An actinomycete strain P-13, with antimicrobial activity against muskmelon bacterial spot pathogens, was isolated from the muskmelon rhizosphere soil samples in Xinjiang. The strain P-13 was identified as Streptomyces rochei based on morphological, physiological characteristics and 16S rRNA sequence analysis. The agar diffusion bioassay showed that the diameter of inhibition zone against Acidovorax avenae subsp. citrull BFB and Pseudomonas syringae pv. Lachrymans P4 was above 19 mm and 17 mm, respectively. The antimicrobial substances obtained from strain P-13 were demonstrated to be alkaline and water-soluble compounds according to paper chromatogram analysis and exocellular metabolites. Furthermore, it was stable to be treated by 100?C for 10 min, pH 6 for 6 h, or ultraviolet treatment for 7 h. Moreover, it was insoluble in organic solvents, such as petroleum benzine, diethyl ether, and acetic ether.

Objective To investigate the expressions of receptor for advanced glycation endproducts (RAGE) and nuclear factor κB(NF-κB) in brain hippocampus of rat with chronic fluorosis,and to reveal the mechanism of brain damage resulted from chronic fluorosis.Methods Sixty clean grade SD rats were randomly divided to three groups(20 rats in each group,10 female and 10 male) fed with different contents of fluoride,control group with normal tap-water(＜ 0.5 mg/L fluoride),small dosage of fluoride exposure group(10 mg/L fluoride in tap-water) and large dosage of fluoride exposure group(50 mg/L fluoride) for six months.Then the rats were killed by femoral artery bleeding and hippocampus was removed.Protein and mRNA levels of RAGE and NF-κB in the hippocampus were determined by Western blotting and quantitative real time PCR,respectively.Results As compared to the control groups[(100.00 ± 2.60)％,(100.00 ± 7.80)％],the expressions of RAGE and NF-κB at protein level in the hippocampus were significantly increased in the small dosage of fluoride exposure groups [(205.00 ± 15.30)％,(156.00 ± 12.20)％] and the large dosage of fluoride exposure groups[(232.00 ± 10.90)％,(162.00 ± 9.80)％,all P ＜ 0.05]; for the mRNA level of RAGE and NF-κB,the expressions were higher in the small dosage of fluoride exposure groups(1.27 ± 0.09,0.83 ± 0.15) and the large dosage of fluoride exposure groups (2.60 ± 0.19,1.27 ± 0.19) than those of the control groups(0.66 ± 0.18,0.32 ± 0.08,all P＜ 0.05).Conclusions The increased expressions of RAGE and NF-κB in the hippocampus of rat brain are caused by chronic fluorosis,and these changes may be associated with the mechanism of nerve injury.

Objective Aim of the study is to investigate the expression of syndecan-4 and nuclear factor κB(NF-κB) in the kidney of rat with chronic fluorosis,and to reveal the mechanism of kidney damage resulted from the toxicity of excessive amount of fluoride.Methods According to body mass and sex,sixty SD rats were randomly divided to three groups according to body mass and fed with different contents of fluoride:control group with normal tap-water(＜ 0.5 mg/L fluoride),small dosage of fluoride exposure group (adding 10 mg/L fluoride in tap-water) and large dosage of fluoride exposure group (50 mg/L fluoride) for six months.The protein level of syndecan-4 and NF-κB in the kidney was detected by Western blotting and syndecan-4 mRNA level by quantitative real time PCR.Results As compared to the control group[(100.0 + 8.1)％],the expression of syndecan-4 at protein level in the kidney of rat was significantly increased in the small dosage of fluoride exposure group [(198.5 + 5.6)％,P ＜ 0.05] and large dosage of fluoride exposure group [(209.2 + 13.0)％,P ＜ 0.05]; the protein levels of NF-κB in the small dosage of fluoride exposure group[(284.4 + 11.1)％,P ＜ 0.05] and in the large dosage of fluoride exposure group[(343.2 + 2.9)％,P ＜ 0.05] were significantly increased than that of the control group[(100.0 ± 10.7)％].The mRNA levels of syndecan-4 in the kidney in the small dosage of fluoride exposure group and large dosage of fluoride exposure group(0.431 + 0.058 and 0.453 ± 0.065,both P ＜ 0.05,respectively) were significantly increased than that of the control(0.128 + 0.026).Conclusions The increased expression of NF-κB in the kidney is induced by increased expression of syndecan-4,which may be involved in kidney damage of chronic fluorosis.

