Objective:To investigate the m echanism of D- amino acid oxidase/D- Alanine system in killing K5 6 2 e cells in vitro.Methods:The killing effects of D- Ala on K5 6 2 e cells stably expressing DAAO and GFP were observed.H2 O2 production by DAAO+ cells were m easured by the phenol red oxidation assay.L owry method was used to determ ine the protein quantities of cells and fluorescent intensities of GFP+ cells were assayed by flow cytom eter.Results:KDf Gd cells were killed completely after treated with 2 5 mm ol/L D- Ala for 2 4 h.The effect of D - Ala at 2 0 m mol/L on KDf Gd cells increased apparently within 4 8h,but the same effect was not observed if D - Ala was below 15 m mol/L .The cytotoxicity of D- Ala in KDf Gd cells was more sensitive than in parental K 5 6 2 e cells.The H2 O2 levels in the medium were consistent with the killing effects of D- Ala.Conclusion:The killing effects of DAAO/D- Ala system is m ediated by H2 O2 . [

BACKGROUND: Recently, trans-pedicle screw internal fixation has markedly improved the rigidity of spinal fixation and hence the fusion rate. But when placed incorrectly, the pedicle screw can injure the spinal cord and/or nerve roots, resulting in serious complications.OBJECTIVE: To evaluate the application value of preoperative CT scans-based navigation technique in the spinal pedicle screw internal fixation surgery.DESIGN, TINE AND SETTING: A prospective, randomized, and controlled observation was performed at the Department of Orthopedics, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences between January 2006 and December 2008.PARTICIPANTS: A total of 95 patients who underwent pedicle screw internal fixation due to spine diseases were randomly assigned to a navigation group (n = 45) and a conventional group (n = 50).METHODS: In the navigation group, patients were subjected to pedicle screw insertion with the assistance of computer navigation technique and while in the conventional group, patients underwent pedicle screw insertion using the conventional anatomic landmark combined with fluoroscopy.MAIN OUTCOME MEASURES: Screw channel preparation time, excellent and good rate of screw position, and postoperative complications.RESULTS: In the navigation group, totally 206 pedicle screws were inserted under navigation guidance, with an excellent and good rate of 96.1%; and navigation could not be continued in 9 patients for a three-dimensional registration error. In the conventional group, altogether 285 pedicle screws were inserted, and the excellent and good rate was 100%. No significant difference was found between the two groups (P > 0.05). The navigation group exhibited longer screw channel preparation time than the conventional group [(360±22) seconds vs. (56+8) seconds, P < 0.01]. No postoperative complications were found in each group.CONCLUSION: The preoperative CT scans-based navigation technique produces similar accuracy of pedicle screw insertion, but markedly prolonged operation time, compared with the conventional anatomic landmarks, exhibiting limited application value in the spinal pedicle screw internal fixation.

The internal ribosome entry site (IRES) sequence was derived from encephalomyocarditis virus. It allows to translate two open reading frames at one mRNA, so two genes conjoined by IRES have the same expression rate. K(DfGC) and K(DfGd) cell lines, stably expressing D-amino acid oxidase (DAAO) gene and green fluorescence protein (GFP) genes, were obtained by transfection of K562e cells with retroviral vector pLDfG containing IRES sequence, DAAO cDNA and GFP gene. Fluorescence positive rate and fluorescence intensity of the two cell lines were measured with flow cytometry. H(2)O(2) production by K(DfGC) and K(DfGd) cells treated with D-alanine was measured by the phenol red oxidation assay. The fluorescence positive rate and fluorescence intensity in K(DfGC) and K(DfGd) cell were 94.64% and 96.31% and 202 units and 174 units per 2 x 10(4) cells, respectively. There was exponential correlation between fluorescence intensity and H(2)O(2) level. The above-mentioned results demonstrate that DAAO gene and GFP gene were simultaneously expressed in K562e cell line by the regulation of IRES sequence, and DAAO level was correlated with fluorescence intensity of GFP.
Binding Sites ; genetics ; D-Amino-Acid Oxidase ; genetics ; metabolism ; Gene Expression ; Gene Expression Regulation, Enzymologic ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; Humans ; Hydrogen Peroxide ; metabolism ; K562 Cells ; Luminescent Proteins ; genetics ; metabolism ; Retroviridae ; genetics ; Ribosomes ; metabolism ; Transfection

OBJECTIVETo investigate the potential role of Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway in regulation of gene expression of high mobility group box-1 protein (HMGB1) in various tissues in rats with sepsis.

