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Objective: To observe the clinical efficacy of spinal cord stimulation (SCS) in the treatment of diabetic foot (DF). Methods: Sixteen patients who were diagnosed with DF and treated with SCS from the Department of Pain Medicine of the First Affiliated Hospital, China Medical University from October 2015 to October 2018 were retrospectively analyzed. Visual analogue scale (VAS), 36-item short form health survey (SF-36), transcutaneous oxygen partial pressure (TcPO 2 ) and skin temperature of lower limbs were compared before and after treatment, and the complications were recorded. Results: Among the 16 patients, 14 were equipped with implantable pulse generator (IPG). The VAS scores decreased significantly from 7.5±1.2 before treatment to 2.6±0.8, 2.0±0.7, 1.6±0.6, 1.0±0.9, 0.9±0.9 at 1 day, 1 week, 3 months, 6 months and 12 months after electrode implantation respectively (all P< 0.05). At 12 months after treatment, the parameters of SF-36 were significantly different from those before treatment (all P< 0.05). The TcPO 2 was (23±5) mmHg before treatment and (38±6) mmHg at 1 week after treatment, with a statistical difference ( P< 0.05), and the temperature of lower limbs increased significantly ( P <0.05). No serious complications were observed in all patients. Conclusion: SCS can relieve the pain of patients with DF, improve the microcirculation and blood supply of lower limbs, and thus promote the quality of life, with rare serious complications.

Objective: To evaluate the influence of preoperative fasting duration on blood volume status of pediatric patients during induction based on ultrasonic technique. Methods: One hundred and ten pediatric patients, scheduled for elective operation in the Second Affiliated Hospital & Yuying Children's Hospital, were recruited during January and October in 2018. After sedation by inhalation of sevoflurane, the maximum (expiratory) and minimum (inspiratory) diameter of inferior vena cava (IVC(max), IVC(min)) and aorta velocity-time integral (VTI) in apical five-chamber cardiac view were measured with an ultrasound machine. Respiratory variabilities of these parameters were further calculated. Furthermore, passive leg raising (PLR) test was performed and above measurements/calculations were repeated. The correlation between the duration of fasting and IVC respiratory variations index (IVC(RVI)) or aortic VTI variability (ΔVTI) was then analyzed. Results: Before PLR, IVC(max), IVC(min) and IVC(RVI) were (0.78±0.19), (0.43±0.15) cm and 0.45±0.12, respectively. After PLR, IVC(max) and IVC(min) increased to (0.94±0.20), (0.55±0.18) cm, while IVC(RVI) decreased to 0.42±0.13, the differences were statistically significant ( t= 15.66, 10.85, 3.14, all P< 0.05). However, IVC(max), IVC(min) and IVC(RVI) were not significantly correlated with the duration of fasting analyzed by linear regression (before PLR: r= 0.052, 0.163, 0.171; after PLR, r= 0.062, 0.169, 0.165, all P> 0.05). Before PLR, expiratory aortic VTI (VTI(max)), inspiratory aortic VTI (VTI(min)) and ΔVTI were 21±5, 17±4 and 17±8, respectively. After PLR, the VTI(max) and VTI(min) significantly increased to 23±5 and 19±4 ( t= 13.60, 10.43, all P< 0.05), but ΔVTI was not changed significantly, which was 17±8( t= 0.34, P> 0.05). Linear regression analysis showed that VTI(max), VTI(min) and ΔVTI were not significantly correlated with the duration of fasting (before PLR: r= 0.111, 0.100, 0.047; after PLR: r= 0.003, 0.033, 0.073, all P> 0.05). Further multiple linear regression analysis indicated that, age and body weight were independent factors influencing IVC(RVI) and ΔVTI before and after PLR (IVC(RVI): β=-0.441, 0.515, -0.451, 0.507; ΔVTI: β=-0.442, 0.545, -2.422, 2.850; all P< 0.05). However, the duration of fasting was not correlated with IVC(RVI) and ΔVTI after adjusting the age and weight (IVC(RVI): β=0.177, 0.160; ΔVTI: β=0.037, 0.054; all P> 0.05). Conclusion: Age and weight, but not preoperative fasting duration, are correlated with respiratory variabilities of inferior vena cava diameter and aortic VTI in pediatric patients.

