ObjectiveTo analyze the surveillance results at the national surveillance spots of brucellosis from 2006 to 2010,to know the epidemic status of national brucellosis,and to provide scientific evidences for evaluating of surveillance quality and formulating of surveillance strategies and measures.Methods The brucellosis surveillance data collected from the Surveillance Reports of Major Infectious Diseases and Vector-Biological Monitoring Report in China from 2006 to 2010,and the National Diseases Surveillance Information Management System were classified and analyzed using two stage clustering statistical analysis.Results According to national routine surveillance,the incidence of brucellosis increased from 1.55 per 100 000 in 2006 to 2.62 per 100 000 in 2010.Reported brucellosis cases were mainly in Inner Mongolia,Shanxi,Heilongjiang and other regions,and reported time was from March to August,which accounting for 79.87％(114 265/143 064).The brucellosis cases were mainly youth,male,farmers and herdsman.In the national surveillance spots of the brucellosis,217 648 cases received epidemic survey,of which 61 905 cases received serological test,the positive rate was 19.66％(12 169/61 905),and 10 318 new cases were found; the highest positive rate of serological test was 41.19％(2757/6694) which was found in Zhalantun of Inner Mongolia; the detection rate of pathogen culture was 2.49％(29/1165); the positive rates of serological test in cattle and sheep were 1.07％(2355/219 352) and 0.93％ (2766/296 176),respectively.The surveillance spots were classified into four grades according to cluster analysis.Conclusions The epidemic continues to rise in human surveillance spots of brucellosis.Although national surveillance spots play an important role in the surveillance of brucellosis,the quality and quantity of surveillance work of national brucellosis surveillance spots should be improved,different management measures should be taken in different surveillance spots according to classified results of the cluster analysis.This study provides some basis to improve the utilization of health resources and the level of Brucella disease prevention and control work.

Objective To investigate the clinical significance of procalcitonin(PCT) in early diagnosis of neonatal infections.Methods Rapid hemi-quantitative immunoassay was used to measure PCT levels in 196 hospitalized neonates, who were divided into sepsis group,(local-)infection group, virus-infection group and non-infection group.Results If plasma PCT≥0.5 ?g/L was taken as positive,the positive rates in sepsis group,local-infection,virus-infection,non-infection group were 87.87%,41.66 %,12.0%,11.11%,respectively.The positive rate of sepsis group was significantly higher than that of the other 3 groups(all P

OBJECTIVETo investigate the expression of OPCML gene in ovarian epithelial carcinoma and determine the relationship between mRNA expression and methylation of their promoters.

METHODTwenty normal ovarian tissues and 89 ovarian epithelial tumor specimens (72 malignant, 17 benign), as well as 3 ovarian carcinoma cell lines (SKOV-3, CAOV3, and 3AO), were collected for detection of OPCML gene expression by reverse transcription-polymerase chain reaction and for detection of promoter methylation by restriction enzyme cut analysis from 7. 1999 to 7. 2003.

RESULTSAmong ovarian epithelial carcinoma 19.4% expressed OPCML mRNA, while 85% of normal ovarian tissue and 76.5% of benign ovarian tumor. The ratio of expression of OPCML mRNA in ovarian epithelial carcinoma was significantly lower than those of normal (chi2 = 30.108, P = 0.0000) and benign tumors (chi2 = 21.162, P = 0.000). No OPCML mRNA expression was found in SKOV-3 and CAOV3, but was found in 3AO. Methylations were detected in 44.4% of cancer cells promoter, while 0% in normal ovarian tissue and benign ovarian tumors. The ratio of methylation of ovarian epithelial carcinoma was significantly higher than those of normal (chi2 = 13.630, P = 0.0000) and benign tumors (chi2 = 11.797, P = 0.000). Methylation was found in SKOV-3 and CAOV3, but not in 3AO. The relationship between gene expression and promoter methylation was correlated (r = 11.589, P = 0.002), especially at Hap I1 site (r = 11.640, P = 0.004). Methylation was also found in SKOV-3 and CAOV3 cell lines, but not in 3AO cell line.

CONCLUSIONDeletion of OPCML gene exists in ovarian epithelial carcinoma cell. The gene promoter methylations, especially Hap II motif, may be one of pathways that contribute the inhibition of OPCML expression.

Adult ; Aged ; Cell Adhesion Molecules ; genetics ; Cell Line, Tumor ; CpG Islands ; genetics ; DNA Methylation ; Female ; GPI-Linked Proteins ; Gene Deletion ; Humans ; Middle Aged ; Ovarian Neoplasms ; genetics ; pathology ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the activation pattern of signal transducers and activators of transcription (STAT) induced by vascular endothelial growth factor (VEGF) in CD34+ hematopoietic progenitor cells, and gain an insight into the molecular mechanism and signal transduction pathway of VEGF that has an effect on CD34+ hematopoietic progenitor cells.

METHODSAfter isolated from umbilical cord blood by using a high-gradient magnetically activated cell sorting system (MACS), CD34+ cells were stimulated by VEGF (50 ng/ml) for different time (0, 15, 30, 45, 60, 90 min) to detect the tyrosine phosphorylation and nuclear translocation of STAT-3 and STAT-5 with Western blot and immunocytochemistry methods. The expression of VEGF receptor-2 (VEGFR2) on the membrane of CD34+ progenitor cells was examined by immunocytochemistry. ATWLPPR, an effective peptide screened from phage epitope library by affinity for membrane-expressed VEGFR2 and blocking the binding of VEGF to VEGFR2, was used to determine whether the activation of STAT pathway induced by VEGF was blocked.