METHODSA sepsis model reproduced by cecal ligation and puncture (CLP), and 128 male Wistar rats were randomly divided into normal control group (n = 10), sham operation group (n = 10), CLP group (n = 60), AG490 treatment group (n = 24), and rapamycin (RPM) treatment group (n = 24). At serial time points animals in each group were sacrificed after CLP, then tissue samples were harvested to determine HMGB1 mRNA expression and STAT1/3 DNA binding activity.

RESULTSSTAT1 activities increased rapidly in the liver, lungs and small intestine after CLP, peaking at 6 - 12 h, while it increased slowly, and still kept at mild level from 2 to 48 h in the kidneys. Compared with STAT1, lower STAT3 activities were detected only in the liver and lungs, with negative detection in the small intestine and kidneys. HMGB1 mRNA levels significantly increased in liver, lungs and small intestine at various time points after CLP respectively (P < 0.05 or P < 0.01), while they didn't change in the kidneys. Treatment with AG490 could markedly inhibit HMGB1 mRNA expression in the liver and small intestine at 24 and 48 h (P < 0.05 or P < 0.01), and in lungs at 2 h following CLP (P < 0.01). Similarly, treatment with RPM significantly decreased HMGB1 mRNA expression in the lungs at 2, 6, 24 and 48 h, in the liver at 6 and 24 h, and in the small intestine at 24 and 48 h (P < 0.05 or P < 0.01). In addition, STAT1 and STAT3 activities in the liver and lungs were significantly correlated with corresponding tissue HMGB1 mRNA expression.

CONCLUSIONSPeritoneal infection could extensively activate STAT1 and limitedly activate STAT3 in vital organs. Activation of JAK/STAT pathway might be involved in up-regulating the gene expression of HMGB1 and systemic inflammation secondary to severe septic challenge.

Animals ; Disease Models, Animal ; Gene Expression ; physiology ; HMGB1 Protein ; genetics ; Janus Kinases ; metabolism ; Male ; RNA, Messenger ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; STAT Transcription Factors ; metabolism ; Sepsis ; genetics ; physiopathology ; Signal Transduction

The efficiency of the exogenous DNA transfecting mouse sperm was studied by the DIG end labeled and immunohistochemistry technology. The results suggested that: the efficiency of transfecting positive rate of individual mouse sperm was distinct difference (P < 0.01), and the average rate was 13%. The acrosomal reaction was evaluated using the technology of Coomassie brilliant blue stained, and the appropriate in vitro fertilization (IVF) medium TYH was elected. Mouse sperms were transferred with GFP gene in vitro, and the mature oocytes were fertilized using IVF, and then the zygotes were cultured in vitro. The embryos were observed using the fluorescence microscopy, and the transgenic rate was 4.7%. The results suggested that sperm mediated gene transfer (SMGT) was an effective and feasible method.
Animals ; Embryo, Mammalian ; cytology ; metabolism ; Feasibility Studies ; Female ; Fertilization in Vitro ; Gene Expression ; Gene Transfer Techniques ; Green Fluorescent Proteins ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred ICR ; Mice, Transgenic ; Microscopy, Fluorescence ; Oocytes ; cytology ; metabolism ; Spermatozoa ; cytology ; metabolism ; Zygote ; cytology ; metabolism

OBJECTIVETo investigate the sanitary status of urban secondary water supply facilities in Haidian district of Beijing.

METHODSAdopting the quantitative classification table drafted by the Bureau for Sanitation Inspection and Supervision of Haidian district, we carried quantitative classification (A, B, C grade) on all 1725 secondary water supply facilities in Haidian district for two times. At the same time, we collected 20 residential areas with stratified random sampling method. As the public points in the first quantitative classification, the effect of level publicity on changing the sanitary grade of the secondary water supply facilities were observed.

RESULTSIn the first two times of quantitative classification, A-level and B-level secondary water supply facilities took up 81.04% (1398/1725) and 89.04% (1536/1725) of all secondary water supply facilities respectively; the ratio of effective sanitary permits achieved 86.14% (1486/1725) and 92.35% (1593/1725) respectively; and the ratio of effective water quality test reports achieved 86.60% (1494/1725) and 97.10% (1675/1725) respectively. There were 52 secondary water supply facilities in 20 collected areas, including 8 A-level, 27 B-level and 17 C-level secondary water supply facilities before level publicity, and 19, 29 and 4 after level publicity. The impact of level publicity on changing the sanitary grade of the secondary water supply facilities was statistically significant (χ(2) = 12.60, P = 0.002).