Objective: We aimed to explore the feasibility and perioperative safety of performing catheter ablation and left atrial appendage closure (LAAC) in a single (one-stop) session in patients with atrial fibrillation (AF). Methods: This study is an observational study. Consecutive AF patients who underwent the combined procedure of catheter ablation and LAAC with Watchman device of Xinhua Hospital in Shanghai between March 2017 and May 2019 were prospectively enrolled. Baseline, intra-and peri-procedural parameters were evaluated. Results: A total of 358 AF patients (189 males, (69.0±8.0) years) underwent the one-stop procedure. The CHA 2 DS 2 -VASc score was 3.2±1.5 and HAS-BLED score was 2.4±1.1, respectively in this patient cohort. Pulmonary vein isolation was achieved in all patients, while additional linear ablation was applied in 180 (50.3%) patients, yielding immediate success rate of 99.7%. Successful Watchman implantation was achieved in all patients. The perioperative serious adverse event occurred in 14 cases (3.9%). including 6 pericardial effusions (1.7%), 1 stroke (0.3%) and 5 vascular complications (1.4%), yielding procedure-related complication rate of 3.4%. In addition, 2 (0.6%) new-onset heart failures occurred postoperatively. There was no major bleeding or death during the perioperative period. Conclusions: Combined catheter ablation and LAAC can be successfully and safely performed in AF patients with high stroke risk. Follow-up data are needed to evaluate the outcome of this one-stop procedure.

Objective: To develop and prospectively validate a risk score for acute chest pain patients with normal high-sensitivity troponin I (hs-TnI) levels and without obvious ST-segment deviation in China. Methods: Chest pain patients admitted to the emergency department of Beijing Anzhen Hospital from September 2014 to July 2015 were enrolled. Baseline characteristics of patients met inclusion criteria including normal hs-TnI levels and without obvious ST-segment deviation were included. The endpoint (major adverse cardiovascular events) was a composite of acute myocardial infarction, percutaneous coronary intervention, coronary artery bypass graft, and all-cause death within 3 months after initial presentation. Predictors were screened and used to develop the risk score model by logistic regression analysis in a retrospective cohort. Then, the risk score model was evaluated in a prospective validation cohort. Results: The study population of derivation cohort included 1 735 consecutive chest pain patients. Finally, 1 030 eligible patients were enrolled. Multivariate regression analysis defined five independent predictors: male gender (β=0.88); history of chest pain (β value of moderate and high suspicion of coronary heart artery was 2.70 and 3.51 respectively); electrocardiogram (β=0.84); ≥60 years old (β=0.51) and ≥3 risk factors (β=0.85).The range of weighted score was set as 0-13. The area under a receiver operating characteristic (ROC) curve was 0.75 (95% CI 0.72-0.78) in the final model. Major adverse cardiovascular events rates increased in proportion to score increase ( P< 0.01). The internal validity used bootstrap technique showed the same predictor factors as the final model, and its area under a ROC curve was 0.75(95% CI 0.72-0.78).MACE rates in the low risk group (score 0-3), intermediate risk group (score 4-7), and high risk group (score 8-13) were 1.3% (1/77) ,19.0% (22/116) ,and 42.2% (122/289) in the prospective validation cohort, respectively ( P< 0.01). Conclusion: The developed ischemic risk score is feasible for risk stratification of acute chest pain patients with normal hs-TnI and without obvious ST-segment deviation, this score might be helpful to the decision making of treatment and management strategies for these patients.

Objective: To explore whether diabetes mellitus (DM) impairs functions of bone marrow-derived endothelial progenitor cells (BM-EPC) and circulating EPC. Methods: Diabetic model of rabbit was induced by Alloxan injection and the rabbits were then randomly divided into three groups: BM-EPC group, circulating EPC group, and DM group, with six rabbits in each group. Another 6 normal rabbits were enrolled as normal control group as well. 8 weeks later, BM-EPC and circulating EPC from diabetic and healthy rabbits were isolated and cultured. Colony number, proliferation, adhesion and tube formation function were detected. Exogenous diabetic BM-EPC and circulating EPC were analyzed for therapeutic efficacy in acute ischemia model of diabetic rabbits. Left ventricular (LV) function was assessed using Echocardiography. Capillary density and fibrosis area were evaluated by confocal laser scanning microscope (CLSM) and Masson-trichrome staining. The mRNA expression of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) was analyzed using real-time quantitive PCR. Results: Colony number, proliferation, adhesion and tube formation function of diabetic circulating EPC were significantly reduced compared with healthy rabbits. DM impaired tube-forming ability of BM-EPC, but did not influence colony number, proliferation and adhesion function. Compared with circulating EPC and control group, BM-EPC group had fewer fibrosis area (6.98%±0.94% vs 13.03%±2.97% and 15.84%±4.74%, both P =0.001), higher capillary density [(792±87) vs (528±71) and (372±77) vessels/mm(2,) both P <0.001], higher mRNA expression of VEGF (6.25±2.33 vs 2.19±1.01 and 1.55±0.52, both P <0.001) and bFGF (6.38±2.65 vs 1.24±0.76 and 1.18±0.82, both P <0.001), higher left ventricular ejection fraction (LVEF) (61%±4% vs 47%±5% and 50%±10%, both P <0.05). Conclusions: DM not only impaired functions of circulating EPC, but also influenced tube formation function of BM-EPC. Auto transplantation of BM-EPC may rescue the ischemic myocardium by neovascularization and paracrine effect in diabetic rabbits.

