Child ; Drug Resistance, Bacterial ; Humans ; Pneumococcal Infections ; drug therapy ; prevention & control ; Pneumococcal Vaccines ; immunology

Globally, pneumonia is the leading cause of childhood mortality. Pneumonia is the second killer of children less than 5 years of age in China. The World Heath Organization and United Nations Children′s Fund launched the integrated Global Action Plan for the Prevention and Control of Pneumonia and Diarrhea (GAPPD) in 2013. The ambitious goal is to end preventable childhood deaths due to pneumonia by 2025. Countries or regions should achieve the following goals: (1) reduce mortality from pneumonia in children less than 5 years of age to fewer than 3 per 1 000 live births; (2) reduce the incidence of severe pneumonia by 75% in children less than 5 years of age compared to 2010 levels. If the implementation of key interventions is accelerated, the mortality rate of childhood pneumonia will drop substantially every year, which makes the goal achievable.
Child Mortality ; trends ; Child, Preschool ; China ; epidemiology ; Humans ; Infant ; Infant, Newborn ; Pneumonia ; mortality ; Time Factors

OBJECTIVE:To establish a method for the determination of related substances in anastrozole. METHODS:HPLC was performed on the column of Welch Materials XB C18 with mobile phase of acetonitrile-water(35:65,V/V)at a flow rate of 1.0 mL/min,detection wavelength was 210 nm,column temperature of 25 ℃,and the injection volume was 20 μL. RESULTS:Only one impurity(impurity A)was detected in anastrozole,the linear range was 0.1-1.6 μg/mL(r=0.9996);limit of quantitation was 0.1 μg/mL,limit of detection was 0.02 μg/mL;RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 98.95%-105.29%(RSD%=1.78%,n=9). CONCLUSIONS:The method is simple,accurate,and can be used for the determination of related substances in anastrozole.

OBJECTIVE:To study the apoptosis of human leukemic cells induced by Dihydroartemisinin and its molecular mechanism.METHODS:Human leukemia K562 cells were treated by Dihydroartemisinin.The inhibitory effect on cell proliferation was assayed by MTT.Fluorescence microscopy was applied to observe the presence of apoptosis.The expression of caspase-3 was assayed with reverse transcription-polymerase chain reaction(RT-PCR).Levels of mitochondrial and cytoplasmic cytochrome C were determined using Western blot.RESULTS:After treatment with Dihydroartemisinin for 48 hours,the IC50 values of human leukemia K562 cells were 8? 10-5mol? L-1 detected at a wavelength of 570nm by MTT.Distinct morphology changes of cell apoptosis such as karyopyknosis and conglomeration were observed by Hoechst33342/PI staining.RT-PCR assay showed the expression of Caspase-3.Western-blot detection showed the decrease of mitochondrial cytochrome C concentration but the positive expression of cytoplasmic cytochrome C concentration.CONCLUSION:Dihydroartemisinin could inhibit proliferation and induce apoptosis of human leakemic K562 cells,this may partially attributed to the promotion of the delivery of cyt-c and the activation of caspase-3.

OBJECTIVE:To detect the expression of genes of leukemia cell line K562 treated by artemisinin using the gene chip technology and to study the mechanism of artemisinin in the inhibition of leukemia cell line K562 on the molecular level. METHODS:K562 cells were treated with artemisinin for 24h,and then the morphological change of K562 cells were observed under invert microscope and fluorescence microscope. The cell cycle state was examined by flow cytometry analysis (FCM). Total RNA samples were extracted and reverse transcribed to cDNA. Cy3-labelled cDNA samples were hybridized with gene chips.The hybridization results were detected by Gene Pix 4100A. RESULTS: Under invert microscope,different degree of shrinkage of K562 cells was noted,karyoschisis was reduced,cell density was decreased and the numbers of drift cells were increased.Under fluorescence microscope,caryotin was highly concentrated,marginalized and agglomerated to relucent clump,i.e.apoptotic body. Flow cytometric analysis showed that ratio of cells in G2 phase increased markedly. Hybridization analysis showed down-regulation of cyclin D1,cdk4,cdk2,cdc2,DNA-PK,DNA-TopoI,mcl-1,erk,jnk and VEGF in the artemisinin-treated K562 cells.CONCLUSION: The mechanism for artemisinin to inhibit the proliferation of leukemia cell line K562 is related to its action to alter the gene expression of certain regulatory substances involved in cell cycle and induce apoptosis of leukemia cell line K562.