CONCLUSIONThe city secondary water supply facilities in Haidian district are overall in good sanitary conditions. Quantitative classification and level publicity can effectively improve the sanitary status of secondary water supply facilities.

China ; Sanitation ; Urban Health ; Water Supply

OBJECTIVETo investigate the effect of nano-hydroxyapatite/collagen (nHA/collagen) composite as a graft extender and enhancer when combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) on lumbar intertransverse fusion in rabbits.

METHODSSixty-four adult female New Zealand white rabbits, aged 1 year and weighing 3.5-4.5 kg, underwent similar posterolateral intertransverse process arthrodesis and were randomly divided into 4 groups based on different grafts: autogenous cancellous bone alone (ACB group), nHA/collagen alone (HAC group), half autogenous cancellous bone and half nHA/collagen (ACB+HAC group) and nHA/collagen combined with rhBMP-2 (HAC+BMP group). The fusion masses were analyzed by manual palpation, radiography, biomechanical testing and histological examination.

RESULTSFusion was observed in 4 cases in the 6th week and in 5 cases in the 10th week after surgery in ACB group. No case showed fusion in HAC group. In ACB+HAC group, there was fusion in 3 cases in the 6th week and in 4 cases in the 10th week after surgery. In HAC+BMP group, fusion in 1 case was found in the 4th week, in 5 cases in the 6th week and in 6 cases in the 10th week after surgery. It suggested that ACB, ACB+HAC and HAC+BMP groups showed similar fusion ratio and mechanical strength in the 6th and 10th week after surgery. According to the microstructure analysis of the samples, nHA/collagen had no negative effect when implanted together with ilium autograft. In HAC+BMP group, new bone-like tissue was observed in the 2nd week postoperatively, and nearly all of the implanted composites were replaced by mature bone matrix and new bones in 10th week postoperatively.

CONCLUSIONSThe nHA/collagen, especially combined with rhBMP-2, is a promising bone substitute, for it has quick biodegradation, fine bone-bending ability, and high osteoconductivity on posterolateral spinal fusion in rabbits.

Analysis of Variance ; Animals ; Biomechanical Phenomena ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; pharmacology ; Disease Models, Animal ; Durapatite ; pharmacology ; Female ; Lumbar Vertebrae ; drug effects ; surgery ; Osseointegration ; drug effects ; Probability ; Rabbits ; Sensitivity and Specificity ; Spinal Fusion ; methods ; Tensile Strength ; Transforming Growth Factor beta

OBJECTIVETo describe the prevalence of body overweight and obesity and its changes among Chinese people during 1992-2002, and to provide scientific basis for developing intervention strategies for obesity in China.

METHODSThe data from "1992 China third National Nutrition Survey" (78,704 subjects, 38,323 male and 40,381 female) and from "2002 China National Nutrition and Health Survey" (209,849 subjects, 101,377 male and 108,472 female) were used in this study. The overweight and obesity were defined by using WHO Z-score criteria for children younger than 7 years old, Chinese age-, sex-specific BMI criteria for children aged 7-17 years, and Chinese BMI criteria for adults.

RESULTSThe overall prevalence of overweight and obesity of Chinese people was increased by 38.6% and 80.6%, respectively during the period of 1992-2002. The prevalence of overweight and obesity of Chinese people aged 0-6 years, 7-17 years, 18-44 years, 45-59 years and above 60 years was increased 31.7%, 17.9%, 66.7%, 45.2% and 43.7%, respectively. The increase of the prevalence of overweight and obesity was higher in girls younger than 6 years than their male counterparts, while among other age groups, that were higher in boys than girls. The prevalence of overweight and obesity among people living in rural areas was lower than that of their urban counterparts, while the increment of overweight and obesity prevalence among rural people was greater than that of their urban counterparts. It was estimated that another 70 million overweight and 30 million obese Chinese people emerged in China from 1992 to 2002.

CONCLUSIONThe prevalence of overweight and obesity of Chinese people was increased rapidly in the past decade, which had affected 260 million Chinese people. It would continue to increase in the near future if effective intervention measures have not been taken.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Body Mass Index ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Obesity ; epidemiology ; Overweight ; epidemiology ; Prevalence ; Rural Population ; statistics & numerical data ; Urban Population ; statistics & numerical data ; Young Adult

OBJECTIVETo analyze the complications and short-term outcomes of patients receiving intraoperative electron radiotherapy (IOERT) for locally advanced and recurrent periarticular soft tissue sarcomas of the extremities.