Objective: To explore the efficacy of laparoscopic surgery in treatment of advanced mid-low rectal cancer following a long-term neoadjuvant chemoradiotherapy. Methods: Clinicopathologic and perioperative data were collected retrospectively from 74 patients with advanced mid-low rectal cancer, who received both neoadjuvant chemoradiotherapy and resections between January 2010 and January 2013 at Xinjiang tumor hospital. Routine follow-up was conducted. The safety and long-term survival of 36 patients who underwent laparoscopic resection were compared with those of 38 patients who received conventional resection. Results: The laparoscopic group had less amount of blood loss during surgery (50 ml vs 100 ml, P <0.05). The time needed for recovery of gastrointestinal function in the laparoscopic group was significantly shorter than that in the open surgery group (2.0 d vs 3.0 d, P <0.05). The rate of postoperative complication was 19.4% and 42.1% ( P <0.05), respectively. In terms of the range of radical surgery and the numbers of dissected lymph nodes (8 and 10, P >0.05), no significant difference were found in the two groups. The operation duration and hospital stay in the laparoscopic group was longer than that in the open surgery group (240.0 min vs 231.5 min , P >0.05) (22.0 d vs 21.5 d , P >0.05), but no significant difference was found between the two groups. There were no significant difference in the incidence of 3 disease-free survival rate (53.0% vs 43.8%, P >0.05) and overall survival rate (70.0% vs 62.9%, P >0.05) between two groups. Conclusion: Laparoscopic surgery is a safe and feasible option for advanced mid-low rectal cancer patients who undergone the neoadjuvant chemoradiotherapy because of the similar rate of radical resection and satisfied long-term outcomes, which will have a better prospect in the future.

Objective: To assess the feasibility, the safety and short-term outcomes of laparoscopic radical rectal surgery for rectal cancer patients with increased body mass index (BMI) ., Methods: Retrospectively data reviews were conducted for 405 consecutive patients undergoing laparoscopic surgery for rectal cancer from June 2008 to June 2012. They were classified as normal-weight (NW, BMI 18.6-22.9 kg/m(2), n = 165), overweight (OW, BMI 23.0-24.9 kg/m(2), n = 125), and obese (OB, BMI ≥ 25.0 kg/m(2), n = 115)groups according to the categories as proposed by 2007 Chinese Obesity Surgery Treatment Guidelines. The differences of oncologic, intraoperative and postoperative status, postoperative complications, number of resected lymph nodes and short-term survival rates were compared among three groups. The data were analyzed by χ(2) or Fisher exact test. The Mann-Whitney U test or analysis of variance (one-way ANOVA) was used for parametric comparisons. The survival curve was drawn by Kaplan Meier method and the survivals of 3 groups were by the Log-rank test., Results: The comorbidity of patients in the NW and OW groups were less than that in the OB group(27.9% (46/165) and 30.4% (38/125) vs 47.0% (54/115), χ(2) = 12.066, P < 0.05). No significant difference existed among the groups in terms of conversion rate (9.1% (15/165), 10.4% (13/125) and 12.2% (14/115)), the rate of postoperative complications (20.6% (34/165), 21.6% (27/125) and 24.3% (28/115) ), intraoperative volume of blood loss ((105 ± 30), (110 ± 25) and (115 ± 45) ml), first flatus( (2.8 ± 1.2), (2.9 ± 1.1) and (3.1 ± 1.4) d), postoperative hospital stays ((13.7 ± 5.5), (14.3 ± 7.5) and (14.1 ± 8.5) d, all P > 0.05), and the mean number of retrieved lymph nodes(P > 0.05). While the operation duration in the OB group were longer than that in the NW and OW groups ((250 ± 35) vs (205 ± 20) and (210 ± 30) min, F = 7.216, P < 0.05) . And 368 patients (90.9%) were followed up for a median of 24 months(2-48 months). As for survival curves, no significant difference existed among three groups (P > 0.05)., Conclusions: It is both safe and feasible for obese patients with increased BMI to undergo laparoscopic radical rectal cancer. And there is no effect upon immediate survival.

