Objective To establish an animal behavioral model of tinnitus with step -down test ,and make use of the model to analyze the tinnitus genesis site in the salicylate-rats .Methods Forty wistar rats were includscl in this study ,they were randonly into 2 groups(experinent group and constrot group ,20 rats each group) .Ascertain that the animals treated with sodium salicylate(SS) have the sensation of tinnitus with the method of step -down test behavioral model .After having established a behavioral conditioned reflex ,subjects responded correctly to jump onto a climbing pole after hearing the sound .Then animals were under bilateral auditory nerves section ,to test whether the ablated-animals could still jump onto the pole after being treated with SS .Results Animals with the ab‐lating surgery could seldom jump onto the pole ,only reaching 1 .59 ± 0 .12 times average per 10 tests .Compared with con‐trol group ,there were significant differences with t-Test (P<0 .05) ,suggesting that subjects could not hear the tinnitus after the surgery .Conclusion Sodium salicylate could not make the animals with bilateral auditory nerves section to have the sensation of tinnitus .The peripheral auditory system plays an extremely important role in salicylate-induced tinnitus .

Objective To investigate the mechanism of tumor necrosis factor-α (TNF)-α inhibiting osteo blastdifferentiation of mesenchymal stem cells (BMMSCs) in the pathogenesis of osteoporosis in the mouse model of systemic lupus erythematosus (MRL/lpr). Methods The femurs of MRL / lpr and C3He/HeJ mice were isolated, the bone structure were examined by hematoxylin-eosin (HE) staining. The proteins of TNF-α, NF-κB P50, bone morphogenetic protein -2 (BMP-2) and PSmad1/5/8 were measured by immunohistochemical stain. Bone marrow mesenchymal stem cells (BMMSCs) were isolated. After BMMSCs grew on the cover slips, the proteins on top of it were evaluated by immunohistochemistry stain. Moreover, the alkaline phosphatase (ALP) staining was employed for the measurement of the early osteogenic differentiation. BMMSCs together with hydroxyapatite were embedded subcutaneously in the nude mice and eight weeks later, the ectopic bone formation was evaluated. The recombinant human tumor necrosis factor receptor type Ⅱantibody fusion protein (etanercept) or normal saline was subcutaneous injected to the mice with lupus. After four weeks, the expression of these proteins was observed and the ectopic bone formation was investigated. Image-Pro plus 6.0 software was employed for imagine analysis, and Studentˊs t-test was used to test the differences between 2 independent groups. Results MRL/lpr mice showed decreased volume of cortex and the percentage of cortex to the volume of bone of MRL/lpr mice was significantly lower compared to control groups and with C3He/HeJ mice (13.96±0.25 vs 23.61±0.71, n=3, P<0.01). The protein levels of both TNF-αand NF-κB P50 on the femur of MRL/lprl mice were higher than those of the control group (0.643±0.051 vs 0.405±0.022, 0.917±0.023 vs 0.650±0.032, n=3, P<0.01). The expressions of BMP-2 on the femur of MRL/lpr mice were lower than those of the C3He/HeJ mice (0.52 ±0.03 vs 0.72 ±0.03, n=3, P<0.01). There was no difference in the expression of PSmad1/5/8 on the femur between the two groups by immunohistochemistry detection (1.264 ±0.021 vs 1.301± 0.044, n=3, P>0.05). The expressions of TNF-α and NF-κB P50 in BMMSCs of MRL/lprl mice were higher than those of the C3He/HeJ (0.184±0.021 vs 0.136±0.013, 0.132±0.021 vs 0.097± 0.014, n=3, P<0.01), while BMP-2 and PSmad were lower than those of the control group (0.128±0.013 vs 0.216±0.221, 0.115±0.023 vs 0.196±0.034, n=3, P<0.01). After 7 days of BMP-2 stimulation, the activities of ALP of BMMSCs from MRL/lprl mice were reduced detected by ALP staining and the osteoblast differentiation of these cells were decreased than BMMSCs from the control mice by HE and Masson staining. The percentage of the cortex to the volume of bone of the etanercept injection MRL/lpr mice was higher than that of the control group (21.8±1.0 vs 14.3 ±0.6, n=3, P<0.01). Moreover, the proteins of TNF-α and NF-κB P50 on the femurs of such injected mice were lower than those of the control group (0.540±0.024 vs 0.682±0.031, 0.857±0.023 vs 1.098±0.044, n=3, P<0.05), while the expressions of BMP-2 were higher than the control group (0.99±0.04 vs 0.85±0.04, n=3, P<0.05). There was no difference in the PSmad1/5/8 expression on the bone of the two group of lupus mice (0.88 ±0.08 vs 0.84 ±0.04, n=3, P>0.05). The ectopic bone formation of BMMSCs of the etanercept injected MRL/lpr mice was higher than that of the normal saline injected mice, however, it was lower than that of the C3He/HeJ mice. Conclusion TNF-α inhibits osteoblast differentiation of mesenchymal stem cells by depressing Smad signaling which may contribute to the osteoporosis of the lupus mice.

