Objective To investigate the diagnostic value of T cell enzyme-linked immunospot (T-SPOT.TB) assay on extrapulmonary tuberculosis patients.Methods Thirty patients suffered from extrapulmonary mycobacterium tuberculosis(MTB) infection and 30 with non-MTB infection were recruited this study.T-SPOT.TB assay was used to detect early secreting antigen target-6 (ESAT-6) and culture filtrate protein-10(CFP-10) specific T cells in blood samples.PPD skin test was also used.Results (1)The positive rate of MTB detected by T-SPOT.TB assay was 91.89％ (34/37),higher than that of un-tuberculosis group (6.67 ％ (2/30)),and the difference was significant (x2 =48.403,P ＜ 0.001).(2) The sensitivity,specificity,positive prospective value and negative prospective value of T-SPOT.TB assay were 91.89％,93.33％,94.44％ and 90.32％ respectively,better than those of PPD skin test (67.57％,56.67％,65.79％,58.62％),and the differences were markedly (x2 =6.773,10.756,9.392,8.031 respectively ; P =0.009,0.001,0.002,0.005 respectively).Meanwhile T-SPOT.TB assay has low agreement with means of PPD skin test(Kappa =0.311,x2 =6.801,P =0.009).Conclusion T-SPOT.TB assay has a higher sensitivity and specificity in the rapid diagnosis of extrapulmonary tuberculosis.Therefore,it is with great value and applicability as a screening test.

Objective: To investigate the daily learning time status of elementary school students and understand the implementation of Health Requirements of Daily Learning Time for Secondary and Elementary School Students (GB/T 17223-2012) in schools, so as to provide a reference for strengthening and improving school health and health education in the new era. Methods: A stratified cluster random sampling method was used to select 7 776 primary school students in Shandong Province, and a survey questionnaire was designed based on the Health Requirements for Dayily Learning Time for Secondary and Elementary School Students(GB/T 17223-2012) standard to investigate their daily learning arrangements, sleep and physical activities, and breaks between classes. Comparison of intergroup differences were used by Chi square test and Kruskal-Wallis H test. Results: About 55.88 % of primary school students for daily learning time met the standard. There was a statistically significant difference in the daily learning time achievement rate among primary school students in three grades for first and second grade, third and fourth grade and fifth and sixth grade ( Z=1 629.47, P <0.01), and the fifth and sixth grade had the highest proportion of achieving the standard (85.92%). Specifically, the proportions of students whose class hours, class numbers and morning reading time meeting the standards were 30.07%, 10.20% and 42.19%, respectively. The sleep deficiency rate of primary school studnets was 58.69%, and the physical activity deficiency rate was 65.78%; and there was a statistically significant difference in the rate of insufficient sleep time and physical activity time among primary school students of different grades ( χ 2=56.39, 95.95, P <0.01), with sixth grade students showing the highest rates for both sleep and physical activity deficiencies (64.35%, 73.37%). Additionally, 49.78% of students had recess time below the standard requirements. Conclusion The health status of daily learning time among primary school students in Shandong Province is poor, with insufficient implementation of school standards, and the implementation of standards needs to be further strengthened.

The incidence of lung cancer in China ranks first and second among men and women respectively .Meanwhile, lung cancer is also the leading cause of death in malignant tumors in China .Surgical treatment is of great significance in impro-ving the prognosis of patients with lung cancer, and the number of related clinical trials is increasing year by year.Through a retrospective review of clinical trials in the field of lung cancer and thoracic surgery on the Clinicaltrials .gov platform, this arti-cle finds that relevant clinical trials focus on treatment , perioperative management and basic research.Although the clinical tri-als in China started late, they have developed rapidly and the number of clinical trials ranked second.In addition, there are some shortcomings in the clinical trials of China in terms of trial design and research scope , and there is still a gap between Chinese and foreign trials.

Malignant pleural mesothelioma(MPM) is a kind of rare and aggressive malignant neoplasm. Surgery plays one of the most important roles in the treatment of MPM. However, due to the high morbidity and mortality reported, the survival benefit and indication of surgery are still controversial. This article will review the surgical indications, discuss and compare the roles of extrapleural pneumonectomy(EPP) and pleurectomy / decortication(P/D) which aim to achieve macroscopic complete resection(MCR) in the treatment of MPM. Finally, we summarized the progress of other treatment methods including targeted therapy and immunotherapy.

