Regularity of ventilator-associated pneumonia induced by three common pathogens / 中华危重病急救医学

OBJECTIVE@#To investigate the incidence and infection regularity of ventilator-associated pneumonia (VAP) in patients undergoing tracheal intubation and to provide reference for the prevention and treatment of VAP infection in the future.@*METHODS@#A retrospective study was conducted to collect the microbial data of airway secretion cultures from 72 patients with endotracheal intubation admitted to the emergency ward of Shanghai Fifth People's Hospital from May 2020 to February 2021, and the species of microorganisms and intubation time were statistically analyzed.@*RESULTS@#Among 72 patients with endotracheal intubation, males were more than females (58.33% vs. 41.67%); Patients over 60 years old accounted for 90.28%; pneumonia was the main primary disease, accounting for 58.33%. Pathogenic tests showed that: (1) 72 patients were infected with Acinetobacter baumannii (AB), Klebsiella pneumoniae (KP), and Pseudomonas aeruginosa (PA) 48 hours after intubation, 51.39% (37/72), 27.78% (20/72), and 26.39% (19/72), respectively. The infection rate of AB was significantly higher than that of KP and PA. Within 48 hours of intubation, the infection rates of AB, KP, and PA were 20.83% (15/72), 13.89% (10/72), and 4.17% (3/72), respectively. Of the 42 patients with primary pneumonia, 61.90% (26/42) were infected with one or more of the three pathogenic bacteria AB, KP, and PA 48 hours after intubation, indicating a change in the etiology of the pathogenic bacteria, with the main pathogenic bacteria transitioning from other pathogenic bacteria to AB, KP, and PA. (2) AB, KP, and PA were prone to cause late onset VAP (i.e., intubation ≥ 5 days). Respectively, among VAP patients infected with AB, late onset VAP accounted for 59.46% (22/37). Among patients infected with KP, 75.00% (15/20) had late onset VAP. Among patients infected with PA, late onset VAP accounted for 94.74% (18/19), indicating a higher proportion of late onset VAP caused by PA and KP. (3) Infection was closely related to intubation time, and the pipeline can be replaced according to the peak period of infection. AB and KP infections peaked within 4 days after intubation, reaching 57.69% (30/52) and 50.00% (15/30), respectively. It is recommended to replace the tubes or undergo sensitive antimicrobial therapy around 3-4 days after starting the machine. The proportion of PA infection after 7 days of intubation was 72.73% (16/22), and it was considered to replace the pipeline after 7 days. (4) Most of the three pathogenic bacteria, AB, KP, and PA were carbapenem resistant pathogens with multiple drug resistance. Except for PA, the infection rate of carbapenem resistant bacteria (CRAB, CRKP) was significantly higher than that of non-carbapenem resistant bacteria (AB, KP), accounting for 86.54% (45/52) and 66.67% (20/30) of the corresponding infection cases, respectively, while CRPA only accounts for 18.18% (4/22).@*CONCLUSIONS@#The main differences in VAP infection caused by AB, KP, and PA pathogens are infection time, infection probability, and carbapenem resistance. Targeted prevention and treatment measures can be implemented for patients with intubation.

Different strategies of reducing non-viral nucleic acid of meta-virome in plasma: a comparative study / 中国输血杂志

