Objective To study the comparison of the therapeutic effect of Ultrasound guided and freehand botulinum toxin type of the treatment in children with cerebral palsy.Methods From March 2014 to September 2016,98 cases of children with cerebral palsy in our hospital by injection of botulinum toxin were treated.According to the way of injection,the children were divided into ultrasound group and free group(n =49).They were respectively gave botulinum toxin injection by guided Ultrasound guided and freehand ultrasound injection of hotulinum toxin injection.The amount of two groups of children was 3U/kg.Children muscle spasm degree of before and after treatment was assessed by modified Ashworth spasm scale.And exercise capacity of children before and after treatment was assessed by the gross motor function rating scale.The follow-up time was 3 months,the clinical curative effect of two groups of children was observed.Results The total efficiency of ultrasound guided group (98.0％) was significantly higher than the free group (79.6％),with statistically significant (P ＜ 0.05).Both groups before and after treatment in children with Ashworth score and GMFM score were significantly improved,with statistical significance (P ＜ 0.05).Ashworth score and GMFM scores of ultrasound guided group were significantly better than the free group,with statistically significant (P ＜ 0.05).Conclusion Botulinum toxin injection in the treatment of cerebral palsy children can effectively solve the spastic problem.Treatment of children with cerebral palsy by injecting botulinum toxin with ultrasound guided injection of botulinurn toxin is better than manual injection of botulinum toxin.

Objective To realize antimicrobial resistance rate of Pseudomonas aeruginosa (P. aeruginosa)and it’s correlation with antimicrobial use density(AUD),and to provide reference for control of healthcare-associated infec-tion.Methods From July 2011 to December 2013,antimicrobial resistance rate of P. aeruginosa isolated from hospi-talized patients and AUD of patients were monitored,and the correlation between them was analyzed.Results AUD of patients decreased from 73.61 in the third and fourth quarters of 2011 to 41.33 in the same quarters of 2013. Corre-lation coefficient of AUD and antimicrobial resistance rate of P. aeruginosa was -0.32~0.88,correlation coeffi-cient of resistance rate of P. aeruginosa to aztreonam and aztreonam use density was 0.88,there was statistical sig-nificance. Conclusion AUD of hospitalized patients revealed a decreasing tendency,suggesting antimicrobial selec-tive resistance should be considered in clinic.

qualitative and quantitative analysis of solid depolymerization products for supercritical methanolysis of ploy(ethylene terephthalate) polyester (PET) were studied by means of reversedphase high performance liquid chromatogr aphy on ZorbaxC8(4.6mm×250mm) with 70%(V/V) methanol as mobile phase. The results showed that the solid depolymerization products were mainly composed of dimethyl terephthalate (DMT), methyl(2hydroxyethyl)terephthalate (MHET), bis(hydroxyethyl)tere phthalate (BHET), dimers and oligomers, which could be separated effectively under the above chromatographic conditions. The calibration curves of DMT and BHET were linear in the range of 1.0～45 mg/L (n=8),r=0.9998～1.0000 and RSD＜2.8%. The determination limits of DMT and BHET were 4.0×10-4μg and 6.0×10-4μg respectively. 

Objective To discuss the clinical characteristics and treatment methods of lumbar disc herniation in adolescent and to in-crease the awareness of lumbar disc herniation among orthopedists.Methods Retrospectively reviewed the clinical data of 201 adolescent pa-tients(aged 20 years or less)with lumbar disc herniation who were admitted in General Hospital of Shenyang Military Area Command from January 2001 to December 2010.Analyzed the clinical characteristics and treatment methods,and compared the VAS score and ODI score be-fore operation,one week after operation and at the last follow up.Meanwhile,the complications were observed and compared.Results Among the 201 patients,187 patients(93.0%)presented with low back pain with or without radiating pain,20 patients(10.0%)presented with leg pain for the first symptom.There were only 97 patients(48.3%)were diagnosed with lumbar disc herniation for their first diagnosis in other hospital.Totally 70 patients(34.8%)had a history of trauma before the onset.The most common segments were L4 /L5 (101 cases,50.2%) and L5 /S1 (70 cases,34.8%).There were 140 patients treated by microendoscopy discectomy(MED),25 patients treated by percutaneous endoscopic lumbar discectomy(PELD),and 36 patients treated by open lumbar discectomy(OLD).All the patients were followed up for more than 12 months with averagely followed up for(22.9 ±11.7)months.There was significant improvement one week after operation and at the last follow up compared with the preoperative VAS score and ODI score,and the differences were statistically significant(P <0.05).The rate of postoperative complications was 3.6%,4.0%,8.3% in patients with MED,PELD and OLD respectively.Conclusion Diagnosis of LDH in adolescent is usually delayed.Increase the awareness of lumbar disc herniation could help orthopedists to get a exact medical history,per-form a directed physical examination,and order appropriate imaging studies.Minimally invasive surgery is an effective method to treat lumbar disc herniation in adolescent.

