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Objective: To investigate the effect of the AML1-ETO (AE) fusion gene on the biological function of U937 leukemia cells by establishing a leukemia cell model that induces AE fusion gene expression. Methods: The doxycycline (Dox) -dependent expression of the AE fusion gene in the U937 cell line (U937-AE) were established using a lentivirus vector system. The Cell Counting Kit 8 methods, including the PI and sidanilide induction, were used to detect cell proliferation, cell cycle-induced differentiation assays, respectively. The effect of the AE fusion gene on the biological function of U937-AE cells was preliminarily explored using transcriptome sequencing and metabonomic sequencing. Results: ①The Dox-dependent Tet-on regulatory system was successfully constructed to regulate the stable AE fusion gene expression in U937-AE cells. ②Cell proliferation slowed down and the cell proliferation rate with AE expression (3.47±0.07) was lower than AE non-expression (3.86 ± 0.05) after inducing the AE fusion gene expression for 24 h (P<0.05). The proportion of cells in the G(0)/G(1) phase in the cell cycle increased, with AE expression [ (63.45±3.10) %) ] was higher than AE non-expression [ (41.36± 9.56) %] (P<0.05). The proportion of cells expressing CD13 and CD14 decreased with the expression of AE. The AE negative group is significantly higher than the AE positive group (P<0.05). ③The enrichment analysis of the transcriptome sequencing gene set revealed significantly enriched quiescence, nuclear factor kappa-light-chain-enhancer of activated B cells, interferon-α/γ, and other inflammatory response and immune regulation signals after AE expression. ④Disorder of fatty acid metabolism of U937-AE cells occurred under the influence of AE. The concentration of the medium and short-chain fatty acid acylcarnitine metabolites decreased in cells with AE expressing, propionyl L-carnitine, wherein those with AE expression (0.46±0.13) were lower than those with AE non-expression (1.00±0.27) (P<0.05). The metabolite concentration of some long-chain fatty acid acylcarnitine increased in cells with AE expressing tetradecanoyl carnitine, wherein those with AE expression (1.26±0.01) were higher than those with AE non-expression (1.00±0.05) (P<0.05) . Conclusion: This study successfully established a leukemia cell model that can induce AE expression. The AE expression blocked the cell cycle and inhibited cell differentiation. The gene sets related to the inflammatory reactions was significantly enriched in U937-AE cells that express AE, and fatty acid metabolism was disordered.
Subject(s)
Humans , U937 Cells , RUNX1 Translocation Partner 1 Protein , Leukemia/genetics , Core Binding Factor Alpha 2 Subunit/genetics , Oncogene Proteins, Fusion/genetics , Leukemia, Myeloid, Acute/geneticsABSTRACT
AIM: To investigate the expression and clinical significance of interferon regulatory factor 4(IRF4)and soluble suppression of tumorigenesis 2(sST2)in conjunctival epithelial cells and tears of patients with dry eye.METHODS: A total of 94 patients with dry eye who admitted to our hospital from January 2019 to December 2021 were selected as the dry eye group, and 97 physical examiners who underwent ophthalmic examination were selected as the control group at the same time. The conjunctival epithelial cells and tears of the subjects were collected, and the clinical indicators, including tear film break-up time(BUT), corneal fluorescein staining(CFS)score, and Schirmer Ⅰ test(SⅠt)were recorded. The levels of IRF4 and sST2 in conjunctival epithelial cells were detected by quantitative real-time polymerase chain reaction(qRT-PCR), and the levels of IRF4 and sST2 in tears were detected by enzyme-linked immunosorbent assay(ELISA). Pearson method was used to analyze the correlation between IRF4 and sST2 levels in conjunctival epithelial cells and tears and clinical indicators of dry eye patients.RESULTS: The levels of IRF4 and sST2 in conjunctival epithelial cells and tears in dry eye group before treatment were significantly higher than those in control group(P<0.001). The levels of IRF4 and sST2 in conjunctival epithelial cells and tears of dry eye patients at 4wk after treatment were significantly lower than those before treatment(P<0.001). The BUT and SⅠt of dry eye patients increased significantly at 4wk after treatment, and the CFS score decreased significantly(P<0.001). The levels of IRF4 and sST2 in conjunctival epithelial cells and tears of dry eye patients before treatment were positively correlated with CFS score before treatment and negatively correlated with BUT and SⅠt before treatment(P<0.001).CONCLUSION: The levels of IRF4 and sST2 in conjunctival epithelial cells and tears of patients with dry eye are increased, and are significantly correlated with BUT, SⅠt and CFS scores, which has potential to become a new therapeutic target for dry eye.