Objective: To establish a lapatinib resistance cell line for elucidating the mechanisms of drug resistance of lapatinib in human breast cancer cells., Methods: The human breast cancer MDA-MB-231 cells were exposed in an incremental dose of lapatinib to establish a lapatinib resistance rMDA-MB-231 cell line. The assay of methyl thiazolyl tetrazolium (MTT) was used to detect the cytotoxic activity of lapatinib against MDA-MB-231 and rMDA-MB-231 cells. The protein expression was detected by Western blot. Small interfering RNA was used to specifically knock down mammalian-target-of-rapamycin (mTOR) in rMDA-MB-231 cells. Apoptosis was determined by fluorescein isothiocyanate (FITC)-annexin V/PI staining and flow cytometry., Results: The human breast cancer lapatinib resistance cell line rMDA-MB-231 was induced by lapatinib. The half maximal inhibitory concentration (IC50) values of lapatinib against MDA-MB-231 and rMDA-MB-231 cells were (6.1 ± 0.6) and (34.9 ± 2.7) µmol/L respectively (P < 0.01). Compared with MDA-MB-231 cells, the protein expression of mTOR in rMDA-MB-231 cells was significantly up-regulated. The protein expression of mTOR was significantly down-regulated by specific siRNA duplexes in rMDA-MB-231 cells. After siRNA interference, 20 µmol/L lapatinib was added into control, negative siRNA control and mTOR-targeted siRNA groups respectively. The percents of cell apoptosis in control, negative control and targeted siRNA groups were 13.4% ± 2.5%, 14.2% ± 2.8% and 34.6% ± 5.8% respectively, there was no significance between the first two groups (P > 0.05) , and there was significant difference between the control and targeted siRNA group (P < 0.01) ., Conclusions: The up-regulation of mTOR plays an important role in the lapatinib-resistant phenotype of human breast cancer rMDA-MB-231 cells. And the down-regulation of mTOR increases the apoptotic death of lapatinib against rMDA-MB-231 cells.

Objective: To evaluate the efficacy and safety of unmanipulated haploidentical allogeneic peripheral blood stem cells transplantation (PBSCT) on hematologic diseases., Methods: Patients who underwent unmanipulated HLA-mismatched/haploidentical PBSCT from July 2007 to December 2011 were investigated retrospectively., Result: Forty-nine patients with hematologic diseases underwent unmanipulated human leukocyte antigen (HLA)-mismatched/haploidentical PBSCT with myeloablative conditioning. All patients were mismatched at the allele level for HLA-A, HLA-B, HLA-Cw, HLA-DRB1 and HLA-DQ1. Fifteen patients were mismatched in 5 loci, 11 patients in 4 loci, 7 patients in 3 loci, 5 patients in 2 loci, and 11 patients in 1 locus. The median numbers of mononuclear cells and CD₃₄⁺ cells infused at transplantation were 10.01(7.05-25.34) × 10⁸/kg and 4.51 (2.01-11.47) × 10⁶/kg, respectively. Patients achieved myeloid and platelet engraftment at a median of 14 (10-25) days and 22 (10-135) days, respectively. The cumulative incidence of acute graft versus host disease (aGVHD) on day 100 was (61.6 ± 7.3)%, and the 2-year cumulative incidence of chronic graft versus host disease (cGVHD) was (42.6 ± 8.5)%. One hundred-day transplantation related mortality (TRM) rate and 2-year cumulative TRM rate were (14.7 ± 5.1)% and (30.9 ± 8.8)%, respectively. The 2-year cumulative incidence estimate of relapse was (25.4 ± 7.0)%. The 2-year cumulative overall survival rate was (58.1 ± 8.8)% and 2-year disease-free survival rate was (53.9 ± 8.4)% with an 11.5-months median follow-up., Conclusion: Unmanipulated PBSCT is a promising protocol for patients with hematologic diseases in HLA-mismatched/haploidentical transplant settings.

Objective: To evaluate the apparent diffusion coefficient (ADC) value features of the lesions after transcatheter arterial chemoembolization (TACE) plus radiofrequency ablation in hepatocellular carcinoma (HCC) with 3.0T magnetic resonance imaging (MRI) and diffusion-weight imaging (DWI) and analyze the value of 3.0T DWI in detecting the pathological lesion features of post-TACE plus radiofrequency ablation in HCC., Methods: Twenty-eight HCC patients were enrolled to receive TACE firstly. Then all viable tumors around the lesions underwent radiofrequency ablation. At 1-4 months after radiofrequency ablation, 3.0T MRI and DWI (b = 600 sec/mm(2)) were performed to measure the ADC values of different lesions of post-TACE plus radiofrequency ablation. The features of MRI and ADC values of different lesions, the difference of contrast enhancement sequence and DWI in evaluating the lesions of post-TACE plus radiofrequency ablation were analyzed., Results: Viable tumors occurred in 14 of 28 HCC patients after TACE plus radiofrequency ablation. The ADC values of necrotic tissues with lipiodol, necrotic tissues without lipiodol, viable tumors and normal liver tissues were 1.905 ± 0.487, 0.726 ± 0.116, 1.449 ± 0.054 and 1.777 ± 0.094 (10(-3) mm(2)/sec) respectively. There was no significant difference of ADC values between necrotic tissues with lipiodol and normal tissues (P = 0.115). But there were significant differences of ADC values among necrotic tissues with lipiodol, necrotic tissues without lipiodol and viable tumors (P < 0.05). The viable tumor tissues after TACE plus radiofrequency ablation appeared as nodular lesions with slightly heightened signal intensities around the necrotic tissues, the lesions with heterogeneous enhancement during arterial phase, portal vein phase and parenchymal phase. Necrotic tissues without lipiodol occurred outside necrotic tissues without lipiodol, around normal liver tissues, with low signal intensities on T2WI, without enhancement during arterial phase, portal vein phase and parenchymal phase. There were no significant difference between contrast enhancement and DWI sequence in detecting viable tumors after TACE plus radiofrequency ablation (P > 0.05)., Conclusion: The ADC values of 3.0T MR DWI may be used to distinguish the viable residue or recurrent tumor tissues, necrotic tissues in HCC after TACE plus radiofrequency ablation.