OBJECTIVE:To optimize the formulation and preparation technology of Propylthiouracil(PTU)solid lipid nanopar-ticles(PTU-SLN)and to evaluate the quality of PLN-SLN. METHODS:PTU-SLN was prepared by emulsion ultrasound dispersing method. The formulation of PTU-SLN was optimized by orthogonal design with the entrapment efficacy and particle size as index, using the amount of lipid material,soybean lecithin and poloxamer 188 and ultrasonic time as factors. The quality of prepared nanoparticles was evaluated with particle size,Zeta potential,entrapment efficiency,stability and in vitro drug release rate as in-dex. RESULTS:The optimal formulation and technology was as follows as lipid material 0.6 g,soybean lecithin 1.0 g,poloxamer 188 0.8 g,ultrasonic time 10 min. The obtained PTU-SLN was round and smooth in appearance and distributed evenly in particle size with average particle size of 93.5 nm,Zeta potential of -30.8 mV and average entrapment efficiency of 74.9%. Prepared nanoparticles had no significant change after placing for 15 d at 4 ℃. Accumulative release rate of PTU-SLN was 56.1% at 4 hour in vitro and reached 98.4% at 24 hour. CONCLUSIONS:PTU-SLN is prepared successfully and reasonable in technology,and can reach sustained-release effects.

Objective:To observe influence of up‐regulated expression of myocardial heat shock protein (HSP) 70 in‐duced by heat stress on myocardial calcium‐activated potassium channel (KCa ) 3.1 expression in rabbits with atrial fibrillation (AF) caused by rapid atrial pacing (RAP) .Methods :A total of 24 New Zealand white rabbits were ran‐domly divided into sham operation group (n=8 ,only implant electrode without pacing ) ,pacing group (n=8 ,right atrium (RA) received RAP at 600 times/min for 6h) and heat stress pacing group (heat stress group ,n=8 ,received heat stress preconditioning ,then the same RAP as pacing group ) .Results:Compared with sham operation group and pacing group ,there were significant up‐regulation of HSP70 mRNA and protein expression in different sites of heart [HSP70 protein ,left atrium (LA):(39.00 ± 3.21) vs .(39.75 ± 2.82) vs .(69.75 ± 3.45) ,RA: (38.38 ± 2.92) vs .(39.50 ± 3.89) vs .(69.00 ± 2.93) ,left atrial appendage (LAA):(37.75 ± 3.28) vs .(39.00 ± 3.89) vs . (68.63 ± 3.23) ,right atrial appendage (RAA): (37.00 ± 3.85) vs .(38.38 ± 3.74) vs .(68.75 ± 2.82)] in heat stress group , P<0. 01 all ,but there were no significant difference between pacing group and sham operation group , P>0.05 ;compared with pacing group with down‐regulation of KCa3.1 mRNA and protein expressions ,there were significant up‐regulation of KCa3.1 mRNA and protein expressions in different sites of heart [KCa3.1 protein ,LA:(21.25 ± 1.67) vs .(24.00 ± 2.62) ,RA :(21.13 ± 1.96) vs .(23.75 ± 1.83) ,LAA :(21.00 ± 2.07) vs .(23.75 ± 1.67) ,RAA:(20.88 ± 2.03) vs .(23.50 ± 2.45)] in heat stress group ,P<0.05 all ,and there were no significant difference between heat stress group and sham operation group , P>0. 05. Conclusion:Heat stress may induce up‐regulated expression of myocardial HSP 70 of myocardium ,and HSP 70 may inhibit down‐regulation of KCa 3. 1 mR‐NA and protein expressions in rabbits with atrial fibrillation.

