Objective: To investigate the effects of colony-stimulating factor 1 receptor (CSF-1R) inhibitor pexidartinib (PLX3397) on the senescence of bone marrow-derived macrophages (BMDM) stimulated by lipopolysaccharide (LPS). Methods: BMDM were isolated and cultured from femurs and tibiae of 10 male C57BL/6 mice aged 6-8 weeks (obtained from Laboratory Animal Center of Guizhou Medical University). They were divided into blank control group, LPS group (treated with 1 μg/ml LPS for 24 h) as well as low, medium and high concentration PLX3397 pretreatment groups (treated with 100, 500 and 1 000 nmol/L PLX3397 for 4 h respectively followed by 1 μg/ml LPS for 24 h). The corresponding markers of macrophages were detected by flow cytometry. Cell viability was detected by cell counting kit-8 and cellular senescence was detected by senescence-associated-β-galactosidase (SA-β-gal) staining. Meanwhile, protein expressions of cycle-dependent kinase inhibitor p16, p21 and CSF-1R were detected by Western blotting, and the expressions of p16 and p21 were detected by intracellular immunofluorescence. Real-time fluorescence quantitative PCR (RT-qPCR) was used to investigate the mRNA levels of senescence-associated secretory phenotype (SASP) genes including interleukin (IL), IL-1β, chemokine-1/10 (CXCL-1/10), matrix metalloproteinase-8 (MMP-8), and transforming growth factor-β (TGF-β). Results: The rate of SA-β-gal positive staining in medium and high concentration PLX3397 pretreatment groups [(39.33±4.93)% and (36.33±3.06)% respectively] were significantly downregulated compared with LPS group [(52.00±3.00)%] (P=0.020, P=0.005). The expression of CSF-1R protein in low, medium and high concentration PLX3397 pretreatment groups were (0.74±0.18, 0.61±0.07, 0.54±0.06), all of which were significantly lower than that in LPS group (1.16±0.08) (P=0.013, P=0.002, P<0.001). The expression levels of CSF-1R mRNA in low, medium and high concentration PLX3397 pretreatment groups (1.04±0.06, 0.90±0.05, 1.18±0.08) showed similar trend (2.90±0.25) (P<0.001). The average fluorescence intensity of p16 in all PLX3397 pretreatment groups were 49.76±3.65, 48.21±1.72, 47.99±1.26 respectively, which were significantly lower than that in LPS group (66.88±5.85) (P=0.001, P<0.001, P<0.001). The average fluorescence intensity of p21 in medium and high concentration PLX3397 pretreatment groups were (34.43±3.62, 30.13±0.86), significantly lower than that in LPS group (46.82±5.33) (P=0.043, P=0.007). The expression of p16 protein in low, medium and high concentration PLX3397 pretreatment groups (0.56±0.04, 0.55±0.04, 0.35±0.19) were significantly lower than that in LPS group (0.98±0.10) (P=0.003, P=0.002, P<0.001), as well the expression of p21 protein (0.69±0.20, 0.42±0.08, 0.26±0.14) (P=0.032, P=0.002, P<0.001). According to the results of RT-qPCR, the expressions of IL-6, IL-1β, CXCL-1, CXCL-10 and MMP-8 in PLX3397 pretreatment groups were significantly lower than those in LPS group (P<0.001), while the expression of TGF-β increased (P<0.001). Conclusions: LPS could induce the cell senescence, increase the secretion of SASP and aggravate local inflammation by activating the CSF-1R on the cell surface of bone marrow-derived macrophages. CSF-1R inhibitor PLX3397 might attenuate CSF-1R activation associated with LPS and inhibit the senescence of bone marrow-derived macrophages induced by LPS.
Mice ; Animals ; Male ; Lipopolysaccharides/pharmacology* ; Macrophage Colony-Stimulating Factor/metabolism* ; Matrix Metalloproteinase 8/metabolism* ; Mice, Inbred C57BL ; Macrophages ; Transforming Growth Factor beta/metabolism* ; RNA, Messenger/metabolism*

