Objective To explore the effects of chronic electrical stimulation (ES) on Nogo-A expression in the hippoeampus of rats.Methods Thirty male Wistar rats were randomly divided into a control group and 4 exper- imental groups.The rats in the control group were given sham ES,while those in the experimental groups received 1, 3,6,or 9 days of ES before being sacrificed for the detection of Nogo-A expresson in the hippoeampus by immunohis- tochemistry and Western plotting.Results There was a positive correlation between the level of Nogo-A expression and the duration of ES,as shown by the immunohistochemistry technique.Western blotting showed the same result. Conclusion In general,seizure occurred 8 days after electrical stimulation began.Elevated Nogo-A expression in the hippocampus began earlier than seizures in the epilepsy model groups.

Objective To understand the specimen sources,clinical characteristics,and antimicrobial resistance of Burkholderia cepacia (B .cepacia )isolated from infected patients in intensive care unit(ICU),so as to provide reference for guiding rational use of antimicrobial agents.Methods Clinical data of patients with B .cepacia infec-tion in an ICU between 2011 and 2014 were analyzed retrospectively,antimicrobial resistance of strains was ana-lyzed.Results A total of 267 B .cepacia strains were isolated,the major specimen sources were sputum (80.15%, n=214),blood(14.23%,n =38),and urine(3.37%,n =9).Antimicrobial susceptibility testing results revealed that B .cepacia had multiple resistance,and was naturally resistant to multiple clinically used antimicrobial agents, such as ampicillin,cefazolin,ampicillin/sulbactam,nitrofurantoin,and cefuroxime,resistant rates were all 100%;resistant rates to ceftazidime and levofloxacin were 4.12% and 3.00% respectively;resistant rate to compound sulfa-methoxazole had increased tendency(χ2 =5.885,P =0.015).Conclusion Isolation of B .cepacia in ICU increased year by year,antimicrobial resistance is serious,management and targeted monitoring of prevention and control of healthcare-associated infection should be strengthened,antimicrobial agents should be chosen according to antimi-crobial susceptibility testing results.

Objective To understand colonization of pathogens in nasal vestibular of health care workers (HCWs) in intensive care unit (ICU),and provide evidence for strengthening the prevention and control of healthcare-associated infection (HAI)in ICU.Methods On may 2015,colonization status of pathogens in nasal vestibular of uninfected HCWs in ICU were actively screened,bacterial culture,isolation and identification were performed.The surveyed results were analyzed and compared with antimicrobial resistance of pathogens from patients at the same stage.Results A total of 96 HCWs were surveyed,43 pathogenic strains were isolated from different HCWs’na-sal vestibular,isolation rate and carriage rate were both 44.79%.The main pathogenic bacteria was Staphylococcus aureus(n=15,34.88%),followed by Enterobacter aerogenes (n =9,20.93%)and Klebsiella pneumoniae (K . pneumoniae ,n=7,16.28%).There was a high detection rate of pathogens from nasal vestibular of doctors,HCWs who smoked frequently and those who never exercised (all P <0.05).There were 1 strain of imipenem-resistant K . pneumoniae among 43 pathogenic strains.Resistance rate of 7 K .pneumoniae from HCWs to ampicillin/sulbactam, cefazolin,and furantoin were all >50.00%,resistance rates to cefotaxime and imipenem were 28.57% and 14.29%respectively;resistance rates of 11 strains of K .pneumoniae from patients to furantoin was 100.00% during the same stage,but were sensitive to other commonly used antimicrobial agents.Resistance rate of 4 strains of Esche-richia coli (E.coli)to ampicillin was 75.00%,to gentamicin,tobramycin,levofloxacin,ciprofloxacin,and com-pound sulfamethoxazole were all 50.00%,6 strains of E.coli isolated from patients during the same period were found to be resistant to most commonly used antimicrobial agents.Conclusion Colonization rate of pathogens is high in nasal vestibular of HCWs in ICU,active screening and monitoring on colonization of pathogens in HCWs’ nasal vestibular is significant for preventing the occurrence and cross transmission of HAI among HCWs and pa-tients.

