Objective To find humoral protein markers to develop new experimental diagnostic methods for tuberculous pleurisy. Methods Proteomes of 7 d and 14 d culture filtrate of Mycobacterium tuberculosis growing in Middlebrook 7H9, serum and pleural effusion from five patients with tuberculous pleurisy were detected by surface-enhanced laser desorptionionization time-of-flight massspectrometry (SELDI-TOF-MS). And the data were analyzed with descriptive statistics method to observed the protein components all owned by three kinds of proteome. Results From protein species, proteins of all culture filtrate were far more than that of pleural effusion and serum while proteins of pleural effusion in four cases were more than that of serum. The kinds of common proteins between culture filtrate and pleural effusion, between culture filtrate and serum, between serum and pleural effusion, among culture filtrate and pleural effusion and serum were different. But the protein of relative molecular mass of 2 660 depending on the ratio of mass to charge existed in all samples of culture filtrate, pleural effusion and serum. Conclusion The protein of relative molecular mass of 2 660 possess the latent quality as a specific humoral protein marker for tuberculous pleurisy. But it is essential that must be further confirmed among large samples.

Objective To assess the efficacy of using four kinds of proteins / peptides to distinguish the tuberculosis patients from healthy people. Methods A, B, C and D were used to represent four proteins /peptides with 1 060, 1 944, 2 081 and 3 954 of mass to charge ratio (m / z) in serum, respectively. Levels A, B, C and D in serum of 57 patients with tuberculosis and 30 healthy people were determined by using the surface-enhanced laser desorption-ionization time of flight mass spectrometry (SELDI-TOF-MS). Then the differences of levels of f A, B, C and D were anlyzed between tuberculosis patients and healthy people. The efficacy of distinguishing tuberculosis patient from healthy people were evaluated by using diagnostic test evaluation method. Results (1) The levels of A, B, C and D were 1 ± 11, 1 597 ± 3 102, 460 ± 765 and 1 208 ± 1 003 in tuberculosis patients, while they were 123 ± 201, 47 ± 98, 36 ± 93 and 397 ± 355 in healthy people. (2) The area under the receiver operator characteristic (ROC) curve was 0.644, 0.848, 0.735 and 0.810 respectively. The serum levels of A, B, C and D could be used to distinguish tuberculosis patient from healthy people and the cut-off values of A, B, C and D were ≤166, ≥318, ≥48 and ≥728, respectively. Conclusions B, C and D have better performances to distinguish tuberculosis patients from healthy people , which may be regarded as new promising candidate markers for diagnosis of tuberculosis.

In the practice of building its humanistic environment of a research hospital,Nanjing Drum Tower hospital adheres to such humanistic characteristics of the hospital as humanistic concept,planning, environment,management,service,and quality.Furthermore,the hospital upholds such keys as learning, innovation, cooperation, undertaking, competition and development. High focus, high starting point planning,persistence,and down-to-earth efforts,effectively promoting hospital development.

To identify spatiotemporal expression patterns of vernalization genes in common wheat, we analyzed expression characteristics of several vernalization genes (VRN1, VRN2 and VRN3) in the wheat cultivars 'Chinese spring' and 'Luohan 2' by RT-PCR. The VRN1 gene was expressed at different levels in the leaves and roots at the 3-leaf stage, stems, flag leaves at the grain-filling stage, anthers, ovules, and developing seeds in 'Chinese spring'. Expression of VRN1 increased before flowering date, then decreased after flowering time. Expression of VRN1 was not detected in dry seeds or seeds germination. Expression patterns of VRN1 in 'Luohan 2' were similar to those in 'Chinese spring', except that it was not expressed in roots or in the leaves at the 3-leaf stage in 'Luohan 2'. Expression of VRN2 was only detected in the leaves at the 3-leaf stage and in the embryo buds during seeds germination. The Spatiotemporal expression of VRN3 was similar to that of VRN1, except that VRN3 was not expressed in roots. These results improved our understanding of the molecular regulation of vernalization genes in common wheat.
Flowers ; genetics ; physiology ; Gene Expression Regulation, Plant ; genetics ; Genes, Plant ; genetics ; Triticum ; genetics