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OBJECTIVES@#To study new biomarkers for the early diagnosis of retinopathy of prematurity (ROP) by analyzing the differences in blood metabolites based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) and metabolomics.@*METHODS@#Dried blood spots were collected from 21 infants with ROP (ROP group) and 21 infants without ROP (non-ROP group) who were hospitalized in the Sixth Affiliated Hospital of Sun Yat-sen University from January 2013 to December 2016. LC-MS/MS was used to measure the metabolites, and orthogonal partial least squares-discriminant analysis was used to search for differentially expressed metabolites and biomarkers.@*RESULTS@#There was a significant difference in blood metabolic profiles between the ROP and non-ROP groups. The pattern recognition analysis, Score-plot, and weight analysis obtained 10 amino acids with a relatively large difference. Further statistical analysis showed that the ROP group had significant increases in blood levels of glutamic acid, leucine, aspartic acid, ornithine, and glycine compared with the non-ROP group (P<0.05). The receiver operating characteristic curve analysis showed that glutamic acid and ornithine had the highest value in diagnosing ROP.@*CONCLUSIONS@#Blood metabolites in preterm infants with ROP are different from those without ROP. Glutamic acid and ornithine are the metabolic markers for diagnosing ROP. LC-MS/MS combined with metabolomics analysis has a potential application value in the early identification and diagnosis of ROP.
Subject(s)
Infant, Newborn , Infant , Humans , Tandem Mass Spectrometry , Infant, Premature , Chromatography, Liquid , Retinopathy of Prematurity/diagnosis , Glutamic Acid , OrnithineABSTRACT
Objective:To study the genetic profile of neonatal hyperbilirubinemia with unknown etiology in Guangdong Province and the clinical significance of jaundice-related genetic screening.Methods:From July to September, 2021, neonates with hyperbilirubinemia of unknown etiology born in different hospitals in Guangdong Province were studied. 24 neonatal jaundice-related exons were sequenced using targeted capture and high-throughput sequencing technology. The pathogenic variants were analyzed.Results:A total of 331 cases, 139 (42.0%) cases showed positive screening results with five diseases, including 65 (19.6%) cases of Gilbert syndrome, 48 (14.5%) cases of glucose-6-phosphate dehydrogenase (G6PD) deficiency,18 (5.4%) cases of sodium taurocholate cotransporting polypeptide deficiency, 4 (1.2%) cases of Citrin deficiency and 4 (1.2%) cases of Dubin-Johnson syndrome. 149 (45.0%) cases carried one or more genetic variants and 43 (13.0%) cases showed no clinically significant variants. The 8 high-frequency mutation loci (carrier rate >1%) are UGT1A1 gene c.211G>A and c.1091C>T, G6PD gene c.1466G>T and c.1478G>A, SLC10A1 gene c.800C>T, SLC25A13 gene c.852_855del TATG, HBB gene c.126_129delCTTT and c.316-197C>T.Conclusions:Genetic factors are important for neonatal hyperbilirubinemia with unknown etiology in Guangdong. The common pathogenic genes are UGT1A1, G6PD, SLC10A1, and SLC25A13 and the population carries high-frequency mutation loci. Therefore, genetic screening in neonates with hyperbilirubinemia of unknown etiology has important clinical significance.
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Objective:To detect the genes of common genetic diseases in newborns with the high-throughput sequencing technology based on target gene capture, to study the incidence rate of such diseases, the carrying rate and variant types of pathogenic mutations related to such diseases, and to explore the application value of the high-throughput sequencing technology in screening genetic diseases of newborns.Methods:The heel blood of 1 793 newborns born in Guangdong province from June 2019 to April 2020 were collected, and the exon regions of 138 common genetic disease-related genes in neonates were detected using the high-throughput sequencing technology based on target gene capture.The pathogenicity of the mutations was interpreted according to the " Classification Criteria and Guidelines for Genetic Variation(2017)" , in which known disease and probable disease were considered as positive mutations.The positive mutations were verified by Sanger sequencing technology, and the test results were analyzed with statistical methods.Results:Among the 1 793 newborns, 978 were male and 815 were female.A total of 158 positive cases were screened(8.81%), and 11 positive diseases were detected.Among the positive diseases, there were 41 cases(2.29%)of autosomal recessive deafness type 1A, 40 cases(2.23%)of Gilbert syndrome or Crigler-Najjar syndrome, and 33 cases(1.84%)of glucose-6-phosphate dehydrogenase deficiency(1.84%), 19 cases(1.06%)of familial hypercho-lesterolemia, 18 cases(1.00%) of sodium taurocholate cotransporter peptide deficiency disease, 2 cases(0.11%)of mitochondrial non-syndromic deafness, 2 cases(0.11%)of Citrin deficiency, 1 case(0.06%)of holocarboxylase synthase deficiency, 1 case(0.06%)of β-thalassemia and 1 case(0.06%)of metachromatic leukodystrophies.Of all studied cases, 972 carried one or more positive mutations, involving 85 kinds of diseases in total.The diseases with a high carrying rate were Gilbert syndrome or Crigler-Najjar syndrome(359 cases, 20.02%), autosomal recessive deafness type 1A(302 cases, 16.84%), and sodium taurocholate cotransport peptide deficiency disease(291 cases, 16.22%). The high-frequency mutation sites were UGT1A1 gene c. 211G> A, GJB2 gene c .109G> A and SLC10A1 gene c. 800C> T. Conclusions:The common genetic diseases detected in neonates from Guangdong province are autosomal recessive deafness type 1A, Gilbert syndrome or Crigler-Najjar syndrome, glucose-6-phosphate dehydrogenase deficiency, familial hypercholesterolemia, and sodium taurocholate cotransport peptide deficiency.There are high-frequency carrying mutation sites in the population.Preliminary genetic screening of common neonatal genetic diseases can accumulate data and experience for the development of newborn genetic screening.
