Objective To investigate whether pregnant rats exposure to ketamine cause offspring changes in space cognitive abilities and exploration abilities.Methods 3-month Sprague-Dawley female rats ( n =24)were randomly divided into four groups:group N (control group),group K1 (small doses of ketamine group),group K2 ( clinical anesthesia dose of ketamine group),group K3 ( large doses of ketamine group).3-month Sprague-Dawley male rats ( n =4) and female rats were mated at the same cage by the proportion of 2∶ 1.Pregnant mice were treated at tenth day:group N were treated saline with equal-volume to ketamine vein injection; group K1,group K2,group K3 administered vein injection 3,8,20mg/kg of ketamine.Then the 20-day offspring rats'learning and memory were assessed used Open Field Test ( record the time of the offspring in the central case through the number of grid within 2 min ) and Hole Board Test ( Counting the times of offspring stretch into the hole in 5 min) at postnatal days 20.Results In the Open Field Test,the retention time in central check of group N,group K2 and group K3 were (2.45 ± 1.23)s,(6.42 ±2.50)s,(6.41 ±2.19)s.Compared with group N,the retention time in central check of group K2 and group K3 were significantly higher (F=13.42,P＜0.01 ),and group K1 were not significant different ( t =1.33,P＞0.01 ),and the locomotion of group K1,group K2,group K3 were significantly reduced( ( 15.33 ± 6.81 ),( 13.75 ± 5.93 ),( 16.92 ± 6.54 ),F =4.24,P ＜ 0.05 ).In the Hole Board Test,the times of offspring stretch into the hole were not significant different comparing with the control group(F=2.17,P ＞ 0.05 ).Conclusion The dose of ketamine that equivalented clinical anesthesia can affect offspring rats' space cognitive abilities; but the exploring cognitive ability were not significantly influenced.

Objective To observe the effects of bolus volume on pharyngeal and upper esophageal sphincter pressures and durations in healthy volunteers by using high-resolution manometry (HRM).Methods Twentyfour health subjects were recruited and asked to swallow three volumes of bolus (3 ml,5 ml and 10 ml) in the neutral head position.Pressure and duration measurements were acquired by utilizing a high-resolution solid-state manometer,with an emphasis on the hypopharynx and upper esophageal sphincter (UES).Variables including UES residual pressure,UES relaxation duration,maximum hypopharygeal pressure and hypopharyngeal pressure duration were analyzed across bolus volumes and consistencies by using three-way repeated measures analysis of variance (ANOVA) to investigate influence of bolus volume.Results UES residual pressure [-1.71 mmHg(3 ml thick liquid)vs.-4.68 mmHg(10 ml thick liquid)],UES relaxation duration[590.45 ms(3 ml thick liquid) vs.702.49 ms (10 ml thick liquid)],maximum hypopharygeal pressure [169.91 mmHg (3 ml thick liquid) vs.204.42 mmHg (10 ml thick liquid)] and hypopharyngeal pressure duration(P ＜0.05) varied significantly across bolus volumes when swallowing water or thick liquid.The UES relaxation duration,UES residual pressure and maximum hypopharyngeal pressure had a direct positive relationship with bolus volume.There was significant differences with regard to UES relaxation duration [685.75 ms(3 ml paste)vs.772.27 ms (10 ml paste)] but not to UES residual pressure (P ＞ 0.05) and maximum hypopharyngeal pressure (P ＞ 0.05) across bolus volume when swallowing paste.Conclusions Difference in hypopharyngeal pressure and duration,UES residual pressure and duration were detected across varying bolus volumes.Consideration of these variables is paramount in understanding normal and pathological swallowing.

Objective To study the influence of unilateral and bilateral amplification on the effect of hearing aid evaluation .Methods Using the subjective method that International Outcome Inventory for Hearing Aids (IOI-HA) and objective method that medium acoustic intensity (65 dB SPL) word recognition score(WRS) to evaluate the effect of unilateral and bilateral hearing aid fitting of middle -aged severe sensorneural hearing loss .Results Hearing aid were used for severe sensorneural hearing loss and the improvement of monosyllables and sentences in quiet and noise test of unilateral were 35 .73% ,43 .15% ,43 .23% ;the improvement of monosyllables and sentences in quiet and noise test of bilateral were 37 .90% ,51 .33% ,54 .86% .The WRS of bilateral was higher than unilater-al .The score of IOI-HA was 15~37 ,meaning patients with severe sensorneural hearing loss were satisfied with hearing aid ,and there was no statistical significance between unilateral and bilateral fitting .Conclusion The bilat-eral hearing aid fitting was better than unilateral .Binaural hearing loss are recommended to fit bilaleral hearing aids .