Objective To study the alteration of microbial population distribution and resistance of clinical bacterial isolates in patients with severity muhiple injuries. Methods The distributed Features of 432 strains of infection germs detected among the patients with severe muhiple iniuries admitted into hospital from January 2004 to December 2006 were statistically analyzed during. Results In the total 432 strains,the G accounted for 62.9%(272/432),dominated mainly by pathogens including Pseudomonas aeruginosa,Acinetobacter baumanni I and Escherichia Coli.The G+accounted for 37%(160/432),mainly including Staphylococcus anreus,enterococci and coagulase negative staphylococcus (CNS).Mixed infection rate was 41.1%.The isolating rate of enterococci.CNS and Sten Matophilia was obviously upgraded. Conclusions The source of infection in patients with severity multiple injuries is Gram-negative bacterium,suggesting that surveillance of bacterial resistance and rational use of antimicrobial agents should be emphasized during clinical therapy.

Objective To explore the impact of chronic cough on children’s life quality, and to observe their life quality after drugs and psychological intervention. Methods One hundred 9 to 12 years old children with chronic cough were randomly selected. The drugs and psychological intervention were administrated. The children had been follow-up. The children’s quality of life was assessed by“Inventroy of Subjective Life Questionnaire”before and after treatment. Meanwhile 100 healthy children were randomly selected as a control group. Results With the prolonged treatment, the recovery rate and effective rate in children with chronic cough increased. Before the treatment, the scores of family life, peer interaction, self cognitive, experience of depression, experience of anxiety, cognitive and emotional component, and overall satisfaction were significantly lower in children with chronic cough than those in healthy children (P<0.05). After the treatment, the scores of family life, peer interaction, self cognitive, experience of depression, experience of anxiety, physical experience, cognitive and emotional component, and overall satisfaction were significantly improved in children with chronic cough (P<0.05), even the scores of physical experience and emotional component were significantly higher in children with chronic cough than those in healthy children (P<0.05). Conclusions The quality of life in children with chronic cough decline, however, drug and psychological intervention can improve their quality of life.

Objective To analyze the value of MRI in the diagnosis of giant cell tumor of tendon sheath(GCTTS). MethodsThe MR images of 21 GCTTS cases including 3 cases of recurrences were retrospectively evaluated.All were confirmed giant cell tumor of tendon sheath by surgery and pathology.All the patiens were examined by MRI,and 19 received contrast enhanced MRI.The characteristics of MRI presentations were explored.Results Of the 21 cases,17 were in diffuse form and 4 in localized form.On T1WI,the signal intensities of the giant cell tumor of tendon sheath almost equalled to those of skeletal muscle in 18 cases and were slightly lower than those of skeletal muscle in 3 cases.On T2WI,the signal intensities tended to range between those of skeletal muscle and fat in 7 cases,almost equalled to those of skeletal muscle in 12 cases,and were slightly lower than those of skeletal muscle in 2 cases.Of the 19 cases with gadolinium-enhanced images,17 showed inhomogeneous enhancement and 2 homogeneous enhancement,and all were observed with a fat suppression sequence on T1WI. Conclusion MRI is able to depict the characteristic internal signal of giant cell tumor of tendon sheath,which is a valuable for diagnosis,treatment and follow-up.