OBJECTIVE: To carry out prenatal diagnosis for a fetus with mosaicism Yq deletion. METHODS: A fetus with high risk of sex chromosomes indicated by non-invasive prenatal testing (NIPT) at Tianjin Medical University General Hospital in July 2021 was selected as the study subject. Prenatal diagnosis of the fetus was performed with combined G-banded chromosomal karyotyping, fluorescence in situ hybridization (FISH), copy number variation sequencing (CNV-seq), real-time fluorescence PCR (QF-PCR), and ultrasound examination. RESULTS: Analysis of the amniocytes at 23 gestational weeks had yielded a 45,X karyotype. However, FISH had shown signals of Y chromosome. Re-examination by cordocentesis had shown a mosaicism of 46,X,+mar[33]/45,X[17]. FISH showed that 69% of the cells had contained Y chromosome signals. The result of CNV-seq was seq[19]del(Y)(q11.1q12)(mos) chrY: g.13200001_ 28820000del (mosaicism rate = 64%), which suggested mosaicism for a Yq deletion, which encompassed the azoospermia factor (AZF) region. Deletion of the AZF region was verified by QF-PCR. The fetal karyotype was ultimately determined as mos46,X,del(Y)(q11.1)[33]/45,X[17]. Although ultrasound examination had shown no abnormality in the fetus, the couple had opted to terminate the pregnancy, and the induced fetus had a normal male appearance. CONCLUSION The combined use of multiple techniques is beneficial for accurate and rapid prenatal diagnosis. For fetuses with mosaicism chromosomal abnormalities, it may be difficult to accurately predict the postnatal phenotype. It is therefore necessary to further explore their genotype-phenotype correlation in order to provide better guidance upon genetic counseling.
Female ; Pregnancy ; Male ; Humans ; Mosaicism ; DNA Copy Number Variations ; In Situ Hybridization, Fluorescence ; Y Chromosome ; Fetus

OBJECTIVE: To assess the clinical efficacy and health economic value of non-invasive prenatal testing (NIPT) for the prenatal screening of common fetal chromosomal aneuploidies. METHODS: 10 612 pregnant women from October 2017 to December 2019 presented at the antenatal screening clinic of the General Hospital of Tianjin Medical University were selected as the study subjects. Results of NIPT and invasive prenatal diagnosis and follow-up outcome for the 10 612 pregnant women were retrospectively analyzed and compared. Meanwhile, NIPT data for two periods were analyzed for assessing the health economic value of NIPT as the second- or first-tier screening strategy for the prenatal diagnosis of fetal trisomies 21, 18 and 13. RESULTS: The NIPT was successful in 10 528 (99.72%) subjects, with the sensitivity for fetal trisomies 21, 18 and 13 being 100%, 92.86% and 100%, and the positive predictive value (PPV) being 89.74%, 61.90% and 44.44%, respectively. The PPV of NIPT for sex chromosome aneuploidies was 34.21%. Except for one false negative case of trisomy 18, the negative predictive value for trisomy 21, trisomy 13 and other chromosomal abnormalities were 100%. For pregnant women with high risk by serological screening, advanced maternal age or abnormal ultrasound soft markers, NIPT has yielded a significantly increased high risk ratio. There was no statistical difference in the PPV of NIPT among pregnant women from each subgroup. NIPT would have higher health economic value as a second-tier screening until 2019, while compared to 2015 ~ 2017, its incremental cost-effectiveness ratio as a first-tier screening had declined clearly. CONCLUSION The screening efficacy of NIPT for trisomies 21, 18 and 13 for a mixed population is significantly better than conventional serological screening, but it is relatively low for sex chromosomal abnormalities. NIPT can also be recommended for populations with relatively high risks along with detailed pre- and post-test genetic counselling. From the perspective of health economics, except for open neural tube defects, it is possible for NIPT to replace the conventional serological screening in the future as its cost continues to decrease.
Pregnancy ; Female ; Humans ; Trisomy/genetics* ; Retrospective Studies ; Prenatal Diagnosis/methods* ; Down Syndrome/genetics* ; Aneuploidy ; Chromosome Aberrations ; Trisomy 18 Syndrome/genetics* ; Sex Chromosome Aberrations ; Fetus