Objective: To compare the efficacy and adverse effects of bortezomib + adriamycin + dexamethasone (PAD) and vincristine + adriamycin + dexamethasone (VAD) regimens in untreated multiple myeloma (MM)., Methods: There were 26 and 28 new diagnosed MM patients in PAD and VAD groups. Both clinical effects and adverse effects were observed. Patients accepted VAD or PAD regimens for 2 - 4 cycles and followed up for 7 - 27 months., Results: There were 10, 5 and 11 patients accepted 2, 3 and 4 cycles in PAD group, and 6, 11 and 11 in VAD group. In PAD group, there were 2, 14, 9, 1 and 0 patients achieved complete remission (CR), very good partial remission (VGPR), partial remission (PR), stable disease (SD) and progressive disease (PD); in VAD group, the number were 0, 4, 12, 10 and 2. The rate of patients who achieved good efficacy (CR + VGPR) in PAD group was 61.5%, which was higher than that in VAD group (14.3%). The incidences of infection and gastrointestinal symptoms were similar in the two groups, while the incidences of peripheral neuropathy, thrombosis and Herpes Zoster infection in PAD group were higher than those in VAD group., Conclusions: Compared with the conventional VAD chemotherapy, PAD may improve CR and VGPR rates in new diagnosed MM, while it may bring more and severer toxicities in peripheral neuropathy, thrombosis and Herpes Zoster infection. Preventive medical care is necessary in PAD protocol.

Objective: To investigate the efficacy of CARTO-guided stepwise ablation approaches for treatment of patients with nonparoxysmal atrial fibrillation(AF)., Methods: Stepwise ablation approaches were performed in 40 patients with nonparoxysmal atrial fibrillation. Pulmonary vein atrium isolation (PVAI), linear ablation in atria, complex fractionated atrial electrograms (CFAEs) ablation and cardioversion were applied sequentially till sinus rhythm (SR) restoration. All patients were followed up 6 to 18 months., Results: SR was restored in 11 patients after PVAI, in 11 patients after linear ablation and in 6 patients after CFEAs ablation. SR was restored in the remaining 13 patients post cardioversion. During follow-up, 3 atrial fibrillation, 3 atrial tachycardia and 5 atrial flutter were evidenced. Seven out of the 11 patients with reoccurred arrhythmia were treated only by PVAI., Conclusions: CARTO-guided stepwise ablation approaches are safe and effective in the treatment of patients with nonparoxysmal atrial fibrillation. PVAI approach was associated with lower successful rate and high recurrence rate.

Objective: To investigate the roles of signal transducer and activator of transcription 4 (STAT4) and suppressor of cytokine signaling 3 (SOCS3) in differentiation and cytokine secretion of T helper (Th) cells by separate silencing., Methods: Th0 cells were induced to differentiate to Th1 or Th2 cells by interleukin 12 (IL-12) or interleukin 4 (IL-4). The mRNAs of STAT4 and SOCS3 were silenced separately by small interfering RNA (siRNA) in Th0, Th1 and Th2 cells and then the lymphocytes cultured in Th1 or Th2 medium with IL-12 or IL-4 for another 48 h before cells and supernatants were collected for the detection of Th1 and Th2 type cytokines by reverse transcriptase polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). By comparing different amounts of cytokines between before and after silencing the target mRNAs, the roles of STAT4 and SOCS3 in the differentiation and cytokine secretion in T helper cells were evaluated., Results: Th1 type cytokine significantly decreased and Th2 type cytokine increased after silencing STAT4 in Th0 and Th1 cells; minimal changes were detected in both Th1 and Th2 type cytokines after silencing SOCS3., Conclusion: STAT4 mediates the IL-12-induced differentiation of T cells to Th1 subset and maintains the cytokine secretion of Th1 cells; STAT4 directly inhibits the development and cytokine secretion of Th2 cells; STAT4 deficiency results in impaired development and cytokines secretion of Th1 cells and the enhanced development and cytokines secretion of Th2 cells. SOCS3 has minimal effects on IL-12 or IL-4 induced differentiation and cytokine secretion of T helper cells.