Objective: To summarize the clinical characteristics of epileptic seizure associated with neurofibromatosis type 1 (NF1). Methods: From January 2017 to July 2023 at Children's Hospital Capital Institute of Pediatrics, medical records of patients with both NF1 and epileptic seizure were reviewed in this case series study. The clinical characteristics, treatment and prognosis were analyzed retrospectively. Results: A total of 15 patients(12 boys and 3 girls) were collected. Café-au-lait macules were observed in all 15 patients. There were 6 patients with neurodevelopmental disorders and the main manifestations were intellectual disability or developmental delay. The age at the first epileptic seizure was 2.5 (1.2, 5.5) years. There were various seizure types, including generalized tonic-clonic seizures in 8 patients, focal motor seizures in 6 patients, epileptic spasm in 4 patients, tonic seizures in 1 patient, absence in 1 patient, generalized myoclonic seizure in 1 patient and focal to bilateral tonic-clonic seizure in 1 patient. Among 14 patients whose brain magnetic resonance imaging results were available, there were abnormal signals in corpus callosum, basal ganglia, thalamus or cerebellum in 6 patients, dilated ventricles of different degrees in 3 patients, blurred gray and white matter boundary in 2 patients, agenesis of corpus callosum in 1 patient and no obvious abnormalities in the other patients. Among 13 epilepsy patients, 8 were seizure-free with 1 or 2 antiseizure medications(ASM), 1 with drug resistant epilepsy was seizure-free after left temporal lobectomy, and the other 4 patients who have received 2 to 9 ASM had persistent seizures. One patient with complex febrile convulsion achieved seizure freedom after oral administration of diazepam on demand. One patient had only 1 unprovoked epileptic seizure and did not have another seizure without taking any ASM. Conclusions: The first epileptic seizure in NF1 patients usually occurs in infancy and early childhood, with the main seizure type of generalized tonic-clonic seizure and focal motor seizure. Some patients have intellectual disability or developmental delay. Most epilepsy patients achieve seizure freedom with ASM.
Male ; Female ; Humans ; Child, Preschool ; Child ; Neurofibromatosis 1/diagnosis* ; Retrospective Studies ; Intellectual Disability ; Electroencephalography ; Epilepsy/etiology* ; Seizures/etiology*

Objective:To explore the type distribution and severity of central nervous system(CNS) complications in severe hand, foot and mouth disease (HFMD) cases prior to the introduction of human enterovirus A group 71 type (EV-A71) vaccine, and provide scientific data for early clinical intervention.Methods:A total of 3 583 laboratory-confirmed severe HFMD cases in Jiangsu province during 2010-2016 were collected and analyzed retrospectively. Related early warning signs of increased HFMD severity were estimated with logistic regression analyses.Results:The severity-fatality rate, severity-pediatric intensive care unit(PICU) admission rate, and sequelae rate were 8.09‰(29/3 583), 11.75% (421/3 583)and 5.30‰(19/3 583). Of them, 39.02% (1 398/3 583) patients suffered from mild CNS involvement, 59.22% (2 122/3 583) patients suffered from severe CNS involvement, 1.76%(63/3 583) suffered from critical CNS involvement. The rates of the cases whose age of onset was 6-11 months, the rates of cases with atypical rash, respiratory-related signs and symptoms (shortness of breath, slowed breathing, dyspnea, etc), neurological-related signs and symptoms [(hand and foot shaking, convulsions, lethargy(sleepiness), etc], circulatory-related system signs and symptoms (faster heart rate, abnormal skin color, arrhythmia, cold limbs), laboratory-related indicators (increased white blood cell count, increased lymphocyte count, increased platelet count, increased C-reactive protein, etc), clinical auxiliary examination [electroencephalogram(EEG), brain CT, chest X-ray)] were highest in the critical CNS involvement group, and the differences were significant ( P<0.05). Multivariate logistic regression analysis showed that with the increase of proportion of convulsions, slowed breathing, vomiting, meningeal irritation and other 7 variables, the severity of CNS complications increased ( P<0.05). Conclusions:The indicators such as easily startled, slow breathing, vomiting, elevated lymphocytes, abnormal EEG and other indicators have important clinical significance for children with severe HFMD to progress to CNS complications of different severity.