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Objective:To compare the difference of blood amino acids and acylcarnitine levels between small-for-gestational-age (SGA) and appropriate-for-gestational age (AGA) full-term newborns, and to explore the changes of the blood metabolism spectrum of full-term SGA, so as to provide evidence for clinical intervention.Methods:Seventy-nine full-term SGA newborns born in the Sixth Affiliated Hospital of Sun Yat-Sen University from January to December 2018 were selected as the study objects.Seventy-nine gestational age-and gender-matched healthy full-term AGA newborns born in the same hospital at the same time were selected as the control group.The dry blood spot samples were collected and detected by tandem mass spectrometry on the third day after birth.The differences between two groups and considerable biomarkers were explored by the orthogonal partial least squares discriminant analysis (OPLS-DA).Results:The birth weight of SGA newborns was (2.5±0.2) kg, and that of AGA newborns was (3.2±0.3) kg.OPLS-DA model analysis showed that 12 kinds of blood metabolites were identified which possessed the biggest weight discriminating the full-term SGA group from the AGA group, and the ratios of these blood metabolites of two groups were compared as follows: propionylcarnitine (0.34±0.13 vs. 0.42±0.15), tyrosine [0.24(0.18, 0.27) vs.0.28(0.22, 0.37)], free carnitine (0.43±0.14 vs. 0.37±0.12), valine [0.39(0.35, 0.45) vs.0.44(0.36, 0.53)], octanoylcarnitine (0.33±0.13 vs. 0.29±0.09), myristoylcarnitine (0.35±0.12 vs. 0.31±0.10), butylcartine (0.37±0.13 vs. 0.41±0.14), 3-hydroxyisovlerylcartine[0.35(0.25, 0.43) vs.0.35(0.26, 0.45)], decenoylcarnitine (0.26±0.13 vs. 0.23±0.08), isovalerylcarnitine[0.33(0.26, 0.34) vs.0.33(0.30, 0.35)], leucine [0.38(0.30, 0.47) vs.0.40(0.33, 0.48)]and methionine (0.42±0.14 vs. 0.46±0.15). The level of propionylcarnitine ( t=3.920), tyrosine ( Z=3.536) and valine ( Z=2.838) in the full-term SGA group were significantly lower than those in the AGA group, while the levels of free carnitine ( t=-2.863), octanoylcarnitine ( t=-2.266) and myristoylcarnitine ( t=-2.194) in the full-term SGA group were significantly higher than those in the AGA group (all P<0.05). Conclusions:The concentration of amino acids and acylcarnitine in the blood of SGA newborns is different from that in AGA newborns.Aromatic amino acids and branched chain amino acids should be added in full-term SGA nutrition support as they can meet the energy metabolism by mobilizing medium and long chain fatty acids in the early stage.
