Objective To observe the cerebral activation pattern in patients with cervical dystonia (CD) during finger movements and the changes caused by botulinum toxin injection by functional MRI.The possible etiological mechanism of CD and effect of peripheral botulinum toxin treatment on the level of central nerves system are investigated.Methods A designed functional MRI block with complex finger movements was applied and 11 patients with CD as well as 11 age and gender matched controls were scanned.Compare the activation pattern of CD pre/post treatment groups versus health controls.Evaluate and compare the symtom severity with Toronto Western Spasmodic Torticollis Rating Scale (TWSTRS).Make a correlation analysis between activation pattern alteration and TWSTRS change in CD pre/post treatment groups.Results An reduced extent of activation in patients with CD was demonstrated compared to healthy controls in ipsilateral putamen,prefrontal cortex and contralateral somatosensory cortex to the direction of the head deviation,while an elevated extent of activation in ipsilateral precuneus and fusiform with statistic significance.At the time point of 4 weeks after botulinum toxin treatment patients showed no evident difference with healthy controls except for the decreased activation in contralateral precuneus to the direction of the head deviation.TWSTRS of patients with CD decreased from (20.02 ± 5.52) to (4.11 ± 4.34) with statistic significance (t =11.71,P =0.000) after botulinum toxin treatment.There was a positive correlation between cerebral activation pattern change in some cerebral areas (ipsilateral primary somatosensory cortex,premotor cortex,supplementary motor cortex,insula,fusiform,hippocampus with parahippocampa as well as contralateral middle temporal cortex and hippocampus with parahippocampa to the direction of the head deviation) and TWSTRS score decrease.Conclusions There are widespread abnormalities in cortical and subcortical activation pattern in patients with CD,which might due to dysfunction of sensory-motor integration.We speculate a basic pre-dystonic state is present in affected body parts prior to clinical symptoms appear.Botulinum toxin fulfills its subsequent central effect by reorganizing and normalizing the cerebral cortex in patients with CD.

Objective To evaluate the efficacy and safety of combination therapy of Endostar and oxaliplatin plus S-1 ( SOX regimen) in patients with advanced Primary carcinoma of the liver. Methods 32 advanced primary liver cancer patients admitted from February 2012 to August 2014 were assigned to SOX regimen as systemic chemotherapy: oxaliplatin 130 mg/m2 iv d1; S-1 (80 ~ 120 mg, twice-daily) for 14 days; 150 mg Endostar which was dissolved in 210 mL normal saline for 120 h durative transfusion. Treatment was repeated every 21 days. Objective clinical efficacy and adverse effect was assessed every 2 cycles. Serum alpha fetoprotein (AFP) level was also monitored according to the schedule. Results All 32 patients were available to be assessed, the objective response rate (ORR), disease control rate (DCR) ,the clinical benefit response rates (CBR), 1 year survival rate was 15.6%, 46.9%, 56.3%, 58.3% respectively. The serum AFP respond rate was 19.4%. Major adverse effects were myelosuppression and fatigue , mostly graded at 1 ~ 2. There were no treatment-related death. Conclusions These preliminary results suggest that continuous intravenous pumping of Endostar combined with SOX regimen could provide survival benefits with tolerable adverse effects.

Objective To evaluate influence of patient-controlled intravenous analgesia (PCIA) and patient-controlled epidural analgesia (PCEA) on expressions of serum inflammatory cytokines in elderly patients with hemi or whole hip replacement using cemented artificial joint. Methods Elderly patients undergoing hip replacement were selected and were divided into PCIA group and PCEA group. VAS scores were calculated at 12 h postoperatively. Patients whose VAS scores were not more than 2 at postoperative 12h were included. 30 cases in each group were finally included. Fifteen cases were randomly chosen in each group and underwent sample blood drawing for assays. Expressions of serum inflammatory cytokines were detected by ELISA , RT-PCR and Western-blot. Results Gene and protein expressions of TNF-a as well as IL-6 in group PCEA were lower and expression of IL-10 was higher than that in group PCIA. Serum level of TGb-β was higher in group PCEA detected by ELISA. There was no significant difference in expression of IL-8 between groups. Conclusions PCEA may better promote expressions of anti-inflammatory cytokines and inhibit expressions of proinflammatory cytokines. PCEA is superior in maintenance of inflammatory cytokine balance.