Hepatic echinococcosis, also called echinococcosis, is a health-threatening disease commonly found in pasture, and belongs to parasitic zoonoses. The purpose of this study was to investigate the prevalence and the risk factors of echinococcosis in Qinghai province in order to provide fundamental data for prevention and control of echinococcosis in Qinghai province. A total of 23 445 people from 21 counties were enrolled in this study by multi-stage stratified random sampling. Echinococcosis was diagnosed by using B-mode ultrasonography and serological tests. The results showed that the prevalence of echinococcosis was 4.47% (95%CI: 4.21%-4.73%) and serum positive rate (seroprevalence) was 15.47% (95%CI: 14.92%-16.02%) in 2010. The distribution of echinococcosis differed in age, sex, ethnicity, occupation and regions in Qinghai (P<0.05). GLMM analysis revealed that gender (female vs. male), ethnicity (Tibetan vs. other ethnicities), profession (herders vs. other professions) and region (autonomous prefectures vs. cities) were significant risk factors for echinococcosis (P<0.05). It was concluded that the prevalence of echinococcosis in 2010 was about 4% in Qinghai province, and the distribution of echinococcosis in Qinghai was associated with age, sex, ethnicity and profession.
Adolescent ; Adult ; Age Factors ; Aged ; Animals ; Antibodies, Helminth ; blood ; Child ; Child, Preschool ; China ; epidemiology ; Echinococcosis, Hepatic ; diagnostic imaging ; epidemiology ; parasitology ; Echinococcus ; immunology ; physiology ; Epidemics ; prevention & control ; Female ; Host-Parasite Interactions ; Humans ; Male ; Middle Aged ; Occupations ; Prevalence ; Risk Factors ; Seroepidemiologic Studies ; Sex Factors ; Ultrasonography ; Young Adult

OBJECTIVETo investigate the therapeutic efficiency of antiviral treatment with pegylated-interferon (Peg-IFN) for hepatitis B e antigen (HBeAg)-positive chronic hepatitis B (CHB) and to explore whether liver histopathological features or other factors influence the HBeAg seroconversion treatment response.

METHODSEighty HBeAg-positive CHB patients with diagnosis confirmed by liver puncture were treated with Peg-IFN(2a or 2b)body weight dose, once weekly). At treatment week 48, the rate of HBeAg seroconversion was determined and used to analyze the influence of liver histopathological features (liver biopsy assessment of: inflammation, graded G0 to G4; fibrosis stage, graded S0 to S4), sex, age, differential levels (pre-treatment baseline vs. week 48 post-treatment) of serum alanine transferase (ALT), and HBV DNA, by binary logistic analysis.

RESULTSAt week 48, the overall rate of HBeAg seroconversion was 30.0%. The rate of HBeAg seroconversion gradually advanced with increased liver inflammation (X2 = 8.435, P = 0.015): 9.09% of the 22 patients with G1; 31.58% of the 38 patients with G2; 47.30% of the 19 patients with G3; the one patient with G4. In contrast, the rate of HBeAg seroconversion showed a much weaker association with liver fibrosis (X2 = 5.917, P = 0.116). Only baseline HBeAg level, and no other baseline index, was significantly different between the patients who achieved HBeAg seroconversion and those who did not. Liver inflammation and baseline HBeAg level were identified as influencing factors of HbeAg seroconversion in response to Peg-IFN treatment.

CONCLUSIONPeg-IFN therapy induces a higher rate of HBeAg seroconversion in HBeAg-positive CHB patients with severe liver inflammation; histological analysis of pre-treatment liver biopsies may help to identify patients most likely to benefit from the antiviral regimen.

Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; pathology ; Humans ; Interferon-alpha ; therapeutic use ; Liver ; pathology ; Male ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Serologic Tests

OBJECTIVETo explore the role of the phosphatidylinositol 3 kinase/protein kinase B (PI3K/Akt) pathway in silica-induced α-SMA (α smooth muscle actin) expression in HEB (human bronchial epithelial) cell.