Objective:To investigate the molecular pathogenesis of a newly discovered gene mutation in a family with hereditary coagulation factor Ⅺ(FⅪ) deficiency.Methods:The proband was admitted to the First Affiliated Hospital of Wenzhou Medical University in September 2021 due to "calculus of intrahepatic duct". The patient had no symptoms of spontaneous bleeding.The clinical data and blood samples of the proband and her family members (10 persons in 3 generations) were collected.The activated partial thromboplastin time (APTT) and FⅪ activity (FⅪ:C) were performed by the one-stage clotting assay. FⅪ antigen (FⅪ:Ag) were detected by enzyme linked immunosorbent assay (ELISA). Genomic DNA extracted from peripheral blood cells of subjects was used as template to analyze F11 gene mutation by DNA direct sequencing. Bioinformatics software was used to analyze the effects of mutations on protein structure and function. Wild-type and mutant FⅪ protein expression vectors were constructed and transient transfected into HEK293T cells. The total RNA was extracted from positive transfected cells and then reversely transcribed into cDNA. The mRNA expression level of F11 gene in transfected cells was detected by real-time fluorescence quantitative PCR (qRT-PCR). The content of FⅪ:Ag and the expression of FⅪ protein in transfected cell lysates and culture supernatant were detected by ELISA and western blot.Results:The APTT of the proband was significantly prolonged to 107.9s (reference range 29.0-43.0s), while FⅪ:C and FⅪ:Ag were significantly decreased to 2% (reference range 84%-122%) and 5% (reference range 76%-127%), respectively. Gene sequencing analysis indicated that the proband had c.536C>T (p.Thr161Met) heterozygous missense mutation and c.1556G>A (p.Trp501Ter) heterozygous nonsense mutation in exon 6 and 13 of the F11 gene, respectively. Bioinformatics analysis showed that the amino acids at site 161 of FⅪ protein were threonine (Thr) in the matrix composed of five different species, indicating that Thr161 site was highly conserved among homologous genes in different species. p.Thr161Met heterozygous mutation affected the stability of local intermolecular structure of FⅪ protein. In vitro expression experiments of p.Thr161Met mutation showed that FⅪ protein had a normal synthesis in the cells but secretion dysfunction.Conclusions:c.536C>T (p.Thr161Met) heterozygous missense mutation and c.1556G>A (p.Trp501Ter) heterozygous nonsense mutation were mainly responsible for the decrease of FⅪ in this family. p.Thr161Met mutation was first reported in the world and did not affect the normal synthesis of FⅪ protein, but caused secretion dysfunction.

Objectives:To examine the influence of the proportion of pathological subtypes on the prognosis of stage ⅠA lung adenocarcinoma cases, and to explore the association between the presence/absence of solid or micropapillary (S/M) components and survival outcome.Methods:Totally 321 patients with stage ⅠA lung adenocarcinoma who received complete surgical resection at Department of Thoracic Surgery, Tongji University Affiliated Shanghai Pulmonary Hospital from January 2011 to December 2013 were retrospectively analyzed. There were 130 males and 191 females, aging 59(11) years ( M(IQR)) (range: 55 to 66 years). The diagnostic value of the proportion of each pathological growth subtype on relapse-free survival (RFS) and overall survival (OS) were analyzed by using receiver operator characteristic curve. Patients were firstly divided into two groups according to the presence or absence of S/M components. And patients without S/M components were farther divided into two groups according to predominant growth pattern. There were three groups in total: with S/M components (group S/M +), without S/M components and lepidic growth pattern predominant (group S/M -LPA), without S/M components and papillary or acinar growth pattern predominant (group S/M -P/A). Kaplan-Meier method were used to draw the survival curves of the three groups, and Log-rank test were used to compare RFS and OS among the three groups. Cox proportional risk model was used to verify whether the presence of S/M components was a prognostic factor on RFS. Results:The proportion of S/M components had no diagnostic value for recurrence (solid: area under curve (AUC)=0.588, P=0.095; micropapillary: AUC=0.566, P=0.106) and death (AUC=0.589, P=0.104; AUC=0.607, P=0.056). The 5-year RFS rate of group S/M -LPA, S/M -P/A and S/M + were 92.4%, 82.3% and 77.3%, respectively (all P<0.05), while the 5-year OS rate were 97.4%, 94.5% and 83.2%, respectively (all P<0.05). Multivariable analysis showed that the 3 groups were independent predictors of recurrence (S/M -P/A vs. S/M - LPA: HR=2.691, 95% CI: 1.249 to 5.799, P=0.011; S/M +vs. S/M -LPA, HR=6.763, 95% CI: 3.050 to 14.996, P<0.01). Conclusions:The proportion of S/M components in stage ⅠA lung adenocarcinoma with complete resection cases did not affect survival outcome. New grouping method based on the presence or absence of S/M components were significantly associated with patient survival outcomes: S/M + patients had the worst prognosis and S/M -LPA patients had the best prognosis.