OBJECTIVETo observe the effect of electric acupuncture (EA) on the Nogo receptors (NgR) protein expression in the cerebral cortex, the medulla oblongata, and the spinal cord of cerebral ischemia-reperfusion (I/R) stroke-prone renovascular hypertensive rats (RHRSP) with middle cerebral artery occlusion (MCAO) at different time points, and to investigate its possible mechanisms for remote-organ injury of acute cerebral infarction (ACI).

METHODSThe RHRSP model was duplicated in male SPF grade SD rats. Then the MCAO model was prepared by a thread stringing method. Rats were divided into the hypertension group,the sham-operation group, the MCAO group, the EA group, and the sham-acupoint group by random number table method, 60 in each group. Rats in the MCAO group only received MCAO reperfusion treatment. Those in the sham-operation group only received surgical trauma. Baihui (DU20) and Dazhui (DU14) were needled in the EA group, once daily for a total of 28 days.The needles were acupunctured at the skin one cun distant from Baihui (DU20) and Dazhui (DU14) and then the same EA treatment was performed in the sham-acupoint group. At day 1, 7, 14, 28 after treatment, six rats were executed from each group, and their right cortex and medulla oblongata, and the left spinal cord were isolated. The infarct volume was detected by Nissl's staining method. The NgR expression was detect by Western blot.

RESULTS(1) In the cortex area: compared with the hypertension group,the NgR expression increased in the MCAO group at day 1,7,14,and 28 after MCAO (P < 0.05). Compared with the MCAO group, the NgR expression of the EA group and the sham-acupoint group were equivalent at 1 day af ter MCAO (P > 0.05). At day 7, 14,and 28 after MCAO, the NgR expression decreased in the EA group (P < 0.05), it was quite similar to that in the sham-acupoint group (P > 0.05). (2) In the medulla oblongata area: compared with the hypertension group, the NgR expression was equivalent in the sham-operation group. the MCAO group,the EA group, and the sham-acupoint group at 1 day after MCAO (P > 0.05). At day 7.14, and 28 after MCAO, the NgR expression increased in the MCAO group (P < 0.05). Compared with the MCAO group,the NgR expression decreased in the EA group at day 7, 14, and 28 after MCAO (P < 0.05), whereas it was similar in the sham-acupoint group (P > 0.05). (3) In the spinal cord area: compared with the hypertension group, the NgR expression was equivalent in the sham-operation group, the MCAO group,the EA group, and the sham-acupoint group at day 1 and 7 after MCAO (P > 0.05). At day 14 and 28 after MCAO, the NgR expression increased in the MCAO group (P < 0.05). Compared with the MCAO group, the NgR expression decreased in the EA group at day 14 and 28 after MCAO (P < 0.05), whereas it was equivalent in the sham-acupoint group (P > 0.05).

CONCLUSIONSIncreased NgR expression in the cerebral cortex, the medulla oblongata, and the spinal cord of cerebral infarct rats was an important reason for involving remote-organ injury of ACI. The protective effect of EA on hypertensive I/R cerebral injury rats might be closely related to down-regulating central nervous system myelin growth inhibition mediated factors Nogo-A receptor NgR protein expression.

Animals ; Cerebral Infarction ; metabolism ; therapy ; Disease Models, Animal ; Electroacupuncture ; GPI-Linked Proteins ; metabolism ; Hypertension, Renal ; metabolism ; therapy ; Male ; Medulla Oblongata ; metabolism ; Myelin Proteins ; metabolism ; Nogo Receptor 1 ; Rats ; Rats, Sprague-Dawley ; Receptors, Cell Surface ; metabolism ; Spinal Cord ; metabolism