Objective:To examine the expression of CLC-3 in colorectal tissues and the effect of CLC-3 on the viability and invasion of colorectal cancer (CRC) SW480 and SW620 cells. Methods:The mRNA levels of CLC-3 in CRC cell lines were determined by RT-PCR. CLC-3 expression was inhibited by adding DIDS or NPPB to the CRC cells. Subsequently, cell viability and invasion were assessed by CCK-8 assay and Transwell assay, respectively. In addition, the effects of DIDS and NPPB on the Wnt orβ-catenin signaling pathways were de-termined by Western blot analysis. Results:The mRNA level of CLC-3 was remarkably increased in the CRC tissues compared with that in normal colorectal tissues (P<0.05) and was positively correlated with the T stage of CRC. The blockade of CLC-3 inhibited the viability and invasion of CRC cells (P<0.05). The expression ofβ-catenin, C-myc, cyclin D1, Ki-67, and survivin were evidently reduced by the in-hibition of CLC-3 (P<0.05). Conclusion:The inhibition of CLC-3 decreases the cell viability and invasion of CRC cells by reducing the ex-pression of the proteins related to the Wnt orβ-catenin signaling pathway.

Objective To understand the distribution of pathogenic genes in uropathogenic Esche-richia coli (UPEC) from urinary specimen and to analyze the pathogenicity of UPEC and their mechanism of apoptosis to HeLa cells. Methods We have analyzed 6 pathogenic genes among the 28 strains of the clini-cally isolated E. coli from urinary tract infected patients. The 6 pathogenic genes were surveyed by using the PCR amplification of the target genes. The adhesion experiments and tryphan-blue staining was used to screen the phenotype of the pathogenic strains, while Annexin V/PI method was applied to study the strains to cause apoptosis of the HeLa cells, which was further confirmed with electronic microscopy. Results We have detected 6 strains that carried the usp gene. Phenotype screening identified two high virulent isolates (strain 6N and 27N) from the 6 strains. Strain 27N and 6N contained very similar virulence gene profile ex-cept that strain 27N did not contain usp gene. Both strains can destroy HeLa cell within 3 hours causing cell death. Results of apoptosis detected by flow cytometry revealed that strain 6N induced 20.75% of HeLa cells to an early stage apoptosis within 1.5 hours. On the other hand, strain 27N induced only 1.55% of HeLa cells to apeptosis. Conclusion High virulent UPEC strain carrying usp gene can induce HeLa cell rapid early apoptosis.

Objective To evaluate the changes in the activity of extracellular signal-regulated kinase 5 (ERK5) in the distal cerebrospinal fluid contacting neurons (CSF-CNs) in the mid-brain of morphine dependent rats.Methods Forty-eight male adult Sprague-Dawley rats weighing 230-270 g,were randomly divided into 2 groups (n =24 each) using a random number table:control group (group A) and morphine dependence group (group B).Morphine dependence was induced by increasing doses of subautaneous morphine for 5 consecutive days.The initial dose of morphine was 10 mg/kg twice a day and was increased by 10 mg/kg everyday until 50 mg/kg on 5th dav.The equal volume of normal saline was injected subcutaneously instead of morphine in group A.On 3rd day after morphine dependence was induced,the distal CSF-CNs in the mid-brain was labeled with 30％ cholera toxin subunit B and horseradish peroxidase compound (CB-HRP) 3 μl injected in the lateral cerebral ventricle in the morning.At 4 h after the last injection of morphine,the segments in which CSF-CNs were located were removed,and CB-HRP positive neurons,phosphor-ERK5 (p-ERK5) positive neurons and CB-HRP/p-ERK5 positive neurons were counted.Results Compared with group A,the number of p-ERK5 and CB-HRP/p-ERK5 positive neurons in the mid-brain was significantly increased (P ＜ 0.05),and no significant change was found in CB-HRP positive neurons in group B (P ＞ 0.05).Conclusion The enhanced activity of ERK5 in the distal CSFCNs in the mid-brain may contribute to the development of morphine dependence in rats.

