Objective To investigate the effects of special siRNA targeting TLR4 gene (TLR4-siRNA) on hyperoxia-induced secretion of inflammatory factors from human alveolar epithelial cell line A549.Methods Human alveolar epithelial cell line A549 was purchased from Chongqing Children Hospital and cultured in six-well tissue culture plates and randomly divided into 4 gorups:A549 cells + ambient air group (group C) ; A549 cell + hyperoxia group (group H); A549 cells transfected with TLR4-siRNA + hyperoxia group (group TR) and A549 cells transfected with control-siRNA + hyperoxia group (group NR).TLR4-siRNA and control-siRNA were transfected into A549 cells through lipofectamine 2000.The efficiency of transfection was assessed by detection of FAM expression using flow cytometry.The cells were exposed to 95％ O2-5％ CO2 delivered at 1 L/min for 30 min in groups H,TR and NR.TLR4 mRNA and protein expression was detected by RT-PCR and cytometry respectively.The NF-κB activity in A549 cells was measured by EMSA and the IL-6 and IL-8 levels in supernatant were determined by ELISA.Restults Hyperoxia significantly up-regulated TLR4 mRNA and protein expression and increased NF-κB activity in cells and IL-8 and IL-6 concentrations in supematant in group H as compared with group C.Transfection with TLR4-siRNA significantly attenuated hyperoxia-induced up-regulation of TLR4 mRNA and protein expression and increase in NF-κB activity in the cells and IL-6 and IL-8 concentrations in supernatant in group TR as compared with group H,but transfection with control-siRNA did not.Conclusion TLR4-siRNA could significanly inhibit hyperoxia-induced secretion of inflammatory factors from human alveolar epithelial cell line A549.

Objective To investigate the mechanism of brain injury caused by thrombin and the intervention effect of hirudin and nimodipine.Methods Different doses of thrombin or/and hirudin were injected into nucleus caudatus of SD rats,nimodipine was given intraperitoneally.Dry-wet-weighing technique,immunohistochemical method and TUNEL were used to examine brain edema,the changes of cytoskeleton,neuron apoptosis and histological changes.Results(1) High dose of thrombin resulted in severe cerebral edema as early as 4 h after injection and the maximum edema occurred at 24~48 h.The edema gradually decreased and got close to normal within 3~7 d.Cytoskeleton changes were observed at early stage(4h),reversible or irreversible injuries presented at 24~48 h,and neuron necrosis occurred within 3~7 d.Neuron apoptosis started at 4h and peaked at 24~48 h.In contrast,low dose of thrombin and normal saline did not show these effects.(2) The effects of thrombin could be inhibited by hirudin and nimodipine(a calcium-ion antagonist) could relieve or delay cell injury.Conclusions High dose of thrombin may result in severe brain edema,neuron irreversible injury and apoptosis,which all peak at 24~48 h.Early treatments could greatly reduce brain damage and improve prognosis of intracerebral hemorrhage.

Objective To investigate the genotype and drug resistance of extended-spectrum beta-lac-tamases(ESBLs) -producing Shigella in pediatric patients.Methods A total of 59 strains of Shigella were isolated from stool specimens of hospitalized children with shigellosis in Beijing Children's Hospital from January 2004 to December 2008.Phenotypic confirmatory test,which is based on Clinical and Laboratory Standards Institute(CLSI),was used to detect the ESBLs-producing strains.Agar dilution method was used to determine the minimal inhibitory concentration (MIC).PCR amplification was performed for ESBLs producers to determine the genotype.PCR product was sequenced and then analyzed to confirm the subtype of ESBLs.Results Of the 59 isolates,21 (35.6%) strains were identified as ESBLs producers.The 21 strains of ESBLs-producing Shigella all carried the genes of CTX-M as shown by PCR,and CTX-M-1,CTX-M-9 accounted for 6,15,respectively.Among the 21 CTX-M producers,there were 4 strains accompanied by TEM-type and 6 strains accompanied by OXA-type.Nucleotide sequence analysis showed that there were CTX-M-3 (n = 1),CTX-M-15 (n = 2),CTX-M-57(n =3) of the 6 CTX-M-1-producing isolates.The subtypes of CTX-M-9,TEM,OXA were all CTX-M-14,TEM-1,OXA-1,respectively.The sensitive drugs to ESBLs producers were imipenem,meropenem,piperacillin/tazobactam,cefoperazone/sulbactam and cefoxitin,with resistance rate all less than 15%.The resistance to ceftazidime was remarkably variable among different CTX-M producers.Conclusion The prevalence of ESBLsproducing Shigella is in a high level in pediatric patients in this area.The genotypes of ESBLs are all CTX-M.Most of them are CTX-M-14,but some are CTX-M-3,CTX-M-15 and CTX-M-57.Most ESBLs-producing strains are multidrug resistant.Carbopenems should be the first choice for ESBLs producers.