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Objective To analyze the changes in blood metabolites in premature infants with bronchopulmonary dysplasia (BPD) within 36 h and in the 3rd week after birth in order to find new biomarkers for diagnosis of BPD.Methods The BPD group included 20 premature infants (＜32 gestational weeks) hospitalized in the Neonatal Intensive Care Unit (NICU) of the Sixth Affiliated Hospital of Sun Yat-sen University and diagnosed with BPD from January 2014 to October 2016.Another 20 non-BPD premature infants with similar gestational age (within one week) who were admitted during the same period were enrolled in the control group.Blood samples of both groups were collected within 36 h and in the 3rd week after birth.Liquid chromatography-tandem mass spectrometry was used to detect blood metabolites and the obtained data were subjected to metabolomics analysis using orthogonal partial least squares discriminant analysis.Chi-square test (or Fisher's exact test),Mann-Whitney U test or t test was used for statistical analysis.Results (1) Twenty and 11 blood samples were collected within 36 h and in the 3rd week after birth from the BPD and the control group,respectively.Compared with the control group,the interval between premature rupture of membranes and delivery,the average length of hospital stay,non-invasive and invasive mechanical ventilation duration and the total duration of supplemental oxygen during hospitalization in the BPD group were longer [M (P25-P75) or ((x)±s):13.5 (0.0-98.3) vs 0.0 (0.0-0.0) h,Z=3.049;(66.6±20.5) vs (43.9±9.3) d,t=4.574;267.0 (199.5-516.1) vs 110.5 (0.0-238.5) h,Z=-3.428;117.5 (0.0-269.3) vs 0.0 (0.0-72.0) h,Z=-2.785;(1 184.0±386.6) vs (595.9±270.3) h,t=5.576;all P＜0.05].(2) Within 36 h after birth,the levels of glycine,proline,tryptophan and piperamide-C5:1 in the BPD group were decreased obviously compared with those in the control group [(201.59±65.01) vs (290.90± 137.56) μmol/L,t=-2.625;103.55 (72.43-434.57) vs 439.48 (103.80-608.98) μ mol/L,Z=-2.245;29.54 (20.30-41.04) vs 47.42 (29.46-73.57) μ mol/L,Z=-2.326;50.04 (35.29-104.78) vs 95.79 (76.21-129.97) μmol/L,Z=-2.029;all P＜0.05].However,the glutamate level was increased [(224.30±67.40) vs (182.67±40.87) μmol/L,t=2.362,P＜0.05].(3) In the 3rd week after birth,the levels of glycine,proline and tryptophan in the BPD group were lower compared to those in the control group [(185.92±61.51) vs (271.85± 115.85) μmol/L,t=-2.177;(39.41± 18.22) vs (63.92± 17.50) μ mol/L,t=-3.217;90.23 (37.93-146.37) vs 330.15 (47.79-622.90) μ mol/L,Z=-2.134;all P＜0.05].However,the ornithine level was higher [(75.09± 43.21) vs (39.25 ± 16.53) μ mol/L,t=2.569,P＜0.05].Conclusions Glycine,proline and tryptophan in blood are potential biomarkers for early diagnosis of BPD.
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OBJECTIVE@#To construct a W203X-mutant mouse model of cblC type methylmalonic acidemia based on the CRISPR/Cas9 technology.@*METHODS@#At first, BLAST was used to compare the conservative nature of the cblC gene and protein sequences in humans and mice, and then, the CRISPR/Cas9 technology was used for microinjection of mouse fertilized eggs to obtain heterozygous F1 mice. Hybridization was performed for these mice to obtain homozygous W203X-mutant mice. The blood level of the metabolite propionyl carnitine (C3) was measured for homozygous mutant mice, heterozygous littermates, and wild-type mice.@*RESULTS@#The gene and protein sequences of MMACHC, the pathogenic gene for cblC type methylmalonic acidemia, were highly conserved in humans and mice. The homozygous W203X-mutant mice were successfully obtained by the CRISPR/Cas9 technology, and there was a significant increase in C3 in these mice at 24 hours after birth (P<0.001).@*CONCLUSIONS@#A W203X-mutant mouse model of cblC type methylmalonic acidemia is successfully constructed by the CRISPR/Cas9 technology.
Subject(s)
Animals , Mice , Amino Acid Metabolism, Inborn Errors , CRISPR-Cas Systems , Carrier Proteins , Heterozygote , MutationABSTRACT
OBJECTIVE@#To analyze the changes in tumor lymphatic vessel density (LVD) in patients with lung adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA), and invasive adenocarcinoma (IA) and explore the regulatory factors of LVD.@*METHODS@#Complete clinicopathological data were collected form a total of 301 patients with lung adenocarcinoma, including 28 (9.3%) with AIS, 86 (28.6%) with MIA, and 187 (62.1%) with IA. The LVD of all the adenocarcinomas were calculated after D2-40 immunohistochemical staining, and MT1-MMP and VEGF-C expression levels were also evaluated. The differences in LVD among the groups and the correlations of tumor LVD with the expressions of MT1-MMP and VEGF-C and the clinicopathological factors were analyzed.@*RESULTS@#The LVD differed significantly among AIS, MIA, and IA groups (= 0.000). The LVDs was significantly correlated with the level of VEGF-C protein expression (=0.917, =0.009), tumor size (= 0.686, =0.017), lymph node metastasis (=0.739, =0.000), and clinical stage (=0.874, =0.012) of the patients.@*CONCLUSIONS@#Tumor lymphangiogenesis plays an important role in lung adenocarcinoma progression, and VEGF-C may promote this process.
