OBJECTIVETo investigate the clinical features, treatment modalities and the prognosis of advanced neuroblastoma in children.

METHODSThe medical records of 63 children with stage III or IV neuroblastoma from January 1996 to December 2005 were retrospectively reviewed. Sixty patients were treated by tumor resection and (or) chemotherapy and (or) radiation. Fourteen out of the 60 patients received another autologous peripheral blood stem cell transplantation.

RESULTSOf the 63 patients with advanced neuroblastoma, the male/female ratio was 2.7:1 and the median age at diagnosis was 4 years old. Most of the initial symptoms included pyrexia, abdominal pain, abdominal mass, and leg or articular pain. Primary tumor sites were adrenal (38%), retroperitoneal (35%), mediastinal (17%), pelvic (6%) and cervical (2%). The sites of metastasis at diagnosis included local (41%) and (or) distant (37%) lymph nodes, bone marrow (60%), bone (46%) and liver (16%). The median survival time of the 63 patients was 32.7 months. The 2-year survival rate was 44.3%. Statistical analysis demonstrated that unfavorable survival prognostic factors were the following: age > 1 year at diagnosis (P < 0.05); serum neuro-specific enolase > 100 mg/L (P < 0.05); serum lactic dehydrogenase > 1500 U/L (P < 0.01); serum ferritin >150 mg/L (P < 0.05). The overall survival period of the patients was prolonged through total resection of the primary tumor (P < 0.05). Intensive chemotherapy in combination with autologous peripheral blood stem cell transplantation could also result in a prolonged overall survival period (P < 0.01).

CONCLUSIONSNeuroblastoma with advanced stages often presents with various clinical manifestations and has a poor prognosis. It is beneficial to improve the prognosis of neuroblastoma through an early diagnosis and a comprehensive therapy including total resection of the primary tumor, autologous peripheral blood stem cell transplantation and intensive chemotherapy.

Adolescent ; Child ; Child, Preschool ; Combined Modality Therapy ; Female ; Humans ; Infant ; Male ; Neuroblastoma ; mortality ; therapy ; Prognosis

In order to investigate the effect of demethylating treatment on the expression of inhibitory KIR and the cytolytic activity of NK-92MI cells, and to study the possible relationship between the demethylation of inhibitory kir gene and the function of NK cells. NK-92MI cells were treated with 5-azacytidine to induce DNA demethylation. The expression of KIR3DL1, KIR2DL2/KIR2DL3, KIR2DL1 and the viability of NK-92MI cells were detected by flow cytometry. The KIR3DL1 positive and the KIR3DL1 negative NK-92MI cells were also sorted by flow cytometry. The cytotoxicity of NK-92MI against K562 cells was detected by the LDH release assay. The results demonstrated that the expressions of KIR3DL1, KIR2DL2/KIR2DL3 and KIR2DL1 in NK-92MI cells all increased after treating with 1.0, 2.5 and 5 micromol/L of 5-azacytidine for 72 hours. And the cytotoxicity of NK-92MI against K562 cells decreased. In these dose range, 5-azacytidine did not influence the viability of NK-92MI cells. Additionally, the cytotoxicity of KIR3DL1 positive NK-92MI cells was lower than that of the KIR3DL1 negative cells. It is concluded that the demethylating treatment suppresses the cytotoxicity of NK-92MI cells through increasing the expression of inhibitory KIR.
Cytotoxicity, Immunologic ; immunology ; DNA Methylation ; Flow Cytometry ; Gene Expression Regulation ; Humans ; K562 Cells ; Killer Cells, Natural ; immunology ; metabolism ; Receptors, KIR2DL1 ; metabolism ; Receptors, KIR2DL3 ; metabolism ; Receptors, KIR3DL1 ; metabolism ; Receptors, KIR3DL2 ; metabolism

