BACKGROUND: Whether horizontal and vertical impacts cause craniocerebral injury of the same degree, and whether serum myelin basic protein (MBP) changes is associated with the direction of impact have been scarcely addressed in previous studies.OBJECTIVE: To observe the change in the serum MBP levels and morphological changes of the brain tissue rabbits with craniocerebral injury resulting from impacts from different directions.DESIGN: A randomized controlled experiment.SETTING: Department of Neurosurgery of a hospital affiliated to a medical university.MATERIALS: This study was conducted in the Central Laboratory of Shenyang Medical College between October 2003 and January 2004. Totally 30 healthy rabbits of either sex were randomly divided into two equal groups to receive vertical and horizontal impact on the head.METHODS: All the rabbits were anesthetized and fixed in prone position onto a table equipped with a device for inducing impact on the head from different directions. Venous blood of 1 mL was taken from the edge of the ear of each rabbit for detecting MBP before injury. The rabbits in vertical impact group received the impact of a hammer falling directly on the parietal bone of the skull. In the horizontal group, the rabbits were subjected to horizontal impact on the lateral side of the skull. Forty-eight hours after the injury, venous blood were again taken for MBP measurement. The rabbits were then killed for pathological examination of the brain.MAIN OUTCOME MEASURES: Changes in serum MBP and pathomorphology of the brain tissues in the two groups after the impact.RESULTS: ① According to intention to treat analysis, all the 30 animals were involved in the experiment. There was no significant difference in serum MBP levels between the two groups before the impact, nor 48 hours after the impact( P ＞ 0.05). In the vertical impact group, serum MBP levels before the impact was( 1.68 ± 0. 86) μg/L, which was significantly lower than that after the impact[ (5.25 ± 1.96) μg/L, t = 3. 226, P ＜ 0. 05]. ② In the horizontal strike group, serum MBP also significantly increased from the level of( 1.70 ± 0.91 ) μg/L before impact to(5.73 ± 2.07) μg/L after that( t = 3. 080, P ＜ 0.05) . ③ After the impact, vasodilation and congestion in the cortex near the midline of the bilateral parietal region and edema in the deep layer of the white matter were observed in the vertical impact group, along with significant enlargement of the extravascular and extracellular space. In the horizontal group, similar changes except those near the midline of the bilateral parietal region took place after the impact; the same pathological changes were also observed in the basal plane, with nearly identical pathological changes in the deep layer of the white matter.CONCLUSION: Vertical and horizontal impacts can both result in almost identical brain injuries demonstrated by similar changes in serum MBP and the pathomorphology of the brain tissues after the impacts.

Objective To investigate the mechanism of carbapenems resistance in Serratia marces-cens strains isolated from Wenzhou and their epidemiological characteristics.Methods 147 non-duplicated Serratia marcescens isolates were collected from the First Affiliated Hospital of Wenzhou Medical University during 2006 to 2012.The antimicrobial susceptibility test for all isolates was performed by using Vitek2 Compact to screen carbapenems-resistant Serratia marcescens strains.The minimum inhibitory concentrations ( MICs) of 10 commonly used antibiotics against carbapenems-resistant Serratia marcescens strains were de-termined by agar dilution method.The phenotypes of carbapenemase were analyzed by using the modified Hodge test.PCR analysis was used to detect the genes encoding carbapenemase, AmpC enzyme, efflux pump and outer membrane proteins.The changes of MICs before and after using CCCP efflux pump inhibitor were measured by agar dilution method.Outer membrane proteins were detected by SDS-PAGE.Carbapene-ms resistance genes were transferred from carbapenems-resistant Serratia marcescens strains to recipient strains by conjugation.The transconjugants were amplified by PCR and measured for MICs.Pulsed-field gel electrophoresis ( PFGE) was used to analyze homology among strains.Results 11 isolates resistant to car-bapenems were screened out from 147 Serratia marcescens isolates and all of them were resistant to penicil-lins, cephalosporins and ertapenem.10 out of the 11 isolates were both resistant to imipenem and meropen-em, but remained susceptible to fluoroquinolones and aminoglycoside.Among the 11 isolates, 10 carried blaKPC-2 gene, 1 carried blaIMP-1 gene, 8 harbored both blaEBC and blaMOX genes, 1 harbored both blaEBC and blaDHA genes, and 1 carried blaEBC , blaMOX and blaDHA genes.No additional genes were identified by PCR.The MICs of imipenem to 7 isolates and the MICs of ertapenem to 3 isolates were respectively decreased by 4-64 folds and 8-256 folds after using CCCP.CCCP had no effects on the MICs of meropenem.Loss of outer membrane protein was not detected among the 11 isolates.The blaKPC-2 genes were successfully transferred from 7 isolates into recipient strains.The MICs against the transconjugants were higher than those against the recipient strains in varying degrees.PFGE analysis demonstrated that 8 out of 11 Serratia marcescens strains belonged to one clonotype.Conclusion KPC-2 carbapenemase played an essential role in carbapenems re-sistance in Serratia marcescens strains isolated from Wenzhou.Attention should be paid to the clonal spread of KPC-2 and its horizontal transmission in Wenzhou.

