Expression of CD24 gene in human malignant pleural mesothelioma and its relationship with prognosis / 中华劳动卫生职业病杂志

Objective: To investigate the expression of CD24 gene in human malignant pleural mesothelioma (MPM) cells and tissues, and evaluate its relationship with clinicopathological characteristics and clinical prognosis of MPM patients. Methods: In February 2021, UALCAN database was used to analyze the correlation between CD24 gene expression and clinicopathological characteristics in 87 cases of MPM patients. The TIMER 2.0 platform was used to explore the relationship between the expression of CD24 in MPM and tumor immune infiltrating cells. cBioportal online tool was used to analyze the correlation between CD24 and MPM tumor marker gene expression. RT-qPCR was used to analyze the expressions of CD24 gene in human normal pleural mesothelial cell lines LP9 and MPM cell lines NCI-H28 (epithelial type), NCI-H2052 (sarcoma type), and NCI-H2452 (biphasic mixed type). RT-qPCR was performed to detect the expressions of CD24 gene in 18 cases of MPM tissues and matched normal pleural tissues. The expression difference of CD24 protein in normal mesothelial tissue and MPM tissue was analyzed by immunohistochemistry. A Kaplan-Meier model was constructed to explore the influence of CD24 gene expression on the prognosis of MPM patients, and Cox regression analysis of prognostic factors in MPM patients was performed. Results: The CD24 gene expression without TP53 mutation MPM patients was significantly higher than that of patients in TP53 mutation (P<0.05). The expression of CD24 gene in MPM was positively correlated with B cells (r(s)=0.37, P<0.001). The expression of CD24 gene had a positive correlation with the expressions of thrombospondin 2 (THBS2) (r(s)=0.26, P<0.05), and had a negative correlation with the expression of epidermal growth factor containing fibulin like extracellular matrix protein 1 (EFEMP1), mesothelin (MSLN) and calbindin 2 (CALB2) (r(s)=-0.31, -0.52, -0.43, P<0.05). RT-qPCR showed that the expression level of CD24 gene in MPM cells (NCI-H28, NCI-H2052 and NCI-H2452) was significantly higher than that in normal pleural mesothelial LP9 cells. The expression level of CD24 gene in MPM tissues was significantly higher than that in matched normal pleural tissues (P<0.05). Immunohistochemistry showed that the expressions of CD24 protein in epithelial and sarcoma MPM tissues were higher than those of matched normal pleural tissues. Compared with low expression of CD24 gene, MPM patients with high expression of CD24 gene had lower overall survival (HR=2.100, 95%CI: 1.336-3.424, P<0.05) and disease-free survival (HR=1.800, 95%CI: 1.026-2.625, P<0.05). Cox multivariate analysis showed that compared with the biphasic mixed type, the epithelial type was a protective factor for the prognosis of MPM patients (HR=0.321, 95%CI: 0.172-0.623, P<0.001). Compared with low expression of CD24 gene, high expression of CD24 gene was an independent risk factor for the prognosis of MPM patients (HR=2.412, 95%CI: 1.291-4.492, P=0.006) . Conclusion: CD24 gene and protein are highly expressed in MPM tissues, and the high expression of CD24 gene suggests poor prognosis in MPM patients.

Clinical and genetic analysis of a patient with Craniofrontonasal syndrome / 中华医学遗传学杂志

OBJECTIVE@#To explore the clinical feature and genetic etiology of a patient with Craniofacial nasal syndrome (CNFS).@*METHODS@#A patient with CNFS who had presented at the Guiyang Maternal and Child Health Care Hospital on November 13, 2021 was selected as the study subject. Clinical data of the patient were collected. Peripheral venous blood samples were collected from the patient and her parents and subjected to trio-whole exome sequencing (trio-WES). Candidate variants were verified by Sanger sequencing and bioinformatic analysis.@*RESULTS@#The patient, a 15-year-old female, had predominantly featured forehead bulging, hypertelorism, wide nasal dorsum and bifid nasal tip. Genetic testing revealed that she has harbored a heterozygous missense c.473T>C (p.M158T) variant of the EFNB1 gene, which was detected in either of her parents. By bioinformatic analysis, the variant has not been recorded in the HGMD and ClinVar databases, and no population frequency was recorded in the 1000 Genomes, ExAC, gnomAD and Shenzhou Genome Data Cloud databases. As predicted by the REVEL online software, the variant can confer deleterious effects on the gene or its product. Analysis using UGENE software showed the corresponding amino acid to be highly conserved among various species. Analysis with AlphaFold2 software suggested that the variant may affect the 3D structure and function of the Ephrin-B1 protein. Based on the American College of Medical Genetics and Genomics (ACMG) standards and guidelines and recommendation of Clinical Genome Resource (ClinGen), the variant was rated as pathogenic.@*CONCLUSION@#Combining the patient's clinical features and genetic finding, the diagnosis of CNFS was confirmed. The heterozygous c.473T>C (p.M158T) missense variant of the EFNB1 gene probably underlay the disease in this patient. Above finding has provided a basis for the genetic counseling and prenatal diagnosis for her family.