Objective To investigate the effects of the rat serum with chronic renal failure (CRF) on ubiquitin-proteasome pathway,histone acetyltransferase p300 and activation of activating transcription factor 4(ATF4) of rat arterial vascular smooth muscle cells(VSMCs) cultured in vitro,and explore the possible mechanism.Methods To establish the rat model of CRF by 5/6 nephrectomy,VSMCs were incubated in the media with the 10％ of CRF serum or control serum in vitro.The mRNA expressions of ubiquitin(Ub),ubiquitin activating enzyme(E1),ubiquitin ligases enzymes (β-transducin repeat containing protein 1,β-TrCP1),p300 and ATF4 in the rat VSMCs were examined by using realtime PCR.Expressions of E1,β-TrCP1,p300 and ATF4 proteins in response to the CRF serum in VSMCs were determined by Western blotting analysis.The enzyme activities of 20S proteasomes in the total protein were examined by using three special fluorogenic peptide substrates.Results The CRF serum significantly promoted the mRNA expressions of Ub,E1,β-TrCP1,p300 and ATF4 in VSMCs in a time dependent manner.Compared with that in control serum group,the mRNA levels of Ub,E1,β-TrCP1,p300 and ATF4 in CRF serum group increased significantly (P ＜ 0.01).The CRF serum also increased the protein expressions of E1,β-TrCP1 and p300 in a time dependent manner.The expression of ATF4 was decreased,but the difference was not significant (P ＞ 0.05).Compared with that in control serum group,the protein expressions of E1,β-TrCP1,p300 and ATF4 in CRF serum group increased significantly (P ＜ 0.01).The activities of 20S proteasomes in the CRF serum group were significantly increased in a time dependent manner.Compared with that in control serum group,the activities of 20S proteasomes in the CRF serum group increased significantly (P ＜ 0.01).Conclusions The serum of CRF rat can effectively active the ubiquitin-proteasome pathway,but ATF4 ubiquitinylated degradation is blocked.The latter may be associated with increased expression of p300.

BACKGROUND: Much attention has been focused on mandibular condyle fracture, which has long been considered as a severe injury to the temporomandibular joint(TMJ). Restricted by diagnostic methods, many studies have been conducted on transverse fracture whereas little attention has been paid to longitudinal fracture of the mandibular condyle. Even less is known about longitudinal fracture in young children.OBJECTIVE: To evaluate the effects of childhood longitudinal fracture of the mandibular condyle on secondary ankylosis of TMJ.DESIGN: A randomized controlled study.SETTING: Department of Stomatology, General Hospital of Chinese PLA.MATERIALS: The experiment was completed in the Center for Experimental Animals, General Hospital of Chinese PLA. Twelve young Chinese experimental miniature pigs bred by the Institute of Experimental Animals, Beijing Agriculture University, aged 2 -3 months and weighing(5 -5.5) kg, were used and raised with mixed feed, and then divided into 3 groups randomly.METHODS: Food and water was forbidden for the miniature pigs 12 hours before operation. Under anesthesia with(5.0 - 15.0) mg/kg ketamine/xylazine abdominally injected, conventional disinfection was performed in the right lateral decubitus position and layer-by-layer preauricular incisions were made on the left side of the miniature pigs. The joint cyst was incised transversely and the lower cavity of TMJ was exposed and dragged downwards. The mandibular condyle was cleaved into two vertically along the inner 1/3 part of it with a 5.0 mm-wide bone chisel to cause inferior and posterior sagittal fractures. The miniature pigs were executed 3 months and 6 months after operation, and their condyles were cut for observation.MAIN OUTCOME MEASURES:①The morphological changes of mandibular condyle specimens in the three groups observed with the naked eyes.②The pathological changes of mandibular condyle specimens under light microscope.RESULTS: The articular disc was adhered to the mandibular condyle after 3months. Temporal fossa adhered to the articular disc and mandibular condyle could still be observed in one specimen. Bifid condylar deformity was found in two specimens and no disc perforation was found. The adhesions became more severe after 6 months and bifid condylar deformity was formed in all the 3 specimens, with distinctly thickened discs. Light microscope observation:Three months after fracture, the adhesion between disc and condyle was obvious. There was no clear boundary between the surrounding tissues. The adhesion was composed of fibro-connective tissues, with a large number of chondrocytes in it. Fibro-connective tissues tightly adhered to the trabeculae were found in some adhesions. The fiber of disc became deranged and blood vessels and fatty cells were seen. Six months after fracture, blood vessels, a large number of fibroblasts and sparse chondrocytes were seen in the adhesions. The collagen fibers of the discs were arranged in whirlpool shape. There were a large number of fatty cells and blood vessels within it. The fiber-cartilage of the condylar surface was found thickened and the fiber deranged in the unadhered tissues. Proliferative changes were seen on disc surfaces.CONCLUSION: Childhood longitudinal fracture of the mandibular condyle causes severe secondary injury to TMJ, suggesting a close correlation between longitudinal fracture of the mandibular condyle and TMJ ankylosis.