Objective:To explore the value of panoramic teaching in clinical practice teaching of department of respiratory medicine.Methods:A total of 35 clinical interns who had a 3-week rotation in the department of respiratory medicine from October 2019 to June 2020 were selected, and they were divided into a control group ( n=17) and a study group ( n=18) according to the order of admission. The control group used traditional teaching, and the research group used panoramic teaching. After the teaching, the teaching effect was evaluated through theoretical knowledge assessment, practical assessment and questionnaire survey. SPSS 19.0 was used for t test. Results:There was no statistically significant difference between the two groups of interns in theoretical assessment [(89.62±4.05) points vs. (90.84±4.12) points, t=-0.891, P=0.379]. The results of practice assessment (case analysis, physical examination of the department, medical history inquiry and skills manipulation) of the study group were significantly higher than those of the control group ( P<0.05). The interns in study group were better than those in control group in such aspects as learning interest, self-learning ability, self-study ability, doctor-patient communication language expression ability, literature review ability, clinical thinking ability, comprehensive analysis ability, knowledge development ability, system diagnosis ability, teamwork ability and teaching satisfaction ( P<0.05). Conclusion:Panoramic teaching can effectively improve the ability of interns to master the clinical diagnosis and treatment of respiratory medicine, improve the teaching effect, and improve the learning ability and clinical competence of interns in the future.

BACKGROUND: It has been proven that CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated 9) system was the modern gene-editing technology through the constitutive expression of nucleases Cas9 in the mammalian, which binds to the specific site in the genome mediated by single-guide RNA (sgRNA) at desired genomic loci. The aim of this study is that the animal model of EZH2 gene knockout was constructed using CRISPR/Cas9 technology. METHODS: In this study, we designed two single-guide RNAs targeting the Exon3 and Exon4 of EZH2 gene. Then, their gene-targeting efficiency were detected by SURVEYOR assay. The lentivirus was perfused into the lungs of mice by using a bronchial tube and detected by immunohistochemistry and qRT-PCR. RESULTS: The experimental results of NIH-3T3 cells verify that the designed sgEZH2 can efficiently effect the cleavage of target DNA by Cas9 in vitro. The immunohistochemistry and qRT-PCR results showed that the EZH2 expression in experimental group was significantly decreased in the mouse lung tissue. CONCLUSIONS The study successfully designed two sgRNA which can play a knock-out EZH2 function. An EZH2 knockout animal model was successfully constructed by CRISPR/Cas9 system, and it will be an effective animal model for studying the functions and mechanisms of EZH2.
Animals ; CRISPR-Cas Systems ; Enhancer of Zeste Homolog 2 Protein ; genetics ; metabolism ; Female ; Gene Knockout Techniques ; Gene Targeting ; Humans ; Lung Neoplasms ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; RNA, Guide

BACKGROUNDAccording to data from the China Hospital Invasive Fungal Surveillance Net (CHIF-NET) 2010, Candida tropicalis (C. tropicalis) is the third most common pathogen causing invasive candidiasis. Moreover, the majority of fluconazole-resistant C. tropicalis isolates were from a single hospital. Therefore, a molecular epidemiological survey is necessary to investigate the genetic relatedness of C. tropicalis isolates in China.

METHODSIn this study, 48 C. tropicalis isolates causing invasive fungal infections from four tertiary hospitals in China were studied. All the isolates were identified by sequencing the internal transcribed spacer region. Antifungal susceptibility to triazoles, amphotericin B, and caspofungin was determined by the Clinical and Laboratory Standards Institute standard broth microdilution method. Multilocus sequence typing (MLST) was performed, and phylogenetic analysis was further performed by the eBURST and maximum parsimony (MP) methods to characterize the genetic relatedness of isolates.

RESULTSMLST discriminated 40 diploid sequence types (DSTs) among 48 isolates, including 36 novel DSTs, and the XYR1 gene showed the highest discriminatory power. The DSTs obtained from this study were compared with those of previously reported C. tropicalis isolates, and there was poor type alignment with regional strains. Nine groups and 11 singletons were identified by eBURST, whereas two groups and 10 subgroups were clustered by MP analysis. Generally, there were no obvious correlations between clonal clusters generated and the specimen source or hospital origin. Seven fluconazole-resistant isolates were confirmed and assigned to three distinguishable branches.

CONCLUSIONSThe results suggested diverse origins of invasive C. tropicalis isolates in China. Although most invasive C. tropicalis strains in the mainland of China were clustered with previously characterized Asian isolates, major C. tropicalis clusters identified in this study were genetically distinct from those of other geographic regions.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antifungal Agents ; therapeutic use ; Candida tropicalis ; drug effects ; pathogenicity ; Candidiasis ; drug therapy ; China ; Drug Resistance, Fungal ; genetics ; Female ; Humans ; Male ; Middle Aged ; Multilocus Sequence Typing ; methods ; Phylogeny ; Young Adult