OBJECTIVEThis paper aimed to determine the mRNA expression of osteoclast-related factors interleukin-6 (IL-6), interleukin-12 (IL-12) p35, IL-12p40, matrix metalloproteinase-9 (MMP-9), nuclear factor of activated T-cells cytoplasmic 1 (NFATcl), receptor activator of nuclear factor-KB (RANK), and tumor necrosis factor-α (TNF-α) mRNA in murine macrophages infected by a periodontitis patient's own tissue nucleic acid. Another aim was to investigate the effects of a periodontitis patient's own tissue nucleic acid on the differentiation of macrophages into osteoclasts.

METHODSInflammatory periodontal tissue samples of chronic periodontitis patients were taken during periodontal flap surgery, and healthy gingival tissue samples were taken from orthodontic patients during tooth extractions. Total RNA from periodontal tissue was extracted and reversely transcribed into cDNA and then cryo-preserved until further use. First, specific sequence oligodeoxynucleotide MT0I at a concentration of 1 µg · mL⁻¹ was added in murine macrophage RAW264.7, and the cells were incubated for 3 hours. Cells with PBS (1 µg · mL⁻¹) were used as negative controls. The inflammatory periodontal tissue cDNA and healthy periodontal tissue cDNA (1 µg · mL⁻¹) was added subsequently. There were four experimental groups: healthy periodontal tissue cDNA+ RAW264.7, inflammatory periodontal tissue cDNA+RAW264.7, MT01+healthy periodontal tissue cDNA+RAW264.7, and MT01+inflammatory periodontal tissue cDNA+RAW264.7. Real-time quantitative polymerase chain reaction was used to detect the mRNA expression of osteoclast-related factors IL-6, IL-12p35, IL-12p4O, MMP-9, NFATcl, RANK, and TNF-α mRNA after 3, 6, 12, and 24-hours.

RESULTSThe mRNA levels of osteoclast-related factors NFATc1, MMP-9, TNF-a, IL-6, IL-12p40, IL-12p35, and RANK in RAW264.7 were markedly upregulated with the treatment of periodontitis patient's own tissue nucleic acid. However, the mRNA expression of osteoclast-related factors was inhibited by use of an immunosuppressant MT01.

CONCLUSIONThe periodontitis patient's own tissue nucleic acid could promote the differentiation of murine macrophage into osteoclasts.

Animals ; Cell Differentiation ; Cytokines ; metabolism ; Gene Expression ; Gingiva ; Humans ; Interleukin-12 Subunit p40 ; Interleukin-6 ; Macrophages ; Matrix Metalloproteinase 9 ; Mice ; Osteoclasts ; metabolism ; Periodontitis ; RNA, Messenger ; Tumor Necrosis Factor-alpha

Objective To observe the variation of enzymatic activity and areas and bulk of focus of heart injuries by using controllable catheter to ablate epicardial tmsue of rabbits and focus underneath atrioventrieular ring narcosis with 20% urethane(4 ml/kg)and divided into three groups.Each group included 7 rabbits.Anterior wallepieardium of left ventricle was ablated thirty seconds in each group(10,20 and 30 W)with self-made ablationspheroid microwave antenna,refilling with high pressure normal saline at same time.Then all of the rabbits were sacrificed respectively and their ventricular myocardium were taken out to undergo immunohistochemistry in order to display suceinate dehydrogenase(SDH).Also amplitude Wag measured in order to calculate areas of heart injuries.(8F)wag delivered to the pre-selected sites around atrioventricular ring of thirty-two healthy dogs,which had beenin intravenous narcosis with pentobarbital sodium(30 mg/kg).The dogs were divided into four groups(40,50,60 and 80 w) and two time points(60 and 120 s),by the combined method of X-ray and endocardial electrocardiograph,the microwave antenna could be confirmed to be located at the accurate position between anterior and posterior wall close to septum of left/right ventricle.After ventricular myocardium had been taken out,amplitude were measuredin order to calculate bulk of heart injuries by 1/6×3.14 x long×wide×deep.In addition.the histological changesand transmural injury were examined by optic microscope.Results In each group,the centre of injuries wagenzyme deficiency locus.The diameter and areag of heart injuries enlarged significantly(3.99.±0.41),(5.20±0.25),(6.31±0.37)mm and(12.53±2.56),(21.19±3.14),(30.96±3.76)mm2 with the increased microwave power level(10、20、30 W).Group comparison had statisficM significance(F＝76.8,58.5;P＜0.01 or ＜0.05).A total of 116points were ablated.The myocardial lesion showed ellipse in shape,and continuous symmetrical coagulationnecrosis under microscopic examination.There was a clear demarcated line around tlle myocardial tissue and fewparietal thmmbus.There were 16 transmura]injuries and five-with lung damage.The bulk of lesion aroundatrioventrieular ring hag been significantly enlarged(46.7±2.5),(51.1±2.7),(133.2±3.4),(141.8±3.9),(248.5±6.2),(260.3±6.5),(313.7±9.5),(327.4±10.5)with the increased microwave power level(40,50,60and 80 W)and/or distance of microwave ablation(60 and 120 s).Groups comparison had statistical significance(F＝31.16,27.85;all P＜0.01).In each time point,the lesion bulks had conspicuous distinction of statistics.In the same microwave power,the time wag longer,the bulk was larger(P＜0.01).Conclusions The more the microwave power level and time,the severe the heart injuries is.It is possible to use the microwave energy to ablate the deep focus under endocardium around atrioventricular ring.

