Objective To investigate the expression of transcriptional factors zinc finger Krüppel like transcription factor 2 ( Klf2) and NF-E2 related factor 2 ( Nrf2) /BTB CNC homology 1 ( Bach1) in rat bronchial epithelial cells stimulated by cigarette smoke extract ( CSE) , and explore the regulatingmechanism of γ-glutamylcysteine synthetase ( γ-GCS) expression in the oxidative condition. Methods Rat bronchial epithelial cells were harvested using enzyme digestion method, and intervened by 10% CSE for 6 hours. Then γ-GCS activity was detected by two enzymes method, and the nuclear transfer of Nrf2 /Bach1 in cells was detected by immunohistochemistry. Western blot and reverse transcription-polymerase chain reaction ( RT-PCR) techniques were used for detecting the protein and mRNA expressions of Klf2, Nrf2, Bach1, and γ-GCS in the cells. Results The γ-GCS activity was elevated in the CSE group. Immunohistochemical results showed that Nrf2 translocated from cytoplasm to nucleus in response to stimulation by CSE. On the contrary, Bach1 translocated from nucleus to cytoplasm in the same condition. Western blot results showed that protein levels of Klf2, Nrf2, Bach1, and γ-GCS were higher in the CSE group than those in the control group ( Plt;0.05) . RT-PCR results were the same as the Western blot results ( Plt;0.05) . Linear correlation analysis showed that γ-GCS expression was positively correlated with Klf2, Nrf2, and Bach1 ( Plt;0. 05) . Conclusion CSE might regulate the expression of γ-GCS through the transcription factors of Klf2, Nrf2, and Bach1.
ObjectiveTo explore the value of T-SPOT.TB test in the diagnosis of active tuberculosis and its influencing factors. MethodsFrom July 2010 to November 2012, a total of 289 suspected active tuberculosis patients were enrolled in the study and underwent T-SPOT.TB test. All the patients enrolled were from West China Hospital of Sichuan University. The diagnostic value of T-SPOT.TB applied in determining active tuberculosis was then evaluated. ResultsAccording to the diagnostic criteria, 84 patients diagnosed with active tuberculosis were eligible for analysis and enrolled as a tuberculosis group, and 156 patients were enrolled as a control group. The sensitivity of T-SPOT.TB test was 83.3%, while the specificity was 80.1%. Both univariate and multivariate analyses showed the characteristics of patients such as general conditions (eg. age, sex) and basic diseases (eg. immunosuppression condition, malignant tumour) were not the risk factors of false-positive or false-negative result of T-SPOT.TB. ConclusionT-SPOT.TB test for the diagnosis of active tuberculosis has high sensitivity and specificity, with important value referred for diagnosing suspected active tuberculosis patients.
ObjectiveTo evaluate the diagnostic value of interferon-gamma release assay (TB-IGRA) for tuberculosis in the Tibetan. MethodsFrom January 2014 to December 2014, suspected Tibetan tuberculosis patients were enrolled from AVIC 363 Hospital and underwent TB-IGRA test. All patients were also underwent smear test for Mycobacteria. The diagnostic value of TB-IGRA test for Tibetan TB patients was analyzed. ResultsA total of 77 suspected Tibetan tuberculosis patients were included. According to the diagnostic criteria, of the 77 suspected patients, 50 were diagnosed as TB patients, and 27 were diagnosed as not-TB patients. The sensitivity and specificity of TB-IGRA test was 86% and 81.5%. While the sensitivity and specificity of smear test were 22% and 100%, respectively. ConclusionThe TB-IGRA test is superior to smear test, and is the fast and sensitivity test for diagnosing Tibetan TB patients.