Objective To investigate the clinical application of the coblation in the treatment of the infant with sleep-disordered breathing. Methods The clinical data were reviewed from 161 infants, who had the cobtilaon tonsillectomy and/or adenoidectomy sugeries in our hospital from January , 2008 to June, 2012. Among the 161 SDB cases, there were 85 obstructive sleep apnea hypopnea syndrom cases and 76 primary snoring cases. After 12 months, the follow-up visit is cut off in January, 2013. Successful follow-ups had been done to 161 infant with SDB. And the clinical efficacy and the occurrence of the complications were investigated. Results the intra-operative blood loss was 10 mL or less. 2(1.24%) were delayed hemorrhage with less pain afteroperation. One year after the surgery, there were 141 cured (87.6%), 15 with apparent effects (9.3%), 3 with effective results (1.9%), 2 with no effect(1.2%)and the total effective rate is 98.8%. Conclusion It is minimally invasive, safe and effective to use coblation to remove tonsil and adenoid in the treatment of infants with sleep-disordered breathing.

Objective To study the expressions of estrogen receptor(ER)and progesterone receptor(PR)in elderly patients with breast cancer,and the correlation between their expressions and clinicopathological characteristics.Methods The expressions of ER and PR in 110 elderly patients with breast cancer(over 60 years old)from 1995 to 2004 were detected,and their clinical and pathological features were analyzed retrospectively.Another 110 patients aging 30 to 59 years with breast cancer served as control.Results Breast cancer in aged and non-elderly group had similar pathohistological types,but the aged had more tumors with low-grade malignant types.ER and PR were found to be over-expressed in elderly patients but not in the control.No obvious correlation was seen with clinical pathological stages.The expression levels of ER and PR in 0-3 positive nodes group was higher than ≥4 positive nodes group.Conclusion Elderly patients and non-elderly patients have similar pathohistological types of breast cancer.The former have higher expressions of ER and PR,and their expressions are associated with axillary lymph node status.

OBJECTIVEThis study aims to synthesize MTO1 (a kind of oligodeoxynucleotides) and N-isopropylacrylamide-modified polyethylenimines (PEN) complexes (MT01/PEN) as well as to investigate the effect of the complexes on the expression of osteoprotegerin (OPG) and the receptor activator of nuclear factor κB ligand (RANKL) in the human osteoblast-like cell line MG63.

METHODSMG63 cells were transfected by MT01/PEN complexes formed with three different mass ratios (1:2, 1:4, 1:6) of MT01 to PEN. MT01 and MT01-s were used as positive control. Enzyme-linked immunosorbent assay and real-time polymerase chain reaction were performed to estimate the amount of OPG and RANKL released into the culture media and in MG63 at 24, 48, 72 h.

RESULTSMG63 responded to the MT01/PEN complexes by significantly upregulating the OPG on the protein and mRNA levels (P < 0.05). The protein and mRNA levels of RANKL were lower in most of the groups with complexes, and the OPG/RANKL ratio were higher (P < 0.05). MG63 were affected by the MT01/PEN complexes with different mass ratios, particularly when the ratio was 1:6.

CONCLUSIONMT01 can enhance the promotion of ossification by establishing the delivery system with PEN.

Acrylamides ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Humans ; Oligodeoxyribonucleotides ; Osteoblasts ; Osteoprotegerin ; Polyethyleneimine ; RANK Ligand ; RNA, Messenger ; Real-Time Polymerase Chain Reaction