Acute Disease ; Anti-Inflammatory Agents ; therapeutic use ; Budesonide ; therapeutic use ; Child, Preschool ; Graft vs Host Disease ; drug therapy ; Humans ; Intestinal Diseases ; drug therapy ; Male ; Treatment Outcome

This study aims to construct a eukaryotic expression system for envelope gene of Jaagsiekte sheep retrovirus, observes its localization in 293T cells, and investigates the potential in inducing malignant transformation of NIH3T3 cells. By RT-PCR, the full-length cDNA of envelope gene of Jaagsiekte sheep retrovirus (exJSRV-env) was amplified from the extract of naturally infected sheep lung. The clone of target gene was sub-cloned into eukaryotic expression system pEGFP-C1, and validated by PCR, restriction endonuclease, and sequencing. Bioinformatic analysis concerning biological function and cellular localiza tion of exJSRV-env was also performed. The recombinant clone of exJSRV-env was transfected into 293T cells and NIH3T3 cells by Lipofectamine LTX. The expression and celluar localization in 293T cells were validated by confocal microscopy. Soft agar colony formation assay was employed to test the anchorage-independent growth of NIH3T3. DNA sequencing and restriction enzyme digestion with Kpn I and Hind III indicated the correct construction of the recombinant plasmid, which was named pEGFP-C1/exJSRV-env. Amino acid sequence alignment of exJSRV-env with reference sequences found 85%-100% homogeneity. A YRNM motif was discovered at the cytoplasmic tail of envelope gene, which is exclusively found in exogenous viruses. Phylogenetic tree analysis showed that our clone of exJSRV-env clustered closely with pathogenic exogenous Jaagsiekte sheep retroviruses. Fluorescence microscopy indicated typical membrane localization of exJSRV-env protein. NIH3T3 cells transfected with exJSRV-env lost contact inhibition, and acquired colony forming ability in soft agar. This study indicated that envelope protein of Jaagsiekte sheep retrovirus can induce malignant transformation of mouse fibroblast cell NIH3T3. Discoveries of this study provide a basis for further structural and functional research on Jaagsiekte sheep retrovirus envelope protein.
Amino Acid Sequence ; Animals ; Betaretrovirus ; chemistry ; classification ; genetics ; physiology ; Cell Transformation, Viral ; Green Fluorescent Proteins ; genetics ; metabolism ; Mice ; Molecular Sequence Data ; NIH 3T3 Cells ; Phylogeny ; Retroviridae Infections ; veterinary ; virology ; Sequence Alignment ; Sheep ; Sheep Diseases ; virology ; Transformation, Genetic ; Tumor Virus Infections ; veterinary ; virology ; Viral Envelope Proteins ; chemistry ; genetics ; metabolism