Subject(s)
Humans , Adenocarcinoma , Chemistry , Pathology , Adenocarcinoma of Lung , Chemistry , Pathology , Immunohistochemistry , Lymphangiogenesis , Lymphatic Vessels , Chemistry , Pathology , Neoplasm Staging , Prognosis , Tumor Burden , Vascular Endothelial Growth Factor CABSTRACT
Aim To observe the protective effects of cordycepin ( Cor) on dopaminergic neurons in 1-meth-yl-4-phenyl-1 , 2 , 3 , 6-tetrahydropyridine ( MPTP )-in-duced mouse model of Parkinson's disease ( PD) and to explore its mechanism. Methods C57BL/6 mice were administered with MPTP to establish the PD mod-el. Mice in Cor groups were pretreated with Cor (2.5,5.0 and 10.0 mg·kg-1 ) by intragastric administra-tion, respectively. The motor functions of the mice were observed in the open-field test, rotarod test and pole test. The content of DA, the numbers of TH-im-munoreactive cells and apoptotic cells were measured respectively by HPLC-ECD, immunohistochemistry staining and TUNEL staining. The expression of apop-tosis related proteins and MAPK signaling pathway-re-lated proteins ( p38 , p-p38 , ERK1/2 , p-ERK1/2 JNK1/2 and p-JNK1/2 ) were determined by Western blot. Results Cor could significantly improve the mo-tor dysfunction in PD mice. The contents of DA, DOPAC and HVA in the striatum remarkably increased after administration of Cor in MPTP-induced mice. Mo-reover, Cor could obviously reduce both the loss of TH-immunoreactive neurons and the numbers of TUNEL-positive cells in substantia nigra pars compacta ( SNpc) of PD mice. The protein expressions of Bax and cleaved caspase-3 were markedly down-regulated,whereas those of Bcl-2 and the ration of Bcl-2/Bax were significantly up-regulated by Cor pre-treatment followed by MPTP treatment. Furthermore, the protein expressions of p-p38 , p-ERK1/2 and p-JNK1/2 signif-icantly decreased in substantia nigra in Cor groups. Conclusions The results suggest that Cor can protect DA neurons against MPTP-induced injury by inhibiting apoptosis, which may be closely relevant to the inhibi-tion of MAPK signaling pathways.
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<p><b>OBJECTIVE</b>To investigate the effects of nerve growth factor (NGF) in the osteogenic action of implants and the maturation and reconstruction changes in bone tissues in the early stage of osseointegration.</p><p><b>METHODS</b>The mouse implant model was established by placing titanium in the femoral head of the mouse and locally injecting NGF in the implant zone. On 1, 2 and 4 weeks after operation, stain samples were collected from animals using hematoxylin-eosin (HE) staining and Masson staining. The effect of NGF on the bone maturation was compared at different time points of early stage osseointegration.</p><p><b>RESULTS</b>The results of HE and Masson staining indicated that the local injection of external NGF can up-regulate bone mass, amount of bone trabecula, and bone maturity in the mouse model. The mature bone rate in treatment group of 1 week and 4 weeks after operation were significantly higher than those in the control group (P<0.05).</p><p><b>CONCLUSIONS</b>NGF can shorten the period of bone maturation.</p>
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ObjectiveTo investigate the protective effect of microRNA-155 (miR-155) antisense oligonucleotid (ASO) on acute lung injury (ALI) mice by establishing a lentiviral expression vector of ASO of miRNA.Methods miR-155 antisense oligonucleotides amplified by polymerase chain reaction (PCR) from genomic, using BamH Ⅰ and Nhe Ⅰ double digestion, ligated into lentiviral expression vector. Sequence and virus titer were measured. According to the random number table method, 54 male BALB/c mice of 4-6 weeks old were divided into three groups. ALI animal models were prepared by intraperitoneal injection of 10 mg/kg lipopolysaccharide (LPS). The three groups were injected with 200μL phosphate buffered saline (PBS) containing 1×108/mL pmiR-155-ASO virus (pmiR-155-ASO group) or 200μL PBS containing 1×108/mL pSMPUW-miR-GFP empty virus (pmiR-cont group) or the same amount of PBS (PBS group) at 24 hours before the molding. Ten mice in each group were used to observe the 7-day survival rate. Blood samples and lung tissues of the remaining 8 mice were harvested after the model was established, and the levels of serum inflammatory cytokines were determined by enzyme linked immunosorbent assay (ELISA); the expression of miR-155 in lung tissue was detected by real-time reverse transcription-polymerase chain reaction (RT-PCR); histopathological changes of lung and distribution of macrophages were observed under microscope.Results There was no significant difference in each index between pmiR-cont group and PBS group. The mature miR-155 expression in lung tissue in pmiR-155-ASO group was significantly lower than that in pmiR-cont group (2-ΔΔCt: 4.92±0.72 vs. 15.38±0.60,P < 0.05). Compared with pmiR-cont group, the injury degree of ALI mice after pretreatment with miR-155ASO was significantly improved, and the 7-day survival rate was significantly increased (72.1％ vs. 61.9％,P < 0.05 ); gross lung observation showed that congestion in lung tissue was significantly reduced, and the ratio of wet/dry weight (W/D) of lung was significantly decreased (4.50±0.13 vs. 5.64±0.61,P < 0.05);hematoxylin-eosin (HE) staining showed that inflammatory cell infiltration in lung tissue was decreased, while immunofluorescence assay showed that macrophage infiltration in lung tissue was significant decreased; the levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL- 6) in serum were significantly decreased [TNF-α (ng/L):379.8±48.9 vs. 495.9±33.3, IL-6 (ng/L): 262.3±61.8 vs. 355.4±22.6, bothP < 0.05], but the level of IL-10 did not change significantly (ng/L: 143.6±32.5 vs. 140.4±22.3,P > 0.05).Conclusion miR-155 ASO has the effect of inhibiting LPS-induced inflammatory response and improving prognosis in ALI mice.