[ ABSTRACT] AIM: To investigate the differential expression of human leukocyte antigen-G ( HLA-G) isoforms and its receptors in human monocyte line THP-1 after human cytomegalovirus ( HCMV) infection for exploring the role of HLA-G in HCMV escaping the immune response of the organism .METHODS: THP-1 cells were infected with HCMV Towne strain.The expression of HLA-G isoforms at mRNA and protein levels was determined by RT-PCR and Western blot, respectively.The surface expression of HLA-G and its receptors ILT2/ILT4 and the cell viability were analyzed by flow cytometry.The levels of soluble HLA-G (sHLA-G) and IL-10 were measured by ELISA.RESULTS:After infection of the THP-1 cells with HCMV , no obvious apoptosis in the cells was observed , and the viability of the cells was high .A significant up-regulation of HLA-G1,-G3,-G4 and-G5 at mRNA expression level 1 d after infection was found , while the protein expression of HLA-G1 and HLA-G5 isoforms was mainly detected .The expression of HLA-G/ILT2/ILT4 was evi-dently up-regulated 1 d after infection .The level of sHLA-G was significantly increased 1 d after infection as compared with control group (P<0.01).The expression of IL-10 was obviously up-regulated 1 d post-infection as compared with control group.CONCLUSION:The differential expression of HLA-G isoforms and secretion of the receptors ILT 2/ILT4 and IL-10 in the THP-1 cells are induced after HCMV infection .This study provides experimental evidence for evaluating the immune mechanism of HCMV infection .

Objective To identify the clinical features and risk factors of early rheumatoid arthritis (RA)-associated osteoporosis in premenopausal women. Methods A total of 76 premenopausal women with early RA were randomly selected in the Department of Kidney and Rheumatology in the hospital. A total of 84 health cases were randomly selected in our hospital as controls. Bone mineral density (BMD) was determined using dual energy X-ray absorptiometry (DEX). Bone metabolism (CTX, PINP) and inflammatory cytokines (IL-17, IL-6, TNF-α) were examined with quantitative enzyme-linked immune-sorbent assay (ELISA). Quantitative data were expressed as x ±s deviation and the data were compared between groups using non- parametric test (Z value). Multi-group comparison was performed with variance analysis. Qualitative data were compared with Fisher's test. Logistic regression was used to investigate the risk factors. Results ①Compared with the control group, BMD in the premenopausal women with early RA group [neck: (0.802 ±0.193) g/cm2, GT zone: (0.923±0.033) g/cm2, L1: (0.862±0.011) g/cm2] was significantly decreased [(0.981±0.032) g/cm2, (0.771 ±0.023) g/cm2, (0.912 ±0.012) g/cm2, F=14.401, 19.860, 6.560, respectively, both P<0.05). The prevalence of osteoporosis in this group was 7%(5/76), which was higher than controls 1%(1/84). ② According to values of Bone meta-bolism [(CTX: (0.37±0.21) ng/ml] and inflammatory cytokines, TNF-α: (9.8±4.1) pg/ml, IL-6: (33.6±5.7) pg/ml and IL-17: (129±24) pg/ml were markedly increased in premenopausal women in early RA group [(0.24 ±0.09) ng/ml, (6.7 ±1.9) pg/ml, (1.5 ±0.4) pg/ml, (45 ±7) pg/ml, Z=2.722, 5.932, 7.501, 4.370, respectively, both P<0.05]. ③ The premenopausal women with early RA group with osteoporosis were signifi- cantly difference with controls in BMI [(9±3) kg/m2 vs (16±3) kg/m2], bone density of neck [(0.85±0.20) ng/ml vs (0.88±0.14) g/cm2], L2 [(0.75±0.23) g/cm2 vs (0.88±0.14) g/cm2], L3 [(0.87±0.07) g/cm2 vs (0.93±0.14) g/cm2], L4 [(0.92±0.12) g/cm2 vs (0.94±0.16) g/cm2], serum ESR [47.8(22.0, 76.0) mm/1 h vs 18.8(8.7, 35.2) mm/1 h] and DAS28-CRP (5.3 ±1.2 vs 3.8 ±1.2) F=0.68, 14.632, 26.114, 20.931, 36.582, Z=3.21, 6.58, respectively, both P<0.05. ④ Logistic regression showed that IL-6 (Wald χ2=5.78, P=0.021), PINP (Wald χ2=5.12, P=0.031), CTX (Wald χ2=9.17, P=0.003), ESR (Wald χ2=9.24, P=0.011), DAS28-CRP (Wald χ2=17.28, P=0.001) were significantly positively correlated with osteoporosis. Moreover, ordered unconditional Logistic regression analysis of the variables (IL-6, PINP, CTX, ESR, DSA28) described above showed that DAS 28-CRP score [OR=1.58, 95%CI: (1.10, 2.20)] was the most important risk factor for osteoporosis in premenopausal women with early RA. Conclusion The incidence of osteoporosis is high in premenopausal women with early RA than healthy cases. DAS 28-CRP score is the important risk factor for premenopausal women with early RA- associated osteoporosis. Measures relieve symptoms of RA can help to prevent and treatment osteoporosis.