【Objective】 To explore the influence of common methods of reducing non-viral nucleic acid on the abundance of plasma virus group. 【Methods】 Three kinds of library construction, five kinds of centrifugation conditions, two kinds of filters, four kinds of enzymes and four concentrations of chloroform were used to treat plasma samples added quantitatively 2.16 mL of pseudorabies virus(PRV) and 2.16 mL of porcine parvovirus(PPV). A total of 21.6 mL of plasma samples were processed, including 54 samples. Subsequently, nucleic acid was extracted, mitochondrial DNA(mtDNA) and two viruses were quantitated, the library of the next generation sequencing was constructed, Illumina NovaSeq 6000 was used for the next generation sequencing. The sequencing data were compared with Kraken Py 2.0 software, and the species annotation analysis was conducted. The corresponding species classification information of each segment was obtained to analyze the impact of different reducing non-viral nucleic acid methods on the relative abundance of microorganisms and two indicator viruses. 【Results】 After sequencing by Illumina NovaSeq 6000, 306.27 GB raw data and 193.17 GB clean data were obtained, with Q20>90%, Q30>85%, Error Rate of 0.03%, and average GC Content of 45.02%. The DNA library construction process significantly increased the proportion of microbial sequences and the PRV abundance [(91.8±0.5)%](P<0.05); RNA library construction and combined library construction can increase the abundance of Pestivirus, an RNA virus, and the PRV abundance was(17.7±3.3)% and(8.1±1.5)% respectively. The Ct value of mtDNA was increased and the proportion of human sequence decreased to less than(89.5±1)%, while the proportion of microbial sequence increased to (2.4±0.03)% after treatment of five centrifugation conditions(P<0.05); After centrifugation at 4℃, 100 g, 30 min, the PRV abundance was increased to (40.6±6)%, and centrifugation at 4℃, 4 000 g, 45 min reduced the PRV abundance to (4.1±0.01)%(P<0.05). Both of 0.22-μm filter and 0.45-μm filter increased the Ct value of mtDNA to above 25.56±0.13, decreased the proportion of human sequence to less than (86.1±0.6)%, increased the proportion of microbial sequence to (3.1±0.1)% and (3.4±0.2)%, and decreased the PRV abundance to (1.6±0.3)% and (4.1±0.7)%(P<0.05), while there was no statistical difference in the effect on PPV concentration and abundance. DNase Ⅰ and Benzonase increased the Ct value of PPV to 25.65±0.06 and 25.36±0.45, decreased the proportion of human sequence to (81.7±5.6)% and (72.8±6.7)%, and increased the proportion of microbial sequence and PRV abundance to (11.0±4.1)% and (16.1±4.7)%, (55.8±2.3)% and (39.0±8.9)%, respectively(P<0 05); After treatment with RNase A, the Ct value of PRV increased to 25.20±0.11, and the human sequence proportion decreased to (85.4±5.6)%(P<0 05); Lysozyme had no effect on removing non-viral nucleic acid. The chloroform of 1%, 5%, 10% and 20% increased Ct value of PRV and mtDNA to no less than 27.17±0.21 and 25.68±0.04; Only 10% chloroform increased the proportion of microbial sequences to (3.1±1.2)%(P<0.05); The abundance of PRV with 1% and 5% chloroform treatment was increased to (48.7±13.3)% and (42.1±5.5)%(P<0.05), while 10% and 20% chloroform reduced PRV abundance to (1.0±0.5)% and (3.4±2.8)%(P<0.05). There was no statistical difference in the effect of chloroform with four contents on PPV abundance. 【Conclusion】 Centrifugation at 4℃, 5 000 g, 10 min is suitable for increasing the overall abundance of virus, and centrifugation at 4℃, 100 g, 30 min is suitable for increasing the content of virus similar to PRV. 0.45-μm filter, DNase Ⅰ, Benzonase and low concentration chloroform can effectively reduce the proportion of non-viral nucleic acid sequence in plasma to increase the abundance of the indicated virus group. Thus, the enrichment effect of plasma meta-virome is closely related to the nature of the virus, and the appropriate virus enrichment method should be selected according to the research purpose to establish the corresponding enrichment strategy.

Autologous cord blood mononuclear cell infusion for the prevention of bronchopulmonary dysplasia in very preterm monozygotic twins: A study protocol for a randomized, placebo-controlled, double-blinded multicenter trial.

Background: Preterm-associated complications remain the main cause of neonatal death. Survivors face the challenges of short- and long-term complications. Among all complications, bronchopulmonary dysplasia (BPD) remains the first important cause of neonatal mortality and morbidity. Current treatment does not address this main preterm complication. Cord blood is regarded as a convenient source of stem cells. The paracrine bioactive factors of stem cells contribute to tissue repair and immune modulation. Our clinical studies and those of others have shown that cord blood cell infusion is both safe and possibly effective in the prevention and treatment of BPD. The therapeutic use of cord blood has emerged as a promising therapy. However, the genetic heterogeneity between control and intervention groups may reduce the comparability especially among small sample trials. The purpose of this study protocol is to investigate the effects of autologous cord blood mononuclear cell (ACBMNC) infusion on the prevention of BPD in very preterm monozygotic twins of less than 32 gestation weeks. Methods: In this prospective, randomized, placebo-controlled, double-blinded multicenter clinical trial, 60 pairs of monozygotic twin preterm neonates of less than 32 weeks admitted to the Neonatal Intensive Care Unit are randomly assigned to receive intravenous ACBMNC infusion (targeted at 5 × 107 cells/kg) or placebo (normal saline) within 24 h after birth in a 1:1 ratio. The primary outcome will be survival without BPD at 36 weeks of postmenstrual age. The secondary outcomes will include the mortality rate, BPD severity, other common preterm complication rates, respiratory support duration, length and cost of hospitalization, and long-term respiratory and neurodevelopmental outcomes during a 2-year follow-up. Furthermore, we will perform single-cell RNA sequencing for cord blood cells and blood cells 3-10 days after intervention and detect whether reactive oxygen species and inflammatory cytokines are present. Conclusion: This will be the first randomized, placebo-controlled, double-blinded trial to evaluate the efficacy of ACBMNC infusion to prevent BPD in monozygotic twin premature infants and investigate the underlying protective mechanisms. The results of this trial will provide valuable clinical evidence for translational application of cord blood cell therapy in very preterm infants.Trial registration: ClinicalTrials.gov, NCT05087498, registered 10/09/2021, https://register.clinicaltrials.gov/prs/app/action/SelectProtocol?sid=S000BAD7&selectaction=Edit&uid=U0002PLA&ts=2&cx=qvyylv.