Objectives:To examine the influence of the proportion of pathological subtypes on the prognosis of stage ⅠA lung adenocarcinoma cases, and to explore the association between the presence/absence of solid or micropapillary (S/M) components and survival outcome.Methods:Totally 321 patients with stage ⅠA lung adenocarcinoma who received complete surgical resection at Department of Thoracic Surgery, Tongji University Affiliated Shanghai Pulmonary Hospital from January 2011 to December 2013 were retrospectively analyzed. There were 130 males and 191 females, aging 59(11) years ( M(IQR)) (range: 55 to 66 years). The diagnostic value of the proportion of each pathological growth subtype on relapse-free survival (RFS) and overall survival (OS) were analyzed by using receiver operator characteristic curve. Patients were firstly divided into two groups according to the presence or absence of S/M components. And patients without S/M components were farther divided into two groups according to predominant growth pattern. There were three groups in total: with S/M components (group S/M +), without S/M components and lepidic growth pattern predominant (group S/M -LPA), without S/M components and papillary or acinar growth pattern predominant (group S/M -P/A). Kaplan-Meier method were used to draw the survival curves of the three groups, and Log-rank test were used to compare RFS and OS among the three groups. Cox proportional risk model was used to verify whether the presence of S/M components was a prognostic factor on RFS. Results:The proportion of S/M components had no diagnostic value for recurrence (solid: area under curve (AUC)=0.588, P=0.095; micropapillary: AUC=0.566, P=0.106) and death (AUC=0.589, P=0.104; AUC=0.607, P=0.056). The 5-year RFS rate of group S/M -LPA, S/M -P/A and S/M + were 92.4%, 82.3% and 77.3%, respectively (all P<0.05), while the 5-year OS rate were 97.4%, 94.5% and 83.2%, respectively (all P<0.05). Multivariable analysis showed that the 3 groups were independent predictors of recurrence (S/M -P/A vs. S/M - LPA: HR=2.691, 95% CI: 1.249 to 5.799, P=0.011; S/M +vs. S/M -LPA, HR=6.763, 95% CI: 3.050 to 14.996, P<0.01). Conclusions:The proportion of S/M components in stage ⅠA lung adenocarcinoma with complete resection cases did not affect survival outcome. New grouping method based on the presence or absence of S/M components were significantly associated with patient survival outcomes: S/M + patients had the worst prognosis and S/M -LPA patients had the best prognosis.

Integrin alphavbeta3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and roles of alphavbeta3 integrin, a stable CHO-677 cell line expressing the murine alphavbeta3 heterodimer (designated as "CHO-677-malphavbeta3" cells) was established using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits alphav and beta3 were detected at the gene and protein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-malphavbeta3 cell line at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-forming assay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replication levels of Foot-and-mouth disease virus (FMDV) in the CHO-677-malphavbeta3 cell line. After infection with FMDV/O/ZK/93, the cell line showed a significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stable alphavbeta3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of alphavbeta3 integrin, and as a cell model for FMDV research.
Animals ; Animals, Suckling ; CHO Cells ; Cloning, Molecular ; Cricetulus ; DNA, Complementary/genetics/metabolism ; Disease Susceptibility/virology ; Foot-and-Mouth Disease/*genetics/virology ; Foot-and-Mouth Disease Virus/*physiology ; Integrin alphaVbeta3/*genetics/metabolism ; Mice