AIM:To investigate the effect of trans-activator of transcription-Rab guanine nucleotide exchange factor 1 (Tat-RabGEF1) fusion protein by intravenous injection on hepatic ischemia/reperfusion (IR) -induced lung injury, and its underlying mechanism.METHODS:Adult male C57BL/6 rats (n=24) were randomized into 3 groups with 8 rats each:sham group, IR group, and Tat-RabGEF1 group.IR was induced by occlusion of the vessels in the hepatic pedicle for 90 min followed by 4 h of reperfusion.The Tat-RabGEF1 protein was intravenously administered right after reperfusion through the caudal vein.The levels of tumor necrosis factor-α (TNF-α) , interleukin-6 (IL-6) and interleukin-1β (IL-1β) in bronchoalveolar lavage fluid (BALF) were assessed by ELISA.The lung histopathological injury score and lung wet/dry weight ratio were evaluated, and the levels ofβ-hexosaminidase A (β-Hex A) and myeloperoxidase (MPO) in the lung tissues were also measured.The expression of tryptase in the lung was determined by Western blot.RESULTS:The pathological changes after ischemia for 90 min followed by reperfusion for 4 h were significant, along with increased lung wet/dry weight ratio and lung injury score.Intravenous injection of Tat-RabGEF1 protein effectively alleviated the lung pathological injury (P<0.05).Compared with IR group, lower levels of TNF-α, IL-6 and IL-1βin BALF, and lower expression ofβ-Hex A, MPO and tryptase in the lung tissues in Tat-RabGEF1 group were observed (P<0.05).CONCLUSION:Intravenous injection of Tat-RabGEF1 protein attenuates the lung injury after IR, and its mechanism may be related with inhibition of mast cell activation, reduced releases ofβ-Hex A and tryptase, and decreased levels of TNF-α, IL-6, IL-1βand MPO.

OBJECTIVETo characterize the HA1 regions of hemagglutinin gene of influenza viruses (H3N2) isolated from children in Beijing from 1998 - 2004.

METHODSThe HA1 regions of hemagglutinin gene were amplified by RT-PCR from the viruses isolated and identified as A3 (H3N2) from clinical samples collected from infants and children during the peak seasons of influenza between 1998 and 2004. PCR products were sequenced or cloned into T-A vector and were analyzed after being sequenced.

RESULTSThe HA1 regions of hemagglutinin genes amplified from those isolates were 987 bp in length, encoding a protein of 329 amino acids in length. The identities of nucleotides and amino acids among these H3N2 isolates in Beijing and vaccines strains from 1998 - 2004 were 95.5% - 100.0% and 93.0% - 100.0%, respectively. The homology of the HA1 regions were related to the date of virus isolation, meaning the homology was higher among those strains isolated in nearer dates than others. Seven potential N-linked glycosylation sites in the HA1 regions located at amino acid positions 8, 22, 38, 63, 126, 165 and 285 were conserved in all the viruses analyzed. Two sites at 122 and 133 were inserted in those virus isolated after 1997, and another site at 144 appeared in those isolated after 1999. More amino acid substitutions located in the five putative antigenic sites or receptor binding sites were found more in the isolates than the isolates from previous year. Phylogenetic analysis showed new branches appeared continuously during 1998 - 2004. The strains isolated during winter in 2004 belonged to different branches, suggesting the appearance of new variants.

CONCLUSIONAmino acid substitutions continuously occurred in the HA1 regions of hemagglutinin genes in influenza virus (H3N2) isolated from children in Beijing from 1998 - 2004, which might have resulted in antigenic drift and led to the appearance of new variants.

Amino Acid Substitution ; China ; DNA, Viral ; analysis ; Gene Amplification ; Hemagglutinins ; genetics ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; Influenza, Human ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA

OBJECTIVETo establish a rapid, specific and effective technique for identifying subtyping A(1), A(3) and B of influenza virus isolates and clinical specimens as well as to analyze the sequences of nucleotides and deduced amino acids of HA1 regions from isolates of influenza virus A(3) isolated from 1996 to 2002.

METHODSSix inner and outer sets of oligonucleotide primers were designed to detect, type and subtype human influenza A and B. The first two corresponding sets differentiate type A and B of matrix (M) gene while, the second two corresponding sets identify the H(1) and H(3) subtypes of type A virus HA gene. To type and subtype influenza viruses in clinical isolates, a mixture of inner primer sets specific for H(1), H(3) and B were used in a single PCR reaction tube. To detect influenza viruses in clinical specimens, a mixture of the outer primer sets were used in a single primary PCR tube, and the inner ones in a single second PCR reaction tube. Amplified products were visualized in 1.2% agrose gel containing ethidium bromide. HA1 regions of hemagglutinin of 5 field strains (H3N2) isolated from 1996 to 2002 in Beijing were amplified by RT-PCR and sequenced directly.