Objective To compare open versus closed reduction in treatment of children with displaced femoral neck fracture whose reduction is delayed by more than 24 hours.Methods We retrospectively reviewed 51 children who had been treated at our department for femoral neck fracture between May 2005 and May 2012 after their reduction had been delayed for more than 24 hours.They were 37 boys and 14 girls,1.7 to 15.5 years of age (average,9.1 years).According to Delbet's classification,27 cases belonged to type Ⅱ,18 to type Ⅲ,and 6 to type Ⅳ.Of them,18 underwent closed reduction and 33 open reduction.Their complications were recorded in detail.Their therapeutic results were evaluated according to Ratliff's criteria and compared between the 2 methods of reduction.Results The 51 children were followed up for an average of 36.7 months (range,from 17 to 61 months).Anatomic reduction was achieved in 6 cases in the closed reduction group (33.3％) and 29 cases in the open reduction group (87.9％),respectively.All fractures healed after an average of 10.9 weeks (range,from 9 to 13 weeks),with one exception of nonunion in the closed reduction group.The incidence of femoral head necrosis in the closed reduction group (27.8％,5/18) was significantly higher than that in the open reduction group (15.2％,5/33) (P ＜ 0.05).According to the Ratliff's evaluation at the final follow-ups,the good to excellent rate in the closed reduction group(66.7％,12/18) was significantly lower than that in the open reduction group (84.8％,28/33) (P ＜0.05).Coxa vara occurred in 3 children in the closed reduction group but in none in the open reduction group.Conclusion Open reduction with internal fixation may lead to better outcomes than closed reduction in the treatment of children with displaced femoral neck fracture whose reduction is delayed by more than 24 hours,probably because open reduction can result in better reduction quality.

Objective To observe clinical efficacy of probiotics agents in the prevention and treatment of severe pulmonary infection in elderly patients with antibiotic-associated diarrhea (AAD).Methods 60 cases of elderly patients with severe pulmonary infection (more than or equal to 60 years old) were randomly divided into the treatment group ( 31 cases ) and control group ( 29 cases ) , the control group received broad-spectrum antibiotics or using two linked above anti-infection treatment, the treatment group were added with probiotic agent ( lactobacillus complex capsules).The diarrhea, the use of antibiotics and the stool routine, bacteria before and after 5, 10, 15 days of group were observed.Results After 15 days treatment, the number of Escherichia coli in treatment group was lower than that in control group (P<0.05), the number of Bacterium lacticum and Bifidobacterium bifidum in treatment group were higher than those in control group ( P<0.05 ) .The AAD rate in treatment goup was 12.90%, which was lower than 41.38% in control group (P<0.05).There were significant differences in beginning time and duration of diarrhea between two groups (P<0.05). Conclusion The intestinal probiotics reduced induced by antibiotics in elderly patients with severe pulmonary infection , the probiotics agents could redress intestinal flora imbalance, keep the steady state of intestinal flora, and prevent and cure the antibiotic-associated diarrhea.

BACKGROUND: Ghildren refractory clubfoot includes idiopathic multiple contracture, untreated idiopathic clubfoot in older children and failure cases of repeated operations. It is very difficult to treat this type of children patients. It has important significance in the exploration of new therapeutic methods.OBJECTIVE: To improve the therapeutic effectiveness of refractory clubfoot in children with Ilizarov method.DESIGN: a self-controlled study.SETTING: Department of Orthopedics, Nanjing Children' s Hospital, Nanjing Medical University.PARTICIPANTS: Totally 3 cases of children idiopathic multiple contracture, 2 cases of idiopathic clubfoot in children aged more than 10 years old without treatment and 6 cases of postoperative recurrent idiopathic clubfoot were admitted by the Department of Orthopedics of Nanjing Children' s Hospital Affiliated to Nanjing Medical University during 1999 to 2000. All cases manifested abnormal ankylosis and shortening deformity in foot.METHODS: Twelve refractory clubfeet in eleven patients were treated with the Ilizarov method. After achilles tendon lengthening and posterior release,kieschner' wires or olive wires with tension were penetrated through multiple planes of the middle lower 1/3 of tibia, calcaneus and the 1st to the 5th metatarsals, which were fixed respectively to external bone fixer of a ring,half-ring or horseshoe shape and connected with each other by multiple screw bars into a three-dimensional external bone fixing set. Plantarflexion, inversion, adduction and shortening deformities were corrected by the adjustment of the distance between each part of the external bone fixing set through extension, compression, and rotation, etc., and navicular and cuboid osteotomy.MAIN OUTCOME MEASURES: All 11 cases were followed up for 2 to 10 years with an average period of 4. 5 years. Indices like foot morphology,talocrural articualr activity range and deformity correction were observed during the follow up.RESULTS: The results were excellent in 5 cases (6 feet), good in 4 cases (4 feet), fair 2 cases(2 feet) according to the Garceau standard scale. The rate of excellent and good clinical results was 83.3%. Mild forefoot adduction was left in 2 feet of 2 cases and quite obvious tarsal articular ankylosis was found in 4 feet of 4 cases; however, there were no significant differences of gait and load bearing between these patients and normal children.CONCLUSION: The Ilizarov technique is a safe and effective method for the treatment of refractory children clubfoot while its indications should be controlled strictly.