Objective To evaluate the effect of topical anesthesia with compound lidocaine cream coated on the tracheal tube on extubation response in patients undergoing Han-uvulopharyngoplasty (H-UPPP).Methods Eighty-four patients,aged 28-48 yr,weighing 91-108 kg,scheduled for elective H-UPPP,requiring tracheal intubation under general anesthesia,were equally and randomly divided into 2 groups:compound lidocaine cream group (group L) and control group (group C).The compound lidocaine cream 2-3 g were coated on the tracheal tube cuff and exterior before induction of anesthesia in group L,while the paraffin oil was coated in group C.Mean arterial pressure (MAP),HR and pulse oxygen saturation were recorded before induction of anesthesia,at the end of infusion of anesthetics,during extubation and 5 min after extubation (T1-4).Blood samples were taken from the forearm veins on the noninfusion side at T1-4 for detection of plasma adrenergic and norepinephrine concentrations.Cardiovascular events during extubation were recorded.Results Compared with group C,MAP and HR were significantly decreased at T3,4,the incidences of hypotension and tachycardia were decreased,the plasma adrenergic and norepinephrine concentrations were significantly decreased at T2-4 in group L (P ＜ 0.05).Conclusion Topical anesthesia with compound lidocaine cream coated on the tracheal tube can effectively reduce the extubation response in the patients undergoing H-UPPP.

Objective To evaluate the effect of sevoflurane pretreatment on the inflammatory response during renal ischemia-reperfusion (I/R) in rats.Methods Thirty pathogen-free male Sprague-Dawley rats,aged 12-14 weeks,weighing 220-260 g,were randomized into 3 groups (n =10 each) using a random number table:sham operation group (group S),I/R group and sevoflurane pretreatment group (group SP).Renal I/R was induced by clamping the left renal pedicle for 45 min followed by reperfusion in I/R and SP groups.In group S inhalation of 3％ sevoflurane in O2 was started at 30 min before I/R and maintained throughout the experiment.Venous blood samples were taken at 3 h of reperfusion for determination of serum blood urea nitrogen (BUN) and creatinine (Cr) concentrations.The animals were then sacrificed and the left kidneys were removed for microscopic examination and for measurement of the content of tumor necrosis factor-apha (TNF-α),interleukin-6 (IL-6) and intercellular cell adhesion molecule-1 (ICAM-1) in renal tissues (by ELISA).Results Compared with group S,the serum BUN and Cr concentrations,severity of necrosis of renal proximal convoluted tubules (0 =normal,4 =necrosis of whole segment of proximal convoluted tubules),and contents of TNF-α,IL-6 and ICAM-1 were significantly increased in I/R and SP groups (P ＜ 0.05).Compared with group I/R,the serum BUN and Cr concentrations,severity of necrosis of renal proximal convoluted tubules,and contents of TNF-α,IL-6 and ICAM-1 were significantly decreased in SP group (P ＜ 0.05).Conclusion Sevoflurane pretreatment can protect kidney against I/R injury by inhibiting the inflammatory responses in the renal tissues of rats.

Objective To evaluate the roles of Toll-like receptor 2 (TLR2),TLR4 and myeloid differentiation factor 88 (MyD88) in immune recognition and immune mediation in sporotrichosis by detecting their expressions in skin lesions of sporotrichosis.Methods Biopsy specimens were obtained from the skin lesions of 19 patients with sporotrichosis and normal skin of 12 healthy human controls.Immunohistochemistry was performed to observe the expressions of TLR4 and MyD88,and real-time fluorescence-based quantitative PCR to quantify the expressions of TLR2 and MyD88 mRNAs.Data were expressed as mean ± standard error.Statistical analysis was done with the SPSS17.0 software.Independent samples t-test was conducted to compare the parameters between the lesional and control specimens.A p-value of less than 0.05 was considered to be statistically significant.Results TLR4 and MyD88 were mainly observed in the whole epidermis except the stratum corneum as well as plasma cells and lymphocytes in the dermis and subcutaneous tissue in lesional skin.However,TLR4 was nearly absent and MyD88 was weakly expressed in the dermis and subcutaneous tissue in normal skin.The expression levels of TLR4 and MyD88 were significantly higher in the lesional skin than in the control skin (TLR4,63.767 ± 3.829 vs.5.167 ± 3.246,t =4.82,P ＜ 0.05; MyD88,57.236 ± 4.744 vs.10.588 ± 1.640,t =3.30,P ＜ 0.05).Similarly,the lesional skin showed significantly stronger expressions of TLR2 and MyD88 mRNAs compared with the normal skin (TLR2,1.974 ± 1.452 vs.1.430 ± 1.073,P ＜ 0.05; MyD88,2.028 ± 2.061 vs.0.688 ± 0.422,P ＜ 0.05).Conclusion Sporothrix may induce the development of sporotrichosis by interacting with host immune system via TLR signaling pathways.