Objective: To evaluate the efficacy of Endosteal(TM) (rh-endostatin, YH-16) combined with docetaxel and carboplatin (TP) regimen for the adjuvant treatment of non-small lung cancer (NSCLC) and its impact on circulating blood markers., Methods: 36 patients with stage Ib-IIIa postoperative NSCLC, were randomly divided into the treatment group, Endosteal(TM) plus TP regimen, and the control group, TP regimen only, respectively. DFS and toxicities of patients were observed. The numbers of CEC and the levels of tumor marker CEA, NSE and CYFR21-1 were measured., Results: The numbers of CEC and the levels of CEA, NSE and CYFR21-1 decreased after treatment. There were significant differences in CEC and NSE between treatment group and control group after four cycles of treatment, respectively (P = 0.016 and 0.013). Disease-free survival time (DFS) was longer in treatment group than control group but without significant difference. CEC was significantly increased in recurrent and metastasis cases and decreased after effective treatment., Conclusion: Endosteal(TM) combined with TP regimen seem to be superior to TP alone in some short term index for the treatment of postoperative NSCLC even though long-term survival is still anticipated. CEC, as a biomarker, may be useful in predicting the efficacy of the such synergistic treatment.

Objective: To investigate the genetic differences and their relativity with multi-drug resistance of Pseudomonas aeruginosa isolated., Methods: Forty-nine Pseudomonas aeruginosa isolates were characterized by antimicrobial susceptibility and four-enzyme (I-CeuI, SpeI, SwaI, PacI) pulsed-field gel electrophoresis (PFGE)., Results: 85.7% of the P.aeruginosa isolates were MDR strains. Strains with PFGE pattern A were all susceptible to amikacin and cefepime, but were resistant to levofloxacin and meropenem. Strains with PFGE Patterns H and P had resistance to 6 - 8 different kinds of antibiotics. Strains with PFGE Patterns I and J were susceptible to all antibiotics tested in this study. Strains with other PFGE Patterns had intermediate resistance. PFGE pattern A was the dominant pattern, which accounted for 61.2% of all P.aeruginosa strains, 100% (2/2) in 2001, 65% (13/20) in 2002, 44.4% (8/18) in 2003 and 77.8% (7/9) in 2004., Conclusion: Four-enzyme combined PFGE analysis is highly discriminatory for the subtyping of MDR P.aeruginosa isolates.

Objective: Overexpression of SERCA2a could improve cardiac function in human and experimental heart failure (HF) models. We observed the proteomics changes post SERCA2a overexpression in a pacing induced HF model in dogs., Methods: Beagles were divided into four groups: control group, HF group (230 beats/min for 4 weeks), HF + EGFP group (myocardial injection of 1 x 10(12) v.g recombinant adeno-associated virus carrying enhanced green fluorescent protein gene, rAAV2/1-EGFP) and HF + SERCA2a group (myocardial injection of 1 x 10(12) v.g recombinant adeno-associated virus carrying SERCA2a gene, rAAV2/1-SERCA2a). Thirty days after gene transduction, left ventricular systolic and diastolic functions were measured by echocardiography and invasive hemodynamics in all animals. By use of 2-dimensional gel electrophoresis (2-DE), -500 distinct protein spots were detected in myocardium of all animals. Protein spots observed to be altered between failing and SERCA2a overexpressed hearts were subjected to tryptic peptide mass fingerprinting for identification by MALDI-TOF mass spectrometry in combination with LC/MS/MS analysis., Results: At 30 day after gene transfer, HF signs were significantly reduced, cardiac function [LVSP: (214.72 +/- 31.74) mm Hg (1 mm Hg = 0.133 kPa) vs. (139.32 +/- 36.79) mm Hg, +dp/dt(max): (6779.43 +/- 217.58) mm Hg/s vs. (2746.85 +/- 931.23) mm Hg/s and -dp/dt(max): (-4341.42 +/- 322.02) mm Hg/s vs. (-2531.14 +/- 616.15) mm Hg/s, LVEDP: (21.86 +/- 6.95) mm Hg vs. (59.78 +/- 6.92) mm Hg] significantly improved in HF + SERCA2a dogs than those in HF + EGFP group(all P < 0.05) and parameters were comparable between HF + SERCA2a and control groups. We identified alterations in the expression level of more than 10 proteins in myocardium. These protein changes were observed mainly in two subcellular compartments: the cardiac contractile apparatus and metabolism/energetics., Conclusion: These results showed that overexpression of SERCA2a could improve cardiac function accompanied with numerous alterations in protein expressions involved in calcium handling, myofibrils, and energy production in this dog model of chronic heart failure.