Objective To investigate the growth inhibition and radiosensitization of Celecoxib in hu-man nasopharyngeal carcinoma cell line CNE-2. Methods CNE-2 growth inhibition by Celecoxib was eval-uated by MTT method. Apoptosis-related changes in morphology were observed by transmission electron mi-croscopy (TEM). Cell cycle distribution and apoptosis rate were measured by flowcytometry (FCM). The ex-pression of COX-2 protein was observed by SP method after the treatment of Celecoxib. Cells were randomly planted into four groups: irradiation control(Ci), drug group(Cd), irradiation group(R), and Celecoxib plus irradiation group(D+R). Single irradiation of 2,4,6,8,and 10 Gy were administered for colonogenic assay. Cell cycle distribution and apoptosis rate were analyzed at 6 Gy irradiation. Results The growth of CNE-2 cell was inhibited by celecoxib in a dose-and time-dependent manner, the IC50 was 80 μmol/L After the treatment, cell ratio of GO and G, phases was increased (47.03±2.76 vs 56.17±1.95, t=4.68, P= 0.010), whereas the ratio of S and G2/M phases was decreased (33.07±1.86 vs 24.87±1.76, t=5.54, P = 0.010; 19.30±0.53: 17.73±0.83, t=2.75, P=0.050), and the apoptosis rate was increased (1.57±0.47:10.47±0.31, t = 27.39, P = 0.000) in a dose-dependent manner. Apoptosis with nuclear chromatin condensation, fragmentation and cell shrinkage was found by TEM. SP method showed that Celeib decreased COX-2 expression (17.48±0.34 vs 12.82±0.51,t=13.20,P =0.00). The sensitivity ratio(D0) was 1.15. FCM showed that the percentage of cells in G2/M phase was significanty more in R and D+R groups than in Ci and Cd groups (68.00±1.65,54.27±5.74,17.60±0.80,14.86±1.23, t=47.70,P=0.000; t=11.63, P=0.000), and also significantly different between R group and D + R group (t=3.99, P= 0.020). The apoptosis rate was higher in R and D + R groups than Ci and Cd groups(4.83±0.97,9.50± 1.35,1.33±0.86 and 2.28±0.42,t=4.67,P=0.010;t=8.81, P=0.000), D + R group than R group(t =4.85,P=0.010). Conclusions Celecoxib can markedly inhibit the growth and induce apoptosis in CNE-2 cells,which may depend on COX-2 pathway. Celeeoxib potently enhances the radiosensitivity of CNE-2 cells,which may due to the repair inhibit of radiation-induced DNA damage, inhibit of cell proliferation,and enhancement of cell apoptosis after irradiation.

Objective To investigate the efficacy and safety of hypertonie saline enhanced radiofrequency ablation (RFA) in the treatment of liver cancer. Methods The clinical data of 42 patients with primary liver cancer (n = 28) or metastatic liver eancer (n = 14) who had been admitted to First Affiliated Hospital of Nanjing Medical University from September 2001 to December 2007 were collected. Forty-eight lesions were detected with a diameter ranging from 1.2 cm to 7.5 cm. RFA electrode and 20G needle were pricked into the target lesion under the guidance of B ultrasound or computed tomography (CT) through percutaneoas puncture or open approaches. An amount of 5-10 ml hypertonie saline was infused through the needle at regular intervals during RFA. All patients were followed up for 3-79 months. Contrast-enhanced ultrasound and CT scanning were performed postoperatively to determine the efficacy of RFA. The levels of alpha-fetoprotein (AFP) before and after treatment were compared using t test, and the survival of the patients were analyzed using a Kaplan-Meier survival curve. Results The AFP expression changed to negative in 14 out of the 18 AFP-positive patients, with statistical difference (t =7.703, P <0.05). The complete necrosis rate of tumors was 94% (45/48), and the necrosis rate of tumors with diameter of ≤4.0 cm reached 100% (35/35). The incidence of complication was 5% (2/42). No perioperative mortality occurred. The 1-, 2-, 3-year survival rates were 91%, 85% and 70%, respectively. Conclusions Hypertonic saline enhanced RFA in the treatment of liver cancer was proved to be safe and effective.