The purpose of study was to evaluate the effect of stopping use of cyclosporine A (CsA) in reversion of donor chimeras of chronic myeloid leukemia (CML) patients relapsed after transplantation. Two CML patients were transplanted with allogeneic peripheral blood stem cells, and relapsed after transplantation, their bcr/abl gene and/or ph1 chromosome showed positive, donor chimeras decreased. For these two CML patients relapsed after transplantation, the use of CsA was stopped immediately, and the patient's body temperature, skin rash, blood picture, liver function and chimeras were planed to observe carefully. The results indicated that acute graft versus host disease (aGVHD) appeared in both patients. A hundred percent (100%) of donor chimeras were then found with bcr/abl gene and/or ph1 chromosome turning to negative in both patients. In conclusion, to stop using of CsA might be effective in the treatment of some CML patients relapsed after transplantation by reversing of donor chimeras and inducing graft-versus-leukemia (GVL) effect accompanied by GVHD.
Adult ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Cyclosporine ; therapeutic use ; Graft vs Leukemia Effect ; immunology ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; genetics ; therapy ; Male ; Neoplasm Recurrence, Local ; genetics ; Peripheral Blood Stem Cell Transplantation ; adverse effects ; Transplantation Chimera ; immunology

OBJECTIVETo determine the current radiographic practices of root canal treatment (RCT) undertaken by endodontists, general dentist practitioners (GDPs) and interns, and to analyze the relationship between the usage of periapical films and the quality of RCT.

METHODSThe patients who had received RCT treatment in West China College of Stomatology were investigated at random and were divided into three groups: Endodontists treating group, GDPs treating group and interns treating group according treating physician. The usage status of periapical film were investigated by the well designed questionnaires. The obturation quality of postoperative films were evaluated. Chi-square analysis was used to compare usage rates of periapical films of three groups. Mann-Whitney test was used to determine the relationship between the usage of periapical films and the quality of RCT.

RESULTS571 teeth of 412 patients were investigated. In endodontists treating group, GDPs treating group and interns treating group, the usage rates of preoperative films were 95.3%, 89.5% and 92.1% respectively. The usage rates of working length films were 5.2%, 1.1% and 5.8% respectively. The usage of master point films was 94.8%, 72.1% and 97.4% respectively. The usage of postoperative films was 97.9%, 76.3% and 95.3% respectively. There were statistically significant differences among the working length films, master point films and postoperative films of three groups (P<0.05). The periapical films of adequate quality RCT and inadequate quality RCT were 3.14+/-0.639 and 2.84+/-0.736 respectively. There was statistically significant difference between periapical films of adequate quality RCT and inadequate quality RCT (P<0.05).

CONCLUSIONRadiographic practices of endodontists and interns are more standardized than GDPs. Quality RCT is related to radiographic practices. Standard radiographic practices would be benefit to RCT.

Dental Pulp Cavity ; Humans ; Root Canal Therapy

Objective To evaluate the clinical outcomes of arthroscopic reconstruction of anterior cruciate ligament (ACL) with femoral tunnel made by three-portal technique.Methods From February 2014 to May 2015,fifty-five ACL rupture patients (38 males and 17 females) with mean age 31.46±7.34 (range,16 to 48) who underwent single-bundle ACL reconstruction with femoral tunnel made by three-portal technique by the same surgeon were followed up.The average preoperative duration was 15.39±18.57 (range,5 to 180) days.All patients presented the positive Lachman test,of which 32 patients had 1 grade pivot-shift test,15 patients 2 grade and 8 patients 3 grade.At final follow-up,the International Knee Documentation Committee (IKDC) and Lysholm scores were used to estimate knee joint function,while the Lachman test and pivot-shift test were used to estimate knee joint stability.Results All patients were followed up for 26.73±5.23 (range,21 to 34) months.The femoral tunnel positions were accurate without fracture of the back walls in all patients.The mean length of femoral tunnel was 38.58±3.76 (range,35 to 42) mm,and the mean angles of femoral tunnel on coronal plane was 49.32°±2.45° (range,45° to 52°).All patients presented negative Lachman test.Fifty-one patients presented negative pivot-shift test,while the remaining 4 patients had 1 grade of the rotational lax-ity.Forty-nine patients showed normal range of motion,while two had 5° flexion deficit,one had less than 5° flexion deficit and three had 5° hyper-extension loss.Furthermore,the IKDC score significantly was improved from 73.31 ±5.83 preoperatively to 94.63±3.16 at the latest follow-up (t=10.14,P＜0.001).The Lysholm score significantly was increased from 66.35±8.91 preoperatively to 95.32±4.25 at final follow up (t=12.76,P＜0.001).Condusion Arthroscopic reconstruction of ACL with femoral tunnel made by three-portal technique is an reliable technique with the advantage of simple procedure and accurate femoral tunnel position in surgery.