Objective To evaluate the role of spinal neuronal extracellular signal-regulated protein kinases 5/cAMP response element binding protein (ERK5/CREB) signaling pathway in withdrawal responses in morphinedependent rats.Methods Ninety-six adult male Sprague-Dawley rats in which intrathecal catheters were successfully placed,weighing 200-250 g,were randomly divided into 4 groups (n =24 each):normal control group (group A),morphine withdrawal group (group B),dimethyl sulfoxide (DMSO) + morphine withdrawal group (group C) and ERK5 inhibitor BIX02188 + morphine withdrawal group (group D).Morphine dependence (MD) was induced by increasing doses of subcutaneous morphine for 6 days.The initial dose of morphine was 10 mg/kg once a day and was increased by 10 mg/kg once a day from 2nd to 5th days until 50 mg/kg on 6th day in B,C and D groups.Morphine withdrawal response (MW) was induced by intraperitoneal naloxone 4 mg/kg at 4 h after last morphine administration in B,C and D groups.In addition,BIX02188 10 μg and 1％ DMSO 10 μl were injected intrathecally at 1 h before naloxone injection in D and C groups,respectively.MW and morphine withdrawal-induced hyperalgesia were scored.The rats were then sacrificed after hyperalgesia was scored and the spinal cord was removed for determination of CREB and phosphorylated CREB (p-CREB) expression.Results Compared with group A,MW and hyperalgesia scores were significantly increased and the expression of p-CREB was up-regulated in B,C and D groups.Compared with group B,MW and hyperalgesia scores were significantly decreased and the expression of p-CREB was down-regulated in D group,and no significant change was found in group C.Conclusion The spinal neuronal ERK5/CREB signaling pathway is involved in withdrawal responses in morphine-dependent rats.

Objective To investigate the effects of melatonin on the changes of superoxide dismutase ( SOD ) , reactive oxygen species (ROS),and malondialdehyde(MDA) in N2A cells under hypoxia conditions.Methods Randomly divided the primary cultured neuroblasto-ma cells of mouse into the control group ,hypoxia group ,and MT treatment group .The MT treatment group were given melatonin 5μg/mL for 24 h to set up the treatment model .Rsbiotech of the hypoxia group and MT treatment group were given gaseous mixture of 95%N2 and 5%CO2 ,The ischemia hypoxia model of N 2A cells was set up with cells in the oxidative stress state and cultured for 24 hours at low concentra-tions of serum .The content of SOD , ROS, and MDA was measured respectively by xanthine oxidase , fluorogenic quantitative detection and thiobarbituric acid chromatometry .Results The expression of SOD in hypoxia group and MT treatment group were significantly decreased compared with that in control group(P<0.05).The level of SOD in MT treatment group significantly recovered compared with hypoxia group with singnificant difference (P<0.05).The expression of MDA and ROS in hypoxia group and MT treatment group were significantly in-creased compared with that in control group (P<0.05).The level of MDA and ROS in MT treatment group significantly recovered compared with hypoxia group with singnificant difference (P<0.05).Conclsion Melatonin provides a protective effect on the secondary damage of nerve cells with hypoxia ischemia .The possible mechanism is melatonin could play the role of free radical scavenging and up -regulate the ex-pression level of antioxidants .

Objective To observe the regulatory effect of bFGF and TGF-β1 for the proliferation of mesenchymal progenitor cells ( MPCs) derived from primary osteoarthritis cartilage, and to provide theoretical evidence in preventing and curing primary OA. Methods Different concentrations of bFGF and TGF-β1 ( alone or combined) were used to treat primary OA cartilage and their effects on proliferation of MPCs were tested by MTT method. Results Either bFGF (10. 0~50. 0 ng/mL) or TGF-β1 (0. 1 ~1. 0 ng/mL) alone can significantly promote the proliferation of MPCs derived from primary OA cartilage (P<0. 05). But with their increased concentration,the proliferation rate was of no significant changes (P>0. 05). The combination of 10. 0 ng/mL bFGF and 1. 0 ng/mL TGF-β1 significantly increased the prolifer-ation of MPCs from primary OA (P<0. 05). Conclusion Both bFGF and TGF-β11 play important roles in the proliferation of MPCs in primary OA cartilage,and they can increase the proliferation in different degree with different concentrations. There must be feasible methods of gene technology to promote cell proliferation and differentiation of MPCs for repairing articular car-tilage injury.