Objective To investigate the effects of ARHI transfection on the chemokines and receptors related gene expression profile of PANC1 cells.Methods Plasmids expressing ARHI and empty plasmid were transfected into PANC1 cells, and the stably expressed cell lines were established by using G418.mRNA expression of chemokines and receptors related genes was detected by PCR Array.Real-time PCR was used to detect mRNA expression of the genes related vascular growth.Results In cells transfected with ARHI gene, the expression levels of mRNA of 36 genes were down-regulated, and 9 were up-regulated.Among the genes related to tumor metastasis and invasion CXCL12 and CXCR4 were significantly down-regulated (<-6 folds), and MMP-2 was slightly down-regulated (<-2 folds).Among the genes related to tumor angiogenesis, pro-angiogenesis genes including CCL2, CXCL1, CXCL2, CXCL8, CXCL12 and CXCR4 were significantly down-regulated, and pro-angiogenesis genes including CXCL3, CCR1 and CCR2 was slightly down-regulated.Anti-angiogenesis genes including CXCL9, CXCL10, CXCL11 and CXCR3 were significantly up-regulated (>6 folds).Among the genes related to the localization of distant organ infiltration and latency, CXCL12, CXCR4 and CCR7 were significantly down-regulated,and CXCR5 was slightly down-regulated.Among the gene with tumor immunity,CXCL8,CXCR1 and CCR7 were significantly down-regulated.Gene expression of CXCL1,CXCL8,CXCR4 and CXCR3 detected by Real-time PCR were consistent with PCR array.Conclusions ARHI gene inhibits the expression of chemokines and receptors related to tumor metastasis,angiogenesis and tumor immunity.

BACKGROUND:Development of new coronary thrombolytic agents is hot in the market. A new drug, mutated recombinant tissue-type plasminogen activator (rtPAm), is the product of mutation of tPA by changing binding loci with plasminogen activator inhibitor (PAI)-1 to reduce the degradation. In vitro test has demonstrated that the activity of rtPAm is much higher than rtPA in the absence of PAI. The present study is to observe the efficacy of mutated recombinant tissue-type plasminogen activator (rtPAm) in coronary thrombolytic therapy. METHODS:A total of 30 adult beagles were equally divided into 5 groups after thrombi:vehicle group, urokinase group, rtPAm low-dose group, rtPAm medium-dose group, and rtPAm high-dose group. Thrombolytic effect and myocardial infarction were observed after thrombolytic therapy. RESULTS:In the urokinase group, time to reperfusion was (15.8±3.8) minutes. TIMI 2 flow was demonstrated in 4 beagles, TIMI 3 flow in 2, and re-occlusion in 4 after 90 minutes respectively. In the low-dose rtPAm group, time to reperfusion was (15±4.5) minutes; TIMI 2 flow was demonstrated in 2 beagles, TIMI 3 flow in 4, and re-occlusion in 2 after 90 minutes. In the high-dose rtPAm group, time to reperfusion was (7.5±2.6) minutes. None of the beagles showed re-occlusion after 90 minutes. The infarction areas were (2.1+0.9)％ in the medium-dose rtPAm group and (0.7+0.4)％ in the high-dose rtPAm group, which decreased significantly than those in the low-dose rtPAm group. The aggregation rate in the medium-dose and high-dose rtPAm groups decreased significantly than that in the urokinase group. CONCLUSION:rtPAm may serve as a thrombolytic agent with platelet-targeted fibrinolysis and antiplatelet aggregation activities.

Objective To summarize the clinical experience of three different treatments for gallbladder and common bile duct stones.Methods The clinical data of 180 cases of gallbladder stones combined with bile duct stones undergoing surgery from May 2010 to May 2013 were retrospectively analyzed.They were divided into three groups,A group of 60 patients underwent a period of endoscopic sphincterotomy (EST),under the second phase of laparoscopic cholecystectomy (LC);Group B 60 patients underwent laparoscopic cholecystectomy (LC) + laparoscopic common bile duct exploration surgery (LCBDE) + T tube drainage;Group C 60 patients underwent conventional open cholecystectomy (OC) + bile duct exploration (OCBDE) + T tube drainage.Results A group of 53 cases completed surgery successfully,5 cases of remaining 7 patients failured in the first phase surgery,2 cases of the 5 patients did LC + LCBDE,3 of the 5 patients underwent conventional surgery.Two patients underwent the conventional surgery in the second phase surgery.B group of 57 cases completed surgery successfully,three cases convert to open surgery.All of the group C completed the surgery successfully.Group A complication was the most in the three groups (P＜0.05);group B had the shortest time of hospitalization (P＜0.05),the complication rate was lower than that in group A (P＜0.05),the complication had no significant difference between A and B.(P and group B ＞ 0.05),group B had shortest operation time (P＜0.05);no statistically significant differences were found among three groups in fasting time.Conclusion Three treatment methods have advantages as well as disadvantages,a reasonable treatment should be selected according to the specific circumstances.