Effect of dehydrocostus lactone on the proliferation of chronic myeloid leukemia K562 cells and its mechanism / 白血病·淋巴瘤

Objective To explore the effect of dehydrocostus lactone on the proliferation of chronic myeloid leukemia cell line K562 and its mechanism.Methods K562 cells in logarithmic growth phase were treated with different concentrations of dehydrocostus lactone.Cell morphology was observed by Wright-Giemsa staining.Cell proliferation was detected by CCK-8 method.Cell cycle,apoptosis and the expressions of cell surface differentiation antigen CD14 and CD1 1b were detected by flow cytometry.JAK-STAT pathway and the expressions of apoptosis and cell cycle related proteins were detected by Western blot.Results Compared with the control group,the proliferation of K562 cells could be inhibited after treatment with different concentrations (4.0,6.0,8.0,10.0 and 12.0 μmol/L) of dehydrocostus lactone for 24 h,and the difference was statistically significant (F =109.510,P ＜ 0.05).After treatment with 5.0 and 10.0 μmol/L dehydrocostus lactone for 24 h,the apoptosis rates of K562 cells were (16.1±3.8)％ and (29.6±4.3)％,which were higher than that of the control group [(3.1±0.5)％] (F =83.255,P ＜ 0.05).After treatment with 5.0 and 10.0 μmol/L dehydrocostus lactone for 24 h,the proportion of G2/M phase cells in K562 cells were (17.0±3.2)％ and (28.8± 3.9)％,which were higher than that of the control group [(9.1±2.3)％] (F =161.598,P ＜ 0.05);the proportion of S phase cells in K562 cells were (48.1±3.9)％ and (61.0±5.4)％,which were higher than that of the control group [(39.6±3.6)％] (F =192.356,P ＜ 0.05).After treatment with 2.5 and 5.0 μmol/L dehydrocostus lactone for 72 h,the expression rates of CD14 in K562 cells were (28.6±3.9)％ and (41.1±4.4)％,which were higher than that in the control group [(3.1±0.5)％] (F =132.811,P ＜ 0.05);the expression rates of CD11b in K562 cells were (42.4±5.0)％ and (61.2±5.7)％,which were higher than that in the control group [(4.2±1.1)％] (F =179.553,P ＜ 0.05).Dehydrocostus lactone could decrease the expressions of JAK2,STAT5,eyclin E,CDK2,cyclin A,CDC25C,cyclin B1,CDK1 and bcl-2 proteins,and up-regulate the expressions of p21 and bax proteins.Conclusion Dehydrocostus lactone can inhibit the proliferation of chronic myeloid leukemia K562 cells,which may be achieved by cell cycle arrest,induction of apoptosis and differentiation.

Pilose antler peptide protects osteoblasts from inflammatory and oxidative injury through EGF/EGFR signaling.

Epidermal growth factor (EGF)/EFG receptor (EGFR) signaling plays an important role in the osteoblastogenesis. The potential effects of pilose antler peptide (PAP) on osteoblast cell damages was investigated in our present study through EGF/EGFR signaling. In MC3T3-E1 osteoblastic cells, PAP treatment significantly inhibited the production of inflammatory cytokines by decreasing the levels of serum proinflammatory cytokines interleukin-1ß (IL-1ß), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). PAP treatment also alleviated the oxidative responses as indicated by increased activities of catalase (SOD) and decreased levels of malondialdehyde (MDA). EGF inhibition, by siRNA knockdown, almost abolished PAP-induced osteoblast cytoprotection against inflammation and oxidant stress. Further, our results showed that PAP stimulated the nuclear erythroid factor 2-related factor 2 (Nrf2)2/heme oxygenase-1(HO-1) signaling, and inhibited the activation of uclear factor kappa B (NF-κB) pathway in MC3T3-E1 cells. On the other hand, EGF siRNA knockdown inhibited PAP-induced cytoprotection, which decreased the expression of Nrf-2, HO-1 and increased the level of p-NF-κBp65, p-IκBα in MC3T3-E1 cells. Thus, our research demonstrated that PAP protects osteoblasts from inflammatory and oxidative injury through EGF/EGFR signaling.

Deep brain stimulation of bilateral nucleus accumbens at high frequency inhibits addiction behavior extinction in rats / 中华神经科杂志

Objective To investigate the influence of deep brain stimulation (DBS) at high frequency to the bilateral nucleus accumbens on morphine-induced conditioned place preference (CPP) and relapse behaviors during extinction phase in rats. Methods Twenty adult SD rats were employed in the experiment. Through stereotactic operation, outer electrode cannula was implanted into rats' bilateral nucleus accumbens. After 5 days of rest, the morphine-dependent rat model with CPP was established through intraperitoneal morphine injection (10 mg/kg). The rats, after being randomly divided into experimental group (morphine+DBS) and control group (morphine+sham DBS), were electrically stimulated using DBS circuits. Rats in the experimental group were given high frequency electrical stimulations while the control group was given sham stimulation. The CPP score of the two groups was recorded the day after stimulation until successful extinction and then the extinction time was compared between the two groups. After successful extinction the rats were given small dose of morphine to trigger relapse within 24 hours, and the CPP score was recorded and compared between the two groups.Results Compared with the control group (six days), the experimental group (26 days) had a longer extinction time. After relapse, the retention time within the drug-paired chamber of the experimental group was (357.01±192.72) s, obviously shorter than that of the control group ((704.91±181.35) s;t=2.370, P=0.034 6). Conclusion High frequency DBS to rats' bilateral nucleus accumbens can prolong extinction time but inhibit relapse behavior.

