 2-year cumulative survival rate in recipients was 81. 2% and 76. 1 %, respectively. No re-transplantation was done. Postoperative complications included PVT, HAT, biliary leakage and refluxing cholangitis, pulmonary infections,surgical site infections and acute rejection. Conclusion LDLT has been the effective treatment for pediatric recipients with BA and provides favorable prognosis. To improve prognosis of recipients, the key points are pre-operative evaluations, surgical technique, and postoperative management

Objective To evaluate the applied value of functional immunity measured by the ImmunKnow assay in the diagnosis of post-transplant infection in Chinese 1iver recipients.Methods Thirty-eight normal adults and 68 adult liver transplant recipients were under investigation.Whole blood samples from either normal volunteers(each sample for one person)or the liver recipients(one or more samples for one person)were collected freshly and cultured within 6h.The CD4+T cells were selected and their ATP value was assayed the next day.The liver recipients were grouped in stable status(n=52)or infection(n=64)according tO their clinieal manifestation.Results The average ATP value in the recipients with infection after liver transplantation was 165.7±100 μg/L,significantly lower(P＜0.05)than that in stable recipients(309±126 μg/L)or normal volunteers (292±83 μg/L).The low ATP levels in post-transplant recipients had fair good correlation to infection clinically(RR=0.5021,P＜0.01).Infectious risk was high when ATP value was less than 165μg/L(OR=11,95%CI 3.9-32.2,P＜0.01).Specificity and sensitivity of low ATP value in post-transplant infection were 86.53%and 73.81% respectively,Conclusion ImmuKnow assay provides a new tool in monitoring immune status in post-transplant recipients,and call helpfully predict and diagnose clinical infection.