METHODSThe cultured HBE cells were divided into 5 groups: control, silica, PI3K inhibitor (Ly294002), both PI3K inhibitor (Ly294002) and silica at the same time and the inhibitor 24 h ahead of silica. The final concentrations of PI3K inhibitor and silica were 10 µmol/L and 100 µg/ml, respectively. Western blots were used to detect protein expressions of Akt, p-Akt, TGF-β and α-SMA. The location and expression of α-SMA were measured by immunofluorescence assay.

RESULTSHBE cell line exposed to silica can induce Akt phosphorylation, in which expressions of p-Akt were up regulated 1 times at 48 and the highest at 72 h. The expressions of TGFβ increased remarkably at 12 h and the peak at 48 h after silica exposure, while the expressions of α-SMA increased at 24 h and the highest at 72 h. However, the PI3K inhibitor (Ly294002) significantly down regulated α-SMA expression. When the cell line exposed to the PI3K inhibitor ahead of silica 24 h, the expressions of p-Akt and α-SMA were more remarkably down regulated which were decreased 1.5 times and 7.6 times respectively compare to silica exposure group. But no significant changes were found for TGFβ expressions. The immunofluorescence assay showed that silica can induce α-SMA expression, which located in cytoplasma, and PI3K inhibitor can decrease the expression.

CONCLUSIONSSilica induced α-SMA expression in HBE cell line is by targeting the PI3K/Akt pathway and PI3K inhibitor can repress α-SMA expression.

Actins ; metabolism ; Cell Line ; Chromones ; pharmacology ; Epithelial Cells ; cytology ; drug effects ; metabolism ; Humans ; Morpholines ; pharmacology ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Signal Transduction ; Silicon Dioxide ; toxicity ; Transforming Growth Factor beta ; metabolism

OBJECTIVETo investigate the clinical value of tumor markers CEA, CA19-9, CA72-4 and CA242 in the diagnosis and prognosis of patients with gastric cancer.

METHODSOne hundred and sixty gastric cancer patients who had received treatment from 2002 to 2007 at the Beijing Cancer Hospital were retrospectively analyzed. Blood samples were taken from patients upon admission to the hospital, and CEA, CA19-9, CA72-4, CA242 levels were detected. Statistical analysis was performed to identify the clinical value of these tumor markers in diagnosis and prognosis.

RESULTSOn initial diagnosis, the positive rates of CEA, CA19-9, CA72-4 and CA242 were 37.7%, 26.7%, 37.6% and 21.3%, respectively, and the positive rate of combined detection was 62.9%. CEA was more frequently positive in patients with lymph node metastasis (P=0.029); CA72-4 was more frequently positive in patients with vascular involvement and advanced stage (P=0.039, P=0.011). Multivaraite analysis showed that CA72-4 was an independent prognostic factor (P=0.012). Patients with positive CA72-4 carried a 2.147-fold increased risk of death than those with negative CA72-4. Kaplan-Meier analysis showed that patients with positive CA19-9 or positive CA72-4 had worse survival than those with negative CA19-9 or CA72-4 (P=0.006, P=0.002).

CONCLUSIONSTumor markers including CEA, CA19-9, CA72-4 and CA242 have clinical significance and prognostic value in patients with gastric cancer. Combined detection of four tumor markers can increase the positive rate. CA72-4 is an independent prognostic factor. CA19-9 and CA72-4 are associated with the prognosis of patients with gastric cancer.

Antigens, Tumor-Associated, Carbohydrate ; blood ; Biomarkers, Tumor ; blood ; CA-19-9 Antigen ; blood ; Carcinoembryonic Antigen ; blood ; Female ; Humans ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Stomach Neoplasms ; blood ; diagnosis ; pathology