Isoalantolactone suppresses proliferation of chronic myelogenous leukemia drug-resistant cell line K562/A02 through bcr-abl-STAT5 signaling pathway / 白血病·淋巴瘤

Objective To explore the effects of isoalantolactone on the proliferation of human chronic myelogenous leukemia drug-resistant cell line K562/A02. Methods K562/A02 cells were treated with 6.25, 12.5, 25, 50 and 100 μmol/L isoalantolactone for 24 and 48 h, cell viability was analyzed with MMT assay. K562/A02 cells were treated with 10, 15, 20 μmol/L isoalantolactone for 24 h. Flow cytometry was used to examine the effect of isoalantolactone on the cell-cycle and apoptosis of K562/A02 cells. The related proteins were analyzed using Western blot. One-way ANOVA was used for statistical analysis. Results Isoalantolactone effectively inhibited the proliferation of K562/A02 cells in a dose-dependent manner (P<0.05) with IC50 value of (15.00 ±1.03) μmol/L at 24 h, respectively; Flow cytometry displayed that isoalantolactone may induce K562/A02 cells apoptosis in a concentration-dependent manner (P<0.05). The apoptotic rate significantly increased from (2.71 ±0.52) % in the control group to (19.10 ±1.55) %, (27.61 ± 2.32)%and (32.01±3.01)%(F=33.901, P<0.05), respectively, after treatment with 10, 15, and 20 μmol/L of isoalantolactone for 24 h. The percentage of cells in the S phase increased from (57.80±2.11) % to (68.62± 2.89)%, (78.41±3.51)%and (80.61±2.90)%, respectively (F=51.328, P<0.05). Western blot indicated that the expression of bcl-2, p-bcr-abl, p-STAT5, CDK2 and cyclin A significantly decreased (P< 0.05), and that of cytochrome C, Bax, and p21 increased with the increasing of isoalantolactone concentration (P< 0.05). Conclusion Isoalantolactone can significantly inhibit the proliferation of K562/A02 cells through bcr-abl-STAT5 signaling pathway.

Polyphyllin D inhibits proliferation of K562 on human leukemia cells by inducing apoptosis and differentiation / 国际肿瘤学杂志

Objective To investigate the inhibitory effect of polyphyllin D on the proliferation of human chronic myelogenous leukemia (CML) cell line K562 cells and its mechanism.Methods Methyl thiazolyl thiazolium (MTT) assay was used to evaluate the inhibition effect of polyphyllin D on the proliferation of K562 cells.Flow cytometry was used to determine the effect of polyphyllin D on the apoptosis of K562 cells.The related proteins were analyzed by Western blotting.Results The MTT assay showed that polyphyllin D obviously inhibited the growth of K562 cells.The hemi-inhibitory concentration (IC50) of 24 hours for the K562 cells was (0.8 ± 0.1) μmol/L.Flow cytometric results demonstrated that polyphyllin D significantly induced apoptosis in a dose-dependent manner,and the apoptosis rates of 0,0.4,0.8,1.2 μmol/L polyphyllin D were (1.73 ± 0.33) ％,(14.47 ± 2.57) ％,(24.10 ± 3.52) ％,(31.51 ± 4.09) ％ after 24 hours,with a significant difference (F =58.231,P ＜ 0.001).Western blotting showed that polyphyllin D significantly down-regulated the expression of Bcl-2,and this was accompanied by up-regulation of Bax,decrease of the Bcl-2/Bax ratio,up-regulation of cytochrome c and activation of caspase-3.Otherwise,polyphyllin D obviously induced monocyte differentiation by increasing CD14 expression on the surface of K562 cells.Conclusion Polyphyllin D can significantly inhibit the proliferation of K562 cells,and the mechanism may be realized by inducing apoptosis and promoting monocyte differentiation.

Inhibitory effect of polyphyllin D on the proliferation of human chronic myelogenous leukemia cell line K562 and its mechanism / 白血病·淋巴瘤

Objective To explore the inhibitory effect of polyphyllin D on the proliferation of human chronic myelogenous leukemia (CML) cell line K562 and its mechanism. Methods K562 cells were treated with various concentrations of polyphyllin D (0, 0.1, 0.2, 0.4, 0.8, 1.2, 2.4 μmol/L) at 24 h, and cell viability was assessed by CCK-8 assay. Flow cytometry was used to detect the effect of polyphyllin D on the apoptosis, and the cell cycle arrest of K562 cells. The relative proteins were analyzed by using Western blot. Results The polyphyllin D could significantly inhibit the proliferation of K562 cells, and the effective inhibitory concentration (IC50) was (0.9 ± 0.1) μmol/L at 24 h. The results of flow cytometry showed that after treatment with 0.9 μmol/L polyphyllin D at 12 h and 24 h, the apoptotic rate of the cells [(11.46 ±1.51) %, (28.87 ± 2.35) %] were significantly higher than that of the control group [(2.05±0.45) %], and the difference was statistically significant (F= 38.637, P< 0.05). The expressions of bcl-2, CDK1, CyclinB1 and bcr-abl fusion protein were down-regulated by polyphyllin D, and the expressions of Bax, cytochrome C, activated caspase-3 and p21 were up-regulated (all P<0.05). In addition, polyphyllin D could arrest cell-cycle at G2/M phase (F=42.355, P<0.05). Conclusion Polyphyllin D can significantly inhibit the proliferation of human CML cell line K562, and its mechanism could play a role by inducing apoptosis and promoting cell cycle arrest.