Vascular endothelial growth factor 121 (VEGF121) was expressed as inclusion bodies by recombinant Escherichia coli. High concentrations of both biomass (46 g dry cell/L) and VEGF121 inclusion bodies (4.5 g/L) were obtained by applying a high-cell-density culture. After the inclusion bodies were washed and dissolved, VEGF121 was refolded at 0.2 mg/ml by ultrafiltration in refolding buffer with a yield of 81%. Renatured VEGF121 was purified by anion chromatography and Sephacry S-100 chromatography with purity higher than 95% and fnal purification yield of 31%. The purified VEGF121 could stimulate the proliferation of human umbilical vein endothelial cells as demonstrated by a biological activity assay.

The aim of this study was to analyze the promoter methylation patterns of inhibitory killer cell immunoglobulin-like receptor (KIR) which gene expression and the effect of demethylation treatment were studied, and to explore the possible regulation mechanism of inhibitory kir gene expression. The promoter methylation levels of kir2DL1 and kir2DL2/kir2DL3 in NK-92MI cell line were detected by bisulfite sequencing technique. Then NK-92MI cells were treated with 5-azacytidine to induce the demethylation of CpG islands. The levels of gene expression of kir were determined by RT-PCR. The results demonstrated that the methylation frequencies of CpG dinucleotides surrounding the promoter regions of kir2DL1 and kir2DL2/kir2DL3 genes were 25% to 88% and 5% to 80% respectively. DNA-demethylating treatment with 5-azacytidine resulted in re-expression of kir2DL1 gene and increased expressions of kir2DL1, kir2DL2 and kir2DL3 genes in NK-92MI cells. In conclusion, the promoter DNA methylation participates in the regulation of kir gene expression in NK-92MI cells.
Cell Line ; DNA Methylation ; Gene Expression ; Humans ; Killer Cells, Natural ; metabolism ; Receptors, KIR2DL1 ; genetics ; metabolism ; Receptors, KIR2DL2 ; genetics ; metabolism ; Receptors, KIR2DL3 ; genetics ; metabolism

The aim of this study was to investigate the effect of microRNA-193b (miR-193b) on C-KIT protein expression and biological behaviors in K562 cells. The FAM-labeled miR-193b mimic and negative control were respectively transfected into K562 cells using HiPerFect transfection reagent. The percentage of FAM-positive cells was monitored by flow cytometry. The levels of C-KIT and phosphorylated C-KIT protein were detected by Western blot. The cell growth was measured by CCK-8 reagent. The apoptosis of cells were analyzed by flow cytometry with Annexin V staining. The differentiation of cells were analyzed by flow cytometry with anti-CD11b or anti-CD15 staining. The results demonstrated that the percentage of FAM-positive cells was about 80% in miR-193b or negative control-transfected K562 cells. Compared with the negative control group, overexpression of miR-193b in K562 cells significantly inhibited the levels of C-KIT and phosphorylated C-KIT protein. Meanwhile, the cell growth was inhibited and the percentages of apoptotic cells, CD11b- or CD15-positive cells increased. It is concluded that miR-193b can reduce C-KIT expression and inhibit cell growth in K562 cells. The growth-inhibitory activity of miR-193b is associated with apoptosis and granulocytic differentiation. This study contributed to further investigate the role of miR-193b in leukemogenesis.
Cell Differentiation ; genetics ; Cell Proliferation ; Humans ; K562 Cells ; MicroRNAs ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; metabolism ; Transfection

BACKGROUNDDiagnosis and treatment for respiratory symptoms (RSs) of gastroesophageal reflux disease (GERD) is more difficult than that for common esophageal symptoms. The goal of this study was to evaluate the efficacy and safety of radiofrequency (RF) treatment on RSs of GERD in a preliminary 12-month follow-up observation.