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The clinical manifestations,diagnosis and treatment process of the Gnathostoma infected patient were collected,and epidemiological investigation was carried out.The investigation results showed that the patients with eating wild boar in stomach nematode,the worms were removed by gastroscopy and examined by microscopy,small spines in the body,the spines of the posterior part and the posterior part of the body are thinner.The patient was confirmed cases of infection by Gnathostoma.
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<p><b>OBJECTIVE</b>To investigate the effects of serum procalcitonin(PCT) levels for predicting the outcome of bacteria bloodstream infection in acute leukemia patients.</p><p><b>METHODS</b>Clinical data from 236 patients with acute leukemia accompanied by bacterial bloodstream infection during July 2014 to November 2017 were retrospectively analyzed, 236 patients were divided into 5 groups (<0.05 ng/ml, 0.05- <0.5 ng/ml, 0.5- <2.0 ng/ml, 2.0- <10.0 ng/ml and >10.0 ng/ml) according to PCT concentrations.</p><p><b>RESULTS</b>The median age of patients was 40(13-73) years old. The male 123 cases(52.1%) and female 113 cases(47.9%) in 236 patients. The incidence of infection-related dealth in 5 groups was 0%, 1.4%, 13.8%, 25.0% and 33.3%, respectively; the incidence of septic shock and other serious complications in 5 groups was 0%, 2.1%, 13.8%, 25.0%, 33.3% and 6.4%, 7.0%, 24.1%, 41.7%, 50.0%, respectively, showing the concentration dependent manner and statistically significant difference (u=2127, P=0.000; u=2234, P=0.000; u=4102, P=0.000). Further analysis showed that with the increase of PCT concentration, the cumulative incidence of septic shock, infection-related death and other serious complications was gradually increased with statistically significance (HR=2.887, P=0.000, 95%CI:1.960-4.260; HR=3.158, P=0.000, 95%CI: 2.100-4.740; HR=2.158, P=0.000, 95%CI:1.550-3.000) respectively. Increased procalcitonin level is an independent risk factor for septic shock and infection-related death (HR=2.517, P=0.000, 95%CI: 1.520-4.168; HR=2.881, P=0.000, 95%CI: 1.692-4.904)respectively.</p><p><b>CONCLUSION</b>Serum procalcitonin level positively correlates with the incidence of serious bacteria bloodstream infection complications in the patients with acute leukemia. Increased procalcitonin level is an independent risk factor for septic shock and infection-related death, indicating that procalcitonin may be an important prognostic factor for infection outcome in acute leukemia patients with bacteremia.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Bacteremia , Biomarkers , C-Reactive Protein , Calcitonin , Calcitonin Gene-Related Peptide , Protein Precursors , Retrospective StudiesABSTRACT
<p><b>OBJECTIVE</b>To evaluate the efficacy of primary prophylaxis of voriconazole against invasive infection of pulmonary aspergillosis (IPA) during remission-induction chemotherapy (RIC) of patients with acute myeloid leukemia (AML).</p><p><b>METHODS</b>Clinical data of 102 de novo AML patients who received primary anti-IPA prophylaxis during the first induction chemotherapy were analyzed retrospectively. All the cases were divided into voriconazole-treated group and posaconazole-treated group according to the prophylactic agent. The incidences of IPA and systemic antifungal treatment during induction chemotherapy were analyzed for both groups.</p><p><b>RESULTS</b>Among 102 enrolled cases, 42 cases received voriconazole and other 60 received posaconazole as primary prophylaxis. IPA occurred in 3 cases of voriconazole group (1 probable, 2 possible); IPA occurred in 4 cases of posaconazose group, and all were possible cases. The incidence of IPA during remission-induction chemotherapy in variconazole group equaled to posaconazose group (7.1% vs. 6.7%) (P=0.925). Beside IPA cases, 2 cases in voriconazole group and 4 cases in posaconazole group received intravenous anti aspergillosis drugs preemptive treatment, and no significant difference of prophylactic success rate was observed between two groups (88.1% vs. 86.7%) (P=0.831). Visual disturbance was the most common adverse event occurred in voriconazole group, but no significant differences of incidences of other adverse effects were observed when compared with posaconazole group.</p><p><b>CONCLUSION</b>According to similar prophylactic effect with posaconazole, voriconazole appears to be a good alternative for primary prophylaxis of IPA during remission-induction chemotherapy in AML patients.</p>