Study of acute lymphoblastic leukemia and acute myeloid leukemia gene mutation in children / 中华实用儿科临床杂志

Acute leukemia is the most common malignant tumor in children in our country.Its pathogenesis includes transformation of proto oncogene,tumor suppressor gene distortion,inhibition of apoptosis etc..Under the action of carcinogenic factors,chromosome mutation,deletion,rearrangement or gene amplification can lead to the structural variation of proto oncogenes and tumor suppressor genes,resulting in a new fusion gene.Some of these genes are tran-scription factors regulating cell proliferation,differentiation,aging and death,when the gene is mutated,directly affect the downstream signaling pathways,leading to cell proliferation,apoptosis and enhanced (or)differentiation disorders, leading to leukemia.In recent years,with the development of gene sequencing technology,more and more leukemia prognostic genes have been displayed in public view and applied in clinical treatment and prognosis,relapse,etc..

High frequency deep brain stimulation of rats bilateral accumbens inhibits relapse behavior in morphine-dependent rats / 中华行为医学与脑科学杂志

Objective To investigate the effect of high frequency stimulation of bilateral nucleus accumbens on relapse behaviors in morphine-dependent rats.Methods Twenty adult SD rats were employed in the experiment.Through stereotactic operation,outer electrode cannula was implanted into rats' bilateral nucleus accumbens.After 5 days of rest,the morphine-dependent rat model with conditioned place preference (CCP) was established through intraperitoneal morphine injection (10 mg/kg).After acquisition of CPP,normal saline was replaced with morphine for CPP extinction training.CPP test was used to exam the effect of extinction.The rats,after being randomly divided into experimental group (morphine+DBS) and control group (morphine+sham DBS),were electrically stimulated using modified DBS circuits.Rats in the experimental group were given high frequency electrical stimulation while the control group was sham stimulation.After consecutive stimulation for 7 days,rats in the two groups were given small dose of morphine (3 mg/kg)to trigger relapse.Results (1) The CPP score increased after the establishment of rat models compared with pre-establishment of the rat models((616.2±74.7) s vs (353.9±84.3) s,P＜0.01).(2) The CPP score after extinction training decreased compared with pre-conditioned CPP score ((456.4± 148.8) s vs (353.9±84.3) s),P=0.0847) and had no statistical difference compared with post-conditioned CPP score ((456.4±148.8) s vs (616.2± 74.7) s,P=0.0219).(3) When the relapse was induced by small doses of morphine within 24h after the last stimulation,the CPP score of the experimental group decreased compared with the CPP score of control group ((330.1 ±212.6) s vs (684.2±230.2)s,P=0.0029),and the relapse was restrained.Conclusion High frequency DBS of bilateral nucleus accumbens can attenuate relapse behavior in rats

Alantolactone inhibits the proliferation of K562/ADR cells through regulating expression of cell cycle-related proteins / 白血病·淋巴瘤

Objective To investigate the effects of alantolactone on cell proliferation,cell-cycle and cell cycle-related proteins in human chronic myelogenous leukemia drug-resistant cell line K562/ADR.Methods K562/ADR cells were treated with 0,1.0,2.0,4.0,6.0,8.0,and 10.0 μmol/L of alantolactone for 12,24 and 48 h,with its cell viability analyzed by MTT assay.Flow cytometry was used to examine the effect of alantolactone on the cell-cycle of K562/ADR cells.The cell cycle-related proteins were analyzed by using Western blot after treatment with alantolactone.Results The results of MTT showed that alantolactone effectively inhibited the proliferation of K562/ADR cells in dose and time-dependent way,and the IC50 value of alantolactone in K562/ADR cells was about 5 μmol/L.Flow cytometric analysis displayed that alantolactone could arrest cell cycle at G2/M phase.The percentage of accumulated cells in the G2/M phase was increased from (15.8±1.7) ％ in the control group to (21.0±2.4) ％,(26.4±2.7) ％,and (30.1±3.9) ％ in cells treated with 2.5,5.0,and 7.5 μmol/L of alantolactone for 24 h,respectively (P ＜ 0.05).Alantolactone significantly decreased the expression of CDK1 and CyclinB1 and increased the expression of cyclin-dependent kinase inhibitor p21.Meanwhile,the treatment of K562/ADR with alantolactone led to a dose-dependent decrease in bcr-abl protein levels.Conclusion Alantolactone can significantly inhibit the proliferation and cell-cycle arrest in G2/M phase of K562/ADR cells,in which mechanism may be associated with the regulation of cell cycle-related proteins and downregulation of bcr-abl protein.