ObjectiveTo observe the effect of betulinic acid(BetA) on the growth of human cytokine induced killer(CIK) cells and the killing activity of CIK cells on the gastric cancer cells in vitro before and after induced by betulinic acid,explore its mechanism.MethodsPeripheral blood mononuclear cell (PBMC) were separated form the healthy and were induced with various of cytokine to become CIK cells in vitro.CIK cells were collected on the tenth day and were induced with betulinic acid in different concentrations,followed by 48 h,the colorimetric methyl thiazolyl tetrazolium(MTT) method assay the proliferation rate of human CIK cells.Flow cytometry (FCM) was used to detect the expression changes of perforin,granzyme B and CD107a of human CIK cells before and after betulinic acid-induced.Lactate dehydrogenase (LDH) release assay was used to measure the influence on cytotoxic activity of CIK cells induced by betulinic acid against gastric cancer cell line SGC-7901 in vitro.Western blot assay was used to measure the extracellular signal-regulated kinase1/2 (ERK1/2),and adapter proteins SH2-domain containing leukocyte protein of 76KD(SLP-76) and linker for activative of T cells(LAT) expression changes of human CIK cells before and after drug-induced.ResultsBetulinic acid can promote CIK cells growth when the concentration were in 0.08-10 μg/ml,the expression of perforin,granzyme B and CD107a of CIK cells were significantly higher than control group(P＜0.05) when the concentration of betulinic acid were in 0.3 μg/ml.In the meanwhile,the cytotoxic activity of CIK cells in vitro against gastric cancer cell line SGC-7901 were also remarkably higher than the control group (P＜0.05).The expression of SLP-76,LAT and ERK1/2 were significantly increased to a certain extent than the control group( P＜0.05 ),when CIK cells were treated with betulinic acid.ConclusionThese results suggest that betulinic acid can promote CIK cells growth in some concentrations and increase the cytotoxic activity of CIK cells against gastric cancer cell line SGC-7901,its mechanism may related with two factors,on the one hand,enhancing the activity of SLP-76,LAT and ERK1/2,on the other hand,increasing the expression of perforin,granzyme B and CD107a on the surface of CIK cells.

Objective To investigate the efficiency and safety of intervention with flexible bronchoscope under general anesthesia by using laryngeal mask in patients with severe tracheal stenosis induced respiratory failure.Methods A total of 16 in-patients with respiratory failure caused by severe tracheal stenosis admitted from September 2009 to March 2012 were retrospectively reviewed.A comprehensive bronchoscopic intervention for the complete patency of airway was successfully performed with various techniques such as cryotherapy,electrocautery,balloon dilatation,and implantation of selfexpanding metal stents under genersl anesthesia by using laryngeal mask.The efficiency of comprehensive bronchoscopic intervention and dyspnea score were evaluated by chest CT scan and bronchoscopic examination before and after the treatment.Data were expressed as ((x-)± s).Paired t test was used for statistical analysis of the data.Results The degrees of tracheal stenosis and dyspnea score before and after intervention were (85.0±8.4)％ vs.(20.9±7.6)％ (P＜0.01) and (3.9±0.3vs.2.4±0.5,P＜ 0.01),respectively.There were no life-threatening complications occurred including massive haemorrhage.Conclusions It is an effective and safe technique to resolve the tracheal stenosis-induced respiratory failure with intervention by using flexible bronchoscope under general anesthesia with laryngeal mask,and it is a promising interventional treatment for clinic application.