Objective To study the effects of fluoride on minichromosone maintenance(MCM)3 mRNA and the bone formation-related gene:bone sialoprotein(BSP),osteocalcin(OC),osteopontin(OP)mRNA expression on human osteoblast cells.The expression of MCM3 was tested for diagnosis and surveillance value on osteoblast treated with excess fluoride.Methods Human osteoblast cell(Saos-2)was cultured in McCoy5A medium and treated with fluoride(sodium fluoride,NaF).There were eight groups including:0(control),0.625,1.250,2.500,5.000,10.000,20.000,40.000 mg/L groups.Expression of MCM3,BSP,OC,OP mRNA were detected by real-time PCR.Dual-standard curve method was used for analysis.ALPase was determined by measuring the absorbance using a micro titer plate reader. Results Expression of MCM3 mRNA was lower in the 0.625,1.250,2.500,5.000,20.000, 40.000 mg/L groups(0.059 ± 0.003,0.027 ± 0.001,0.272 ± 0.004,0.115 ± 0.002,0.137 ± 0.004,0.754 ±0.002, all P > 0.05) and was higher in10.000 mg/L group(21.300 ± 1.200, P < 0.01 ) than control group( 1.000 ±0.020), especially 10.000 mg/L group was higher than groups treated with fluoride(all P < 0.01 ), the differences among groups were significant(F = 305.842, P < 0.01 ). Expression of BSP mRNA was significantly higher in 0.625,1.250,2.500,5.000,10.000 mg/L groups(71.80 ± 3.60,133.00 ± 7.20,85.50 ± 0.60,80.90 ± 1.20,304.00 ± 21.00)than the control group( 1.00 ± 0.04), especially 10.000 mg/L group was higher than others groups treated with fluoride(all P < 0.01 ), the differences among groups were signifieant(F = 159.531, P < 0.01 ). Expressions of OC mRNA were higher in 0.625,1.250,2.500,5.000 mg/L groups(110.00 ± 12.00,143.00 ± 2.10,90.60 ± 4.10,23.70±1.20) than control group(1.00 ± 0.01, all P < 0.01), and the differences among groups were significant (F = 158.734, P < 0.01 ). Expression of OP mRNA were higher in 0.625,1.250,2.500,5.000,10.000,20.000 mg/L groups(167.00 ± 11.20, 111.00 ± 12.10,72.50 ± 3.50,134.00 ± 14.00,42.30 ± 2.40,45.20 ± 3.30) than the control group(1.00 ± 0.04, all P < 0.05 or < 0.01 ), the differences among groups were significant(F = 60.226, P < 0.01 ).Compared with control group(4.2 ± 1.2), the ALPase activity was increased in all groups treated with fluoride (6.0 ± 0.4,5.8 ± 0.1,5.7 ± 0.4,7.7 ± 1.1,19.2 ± 2.4,8.5 ± 3.0,18.1 ± 4.2), but only 10.000 mg/L and 40.000 mg/L groups were higher than control group and other groups treated with fluoride(all P < 0.01 ), the differences among groups were signifieant(F = 7.806, P < 0.01 ). Conclusions Irregular expression of MCM3 mRNA is not suitable as a diagnostic and monitoring biomarker of osteoblasts exposed to excessive fluoride. Fluoride may affect the osteoblast-related gene expression and to promote osteogenic differentiation.