Objective:To explore the sex-based heterogeneity in demographic and pathological trends of lung cancer during the past 30 years.Methods:Patients with primary lung cancer who received surgical treatment in the Department of thoracic surgery, Shanghai Pulmonary Hospital Tongji University from 1989 to 2018 were retrospectively analyzed. The differences between male and female patients in age, smoking history, pathological stage and type were compared. Mann- Kendall trend test was performed for trend analysis. Results:A total of 58 433 patients were included in this study, encompassing 30 729(52.6%) men and 27 , 704(47.4%) women. Compared with male patients, female patients were younger(56.0 years old vs. 59.7 years old), and had a higher proportion of non-smokers(98.3% vs. 52.3%), stage Ⅰ lung cancers(60.6% vs. 49.3%), and adenocarcinoma(93.7% vs. 56.1%, all P-values <0.001). Trend analyses revealed that the proportion of female patients increased year by year, and surpassed males in 2015, with the current ratio of male to female being 1∶1.5. After 2013, the age of onset in females was getting younger, and the average age decreased from 58.7 years old to 54.7 years old( P=0.02). The decrease in the proportion of smoking patients was mainly reflected by male patients(from 68.5% to 31.1%, P<0.01). Stage Ⅰ lung cancers in male and females outnumbered advanced stage in 2012 and 2010, respectively, with a much higher proportion in female patients. Among male patients, adenocarcinoma has replaced squamous cell carcinoma as the most common pathological type since 2012, while in female patients adenocarcinoma remained the most common pathological type of lung cancer, and its proportion continued to increase reaching over 98%. Conclusion:A dramatic change in gender distribution was noticed during the past 30 years. Female patients became the primary population in surgically-treated lung cancers, with a trend of getting younger. The proportion of smokers and squamous cell carcinoma decreased significantly in male patients, and adenocarcinoma has become the most common pathological type of lung cancer. The proportion of stage Ⅰ lung cancers was on a dramatic rise, with the popularization of CT screening for lung cancer.

Objective:To analyze the phenotype and genotype of two propositi with inherited hypodysfibrinogenaemia caused by compound heterozygous mutations, and investigate the molecular mechanism.Metheds:Two propositi and their family members(7 person in 3 generations and 10 person in 3 generations,respectively) were investigated. The activity of plasma fibrinogen (Fg:C) and thrombin time (TT) were analyzed by coagulation method, the antigen of plasma fibrinogen (Fg:Ag) was detected by immunoturbidimetry. All of the exons and flanking sequences of FGA,FGB,FGG of two propositi were amplified by PCR, followed by direct sequencing. The ClustalX-2, 1-win software was used to analyze the conservatism of mutated gene locus. PROVEAN and Mutation Taster were applied to analyze the pathogenicity of mutated amino acid. The changes of the protein spatial structure and intermolecular interaction were analyzed by Pymol.Results:Fg:C and Fg:Ag of proposita A and B were both significantly decreased (0.74 and 0.78 g/L, 0.96 and 0.94 g/L, respectively). Gene analysis revealed that proposita A and B both carried a heterozygous mutation c.2185G>A(p.AαGlu710Lys) in exon 6 of FGA. Furthermore, proposita A also carried a heterozygous mutation c.701G>T(p.γTrp208Leu) in exon 7 of FGG, and proposita B carried a heterozygous mutation c.1015A>C(p.γSer313Arg) in exon 8 of FGG. Phylogenetic analysis suggested that p.AαGlu710,p.γTrp208 and p.γSer313 were highly conserved among homologous species. All variants were predicted to be deleterious by two online bioinformatic softwares. The protein model analysis indicated that protein spatial structure and intermolecular hydrogen bonds were changed by these variants, which destroyed the stability of Fg.Conclusion:The compound heterozygous mutations of p.AαGlu710Lys and p.γTrp208Leu,p.AαGlu710Lys and p.γSer313Arg might account for the hypodysfibrinogenemia in two propositi.