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

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

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

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.

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

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

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 study the clinical effects of the Chinese drug Yimusake, used alone or in combination with trazodone hydrochloride, on primary premature ejaculation (PE).

METHODSSixty-eight primary PE patients were randomized to a control (n=32) and an experimental group (n=36), the former treated with Yimusake 1 tablet (50 mg) pre day, and the latter with 1 tablet of trazodone hydrochloride (50 mg) pre day in addition, both given orally after supper and for 4 weeks, followed by observation of the therapeutic effects.

RESULTSEighteen cases (56.25%) responded in the control and 25 (69.44% ) in the experimental group, with statistically significant difference between the two groups (P<0.05).

CONCLUSIONYimusake combined with trazodone hydrochloride is highly efficacious for primary PE.

Adult ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Integrative Medicine ; Male ; Phytotherapy ; Premature Ejaculation ; drug therapy ; Trazodone ; therapeutic use ; Treatment Outcome ; Young Adult

BACKGROUNDThe extended spectrum β-lactamase (ESBL)-producing Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are the major pathogens causing pneumonia and have a significant impact on the clinical course. Limited data exist on molecular characterization of ESBL-producing E. coli and K. pneumoniae that cause pneumonia. The aim of this study was to investigate the comprehensive multilevel characteristics of E. coli and K. pneumoniae causing pneumonia in China for the first time.

METHODSE. coli (17) and K. pneumoniae (21) isolates responsible for pneumonia were isolated from 1270 specimens collected in a prospective multi-center study in eight teaching hospitals in China from June to December in 2007. The susceptibilities, ESBL confirmation, sequence typing, blaCTX-M and blaSHV genes, their genetic environment and plasmid Inc/rep types were determined.

RESULTSSixteen E. coli (94.1%) and eleven K. pneumoniae (52.4%) isolates were ESBL producers. About 77.8% and 66.7% of them were resistance to ciprofloxacin and levofloxacin, and 100% were susceptible to imipenem. The most prevalent ESBL gene was CTX-M-14, followed by SHV-2, CTX-M-15, CTX-M-3, CTX-M-65, SHV-12, SHV-26 and SHV-28. SHV-1 and SHV-11 were also detected and coexisted with blaCTX-Ms in five strains, and three strains contained only SHV-1. All CTX-M-14 were detected ISEcp1 upstream and nine were found IS903 downstream and the majority of them (64.3%) were carried by IncF plasmids. All blaSHV were flanked by recF and deoR, located on IncF, IncN, IncX and IncH plasmids. Two SHV-2, one SHV-1 and the only SHV-28 were further preceded by IS26. Genes lacY and lacZ were detected at further upstream of two blaSHV-1. The K. pneumoniae carrying SHV-28 was susceptible to β-lactams, and no mutations or deletions in gene or promoter sequences were identified to account for susceptibility. Multilocus sequence typing experiments showed the ESBL-producing strains were genetically diverse.

CONCLUSIONSThe rate of occurrence of blaESBL in E. coli and K. pneumoniae causing pneumonia was high, and blaCTX-M-14 was dominant and probably mobilized by ISEcp1 mainly on IncF plasmids. Importantly, unexpressed blaESBL genes may occur in susceptible isolates and hence may have clinical implications.

Blotting, Southern ; Escherichia coli ; enzymology ; genetics ; Klebsiella pneumoniae ; enzymology ; genetics ; Plasmids ; genetics ; Pneumonia ; microbiology ; Promoter Regions, Genetic ; genetics ; Prospective Studies ; beta-Lactams ; metabolism