Objective The aim of the study was to investigate the expression of P-glycoprotein (P-gp)and nuclear factor κB (NF-κB) in the cerebral cortex of rat with chronic fluorosis,and to reveal the mechanisms of damaged nervous system resulted from the toxicity of fluoride.Methods Sixty SD rats were randomly divided into three groups.The rats in each group were given drinking water containing different levels of fluoride:control group less than 0.5 mg/L,small amount of fluoride exposure group 10.0 mg/L and large amount of fluoride exposure group 50.0 mg/L.The animals were examined at the sixth month after initiating the experiment.Protein levels of P-gp and NF-κB in brain tissues were detected by immunocytochemistry and Western blotting,and the P-gp protein and mRNA level by quantitative real time PCR method.Results As compared to the control group(28.21 ±6.13),the numbers of positive staining cells by P-gp antibody in the cortex of rat brains were significantly increased in both the small and the large amount of fluoride exposure groups[(48.46 ± 8.00),(53.72 ± 9.15),respectively,all P ＜ 0.05] ; the protein levels in the control group(100.00 ± 3.86)％ detected by Western blotting were significantly increased in the cortex of rat brains treated with fluoride in both the small and the large amount of fluoride exposure groups[(189.47 ± 3.14)％,(191.36 ± 11.09)％,respectively,all P ＜ 0.05].The significantly increased expression of NF-κB at the protein level was observed in the cortex of rat brains of the small and the large amount of fluoride exposure groups[(365.97 ± 6.04)％ and (417.15 ± 10.89)％,respectively] as compared with the control group[(100.00 ± 10.07)％,all P ＜ 0.05].The mRMA level of P-gp in the cortex of rat brains of the small and the large amount of fluoride exposure groups(2396 ± 427,3479 ± 371,respectively) were higher than that of the control group(260 ± 106,all P ＜ 0.05).Conclusion The increased expressions of P-gp and NF-κB in the cortex of rat brains are induced by chronic fluorosis,which might be connected with the mechanism of brain damages.

Objective To investigate the changes of reactive oxygen species(ROS) level and mitochondria fission-fusion-balance in cortical neurons of rats with chronic fluorosis and reveal the correlation between these two factors. Methods One hundred and twenty rats were randomly divided into 3 groups(control group, low-dose fluorosis group, high-dose fluorosis group) and 40 rats were in each group according to body weight and the experiments were carried out for 3 months or 6 months. The rats were fed with different concentrations of fluoride (NaF) to establish fluorosis models. Controls were fed with tap water( < 0.5 mg/L), experimental animals in low- or high-dose group were fed with water containing NaF 10.0,50.0 mg/L, respectively. The level of ROS and the morphology in mitochondria fission-fusion balance in neurons of the cortex of rat brains prepared with cortical frozen sections were detected with ROS fluorescent probe and MitoTracker RED probe, respectively. Results Significant differences of the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were found between 3 groups at the experiment period of 3 month and 6 month(F= 3.07,3.06,3.05,3.07, all P < 0.05). As compared with control group(10.43 ± 5.98,4.12 ± 3.86) at the experiment period of 3 month, the level of ROS and the numbers of abnormal mitochondria in morphology in the cortical neurons were obviously increased in high-dose fluorosis group(25.48 ± 6.09,20.47 ± 6.09, all P < 0.05), whereas no significant changes were found in low-dose fluorosis group(11.67 ± 3.49,6.68 ± 3.48, all P> 0.05). Furthermore, the increases in both ROS level and abnormal numbers of mitochondria were significant observed in the cortical neurons of low-dose fluorosis group (63.02 ± 8.15, 49.33 ± 8.61) and high-dose fluorosis group(65.60 ± 7.40,53.10 ± 6.95) as compared with the control group (25.26 ± 6.41,20.26 ± 6.41) at the experimental period of 6 month (all P < 0.05). The abnormal numbers of mitochondria correlated with ROS level(r = 0.93,0.81, all P < 0.05). Conclusions Taking excessive amount of fluoride results in high level of oxidative stress and impaired the balance of mitochondrial fission-fusion,which is dependent on the feeding times and doses of fluoride. The mechanism of the mitochondrial abnormalities might be associated with the high level of oxidative stress induced by chronic fluorosis.