ObjectiveTo systematically review the diagnosis value of T cell ELISA Spot (T-SOPT.TB) in for bacterium negative pulmonary tuberculosis in Chinese. MethodsWe electronically searched databases including PubMed, CBM, CNKI, WanFang Data and VIP from inception to December 31st, 2015 to collect studies about T-SPOT.TB in diagnosis bacterium negative pulmonary tuberculosis in Chinese. Two researchers independently screened literatures, extracted data, and assessed the risk of bias of included studies, and then meta-analysis was performed by using Stata 12.1 and Meta-DiSc 1.4 software. ResultsA total of 11 studies including 1 413 patients and 1 256 controls were included. The results of meta-analysis showed that the pooled sensitivity, specificity, positive likelihood ratio, negative likelihood ratio, diagnosis odds ratio and area under SROC curve for T-SPOT.TB diagnosis bacterium negative pulmonary tuberculosis was 83% (95%CI 0.81 to 0.85), 86% (95%CI 0.84 to 0.88), 5.73 (95%CI 4.99 to 6.59), 0.20 (95%CI 0.18 to 0.23), 31.09 (95%CI 25.11 to 38.49), and 0.92, respectively. ConclusionAs a rapid and accurate method for diagnosis of bacterium negative pulmonary tuberculosis, T-SPOT.TB has a high diagnostic efficiency for bacterium negative pulmonary tuberculosis in Chinese.
Objective To explore the role of over-expression of TBX3 and TBX18 in inducing human induced pluripotent stem cells (HiPS) to enrich and differentiate into sinoatrial node-like cells. Methods The expression of stemness markers OCT3/4, SOX2, and NANOG in HiPS was detected by real-time fluorescence quantitative PCR (qRT- PCR), and compared with human embryonic stem cells (hESCs). Immunofluorescence staining was used to observe the expression of HiPS stemness markers OCT3/4, NANOG, SSEA4, and TRA-1-60. The HiPS were directional differentiated into cardiomyocytes, the expressions of ISL1, NK2 homeobox 5 (NKX2-5), ACTN1, and TNNT2 were detected by qRT-PCR, and human adult cardiomyocytes (hACM) were used as positive control. Immunofluorescence staining was used to observe the expressions of NKX2-5, cardiac troponin (cTnT), α-actinin, atria myosin light chain 2A (MLC-2A), and ventricular myosin light chain 2V (MLC-2V). The positive rate of α-actinin was detected by flow cytometry. On the 3rd day after HiPS were differentiated into cardiomyocytes (mesodermal stage), lentiviral over-expressions of sinoatrial node-related genes TBX3 and TBX18 were carried out for 21 days. The relative expressions of specific markers TBX3, TBX18, SHOX2, NKX2-5, HCN4, and HCN1 in sinoatrial node cells were detected by qRT-PCR, and compared with enhanced green fluorescent protein blank virus. Results OCT3/4, SOX2, and NANOG were highly expressed in HiPS and ESCs, and there was no significant difference in the relative expression of each gene (P>0.05); OCT3/4 and NANOG were specifically distributed in the nucleus of HiPS, while SSEA4 and TRA-1-60 were distributed in the cell membrane. The relative expressions of ISL1 gene at 5, 7, 21, and 28 days and NKX2-5 gene at 7, 21, and 28 days of HiPS differentiation into cardiomyocytes were significantly higher than those of hACM (P<0.05), and the relative expressions of ACTN1 and TNNT2 genes at 3, 5, 7, and 21 days of HiPS differentiation into cardiomyocytes were significantly lower than those of hACM (P<0.05). NKX2-5 was expressed in most of the nuclei, cTnT and α-actinin, MLC-2A and MLC-2V signals were localized in the cytoplasm, presenting a texture-like structure of muscle nodules. Flow cytometry results showed that HiPS was successfully induced to differentiate into cardiomyocytes. The expressions of TBX18, SHOX2, HCN4, and HCN1 in the over-expression TBX3 group were up-regulated when compared with the control group, and difference in the relative expression of SHOX2 gene was significant (P<0.05); the relative expression of NKX2-5 gene was lower than that in the control group, but there was no significant difference (P>0.05). There was no significant difference in the relative expression of each gene between the over-expressed TBX18 group and the control group (P>0.05). Conclusion HiPS and hESCs have similar pluripotency, and we have established a stable method for maintaining and culturing the stemness of HiPS. A technological platform for the efficient differentiation of HiPS into cardiomyocytes has been successfully established. Although TBX3 and TBX18 do not play a significant role in promoting the enrichment and differentiation of HiPS into sinoatrial node-like cells, TBX3 shows a certain promoting trend, which can be further explored in the future.