METHODSTwenty-one adult patients with locally advanced and/or recurrent periarticular soft tissue sarcomas of the extremities treated with IOERT were included in this study. Among them 14 patients had recurrent diseases after prior operation and 7 patients with locally advanced disease with primary treatment. The total dose of radiation ranged from 11 Gy to 21 Gy of 6 - 12 MeV beta ray given by intraoperative radiotherapy after complete tumor resection with negative margins. Five patients were given external beam radiotherapy (EBRT) with a total dose of 40 - 50 Gy, and 10 patients received chemotherapy.

RESULTSThe median follow-up time was 15 months. Five patients (23.8%) had tumor relapse, including one patients with local recurrence, one patient with regional recurrence, two patients with distant metastasis and one patient with local recurrence and distant metastasis. The actuarial local control rate at 1 year was 95.2% (20/21), and at 2 years was 90.5% (19/21). Acute skin toxicity (RTOG) within 3 months after surgery included grade I in 6 patients and grade II in 3 patients. Two year late skin toxicity (RTOG/EORTC) included grade I in 4 patients and grade II in 2 patients. Fibrosis included grade I in 3 patients and grade II in 3 patients. Six patients had joint dysfunction (3 patients with grade I and 3 with grade II) and 6 patients had healing problems. One patient got ipsilateral schiatic neuritis 9 months after IOERT. No adverse events occurred during surgery.

CONCLUSIONSIOERT brings tolerable complications of acute and late toxicities and favorable local control rate. IOERT should be followed by postoperative radiotherapy or increase the intraoperative radiation dose for locally advanced and recurrent sarcomas to get a better local control. Otherwise, distant metastasis is one of the main reasons of treatment failure, so chemotherapy should be added to the treatment regimen.

Adolescent ; Adult ; Aged ; Atrophy ; etiology ; Electrons ; adverse effects ; therapeutic use ; Extremities ; Female ; Fibrosis ; etiology ; Follow-Up Studies ; Humans ; Intraoperative Period ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; radiotherapy ; surgery ; Neoplasm, Residual ; radiotherapy ; Radiotherapy Dosage ; Radiotherapy, Adjuvant ; adverse effects ; Sarcoma ; drug therapy ; radiotherapy ; surgery ; Skin ; pathology ; Young Adult

OBJECTIVETo compare the enhanced recovery program after surgery (ERAS) with conventional perioperative management in patients undergoing radical resection for colorectal cancer.

METHODSThe ERAS protocol included a combination of evidence-based and consensus methodology. A total of 597 consecutive patients undergoing elective colorectal resection were randomized to either the ERAS(n=299) or the control group(n=298). Outcomes related to nutrition and metabolism index, stress index, and recovery index were measured and recorded.

RESULTSDemographics and operative parameters were similar between the two groups(P>0.05). The nutritional status of patients in the ERAS group was improved after surgery compared with that of the control group. On postoperative day (POD) 1, the HOMA-IR in the ERAS group was significantly lower than that in the control group(P<0.01). The cortisol level in the control group was elevated on both POD 1(P<0.01) and POD 5(P<0.01) compared to the preoperative level. However, the cortisol level was not increased until POD 5(P<0.01) in the ERAS group. The levels of TNF-α, IL-1β, IL-6, and IFN-γ were reduced in the ERAS group, indicating less postoperative stress responses compared with the control group. In addition, ERAS group was associated with accelerated recovery of gastrointestinal function. The postoperative length of stay [(5.7±1.6) d vs. (6.6±2.4) d, P<0.01] and expense[(15 998±2655) RMB vs. (17 763±3059) RMB, P<0.01] were reduced in the ERAS group. Twenty-eight patients(9.4%) in the control group and 29(9.7%) in the ERAS group developed complications, while the difference was not statistically significant(P>0.05).

CONCLUSIONERAS protocol alleviates surgical stress response and accelerates postoperative recovery without compromising patient safety.

Adult ; Aged ; Aged, 80 and over ; Colorectal Neoplasms ; surgery ; Female ; Humans ; Male ; Middle Aged ; Perioperative Care ; methods ; Prospective Studies ; Young Adult