Objective:To investigate the effect and mechanism of 6-formylindolo[3,2-b]carbazole (FICZ) on lipopolysaccharide (LPS)-induced acute lung injury (ALI) in mice.Methods:Male C57BL/6J mice aged 8-12 weeks were divided into 4 groups with 8 mice in each group, according to the method of simple random sampling. Sepsis-induced ALI mice model was established by intraperitoneal injection of LPS 5 mg/kg (LPS group), and phosphate buffer saline (PBS) control group (PBS group) was injected with equal volume of PBS. The LPS+FICZ group was intervened by intraperitoneal injection of 1 μg FICZ 1 hour after LPS stimuli, while the FICZ control group (FICZ group) was given the same amount of FICZ 1 hour after intraperitoneal injection of PBS. Serum and lung tissue were collected 24 hours after LPS stimuli, and the pathological changes of lung tissue were analyzed by hematoxylin-eosin (HE) staining and wet/dry weight (W/D) ratio of lung tissue. The concentrations of inflammatory factors in serum and lung tissue were detected by enzyme linked immunosorbent assay (ELISA). The expression levels of endoplasmic reticulum stress signaling pathway related molecules were detected by real-time fluorescent quantitative polymerase chain reaction (RT-qPCR) and Western blotting.Results:Compared with PBS group, inflammatory cell infiltration, alveolar collapse and obvious alveolar exudative lesions had increased, lung tissue W/D ratio was significantly increased, serum interleukin-6 (IL-6) level, lung tissue IL-6 mRNA expression, and the mRNA expressions of glucose-regulated protein 78 (GRP78), protein kinase R-like endoplasmic reticulum kinase (PERK), CCAAT/EBP homologous protein (CHOP), and the protein expressions of GRP78, PERK, activating transcription factor 6 (ATF6), CHOP in lung tissue were significantly increased in LPS group. However, the indexes of FICZ group were not affected. Compared with LPS group, LPS+FICZ group had less inflammatory cell infiltration, relatively intact alveolar structure. Lung W/D weight ratio in LPS+FICZ group was significantly decreased (5.38±0.10 vs. 6.60±0.30, P < 0.01), so as serum IL-6 (ng/L: 15.55±3.77 vs. 32.22±3.84) and lung IL-6 mRNA expression (2 -ΔΔCt: 0.79±0.21 vs. 6.89±0.92, both P < 0.01). The mRNA expressions of GRP78, PERK and CHOP were also significantly decreased [GRP78 mRNA (2 -ΔΔCt): 1.90±0.16 vs. 7.55±1.29, PERK mRNA (2 -ΔΔCt): 1.68±0.20 vs. 4.54±0.89, CHOP mRNA (2 -ΔΔCt): 1.13±0.24 vs. 4.44±1.13, all P < 0.05], and the protein expressions of GRP78, PERK, ATF6 and CHOP were significantly decreased (GRP78/GAPDH: 0.59±0.02 vs. 0.77±0.01, PERK/GAPDH: 0.48±0.03 vs. 1.04±0.05, ATF6/GAPDH: 0.51±0.03 vs. 0.65±0.01, CHOP/GAPDH: 0.91±0.05 vs. 1.11±0.07, all P < 0.05). Conclusion:FICZ protects LPS-induced ALI possibly via suppressing endoplasmic reticulum stress and reducing IL-6 expression in blood and lung tissue.

OBJECTIVETo screen genes differentially expressed in mouse hepatocarcinoma ascites cell line with high potential of lymphatic metastasis.

METHODSA subtracted cDNA library of mouse hepatocarcinoma cell line with high potential of lymphatic metastasis Hca-F and its synogenetic cell line Hca-P with low metastatic potential was constructed by suppression subtractive hybridization (SSH) method. The screened clones of the subtracted library were sequenced and GenBank homology search was performed.

RESULTSTen differentially expressed cDNA fragments of Hca-F with high potential of lymphatic spreading were obtained, two of which were newly identified ones.

CONCLUSIONSSH is a useful technique to detect genes of differential expression and an effective method to clone novel genes.

Animals ; Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Gene Library ; Liver Neoplasms, Experimental ; genetics ; pathology ; Lymphatic Metastasis ; genetics ; Mice ; Mice, Inbred Strains ; Nucleic Acid Hybridization ; methods ; RNA, Messenger ; genetics

AIMTo investigate the effects of fungal elicitors on inophyllums production in suspension cultured cell of Calophyllum inophyllum Linn.

METHODSThe pathogen of leaf spot disease of C. inophyllum L. was isolated and prepared as fungal elicitor. The fungal elicitor was added to the medium with different concentrations and culture period. Their effects on biomass and inophyllums content of the suspension of cultured cells were studied.

RESULTSThe optimum effects of S-I fungal elicitor concentrations on inophyllums content was 60 mg GE x L(-1). Adding the fungi elicitor into the cell suspension culture system at stationary phase (being cultured for 18 days) resulted in a highest inophyllum content of 59.174 mg x L(-1) at the 3rd day with 27% higher than control. Fungal elicitor treatment promoted the inophyllums accumulation in medium.

CONCLUSIONAdding the Stagonospora curtisii (Berk.) Sacc. to the medium was effective approaches to enhance inophyllums yield in the suspension of C. inophyllum L culture cell.

Calophyllum ; cytology ; metabolism ; Cell Division ; Cells, Cultured ; Coumarins ; metabolism ; Culture Media ; Mitosporic Fungi ; physiology