Objective: To evaluate the superiority in diagnosing DVT between venography and duplex ultrasound, and the effectiveness of C-reactive protein (CRP) as a biomarker. Methods: Firstly,the iliac - femoral vein of the dog left hind leg was isolated,and then, the thrombosis model was established by infusing the thrombin after breaking endangium.The recanalization of thrombosis was assessed by duplex ultrasound and venography, and the expression of serum hsCRP was also examined. From 2006 to 2008, 77 patients with acute DVT proximal to the knee joint were admitted. The interval between the onset of DVT and admission was 1-21 days. They were treated mainly with urokinase and low molecular weight heparin for 2 weeks. The assessment of each patient including clinical manifestation, venography, duplex ultrasound and serum highly sensitive C-reactive protein (hsCRP) were performed immediately after admission and 4 weeks after discharge. Results: After medical therapy for 2 weeks, the clinical features prominently subsided in 49 patients, improved in 23 and didn’t ameliorate in 5.4 weeks after discharge, venography showed clot regression in 15 patients; while in the remaining 62 patients the occluded venous lumen were not visualized, duplex ultrasound showed partial lysis of the thrombosis. At admission, the hsCRP was 28.91?29.4mg/L, and it dropped to 8.13?12.7mg/L at 4 weeks after discharge. Conclusion:Duplex ultrasound was effective to assess DVT. The hsCRP was positively related to the severity of DVT.

Objective To investigate the mechanisms of tigecycline nonsusceptibility in carbapen-ems-resistant Acinetobacter baumannii ( CRAB) strains in order to provide a theoretical basis for a reasonable use of antibiotics and the control of nosocomial infection .Methods Susceptibility testing of 120 non-dupli-cate CRAB strains to tigecycline was performed by using the broth microdilution method .Minimal inhibitory concentrations ( MIC) of tigecycline against the A.baumannii strains were determined by using the broth mi-crodilution method before and after exposing the strains to Carbonylcyanide-m-chlorophenylhydrazone (CCCP), which was the efflux pump inhibitor .Polymerase chain reaction (PCR) was used to amply the ef-flux pumps genes including adeB, adeJ, adeG, abeM, adeE, adeRS, tetX and tetX1.The real-time PCR was performed to measure the expression of efflux pumps genes including adeB, adeJ, adeG, abeM and adeE.Results A total of 120 CRAB strains were collected including 13 (10.8%) tigecycline non-suscep-tible A.baumannii (TNAB) strains and 107 (89.2%) tigecycline susceptible A.baumannii (TSAB) strains.The MIC values of tigecline to the 120 CRAB strains were in a range of 0.25 μg/ml to 8 μg/ml. The adeR and adeJ genes were detected in 90.0%and 92.5%of the 120 CRAB strains, respectively.The positive rates of adeB, adeS, adeG and abeM genes among the 120 CRAB strains were all 94.2%.None of the three genes including adeE, tetX and tetX1 were detected .The mean expression levels of adeB and adeJ in TNAB strains were respectively increased by 18.69 folds and 5.46 folds as compared with those in sensi-tive strains.No significant increase in the expression of adeG and abeM genes was observed in TNAB strains . A 4-fold decrease in the MIC was observed in 8 out of 13 TNAB isolates treated with 10 μg/ml of CCCP .The CCCP could partially reverse the resistance pattern of tigecycline .Conclusion The efflux pump sys-tems of adeABC and adeIJK rather than the abeFGH and abeM systems might play an important role in reduc-ing the tigecycline susceptibility in carbapenems-resistant A.baumannii strains.