RESULTSThere was 100% correlation between multiplex RT-PCR and culture to type and subtype influenza viruses from clinical isolates. For typing and subtyping, 76.9%, 57.1% and 86.5% were positive for A(1), A(3) and B by multiplex nested-PCR compared within virus isolation on culture, respectively. The sequence data of HA1 of A(3) strains showed that there was a high homology of nucleotide and amino acid, and the closer the date of isolating was, the higher homology showed.

CONCLUSIONSMultiplex RT-PCR and nested-PCR for influenza viruses could provide a useful alternative to existing methods of influenza detected and identified from clinical isolate and specimens. There were certain, continuous mutations and increasing glycosylated sites which might cause the antigen drift in the A(3) strains during 1996-2002 in Beijing area.

Amino Acid Sequence ; Child ; China ; epidemiology ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A virus ; classification ; genetics ; isolation & purification ; Influenza B virus ; classification ; genetics ; isolation & purification ; Influenza, Human ; epidemiology ; virology ; Molecular Sequence Data ; Point Mutation ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To explore the relationship between the expression of nerve growth factor (NGF) in bladder tissue of Sprague-Dawley rats with diabetes mellitus and the changes of bladder structure and urodynamics.Methods Sixty-three Sprague-Dawley rats were selected and divided into diabetic group (n =33) and control group (n =30).The diabetes mellitus model was made by injecting of steptozotocin (STZ) into the tail vein of rats in the diabetic group;the rats in control group were injected the same volume of citrate buffer solution.The 24 h urinary volume was detected by using metabolic cage;the function of bladder was detected by using Medlab bio-signal collection system;the expression of NGF in the bladder tissue of rats was detected by Western blot;the bladder structure was observed by using light microscope and transmission electron microscope (TEM).Results Four,eight and twelve weeks after the success of diabetic rat model,the level of blood glucose of rats in diabetic group was significantly higher than those in the control group (P ＜ 0.05);the body weight of rats in diabetic group was significantly lower than that in the control group (P ＜ 0.05).The 24 h urinary volume of rats in diabetic group was significantly increased compared with control group (P ＜ 0.05).The maximum capacity of bladder and residual urine volume of bladder and elasticity of bladder of rats in diabetic group were significantly higher than those in the control group (P ＜ 0.05);the maximum bladder pressure of rats in diabetic group was significantly lower than that in the control group (P ＜ 0.05);and the changes of all above indexes of rats in diabetic group were time-dependent(P ＜ 0.05).The expression of NGF in bladder tissues of rats in diabetic group was significantly lower than that in the control group (P ＜ 0.05),and the change of NGF expression in diabetic group was time-dependent (P ＜ O.05).In diabetic group,the bladder smooth muscle was hypertrophic,the organization of smooth muscle was diverse and lose,and the muscle bundle fracture was observed;in control group,the bladder smooth muscle arranged orderly,and the shape was consistent,the structure of fascicle was compact.The condensation and margination of nuclear chromatin and mitochondria swelling were observed in diabetic group;while the bladder tissues in control group showed homogeneous nuclear and mitochondria without fracture and swelling.Conclusion The decrease of the expression of NGF in bladder tissues of diabetic rats can decrease the systolic function of bladder and change the morphology of detrusor and these changes show a time dependence.