Objective To explore the value of ocular fundus artery hemodynamics in diabetes by the correlation study between ocular fundus artery and glycosylated hemoglobin(GHb).Methods GHb was checked in 98 patients (196 eyes) in diabetes,including 49 cases (98 eyes) before treatment and 49 cases (98 eyes) after treatment,and normal group of 100 cases (200 eyes)An all cases,color Doppler ultrasonography was used for monitorind ophthalmic artery(OA),central retinal artery(CRA) and posterior ciliary artery (PCA).Finally,the differences of the parameters vauels of the the OA,CRA,and PCA between diabetic and normal group were studied in order to find out the relationship with GHb.Results Of 98 patients in diabetes,peak systolic velocity and end diastolic velocity of fundus artery decreased,resistance index increased(RI).There was significant difference between diabetic and normal group(P ＜0.05),and a significant difference was exited between diabetic group before and after treatment(P ＜0.05).GHb were negatively correlated with the velocity of fundus artery,and positively correlated with RI.Conclusions Evaluation of ocular fundus artery hemodynamics in diabetes can provide useful information for diabetic retinal perfusion and function change.For evaluation of diabetic retinal disease,it is of great value in clinical application.

Arrhythmias, Cardiac ; etiology ; Coronary Vessels ; pathology ; Humans ; Male ; Mediastinal Cyst ; pathology ; Young Adult

With dwindling number of new antibiotics and inappropriate use of antibiotics, the emergence and spread of antibiotics resistance occurs commonly in healthcare institutions worldwide. In China, antibiotics are commonly overprescribed and misused. This study is to assess the effect of the nationwide special campaign on antibiotic stewardship program (ASP) at specialized hospitals in China by investigating prescription information from 2011 to 2012. Data on the hospital consumption and prescription of systemic antibiotics were obtained from four specialized hospitals, including maternity, children's, stomatological and cancer hospitals. Systematic random sampling was used to select outpatient prescriptions and inpatient cases. A total of 105 specialized hospitals in 2011 and 121 specialized hospitals in 2012 were analysed. The defined daily doses (DDDs) per 100 inpatient days, the percentage of antibiotic use in outpatient prescriptions, and the percentage of antibiotic use in inpatient cases were used as measurements of antibiotic use. The overall antibiotic use density in the selected hospitals decreased between 2011 and 2012 from 39.37 to 26.54 DDD/100 inpatient days (P<0.001). The percentage of antibiotic use in outpatient prescriptions (range: 24.12%-18.71%, P=0.109) and inpatient cases (64.85%-60.10%, P=0.006) also decreased within the two years. Significant changes were observed among regions and different hospitals within the two years. And antibiotic consumption was correlated with the type and size of specialized hospital in 2012, but not with the regions. This analysis of antibiotic consumption of specialized hospitals allows relevant comparisons for benchmarking and shows that national ASP has improved antibiotic rational use in China. The data will assist policymakers in formulating effective strategies to decrease antibiotic overuse and identify areas that require further work.