Progress on the effect of mesenchymal stem cell derived exosomes on multiple organ dysfunction in sepsis / 中华危重病急救医学

Mesenchymal stem cell derived (MSC) exosomes are extracellular vesicles with a diameter of about 50-200 nm. Exosomes contain a large number of biologically active substances including mRNAs, miRNAs, cytokines, chemokines, proteins, lipids, etc. MSC exosomes exert biological effects through paracrine and endocrine pathways in vivo. Uncontrolled inflammation and multiple organ dysfunction are the key roles in the progression of sepsis, moreover, heart, lungs, kidneys and brain are the general target organs to be damaged. MSC exosomes regulate the expression of cytokines, the production of inflammatory cell, the levels of inflammatory response and the recovery of damaged tissues or organ function. Therefore, studying the application of MSC exosomes is significant for the clinical treatment of sepsis. This article reviews the systemic inflammation regulation by MSC exosomes and its protective function on targeted organs such as heart, lungs, kidneys, brain, liver, etc. to provide evidences for the treatment of sepsis.

Effect of chloroquine on proliferation of triple negative breast cancer MDA-MB-231 cells and its underlying mechanism / 中国药理学与毒理学杂志

OBJECTIVE To explore the effect of chloroquine(CQ) on the proliferation of triple negative breast cancer MDA-MB-231 cells and its underlying mechanism. METHODS MDA-MB-231 cells were treated with CQ 20, 40 and 80 μmol • L-1 for 24 and 48 h, respectively. CCK-8 and colony formation assays were used to detect the proliferation of MDA-MB-231 cells. Flow cytometry was employed to analyze cell cycle and apoptosis. Western blotting was used to detect the expressions of cell cycle proteins, apoptosis-related proteins and autophagy-related markers. RESULTS CQ 20, 40 and 80 μmol • L-1 could effectively inhibit the proliferation of MDA-MB-231 cells both at 24 and 48 h. Compared with the cell control group, the percentage of G0/G1 cells significantly increased when exposed to CQ 20 and 40 μmol•L-1 for 24 h (P<0.01), the percentage of G2/M cells increased (P<0.01) and the apoptosis was relatively up-regulated (P<0.01) after treatment with CQ 80 μmol • L-1 for 24 h. The apoptosis was significantly elevated after exposure to CQ 20, 40 and 80 μmol•L-1 for 48 h in a concentration-dependent manner (P<0.05). After treatment with CQ, the expression of cell cycle proteins, such as cyclin-dependent kinase-2 (CDK2), CDK4 and cyclin D3, was decreased (P<0.01), the expression of apoptosis-related proteins cleaved-caspase 3 and cleaved- poly-ADP-ribose polymerase was both elevated (P<0.01), and the expression of autophagy-related markers microtubule-associated proteinl light chain 3 and SQSTM1 was both increased (P<0.01). CONCLUSION CQ can effectively suppress the proliferation of TNBC MDA-MB-231 cells by inhibiting autophagy, inducing cell cycle arrest and promoting apoptosis.

Cassane diterpenes from the seeds of Caesalpinia decapetala / 药学学报

Four casssane diterpenes were isolated from the ethanol extract of the seeds of Caesalpinia decapetala (Fabaceae), with a combination of various chromatographic approaches, including silica gel, Sephadex LH-20, reverse phase C18 and so on. On the basis of spectroscopic data analysis, they were identified as caesalpinin MQ (1), neocaesalpin N (2), caesalmin F (3) and α-caesalpin (4). Among them, compound 1 is a new diterpene, compounds 2-4 were isolated for the first time from the plant of C. decapetala.

Prokaryotic soluble expression of protein D of Haemophilus influenzae type b / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To express the recombinant D protein in prokaryotic expression system solubly and make preparation for producing D-carrier conjugate vaccine next step.</p><p><b>METHODS</b>The hpd gene fragment removed of signal peptide from genomic DNA of Hib CMCC was inserted into pET43. 1a. The recombinant plasmid was transformed to competent E. coli BL21 (DE3) for expression under induction of IPTG. The expressed recombination protein was precipitated with ammonium sulfate, purified by DEAE anion exchange column chromatography and identified for reactogenicity by Western Blot.</p><p><b>RESULTS</b>The expressed recombination protein, in a soluble form, constained about 50% of total somatic protein and showed specific reaction with the HIB antisera after preliminary purification.</p><p><b>CONCLUSION</b>The D protein recombined expression plasmid was constructed successfully and expressed D protein in prokaryotic cells in a solube form.</p>

Myocardial uptake of thiopental in the isolated rabbit heart:the study of pharmacokinetics and pharmacodynamics / 中华麻醉学杂志

To study the dynamic process of myocardial uptake of thiopentai in the isolated rabbit hearts. Method: Thiopental at doses of 500?mol, 1500?mol and 500?mol was given sequentially to the perfused rabbit heart in a total time of 15 min. The outflow concentration of thiopental was measured with high performance liquid chromatography and the left ventricular +dp/dtmax served as a effective parameter. Resuh: The disposition and elimination of thiopental can be best described hy a two-compartment open model. It can disposed into myocardium rapidly (T_(1/2)?=0.5?0.1 min), but elimination was relatively slow (T_(1/2)?=25.3?10.1 min). The transfer rate was slower from peripheral to central compartment than from central to peripheral compartment. The tbeoritical maximum depressant effect of thiopental on + dp/dt (Emax) was 19.0 4-11.2 kPa.s~(-1) corresponding to 1/10 E_0. Conclusion: The myocardial uptake of thiopental can be fitted to a two-compartment open model with rapid disposition and relative slow elimination process.