Objective To investigate the expression and significance of malonaldehyde (MDA), superoxide dismutase (SOD) and endotoxin (ET) in liver injury model of Budd-Chiari syndrome (BCS) in rats. Methods The animal model of BCS was established by partially ligating the inferior vena cava of the posterior segment of liver in rats. The experimental animals were divided into three groups: control group (12 rats), model group (48 rats) and sham operation group (48 rats). The model group and sham operation group were divided into four subgroups (1, 3, 6, 12 weeks) of 12 rats each. After the success of modeling,being confirmed by digital subtraction angiography (DSA), nine rats in each group were sacrificed at random respectively, where their serums and liver tissues was collected. The levels of MDA, SOD and ET in both liver homogenate and serum were examined respectively. ANOVA was used to compare the total difference between groups and within group of each measurement data. The LSD method was used to do multiple comparison within group and between groups. Pearson method was used to do correlation analysis of hypoxia markers. Results The levels of MDA, SOD and ET in liver homogenate and serum at different time points in model group were significantly different from those in control group and sham operation group (MDA: liver homogenate (F=52.906, 219.016), serum (F=21.573, 43.878); SOD: liver homogenate (F=22.927, 19.317), serum (F=10.841, 31.643);ET: liver homogenate (F=33.588, 105.515), serum (F=40.832, 46.323);P＜0.05). The total difference of the MDA level in serum at each time point after the operation was not statistically significant in model group(F=1.965,P=0.139), but that of liver homogenate in the model group was statistically significant (F=7.716, P=0.001). The SOD and ET levels in both liver homogenate and serum of model group were compared within groups at different time points after operation respectively, and the overall difference was statistically significant (SOD: F=17.053, 7.903; ET: F=19.870, 39.372; P＜0.05). The time-varying curves of MDA and ET in liver homogenate and serum in model group were similar, which both increased from 1 week after operation,peaked at 6th week and slightly decreased at 12th week. The increase levels of MDA and ET in liver homogenate were significantly higher than those in serum. There was a negative correlation between MDA and SOD in liver homogenate and serum (r=－0.814,－0.591;P=0.001, 0.001), a positive correlation between MDA and ET (r=0.761, 0.422; P=0.004, 0.001), and a negative correlation between SOD and ET (r=－0.726,－0.490;P=0.001, 0.001). Conclusions The levels of hypoxia related markers, such as MDA, SOD and ET in liver and serum of BCS animal model, change to varying degrees in the early stage, and will be aggravated as the disease continues to advance. In the later stage, with the establishment of collateral circulation, hypoxia will be slightly eased, but is still significantly higher than normal, which indicates that congestion and hypoxia run through the whole process of BCS, and could be the key and initiating factors.

Objective To analyze pathogenic factors and etiological characteristics of suppurative endophthalmitis.Methods A total of 531 consecutive patients (531 eyes) with suppurative endophthalmitis who were hospitalized in Qingdao Eye Hospital of Shandong Eye Institute from January 2006 to December 2015 were included in the study.Among them,410 patients with 410 eyes were males (77.2％),121 patients with 121 eyes were females (22.8％).The average age of the patients was 38.62± 15.36 years.The relevant medical records were collected to analyze the pathogenic factors.Samples of aqueous humor,vitreous or other intraocular samples were taken under aseptic conditions for bacterial and fungal culture and in vitro drug sensitivity test.Results Ocular trauma was the primary pathogenic factor of suppurative endophthalmitis (60.1％),other factors included postoperative endophthalmitis (19.0％),suppurative keratitis-related endophthalmitis (17.1％) and endogenous endophthalmitis (3.8％).Postoperative endophthalmitis mainly occured after cataract surgery.A total of 224 strains of organisms were isolated,among which the predominant organisms isolated were gram-positive bacteria (54.0％) and staphylococcus epidermidis was the most common (25.0％).The other pathogenic organisms were fungi (29.5％) and gram-negative bacteria (16.5％).Among the fungi,aspergillus (10.7％) was the dominant genus,followed by fusarium (9.8％).For gram-positive organisms,susceptibilities were vancomycin 97.4％,gatifloxacin 91.8％,fusidate acid 77.9％ and levofloxacin 54.6％.For gram-negative organisms,susceptibilities were gatifloxacin 85.7％,levofloxacin 77.8％,tobramycin 71.4％ and ceftazidime 62.5％.For fungal isolates,sensitivities were voriconazole 88.2％ and amphotericin B 84.8％.Conclusions Ocular trauma is the main pathogenic factor of suppurative endophthalmitis,followed by postoperative endophthalmitis and suppurative keratitis-related endophthalmitis.Gram-positive bacteria are the major pathogenic organisms,especially staphylococcus epidermidis followed by fungal species,among which aspergillus and fusarium were the dominating pathogenic genus.