Objective In order to extrapolate the respiratory dysfunction of patients in early stage of acute organophosphorus pesticide poisoning (AOPP),transpulmonary thermodilution technique was used in swine models of severe acute dichlorvos poisoning (SADP) to evaluate respiratory function.Methods Twenty healthy female swine were randomly divided into dichlorvos ( n =7 ),atropine ( n =7 ) and control (n =6) groups.In the dichlorvos group,the swine were administered with 80％ emulsified dichlorvos (100mg/kg) via the gastric tube toinduce SADP.In the atropine group,swinewere administered with dichlorvos,and 0.5h later,atropine was injected to obtain and maintain atropinization.The swine of control group were administered with saline solution instead.Arterial and venous blood samples were collected 0,0.5,1,2,4 and 6 hours after modeling for blood gas analysis and detecting acetylcholinesterase levels.Both extravascular lung water index (EVLWI) and pulmonary vascular permeability index ( PVPI ) were measured by using PiCCO (pulse indicator continuous cardiac output ). At the termination of the experiment,the animals were sacrificed and the lung wet/dry weight ratio was determined and histopathological changes of lung tissue were also observed under microscope.Results In the dichlorvos group,EVLWI and PVPI were substantially increased from 0.5 h to 6 h after modeling but PaO2/FiO2 decreased from 0-6 h after modeling.In the atropine group,EVLWI and PVPI increased initially,but then they decreased 1 h afterwards and PaO2/FiO2 was also gradually decreased from 0-1 h.In both dichlorvos group and atropine group,the EVLWI was negatively correlated with PO2/FiO2 and positively correlated with PVPI.Compared with the control group,the lung wet/dry weight ratio increased markedly in the dichlorvos group and mildly increased in the atropine group.Meanwhile,the histopathological changes of lung tissue were obvious in the dichlorvos group and mild in the atropine group.Conclusions SADP swine experienced substantial changes in respiratory function. EVLWI was a reliable and valuable indicator for evaluating respiratory function in the early stage of AOPP.

Objective To investigate the cytotoxicity of vascular endothelial growth factor (VEGF) siRNA combined with fusion suicide gene yCDglyTK on human gastric cancer cell line in vitro.MethodsThe gastric cancer cell line SGC7901 was transfeeted with blank plasmid pcDNA3.1 (-) null [pcDNA3.1 (-) group], or VEGF-siRNA expression plasmid pGenesil-shVEGF (SGC7901/shVEGF group),or fusion suicide gene plasmid pcDNA3.1 (-)CV-yCDglyTK (SGC7901/CDTK group),or combined gene plasmid pcDNA3.1 (-)shVEGF-yCDglyTK (SGC7901//shVEGF-CDTK group) with calcium phosphate nanoparticles (CPNPs).Un-transfected gastric cells were set as control group.The stable transfected cells were selected by G418.The target gene expression was verified by RT-PCR and Western-blot.After given prodrug 5-fluorocytosine (5-FC), the biologic characters variation, apoptotic morphology and apoptotic rate of cells in each group were observed through cell growth curve by MTT assays, by-stander effect, Hoechst 33258 staining and flow cytometry.The data was analyzed with SPSS 13.0 software and multiple groups’ comparison was analyzed with LSD test.ResultsFour gastric cancer cells lines transfected with different plasmids were successfully established.The expression of gene yCDglyTK was detected both in SGC7901/CDTK cells and SGC7901/shVEGF-CDTK cells.By MTT assays, the cell growth curve indicated that the A570 value of SGC7901/shVEGF cells, SGC7901/CDTK cells and SGC7901/shVEGF-CDTK cells decreased significantly compared with that of SGC7901 and SGC7901/null cells after a 24-hour 5-FC treatment (P＜0.01).When the percentage of stable gene trasfected SGC7901 cells was 60％, 80％and 100％, the cell relative viability was 13.09％±2.40％, 9.74％±2.83％ and 5.68％±1.03％,respectively. A large number of cells in SGC7901/CDTK and SGC7901/shVEGF-CDTK group appeared typical apoptotic morphology under fluorescence microscope.The result of flow cytometry showed that the apoptosis rates in SGC7901/shVEG group、 SGC7901/CDTK group and SGC7901/shVEGF-CDTK group were 16.40％ ±4.68％, 57.63％ ± 4.96％ and 69.07％ ± 4.69％,respectively, and there were significant differences compared with control (P＜0.01).Conclusion VEGF siRNA combined with suicide gene can effectively kill gastric cancer cell line SGC7901.Apoptosis induction may be one of the important mechanisms of killing tumor cells.