Inhibitory effect of alantolactone on the proliferation of K562/ADR cells and its mechanism / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the inhibitory effect of alantolactone on the proliferation of adriamycin-resistant human chronic myelogenous leukemia cell line K562/ADR cells and its mechanism.</p><p><b>METHODS</b>K562/ADR cells were treated with various concentrations of alantolactone (0, 1, 2, 4, 6, 8, and 10 μmol/L) for different time points. Cell viability was analyzed with MTT assay. The effect of alantolactone on the apoptosis of K562/ADR cells was measured by flow cytometry. The expression of apoptosis-related proteins after treatment with alantolactone was analyzed using Western blot.</p><p><b>RESULTS</b>Alantolactone could effectively inhibit the proliferation of K562/ADR cells in dose- and time- dependent manner, the IC50 value of alantolactone treatment of K562/ADR cells for 24 h was 4.7 μmol/L (P<0.05). Flow cytometric analysis displayed that the apoptotic rates were 1.35%, 16.91%, 29.61% and 46.26%, respectively, after treatment with alantolactone at 0, 2.5, 5 and 7.5 μmol/L. Meanwhile, the expression of Bcl-2 and BCR-ABL proteins were significantly decreased and that of Bax, cytochrome C, cleaved-caspase-9, cleaved-caspase-3 and cleaved-PARP increased by alantolactone treatment.</p><p><b>CONCLUSION</b>Alantolactone had obvious inhibitory effect on the proliferation of K562/ADR cells through the caspase dependent mitochondrial(or intrinsic)apoptotic pathway.</p>

The role of serum procalcitonin in evaluating the severity and prognosis in patients with community acquired pneumonia / 中华急诊医学杂志

Objective To analyze the value of serum procalcitonin (PCT) in patients with community acquired pneumonia (CAP),and to evaluate the role of PCT in the therapeutic effect,severity and prognosis.Methods A retrospective analysis of data and laboratory tests of 50 patients with CAP admitted from November 15,2011 to November 15,2012 in GICU was carried out.Patients with infection of other parts of body,surgical treatment and trauma were ruled out.The level of PCT (ng/mL) before and during treatment,and the relationships between PCT and respiratory failure,mechanical ventilation,treatment results were analyzed respectively.Results According to the occurrence of sepsis,50 patients were divided into sepsis group and non-sepsis group.In the non-sepsis group,the PCT level before treatment,the highest and average PCT levels during the treatment were 0.1125 (0.078,0.269),0.1235 (0.078,0.494),and 0.1355 (0.08,0.245) respectively.Correspondingly,the PCT levels in the sepsis group were 8.92 (2.715,16.33),13.53 (6.305,25.625),and 4.26 (2.1415,8.2455),and there were statistically significant differences in three values of PCT between groups (ZIst =-4.743,PIST ＜ 0.05 ; ZMax =-5.783,PMax ＜ 0.05 ; ZMean =-5.644,PMean ＜ 0.05).According to the emergence of respiratory failure during treatment,average PCT level in the patients with respiratory failure was 1.7375 (0.224,5.092),and that in the patients without respiratory failure was 0.081 ng/mL (0.049,0.146),presenting the statistically significant difference between two groups (Z =4.472,P ＜ 0.05).In case of using mechanical ventilation (MV),the average PCT level of the patients with mechanical ventilation was 1.618 ng/mL (0.224,5.092),and that in the patients without MV was 0.086 ng/mL (0.061,0.465),producing a significant difference between the two groups (Z =-3.788,P ＜ 0.05).Grouped according to the outcome of patients,the mean value of PCT level in death group was 7.4585 ng/mL (2.392,16.25),and that in the survival group was 0.1965 ng/mL (0.885,0.618),showing statistically significant difference between two groups (Z =3.857,P ＜ 0.05).The first PCT level in the GICU within 24 h after admission was used to make the receiver operating characteristic curve (ROC),and the area under the curve (AUC) was 0.9867,cutoff point was 1.25 ng/mL.Conclusions In case of CAP,the PCT level in patients with sepsis is significantly higher than that in patients without sepsis,and PCT can distinguish sepsis from pneumonia precisely.In addition,PCT is an important biomarker to judge the severity and outcomes of CAP at early stage.

A correlational analysis of histological chorioamnionitis and brain injury in preterm infants / 临床儿科杂志

Objective To explore the correlation between histological chorioamnionitis (HC) and brain injury in preterm infants. Methods Three hundred and forty-seven cases of infants at the gestational age of 28-31 weeks who were admitted to the neonatology department of our hospital were analyzed retrospectively. They were divided into the HC group and the control group according to the pathological examination. Moreover, HC group was divided into FV group and non-FV group according to the pathological findings of fetal vasculitis (FV). Based on the findings of periodical ultrasonography, the incidences of periventricular leukomalacia (PVL), periventricular-intraventricular hemorrhage (PVH-IVH), and the PVL+PVH-IVH were compared among groups. Results The incidences of PVL in the HC group and the control group were 17.9% and 10.3%respectively. The incidences of PVL+PVH-IVH in the two groups were 5.5%and 1.48%respectively, and the difference between two groups was signiifcant (P0.05). In the HC group, the incidences of PVL in FV group and non-FV group were 28.1%and 9.87%respectively, and the difference between two groups was signiifcant (P0.05). The incidences of PVL+PVH-IVH in FV group and non-FV group were 7.81%and 3.70%respectively, and the difference between the two groups was not have signiifcant (P>0.05). Conclusions HC may increase the ncidences of PVL and PVL+PVH-IVH in the preterm infants, while its effect is minimal on PVH-IVH. FV could increase the incidence of brain injury in preterm infants.