ObjectiveTo investigate the diagnostic value of internal medicine thoracoscope combined with pleural GeneXpert MTB/RIF for tuberculous pleurisy.MethodsEighty patients with tuberculous pleurisy admitted to hospital with pleural effusion were treated as tuberculous pleurisy group, and 20 patients with clinical diagnosis of malignant pleural effusion were used as control group. After admission to the hospital, the pre-operative examination of internal medicine thoracoscope were analyzed. All patients were extracted pleural effusion with thoracic puncture in order to send pleural tuberculosis smear and culture. Patients who had no contraindications were arranged internal medicine thoracoscope to get pleural effusion which will be sent to GeneXpert MTB/RIF and pathological tissue biopsy.ResultsIn the tuberculous pleurisy group, nine patients were positive in pleural tuberculous smear, and the positive rate was 11.3%; 4 patients were positive in pleural tuberculous culture, and the positive rate was 5.0%; 75 patients were diagnosed with pathological biopsy, and the positive rate was 93.8%; 69 patients were positive with pleural GeneXpert MTB/RIF, and the positive rate was 86.3%. The positive rate of internal medicine thoracoscopic pleural biopsy combined with pleural GeneXpert MTB/RIF could reached 96.3%. The pleural GeneXpert MTB/RIF lifampin resistance gene was positive in 5 patients, 4 of them were positive for tuberculosis culture, and the drug sensitivity results showed rifampicin resistance. In the control group, patients had negative result in pleural effusion tuberculosis smear, tuberculosis culture and the pleural GeneXpert MTB/RIF.ConclusionsThe diagnosis of tuberculous pleurisy by the combination of internal medicine thoracoscope and pleural GeneXpert MTB/RIF has high specificity and sensitivity. The diagnosis of tuberculous pleurisy by the combination of internal medicine thoracoscope and pleural GeneXpert MTB/RIF has high specificity and sensitivity, which has the value of rapid and accurate diagnosis and early guidance of anti-tuberculosis chemotherapy based on the early judgment of whether rifampin resistance exists.
ObjectiveTo systematically review the diagnostic value of T-SPOT.TB and QuantiFERON-TB (QFT-GIT/QFT-Plus) in active tuberculosis (ATB). MethodsThe PubMed, Web of Science, Cochrane Library, EMbase, CNKI, WanFang Data, and CBM databases were electronically searched to collect diagnostic accuracy studies comparing QFT-GIT/QFT-Plus and T-SPOT.TB for diagnosing ATB from inception to February 8, 2022. Two reviewers independently screened literature, extracted data and assessed the risk of bias of the included studies, then, meta-analysis was performed by using Stata 16.0 software. ResultsA total of 20 studies were included. The results of meta-analysis showed that the pooled sensitivity of T-SPOT.TB and QFT-GIT were 0.89 (95%CI 0.85 to 0.92) and 0.84 (95%CI 0.79 to 0.89), the pooled specificity were 0.85 (95%CI 0.68 to 0.93) and 0.86 (95%CI 0.72 to 0.94), the area under the curve (AUC) of summary receiver operating characteristic (SROC) were 0.93 (95%CI 0.84 to 0.97) and 0.90 (95%CI 0.56 to 0.99), respectively. The pooled sensitivity of T-SPOT.TB and QFT-Plus were 0.93 (95%CI 0.81 to 0.97) and 0.93 (95%CI 0.89 to 0.96), specificity were 0.99 (95%CI 0.39 to 1.00) and 0.94 (95%CI 0.67 to 0.99), the AUC of SROC were 0.99 (95%CI 0.67 to 1.00) and 0.98 (95%CI 0.65 to 1.00), respectively. ConclusionBoth T-SPOT.TB and QFT have high diagnostic accuracy for ATB, and the diagnostic sensitivity of T-SPOT.TB is better than QFT-GIT. Due to the limited quality and quantity of the included studies, more high quality studies are needed to verify the above conclusion.