To investigate the apoptosis-induction effect of brucine on human chronic myeloid leukemia cell line K562 cells, K562 cells were exposed to various dosages of brucine. MTT method was used to assayed the growth inhibition effect of brucine on K562 cells. The apoptosis of K562 cells was detected by acridine orange/ethidium bromide (AO/EB) double staining, Annexin-V/PI double labeling method and DNA agarose gel electrophoresis. The results showed that brucine could remarkably inhibit the K562 cell growth in a concentration-dependent and time-dependent manners at the range of 50 to 400 µg/ml, and its most significant inhibition was observed at 400 µg/ml for 72 hours and the inhibition rate was 94.0%. Staining of cells with AO-EB revealed that brucine induced nuclear chromatin condensation. After the K562 cells were treated with the brucine of 400 µg/ml for 72 hours, the most of the nucleus were orange stained and condensation-like or bead-like showing apoptotic morphology. The K562 cells treated with brucine of different concentrations (50, 100, 200, 400, 800 µg/ml) for 72 hours, Annexin-V/PI detection showed brucine could induce apoptosis of K562 cells, and apoptosis rate increased gradually with increasing concentration of drugs. The K562 cells treated with brucine of 400 µg/ml for 72 hours displayed typical ladder strap in DNA gel electrophoresis. It is concluded that brucine can efficiently inhibit cell growth and induce apoptosis of K562 cells with dose-dependent manner in concentrations of 50 - 400 µg/ml.
Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Humans ; K562 Cells ; Strychnine ; analogs & derivatives ; pharmacology

Objective To analyze the evolution and genetic characterization of echovirus 11 (Echo11 ) from the acute flaccid paralysis (AFP) cases in Shandong province. Methods Isolation of Enterovirus was performed from stool samples of AFP cases from 1994 to 2009. All positive strains were sero-typed by neutralization test. Entire VP1 coding region from 27 strains typed as Echo 11 was amplified by reverse transcription-polymerase chain reaction(RT-PCR), and subsequently phylogenetic analyse on VP1 sequences from these strains and others published in GenBank were conducted. Results From 1994 to 2009, altogether 915 non-polio enterovirus (NPEV) strains were isolated with 79(8.6% ) isolates serotyped as Echo11. There were 876 nucleotides (nt) in the complete VP1 genes, encoding 292 amino acids (aa). The nt identities were 76.4%-100.0% among those Echo11 Shandong strains with the aa identities as 91.4% -100.0%. The nt and aa identities were 77.7%-80.7% and 90.7%-94.8% between Shandong strains and prototype strains, respectively.Conclusion All Echo11 strains could be divided into four genotypes. Shandong strains that forming three (A1, A2 and C1) new sub-genotypes, with every sub-genotype had several brands.Sub-genotype A1 appeared to be the lately circulating one.

Objective To investigate the serum levels of endothelin-1 (ET-1), tumor necrosis factor- α (TNF-α) and their dynamic changes in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP) and their relation with the condition variation of the patients.Methods The serum ET-1 and TNF-α levels were determined with enzyme-linked immunno-sorbent assay (ELISA) in 31 patients with DEACMP, and the dynamic changes of patients' condition were monitored by use of the activity of daily living (ADL) scale, the information-memory-concentration test (IMCT) and the Hasegawa's dementia scale (HDS). The comparisons between patients with DEACMP and both 30 patients with acute carbon momoxide poisoning (ACMP) but without DEACMP and 30 normal controls were also conducted. Results At the acute stage of the DEACMP group, the serum levels of ET-1 and TNF-α were both significantly higher than those in the normal control group (P＜0.05); that of TNF-α was significantly lower than that in the ACMP group (P＜0.05), but that of ET-1 was not significantly different from that in the ACMP group (P＞0.05). The serum levels of ET-1 and TNF- α in the ACMP group were both significantly higher than those in the normal control group(P＜0.05). In the DEACMP group, the serum level of ET-1 at the convalescent stage was significantly lower than that at the acute stage (P＜0.05), but the serum level of TNF-α was not significantly different from that at the acute stage (P＞0.05). At the acute stage of the DEACMP group, ADL scores were significantly higher than those in norms, and IMCT scores and HDS scores were significantly lower than those in norms (P＜0.05). In the DEACMP group, the ADL scores at the convalescent stage were significantly lower than those at the acute stage (P＜0.05), IMCT scores and HDS scores were significantly higher than those at the acute stage (P＜0.05). Significant correlations between scores of any 2 of 3 scales in patients with DEACMP at both acute and convalescent stage were noted (P＜0.05). Conclusion The dynamic detection of serum ET-1 and TNF-α level variations could be used as an indicator for condition severity in patients with DEACMP.