Effects of Bmi-1-siRNA on proliferation of lung adenocarcinoma SPC-A1 cells and its mechanism / 中国癌症杂志

Background and purpose:The human oncogene B-cell-speciifc moloney murine leukemia virus integration site 1 (Bmi-1) is an important member of the polycomb group family, and it regulates cell proliferation and senescence via INK4a/ARF locus. This study investigated the effects of Bmi-1-siRNA on the proliferation of lung adenocarcinoma cell line SPC-A1 cells with INK4a/ARF locus and clarify the mechanism of Bmi-1-mediated effect on proliferation of lung adenocarcinoma cells. Methods:In this study, we chose the most efifcient siRNA chain the pGeneshl-2-Bmi-1 sense-1 and inserted into a pSUPER-retro-neo retroviral vector. The packaged si-Bmi-1 pSUPERret-ro-neo retroviral vector was stably transfected into lung adenocarcinoma SPC-A1 cell line. The stably transfected cells were cultured and passed. After transfection, the levels of Bmi-1 mRNA and protein expression of SPC-A1 cells were analyzed by RT-PCR and Western blot respectively. Trypan blue, MTT and plate colony forming assay were performed to observe the proliferation capibility of SPC-A1 cells and evaluate the cloning forming ability in vitro. The potency of tumorigenesis was observed in nude mouse through hypodermic inoculation of SPC-A1 cells. Cell cycle distribu-tion was analyzed by lfow cytometry (FCM) in SPC-A1 cells. The expression levels of proliferation proteins including p16INK4a, p53, Cyclin D1, PTEN, Akt and Ser473p-Akt were analyzed by Western blot. Results:The mRNA and protein expression levels of Bmi-1 were signiifcantly reduced in SPC-A1-Bmi-1-siRNA cells transfected with pSUPER-retro-neo retroviral vector. Knockdown of Bmi-1 could inhibit the growth, colony formation in vitro and tumorigenesis in vivo of SPC-A1 cells (P0.05), while Cyclin D1 and Ser473p-Akt were downregulated (P<0.01) and PTEN was up-regulated (P<0.01) in the SPC-A1-Bmi-1-siRNA cells. SPC-A1-Bmi-1-siRNA cells were treated with various concentrations of PTEN inhibitor to determine expression levels of PTEN, Bmi-1 and Ser473p-Akt protein. Ablation of PTEN rescued Bmi-1 and Ser473p-Akt expression in SPC-A1-Bmi-1-siRNA cells. Conclusion:Knockdown of Bmi-1 gene can arrest the proliferation of SPC-A1 cells through G0/G1 phase arrest by inhibiting Cyclin D1 expression indirectly, which may be not associated with p16INK4a signaling pathway.

Regular follow-up on physical growth and mental development of small and appropriate for gestational age preterm infants / 临床儿科杂志

Objective To investigate the physical and mental development of small and appropriate for gestational age preterm infants in their early life. Methods This study recruited 220 preterm infants, who were discharged from our hospital and visited preterm following-up clinic at regular intervals from February 2009 to December 2012. All of those infants were divided into two groups based on whether their birth weight below 10th percentile for their gestational ages or not. Weights, lengths and head circumferences were measured up to seventh month age adjusted by gestational age. Meanwhile, mental tests were conducted by the professional staffs working on the children developmental assessment at their adjusted months of 5th, 6th or 7th. All of physical and mental scores were compared between the two groups. Results The SGA group was statistically less than the AGA group on the Z-score of weights from the ifrst to sixth month adjusted by gestational age (P0.05). The SGA group was statistically less than the AGA group on the Z-score of lengths from the ifrst to iffth month adjusted by gestational age (P0.05). The SGA group was statistically less than the AGA group on the Z-score of head circumferences from the ifrst to seventh month adjusted by gestational age (P<0.05). The SGA babies scored statistically less than the AGA babies with a mean development quotient score of 96.7 and 102.9, respectively (P<0.05). The scores of movement, cognitive, language in the SGA group were statistically less than those in the AGA group(P<0.05). Conclusions Preterm SGA could achieve satisfactory weight catch-up gain, with a decreasing difference from preterm AGA while they were getting older. But the length catch-up growth of preterm SGA seemed unsatisfactory with a big differece from preterm AGA. There was the worst catch-up on head circumference in those preterm SGA, backward in mental development, particularly in their movement, cognitive and language capacity.

Three different bacterial detection systems for platelet concentrates under inter-laboratory conditions.