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<p><b>OBJECTIVE</b>To investigate the tumor inhibition effect of Yangfei Kongliu Formula (YKF), a compound Chinese herbal medicine, combined with cisplatin (DDP) and its action mechanisms.</p><p><b>METHODS</b>C57BL/6 mice with Lewis lung carcinoma were divided into six groups: control group (C), DDP group (2 mg/kg, DDP), low-dose YKF group (2.43 g/kg, L), high-dose YKF group (24.3 g/kg, H), low-dose YKF combined with DDP group (L + DDP) and high-dose YKF combined with DDP group (H + DDP). Transforming growth factor-β1 (TGF-β1), mothers against decapentaplegic homolog 3 (Smad3) and Smad7 levels were measured with quantitative real-time polymerase chain reaction (qPCR), Western blotting and immunohistochemistry. An enzyme-linked immunosorbent assay was used to analyze the expressions of interleukin-2 (IL-2) and tumor necrosis factor-α (TNF-α).</p><p><b>RESULTS</b>YKF combined with DDP significantly inhibited the growth and metastasis of tumors relative to the control group, and YKF groups (P < 0.05). There was no significant difference between high-dose YKF group and low-dose YKF group (P > 0.05). We also found that the expression levels of TGF-β1 and Smad3 were both significantly decreased by YKF relative to the control group (P < 0.05). Furthermore, after treatment with YKF combined with DDP, the expression levels of TGF-β1 and Smad3 were decreased but the expression level of Smad7 was increased relative to the DDP group (P < 0.05). Compared to the DDP group, the combination of YKF and DDP enhanced the effect of tumor inhibition (P < 0.05), showing obvious synergy between YKF and DDP. Treatment with DDP or YKF decreased serum levels of IL-2 and TNF-α relative to the control group (P < 0.05). Furthermore, the expression levels of IL-2 and TNF-α were significantly decreased when treated with YKF in combination with DDP. Co-treatment with YKF and DDP significantly inhibited tumor growth, decreased the expressions of TGF-β1, Smad3, IL-2 and TNF-α and increased the expression of Smad7; these differences were significant relative to both YKF groups and the control group (P < 0.05).</p><p><b>CONCLUSION</b>YKF can inhibit tumor growth synergistically with DDP, mainly through the TGF-β1 signaling pathway.</p>
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Objective To investigate the mechanism of natural carboxyl-terminal truncated HBx(with 31 amino acids deleted at the C-terminal end) (HBxΔ31)-dependent down-regulation of Rho GDP dissociation inhibitor alpha (RhoGDIα) expression and its role in enhancing hepatocellular carcinoma (HCC) metastasis. Methods HepG2 cells with stable expression of wild type HBx and its deletion mutant HBxΔ31 protein were selected as the study subjects. The effects of HBx and HBxΔ31 on RhoGDIα expression in HepG2 cells was detected using real-time quantitative reverse transcription PCR(RT-qPCR)and Western blot analysis.A series of deleted and mutated variants of the RhoGDIα promoter were made and individually co-transfected with HBx-and HBxΔ31-expressing vectors or control empty vector into HepG2 cells. The luciferase reporter assay was used to identify the cis-regulatory elements of the RhoGDIα promoter in response to HBxΔ31 regulation.The interaction between Myc-associated zinc finger protein (MAZ) and HBxΔ31 was examined by co-immunoprecipitation experiment. Eelectrophoretic mobility shift assay (EMSA) was performed to determine interaction of MAZ with the RhoGDIα promoter in HBxΔ31-expressing HepG2 cells. The impact of reduced RhoGDIα expression by HBxΔ31 on cell-invasive activity was analyzed by the Matrigel cell invasion assay. In addition, the effects of silencing of shRNA-mediated RhoGDIα in HepG2 or introduction of RhoGDIα by transfection in HBxΔ31-expressing HepG2 cells on cell-invasion were investigated.Results HBxΔ31,but not HBx, suppressed RhoGDIα expression at transcriptional levels. Analysis of the deletion and mutation of RhoGDIα promoter showed that the HBxΔ31 repressive element localized between nt-460 and -242 bp of RhoGDIα promoter and that the transcription factor MAZ binding sites was required for RhoGDIα promoter inactivation regulated by HBxΔ31. In addition, HBxΔ31 represses RhoGDIα expression by enhancing MAZ binding to its promoter through directly associating with MAZ. The cell-migratory and cell-invasive activity were significantly increased in sh-RhoGDIα-expressing HepG2 cells, as compared to control cells (migrated cells number:58±5 vs.98±7,invaded cells number:55±6 vs.113±6,t=18.91 and t=20.12,both P<0.01). However, ectopic expression of RhoGDIα in HBxΔ31-expressing HepG2 cells significantly decreased cell migration and invasion(migrated cells number:40±4 vs.115±5,invaded cells number:42±4 vs.102±4, t=18.14 and t=16.31, both P<0.001). Conclusion Carboxyl-terminal truncated HBx deregulates RhoGDIα expression through MAZ,in turn which promotes the invasion and metastasis of HCC.