Objective:To evaluate the changes in the blood-cerebrospinal fluid barrier in postoperative delirium rats.Methods:One hundred and forty-seven healthy female Sprague-Dawley rats, aged 3 months, weighing 240-300 g, were divided into 3 groups ( n=49 each) using a random number table method: control group (group C), anesthesia group (group A) and postoperative delirium group (group P). Group C received no treatment.Group A received 2-h anesthesia with 1.4% isoflurane.Group S underwent an exploratory laparotomy under 1.4% isoflurane anesthesia.The behaviors of rats in each group were tested at 24 h before surgery and 6, 9 and 24 h after surgery using buried food test, open field test and Y maze test.Sodium fluorescence was injected through the tail vein at 6, 9 and 24 h after surgery.Then the rats were sacrificed, the choroid plexus (CP) was obtained, and cerebrospinal fluid (CSF) of bilateral cerebral ventricles was collected, and the expression of ZO-1, occludin, claudin1, E-cadherin and VE-cadherin in CP was detected using Western blot.FITC-dextran 10, 40 and 70 kDa was injected through the tail vein at 6 h after surgery, and then CSF was collected for determination of the concentrations of NaFI, 10, 40 and 70 kDa fluorescein isothiocyanate labeled dextran (FITC-dextran) in CSF by fluorescence spectrophotometry.CP was obtained to observe the morphology of choroid plexus epithelial cells (CPECs) of bilateral cerebral ventricles with a transmission electron microscope. Results:Compared with group C and group A, the latency to eat food in buried food test was significantly prolonged, the time of staying at the central region was shortened, the percentage of the number of entries into novel arm and percentage of time of staying at novel arm in Y maze test were decreased, the freezing time in open field test was shortened, the expression of ZO-1, occludin and claudin1 in CP was down-regulated, the concentrations of NaFI and 10 kDa and 40 kDa FITC-dextranin CSF were increased ( P<0.05 or 0.01), the CPECs arranged at random and loose, the microvilli of CPECs were absent, the tight junction was blurred, and the gap became wider in group P. Conclusion:The occurrence of postoperative delirium is related to the change in blood-cerebrospinal fluid barrier.

Objectives To investigate the occipital cortex metabolite alterations in repetitive and severe neonatal hypoglycemia rats treated with sodium pyruvate and to reveal the protective role of sodium pyruvate using high resolution 1H nuclear magnetic resonance spectroscopy.Methods Thirty-six 2-dayold Sprague-Dawley rats were randomly divided into hypoglycemia group and pyruvate group with 18 rats in each group.Rats in both groups received intraperitoneal injections of insulin (40 U/kg body weight) at 2,4 and 6 days of age to induce severe hypoglycemia (blood glucose value ≤ 1.4 mmol/L).In the hypoglycemia group,2.5 hours after insulin injection,intraperitoneal injection of 50％ glucose (2 ml/kg) was administered to terminate hypoglycemia,while in the pyruvate group,50％ glucose (2 ml/kg) and sodium pyruvate solution 2.5 ml/kg (500 mg/kg) were injected.Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay was used to observe the status of injured neurons in six neonatal rats,and metabolite changes in occipital cortex of the other 12 rats were detected by 1H nuclear magnetic resonance spectroscopy.The difference between the two groups was compared by independent-samples t test.Results Neonatal rats of both groups reached severe hypoglycemia level 2.5 hours after insulin injection.Compared with hypoglycemia group,pyruvate group had fewer injured neurons (45±5 vs 113 ± 12,t=0.782,P=0.013) and lower injured index in the occipital cortex (0.15 ± 0.03 vs 0.36 ± 0.06,t=l.143,P=0.020).Pyruvate group showed significant decreases in the concentration of taurine [(13.31 ± 2.06) vs (18.44 ± 3.86) mol/kg,t=8.231],glutamine[(1.50 ± 0.24) vs (2.02 ± 0.40) mol/kg,t=3.137],glutamate[(7.04 ± 0.95) vs (9.40 ± 1.73) mol/kg,t=6.449],aspartate[(1.51 ± 0.28) vs (2.15 ± 0.58) mol/kg,t=2.561] and creatine [(6.37±0.99) vs (8.46± 1.77) mol/kg,t =4.226] in the occipital cortex (all P'＜0.017).Conclusions Simultaneous use of glucose and sodium pyruvate to terminate hypoglycemia in repetitive and severe neonatal hypoglycemia rats can effectively alleviate severe hypoglycemia-induced occipital lobe damage via regulating excitatory amino acid neurotransmitters,energy metabolism and other metabolic pathways.