BACKGROUND: A variety of screening methods are currently used worldwide in order to decrease the risk of transfusion-transmitted sepsis and improve the safety of PCs. METHODS/MATERIALS: PCs inoculated with five different transfusion-relevant species of bacteria at concentrations of 1, 10, and 100 colony-forming units (CFU)ml(-1) were stored at 22°C for 7 days. Flow cytometry (FACS), BacT/Alert automated culturing, and a quantitative real-time PCR (Q-PCR) were then used to detect the presence of bacteria in samples prepared from these PCs. RESULTS: At the initial spiking concentrations of 1, 10, and 100 CFU ml(-1), Q-PCR detected all five bacterial species tested. Screening with the BacT/Alert culture-based system allowed bacterial detection (inoculated on day 0) within a mean time of 15.13 h for all three spiking concentrations. Using FACS, positive signals were obtained for all three concentrations of Escherichia coli and Bacillus cereus on day 1 and for initial spiking concentrations of Pseudomonas aeruginosa and Staphylococcus aureus of 1 CFU ml(-1) on day 2. For Staphylococcus epidermidis, detection of an initial inoculum of 1 CFU ml(-1) was possible only beginning on day 6. CONCLUSION: This study shows that under standard laboratory conditions the sensitivity of FACS in the detection of bacterial contamination of PCs was lower than that of either the BacT/Alert automated culturing method or Q-PCR.

Quantitative evaluation of plasma after methylene blue and white light treatment in four Chinese blood centers.

BACKGROUND: Pathogen reduction technology is an important process in the blood safety system, including solvent/detergent treatment, filtration and methylene blue-photochemical technology (MB-PCT). To investigate the quality of MB-PCT-treated plasma, plasma samples from four Chinese blood centers were analyzed over 12 months of storage to determine the recovery of activities and levels of various plasma proteins. MATERIALS AND METHODS: Ten plasma units each from the Suzhou, Yancheng, Chongqing and Shandong blood centers were divided into two aliquots. One was subjected to treatment with one of two methylene blue-photochemical technology instruments and the other was used as control. The treated and untreated sample pairs were stored at -30°C. The recovery rates of coagulation factors, inhibitor proteins, total protein, immunoglobulins, and complement proteins were measured at different time points after storage. RESULTS: The mean recovery of most proteins exceeded 80% after MB treatment. The exceptions were factor XI and fibrinogen, of which only 71.3-74% and 69.0-92.3% were retained during storage. The two equipment types differed in terms of residual MB concentration in the plasma samples (0.18 µM and 0.29 µM, respectively). They had similar protein recovery rates at 0.5 month but differed at later time points. The four blood centers differed significantly with regard to factor II, V, VIII and fibrinogen activities. Only the samples from the Shandong blood center met the methylene blue treated fresh frozen plasma requirement. The major factor that influenced the quality of the MB-FFP samples was the time taken between blood collection and storage. DISCUSSION: Methylene blue treated plasma showed reduced coagulation factor (CF) activity and protein levels. The MB treatment-induced damage to the proteins was acceptable at the four Chinese blood centers, but the quality of the MB-treated plasma in general was not satisfactory. The main factor affecting plasma quality may be the duration of the collection and processing.

Effect of siRNA-mediated silencing Bmi-1 gene expression on the proliferation of lung cancer cell line A549 in vitro and in vivo / 中国癌症杂志

Background and purpose:The pro-oncogene Bmi-1 is a member of the polycomb-group family, can regulation of the proliferation and self-renewal of normal and tumor stem cells. In recent years, Bmi-1 has been found that it is overexpressed in varieties of human malignant tumors. The study aimed to observe the effects of Bmi-1-siRNA on the growth capacity of lung cancer cell line A549 in vivo and in vivo, and explore its mechanism. Methods:The most effective one as a target sequence was chosen from four Bmi-1 siRNA sequences which were designed by our lab, and one random sequence was chosen as a negative control. In short, the chemically synthesized siRNA and control sequences were connected to a retrovirus expressing vector, pSUPERretro-Neo plasmid, and then transfected into A549 cells. The stably transfected cells were cultured and passed. The level of mRNA and protein of Bmi-1 in A549 cells were assessed by RT-PCR and Western blot respectively. The proliferations of A549 cells in vivo was analyzed with MTT, trypan blue exclusion and plate colony forming methods. Flow cytometry was used for cell cycle analysis. The potency of tumorigenesis was observed in nude mouse through hypodermic inoculation of A549 cells. The expressions of cyclin D, p21/27, p-AKT and PTEN were analyzed by Western blot. Results:Compared to A549-ctr and A549-wt cells, Bmi-1 mRNA and protein levels all signiifcantly reduced in A549-Bmi-1-siRNA cells. Bmi-1-siRNA inhibited the growth, colony formation in vitro and tumorigenesis in vitro of A549 cells, and the interference cells cell cycle arrested in G1 phase. In A549-Bmi-1-siRNA cells, p-AKT and cyclinD1 expression were down-regulated while p21/p27 and PTEN were up-regulated. Conclusion:Silencing Bmi-1 gene inhibits the proliferation of A549 cells through G1 phase arrest, which involves the downregulation of cyclin D/p-AKT and upregulation of p21/p27/PTEN.