ABSTRACT
Objective To investigate the mechanism of natural carboxyl-terminal truncated HBx(with 31 amino acids deleted at the C-terminal end) (HBxΔ31)-dependent down-regulation of Rho GDP dissociation inhibitor alpha (RhoGDIα) expression and its role in enhancing hepatocellular carcinoma (HCC) metastasis. Methods HepG2 cells with stable expression of wild type HBx and its deletion mutant HBxΔ31 protein were selected as the study subjects. The effects of HBx and HBxΔ31 on RhoGDIα expression in HepG2 cells was detected using real-time quantitative reverse transcription PCR(RT-qPCR)and Western blot analysis.A series of deleted and mutated variants of the RhoGDIα promoter were made and individually co-transfected with HBx-and HBxΔ31-expressing vectors or control empty vector into HepG2 cells. The luciferase reporter assay was used to identify the cis-regulatory elements of the RhoGDIα promoter in response to HBxΔ31 regulation.The interaction between Myc-associated zinc finger protein (MAZ) and HBxΔ31 was examined by co-immunoprecipitation experiment. Eelectrophoretic mobility shift assay (EMSA) was performed to determine interaction of MAZ with the RhoGDIα promoter in HBxΔ31-expressing HepG2 cells. The impact of reduced RhoGDIα expression by HBxΔ31 on cell-invasive activity was analyzed by the Matrigel cell invasion assay. In addition, the effects of silencing of shRNA-mediated RhoGDIα in HepG2 or introduction of RhoGDIα by transfection in HBxΔ31-expressing HepG2 cells on cell-invasion were investigated.Results HBxΔ31,but not HBx, suppressed RhoGDIα expression at transcriptional levels. Analysis of the deletion and mutation of RhoGDIα promoter showed that the HBxΔ31 repressive element localized between nt-460 and -242 bp of RhoGDIα promoter and that the transcription factor MAZ binding sites was required for RhoGDIα promoter inactivation regulated by HBxΔ31. In addition, HBxΔ31 represses RhoGDIα expression by enhancing MAZ binding to its promoter through directly associating with MAZ. The cell-migratory and cell-invasive activity were significantly increased in sh-RhoGDIα-expressing HepG2 cells, as compared to control cells (migrated cells number:58±5 vs.98±7,invaded cells number:55±6 vs.113±6,t=18.91 and t=20.12,both P<0.01). However, ectopic expression of RhoGDIα in HBxΔ31-expressing HepG2 cells significantly decreased cell migration and invasion(migrated cells number:40±4 vs.115±5,invaded cells number:42±4 vs.102±4, t=18.14 and t=16.31, both P<0.001). Conclusion Carboxyl-terminal truncated HBx deregulates RhoGDIα expression through MAZ,in turn which promotes the invasion and metastasis of HCC.
ABSTRACT
Cordycepin(3′-deoxyadenosine),a derivative of the nucleoside adenosine,is a major functional component of Cordyceps militaris. It has been found that cordycepin possesses a variety of pharmacological activities,such as antibacterial,antivial, antiinflammatory,antitumor,immunoregulation,hypolipidemic,and hypoglycemic activities. In recent years,the effects of cordyce-pin on the central nervous system(CNS)have attracted great attention,and it has been found that cordycepin could not only affect the function of the CNS but also protect nerves from injuries. This paper reviews the effects of cordycepin on sedation and hypnosis ,the im-provement of learning and memory,and the protection of nerve injuries caused by cerebral ischemia/hemorrhage ,nerve toxin(gluta-mate,β-amyloid,rotenone and 6-hydroxydopamine,etc.),lipopolysaccharide and trauma,along with the in vitro toxicity as well as acute and subacute in vivo toxicity,so as to offer valuable references for future study and application of cordycepin.
ABSTRACT
Objective To investigate the expression of p-MNK1 in non-small cell lung cancer and its relationship with clinic pathological features and prognosis.Methods The level of p-MNK1 in 115 cases of nonsmall cell lung cancer was detected by tissue microarray technique and immunohistochemistry technique.Results The level of p-MNK 1 in non-small cell lung cancer was correlated with cancer tissue type (P ＜ 0.05),clinical stage (P ＜ 0.05),lymph node metastasis (P ＜ 0.01) and prognosis (P ＜ 0.05).Cox multiple regression analysis showed that p-MNK1 expression was an independent prognostic factor for non-small cell lung cancer (P ＜ 0.01).Conclusion The level of p-MNK1 in non-small cell lung cancer is associated with poor prognosis.It can be used as an independent prognostic marker and a new therapeutic target.