RCBTB1 gene associated hereditary retinopathy is an extremely rare inherited retinal disease (IRD) discovered recently. The mutation of RCBTB1 gene can lead to a variety of IRD clinical phenotypes, such as early retinitis pigmentosa and delayed chorioretinal atrophy. The hereditary mode of RCBTB1 gene associated retinopathy is autosomal recessive. RCBTB1 gene plays an important role in maintaining mitochondrial function and anti-oxidative stress defense mechanism of retinal pigment epithelium cells. In the future, it is necessary to further determine whether there is a genotypic and phenotypic correlation in the age of onset of RCBTB1 gene associated retinopathy or multi-organ involvement, and evaluate the safety and efficacy of adeno-associated virus-mediated RCBTB1 gene replacement therapy in animal models, to explore the feasibility of gene replacement therapy and stem cell therapy.
Objective To collect the clinical data of victims with traumatic brain injury (TBI) admitted in the West China Hospital of Sichuan University within 2 weeks after 4.20 Lushan earthquake, and to analyze their clinical characteristics and effects of early rehabilitation, so as to provide baseline data for rescue TBI victims with the early rehabilitation treatment during emergency medical rescue. Methods A total of 392 victims admitted in the hospital from April 20th, 2013 to May 3rd, 2013 were screened, of which the TBI victims were clinically assessed and treated with early rehabilitation. Then both the activities of daily living (ADL) and the Rancho Los Amigos Cognitive Recovery Scale (RLA) before and after the treatment were analyzed. The data were input by Excel software, and the statistical analysis was performed by SPSS softwar. Results A total of 51 TBI victims at age from 3 to 84 years old were included finally. The categories of TBI included subarachnoid hemorrhage (41.2%), intracranial hematoma (33.3%) and mixed type (33.3%), and the severity were associated with the type of TBI. The GCS score of cerebral concussion was higher (13.25 ± 0.62) while that of the diffuse axonal injury was lower (4.50 ± 0.71). All victims (100%) had limited ADL, 74.51% had cognitive dysfunction, 9.80% had speech disorder, and 7.84% had dysphagia. After the early rehabilitation treatment, both ADL (before treatment: 34.82 ± 58.29, after treatment: 69.63 ± 22.29) and RLA (pre-treatment: 4.16 ± 1.24, treatment: 7.20 ± 1.69) were obviously higher than those before treatment, with statistical differences (both P lt;0.05). Conclusion The TBI categories of Lushan earthquake victims are various and mixed, and the severity associated with the type of TBI. All TBI victims are accompanied with more clinical problems and functional limitation. Early rehabilitation treatment is safe and effective to improve ADL and RLA as well.
Objective To explore an improved method of surgical operation for reposition of the articular surface with Type Ⅲ Pilon fractures. Methods From January 1999 to December 2005, 20 patients (22 sides) with Type Ⅲ Pilon fractures were treated with the delayed open reduction and the internal fixation, which took the superior articular surface of the talus as a templet so as to reposition the lower articular surface of the tibia, strengthen the bone transplantation, fasten the internal fixation, and make an early functional exercise possible. Complete data were obtained from 16 of the patients with 18 sides (13 males,15 sides; 3 females, 3 sides; age, 14-48 years). The injury due to a fallingaccident was found in 12 patients (14 sides), and due to a traffic accident in 4patients (4 sides). Results The healing of the first intention was achieved in 14 sides, the delayed healing in 3 sides, and the infection in 1 side. The follow-up of all the 16 patients for 971 months (average, 22 months) including the X-ray examinations revealed that no screw for the internal fixation entering the articular cavity. According to the Teeny’s judging standards of radiology evaluating the result of the surgery for Pilon fractures, the anatomical reduction of the related articular surface was found in 77.8% of the sides (14/18) and thehealing of the first intention (stage Ⅰ) in 94.4% (17/18). According to the Mazur’s criteria, an excellent result was obtained in 5 sides, good in 7, fair in 5, and poor in 1. The excellent and good result was 66.7%. Conclusion Propermanagement of the injured soft tissues, prompt recovery of the tibial distant plateau height, and accurate reposition of the articular surface, enough transplant bone for the solid support, b internal fixation for the distant tibial anatomical structure, and early functional exercise are the key points to the successful operation.