Objective To investigate the protective effect of methylene blue (MB) on blood-brain barrier (BBB) injury after focal cere-bral ischemia-reperfusion in rats. Methods 18 male Sprague-Dawley rats were randomly divided into sham-operated group (n=6), model group (n=6) and MB treatment group (n=6). The left middle cerebral arteries were occluded for 1 hour and reperfused. MB was infused intra-venously immediately after reperfusion (3 mg/kg) and again 2 hours post-reperfusion (1.5 mg/kg), while normal saline was administered in the model group. The sham-operated group was treated as same as the model group without occlusion and infusion. HE staining was used to observe the histological injury in the cortex around the infarcted region 47 hours after reperfusion, while albumin immunohistochemistry was used to evaluate the permeability of the BBB, and immunohistochemistry and double immunofluorescence staining were used to exam-ine the expressions of glial fibrillary acidic protein (GFAP) and aquaporin-4 (AQP-4). Results HE staining showed that cells and blood ves-sels were not intact in the cortex around the infarcted region in the model group and they were better in the MB treatment group. The expres-sions of the albumin, GFAP and AQP-4 were higher in the model group than in the sham-operated group (P<0.01), and were lower in MB treatment group than in the model group (P<0.05). The double immunofluorescence staining showed the colocalization of GFAP and AQP-4 in the astrocytes. Conclusion MB may ameliorate the BBB disruption induced by focal cerebral ischemia-reperfusion through reducing glio-cyte proliferation and down-regulation of AQP-4 expression in rats.

Clavicle ; injuries ; Fractures, Bone ; therapy ; Humans

Objective To investigate the change of the first-phase insulin secretion in impaired glucose regulation and its influencing factors.Methods The investigation was performed in 1024 subjects who were selected by cluster sampling.The body waist circumference (WC) was examined in these subjects.Oral glucose tolerance test was performed,and fasting plasma glucose,fasting insulin as well as 30-,60-,and 120-minute plasma glucose and insulin after glucose intake were also tested.All the 1024 subjects were divided into 5 groups based on the results of oral glucose tolerance test:normal glucose tolerance group,impaired fasting glucose group,impaired glucose tolerance group,impaired fasting glucos/impaired glucose tolerance group,and diabetes mellitus group.First-phase insulin secretion index (the ratio of change in insulin to change in glucose during the first 30 minutes after glucose ingestion was calculated.Results Compared with normal glucose tolerance group (1.96 ± 1.03),the first-phase insulin secretion index significantly decreased in the impaired fasting glucose group (1.79 ±0.91) (P =0.007),the impaired glucose tolerance group (1.81 ± 0.97) (P =0.007),the impaired fasting glucose / impaired glucose tolerance group (1.59 ± 0.85) (P =0.005),and the diabetes mellitus group (1.30 ± 0.60) (P =0.004).Logistic regression analysis showed that WC was the strongest influencing factor of first-phase insulin secretion (β =0.716,P =0.000).Conclusions The firstphase insulin secretion index has already dropped in the stage of impaired glucose regulation.WC can be a useful indicator for evaluating first-phase insulin secretion.

Objective To investigate the clinical skill and experience of laparoscopic herniorrhaphy in the treatment of abdominal hernia. Methods Clinical data of 30 cases of abdominal hernia undergoing laparoscopic her-niorrhaphy in Shengjing Hospital from May 2004 to December 2008 were analyzed retrospectively. There were 23 in-guinal hernia,4 incision hernia and 3 hiatal hernia. TEP in 20 cases,TAPP in 3 cases,lPOM in 4 cases and Nissen hiatal hernia repair in 3 cases were performed. Results The mean operating duration was 72 min for TEP, 127 for TAPP,150 min for IPOM,and ]60 min for hiatal hernia repair and the mean time for hospitalization was 4.6,5.8, 7.5, and 5.3 days. No conversion to open surgery happened among these cases. No recurrence was found. Conclu-sions Laparoscopic herniorrhaphy appears to be an effective and convenient technique for the treatment of abdomi-nal hernia and has many advantages of minimally invasive surgery.