Objective: To explore the neurobiological basis involved in the pathogenesis of the lasting emotionality and cognitive impairment following severe psychological stress., Methods: Ninety-six male Wistar rats were divided randomly into 2 equal groups: group of predator stress (Group PS) put into a cage in the experimental box singly to be exposed to a cat in the box but outside the cage for 23-57 min until tremor, polypnea, and nares flaring appeared for 6 min so as to establish predator stress models, and control group, put into the cage without non-injurious exposure of cat. 1, 12, and 24 hours later 8 rats from each group were killed with the hippocampus taken out. Western blotting was used to detect the protein expressions of cAMP response element-binding protein (CREB), phosphorylated CREB (pCREB) and CREB binding protein (CBP). Twelve hours after the experiment 24 rats from each group were killed with their brains taken out to obtain serial coronary sections. Immunohistochemistry was used to detect the positive immunoreactivities of CREB, pCREB, and CBP., Results: Immunohistochemistry revealed that the absorbance (A) value of CREB-in the tissues of hippocampus and frontal cortex 12h after the cat exposure of Group PS were 0.55 +/- 0.13 and 0.88 +/- 0.20 respectively, both significantly lower than those of the control group (1.78 +/- 0.40 and 1.18 +/- 0.26 respectively, both P < 0.01), the A values of. pCREB in the hippocampus and frontal cortex of Group PS were 1.51 +/- 0.34 and 1.07 +/- 0.24 respectively, both significantly higher than those of the control group (0.47 +/- 0.11 and 0.48 +/- 0.11 respectively, both P < 0.01), and the A values of CBP in the hippocampus and frontal cortex of Group PS were 1.01 +/- 0.23 and 0.81 +/- 0.18 respectively, both significantly higher than those of the control group (0.52 +/- 0.12 and 0.29 +/- 0.07 respectively, both P < 0.01). Western blotting showed that the CREB protein expression levels 1 h and 24 h after the cat exposure of Group PS were 2.82 +/- 0.65 and 5.12 +/- 1.13 respectively, both significantly lower than those of the control group (11.86 +/- 2.47 and 10.56 +/- 2.38 respectively, both P < 0.01), the CBP protein expression levels 1 h and 24 h after the cat exposure of Group PS were 1.77 +/- 0.39 and 2.44 +/- 0.55 respectively, both significantly higher than those of the control group (1.06 +/- 0.24 and 0.86 +/- 0.20 respectively, both P < 0.01), and the pCREB protein expression levels 1 h and 12 h after the cat exposure of Group PS were 2.56 +/- 0.59 and 1.93 +/- 0.41 respectively, both significantly higher than those of the control group (1.04 +/- 0.22 and 0.96 +/- 0.21 respectively, both P < 0.01)., Conclusion: The dysfunction of CREB signaling in the central nervous system, especially in the hippocampal formation after predation stress may play an important role in the long-term neuropsychological sequelae following severe stress.

Objective: To investigate the effect of human beta defensin 2 (HBD-2) on Staphylococcus aureus infection., Methods: A minigene of HBD-2 containing pCMV promoter, full length of HBD-2 cDNA, and BGH polyA tail was generated by PCR amplification and introduced into the fertilized oocytes of C57/ICR hybridized mouse by microinjection. After gestation of 3 - 4 weeks, immunohistochemistry was used to detect the expression of HBD-2 peptide in different tissues of the transgenic young mice. Staphylococcus aureus was cultured and injected intraperitoneally to wild type mice and transgenic mice to observe their surviving status., Results: PCR showed that the HBD-2 fragment had been successfully integrated into the chromosome of the mice. A widespread expression of HBD-2 gene was found in many tissues of the transgenic mice: trachea, lung, intestine, esophagus, testis, spleen, skin, endothelium, brain, etc. Four of the 7 transgenic mice survived the Staphylococcus aureus infection, and 10 wild type mice all died within 24 hours., Conclusion: HBD-2 may play an important role ion the host defense against Staphylococcus aureus infection.

Objective: To investigate the relationship between promoter methylation of inhibitor of DNA binding 4 (Id4) gene and acute leukemia., Methods: We detected the status of promoter methylation of Id4 gene in peripheral blood, peripheral blood stem cell transplantation (PBSC) and bone marrow samples from health donors, bone marrow cells from leukemia patients and leukemia cell lines with MS-PCR methods., Results: The frequency of Id4 promoter methylation was 21/25 in acute myeloid leukemia (84%), 12/14 in acute lymphoblastic leukemia (86%), 8/8 in relapse acute leukemias and 2/2 in leukemia cell lines whereas no methylation was detected in peripheral blood, PBSC and bone marrow samples from healthy donors. In addition, Hek937 cell line was unmethylated., Conclusion: Id4 promoter methylation was detected only in acute leukemia patients and leukemia cell lines and it might play a role in leukemia genesis.