METHODSFrom April 2006 to October 2008, 505 GERD patients with mainly respiratory presentations such as wheezing, chronic cough or hoarseness, were treated by endoscopic RF. A questionnaire was completed before and after treatment, using a six-point scale ranging from 0 to 5 to assess symptom severity and frequency. The symptom score was the sum of frequency and severity.

RESULTSSymptom scores were significantly improved at the end of the follow-up period. The mean heartburn score decreased from 5.31 to 1.79. The mean regurgitation score decreased from 5.02 to 1.64; mean cough score decreased from 6.77 to 2.85; mean wheezing score decreased from 7.83 to 3.07; and mean hoarseness score decreased from 5.13 to 1.81 (P < 0.01). No major complications or deaths occurred. Minor complications included temporary post-procedural retrosternal unease or pain (n = 106; 21.0%), mild fever (n = 86; 17.0%), transient nausea/vomiting (n = 97; 19.2%), and transient dysphagia (n = 42; 9.3%). Thirty-five (6.9%) patients had recurrence of symptoms. Endoscopic RF treatment was repeated in six patients, and laparoscopic fundoplication was performed in seven.

CONCLUSIONEndoscopic RF is an effective and safe means to treat RSs in patients with GERD.

Adult ; Aged ; Cough ; surgery ; Esophagogastric Junction ; physiopathology ; radiation effects ; Esophagoscopy ; methods ; Female ; Gastroesophageal Reflux ; physiopathology ; surgery ; Heartburn ; surgery ; Hoarseness ; surgery ; Humans ; Male ; Middle Aged ; Radio Waves ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the effect of intestinal ischemia/reperfusion (I/R) injury on leptin and orexin-A levels in peripheral blood and central secretory tissues, and investigate the roles of leptin and orexin-A in acute inflammatory responses.

METHODSAn intestinal I/R injury rat model was established, and the rats were grouped according to duration of the reperfusion time following a 60-min ischemia. Radioimmunoassay was used to examine the protein levels of leptin in the serum and adipose tissue, and the protein levels of orexin-A in the plasma and hypothalamus. Reverse transcriptase-polymerase chain reaction was also performed to detect the mRNA expressions of adipose leptin and hypothalamus orexin-A.

RESULTSCompared with that before injury, serum leptin level of 60-min ischemia with 30-min reperfusion (I60'R30') group decreased significantly and that of I60'R360' increased significantly. Compared with the sham-operation group (sham) after injury, serum leptin level of I60'R360' group increased significantly, and adipose leptin protein levels of I60'R30' and I60'R90' groups decreased significantly, whereas that of I60'R360' group increased obviously. Compared with sham group after injury, adipose leptin mRNA expressions of I60'R30', I60'R240' and I60'R360' groups all increased significantly, while that of I60'R150' showed significant decrease. No significant changes were noted in the protein levels of orexin-A either in the plasma or hypothalamus after I/R injury. In comparison with sham group after injury, hypothalamus orexin-A mRNA expressions of I60'R30' and I60'R90' groups showed gradual but significant decrease, and till 150 min of reperfusion, the expression reached its lowest, followed then by slow recovery at 240 and 360 min, though still remaining significantly lower than that of sham group.

CONCLUSIONLeptin and orexin-A have a time-dependent response to intestinal I/R injury, but the former appears to exhibit a faster response, and they may play a certain role in the metabolic disorders of acute inflammation.

Animals ; Female ; Inflammation ; blood ; genetics ; physiopathology ; Intestine, Small ; blood supply ; metabolism ; Intracellular Signaling Peptides and Proteins ; blood ; genetics ; Leptin ; blood ; genetics ; Male ; Neuropeptides ; blood ; genetics ; Orexins ; RNA, Messenger ; biosynthesis ; genetics ; Rabbits ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; blood ; genetics ; physiopathology ; Reverse Transcriptase Polymerase Chain Reaction