Objective: To investigate the role of serum and glucocorticoid induced kinase-1 (SGK(1)) pathways in fibronectin (FN) synthesis in human mesangial cell (HMC) under high glucose condition and the mechanism by which SGK(1) contributes to glomerulosclerosis in diabetic nephropathy (DN)., Methods: HMCs were cultured and transfected with (P)IRES2-EGFP-(S422D) SGK(1) mutant (SD), plasmid containing SGK(1) dominant activation mutant, or blank plasmid. Non-transfected HMCs were used as control group. Then the HMCs were divided into 6 groups: transfected with SD + high glucose (SD-HG, 25 mmol/L D-glucose), transfected with FP + high glues (FP + HG), non-transfected + high glucose (NT-HG), transfeted with SD + normal glucose (SD-NG, 5.5 mmol/L D-glucose), transfected with FP + normal glues (FP + NG), and non-transfected + normal glucose (NT-NG). Eight hours after the glucose stimulation, RT-PCR was used to examine the SGK(1) mRNA expression and fibronectin (FN). Western blotting was used to detect the fibronectin (FN) protein expression., Results: The SGK(1) mRNA expression of the SD + HG group was 0.709, significantly higher than those of the FP + HG and NT + HP groups (0.497 and 0.491, both P < 0.01). The SGK(1) protein expression of the SD + HG group was 1,178,497, significantly higher than those of the FP + HG and NT + HP groups (193,875 and 195,597 respectively, both P < 0.01). The FN mRNA expression of the SD + HG group was 0.749, significantly higher than those of the FP + HG and NT + HP groups (0.463 and 0.475 respectively, both P < 0.01). The FN protein expression of the SD + HG group was 659,550, significantly higher than those of the FP + HG and NT + HG groups (342,354 and 340,428 respectively, both P < 0.01). There were not significant differences in the expressions of FN mRNA and protein among different NG groups., Conclusion: SGK(1) may be involved in the signal transduction leading to the increase of fibronectin production in DN and therefore may play an active part in glomerulosclerosis in DN.

Objective: To investigate the molecular mechanisms of tauopathies. Comparative proteomic analysis of brain proteins was employed to study 4 patients with tauopathies as compared with 4 controls., Methods: The brains of subjects who died without clinical or pathological involvement of nervous system and brains of patients with tauopathies were obtained at autopsy. The brain proteins were run by immobilized pH gradient (IPG) isoelectric focusing electrophoresis as the first dimension, and then run by vertical SDS-PAGE as the second dimension. The maps were visualized by silver staining or colloidal coomassie blue and analyzed with Image Master 2D Elite software. The proteins of interest were in-gel digested and identified using MALDI-TOF mass spectrometry or MALDI-TOF/TOF tandem mass spectrometry., Results: 18 protein spots were differentially expressed as compared with age-matched nondemented control brains which were identified as glyceraldehyde 3-phosphate dehydrogenase, uracil DNA glycosylase, human superoxide dismutase, isocitrate dehydrogenase subunit, synaptotagmin I, thioredoxin peroxidase 1, glial fibrillary acidic protein, p25 alpha, enoyl coenzyme A hydratase short chain 1, pyridoxine-5'-phosphate oxidase, Mn-superoxide dismutase and alpha enolase, antioxidant protein 2, ferritin heavy chain, glutamate dehydrogenase precursor, peptidyl-prolyl cis-trans isomerase A, serum albumin precursor and dihydropyrimidinase-related protein 2., Conclusions: We got a number of related-proteins of tauopathies. Some proteins are quite useful for discovering the molecular mechanisms of tauopathies and may be helpful for diagnosis and of treatment tauopathies.

Objective: To compare the results of proteomics between the cerebellum and frontal lobe of the aged., Method: Proteins were isolated from human cerebellums and frontal lobes and separated by two-dimensional (2D) gel electrophoresis. The proteins were then stained with silver or colloidal coomassie blue to produce a high-resolution map of the proteome. Selected proteins from this map were in-gel digested with trypsin and the resulting tryptic peptides were analyzed by MALDI-TOF mass spectrometry and MALDI TOF/TOF tandem mass spectrometry. The mass spectrometric data were used to identify the proteins through searches of the SWISSPROT protein sequence database., Results: Three up-regulated protein spots in 2D gels of cerebellums were found as compared with those of frontal lobes. These protein spots were identified as antioxidant protein 2, creatine kinase precursor, fructose-bisphosphate aldolase C. Two down-regulated proteins in cerebellums were identified as pyruvate kinase M1 isozyme and glial fibrillary acidic protein. 30 common proteins, the expressions of which were not altered between cerebellums and frontal lobes, were also identified., Conclusions: These data will be used for future studies of differential protein expression in neurological disorders. Some proteins are useful for discovering the molecular mechanisms of degenerative dementia and brain aging.