RESULTSTyrosine phosphorylation of STAT-3 and STAT-5 was undetectable in unstimulated CD34+ cells, but was evident at 15 min in response to VEGF stimulation. VEGF resulted in a rapid and transient tyrosine phosphorylation of STAT-3 and STAT-5. The maximal tyrosine phosphorylation was catched at 30 and 45 min, respectively (P = 0.0001), and returned to basal levels at 90 min. Immunocytochemistry confirmed that increased phosphorylated STAT-3 was translocated into the nuclei at 30 min (P = 0.0001), and mainly in cytoplasms again at 90 min after stimulation with VEGF. However, compared with unstimulated CD34+ cells, there was only increased phosphorylation of STAT-5 appeared mainly in cytoplasms, but no significant nuclear translocation was found after stimulation with VEGF (P > 0.05). The presence of VEGFR2 was confirmed using anti-VEGFR2 antibody staining by immunocytochemistry, moreover, the phosphorylation of STAT-3 and STAT-5 failed to be activated by the co-culture with ATWLPPR and VEGF, suggesting that activation of the STAT pathway be specifically mediated by VEGFR2 in CD34+ progenitor cells.

CONCLUSIONSSTAT signaling pathway participates in the signal transduction of VEGF via VEGFR2 in CD34+ hemopoietic progenitor cells.

Adult ; Antigens, CD34 ; metabolism ; DNA-Binding Proteins ; Endothelium, Vascular ; drug effects ; metabolism ; Female ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; metabolism ; physiology ; Humans ; Milk Proteins ; Phosphorylation ; Pregnancy ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; STAT3 Transcription Factor ; STAT5 Transcription Factor ; Signal Transduction ; Trans-Activators ; metabolism ; Transcription, Genetic ; Tyrosine ; metabolism ; Vascular Endothelial Growth Factor A ; pharmacology

OBJECTIVETo investigate the effects of vascular endothelial growth factor (VEGF) on differentiation and function of dendritic cells derived from CD34+ hematopoietic progenitor cells.

METHODSAfter isolation from umbilical cord blood with a high-gradient magnetic cell sorting system (MACS), the CD34+ cells were cultured with a cocktail cytokines for differentiating into dendritic cells (DC). The cells were stimulated by VEGF (25 ng/ml) either at the beginning or at day 9 of culture. Kinetics analysis of cell proliferation was performed during the process of cell culture, and the expression of DC differentiation antigens including CD1alpha, CD83, CD80, CD54 and HLA-DR was examined by flow cytometry. DC function was evaluated by the ability to induce proliferation of allogeneic T cells in mixed lymphocyte reaction (MLR) assay, and the production of IL-12 by ELISA.

RESULTSVEGF added at day 1 of culture induced an increase of total cell numbers by (1.51 +/- 0.23)-folds (P = 0.001). VEGF added at the initial but not the late stage of culture could dramatically down-regulate the expression of CD1a [(33.00 +/- 2.12)% vs (81.20 +/- 6.93)%], CD83 [(42.23 +/- 1.15)% vs (87.98 +/- 7.97)%], CD80 (42.93 +/- 1.32)% vs (94.53 +/- 0.87)%], and HLA-DR [(37.93 +/- 5.30)% vs (74.15 +/- 3.74)%], while obviously up-regulate the expression of CD14. Moreover, the inhibitory effect of VEGF on DC function was confirmed by a reduced ability to induce proliferation of allogeneic T cells and production of IL-12 (P < 0.01).

CONCLUSIONSVEGF could induce the expansion of hematopoietic progenitor cells and inhibit at the early stage their differentiation into mature DC.

Antigens, CD ; analysis ; Antigens, CD1 ; analysis ; Antigens, CD34 ; analysis ; blood ; B7-1 Antigen ; analysis ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Dendritic Cells ; cytology ; immunology ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Fetal Blood ; cytology ; immunology ; Flow Cytometry ; HLA-DR Antigens ; analysis ; Hematopoietic Stem Cells ; cytology ; immunology ; metabolism ; Humans ; Immunoglobulins ; analysis ; Intercellular Adhesion Molecule-1 ; analysis ; Interleukin-12 ; analysis ; Membrane Glycoproteins ; analysis ; Vascular Endothelial Growth Factor A ; pharmacology

Objective To compare the clinical effects on Gartland type Ⅲ fractures in children by open reduction and external fixation with two parallel Kirschner wires and closed reduction.Methods From February 2013 to October 2016,84 children with Gartland type Ⅲ supracondylar fracture of humerus were selected and randomly divided into a control group and an observation group,each with a total of 42 cases.The control group received closed reduction treatment,and the observation group received open and external fixation with two parallel Kirschner wires.The two groups were compared on intraoperative blood loss,incision length,operating time,decreased degrees of carrying angle,reduced flexion-extension at the fractured side and the rates for good and excellent clinical effects.Results The observation group behaved significantly better than the control group in intraoperative blood loss,incision length and operating time (P＜0.05).The rates for good and excellent effects was 90.47％ in the observation group and 73.81％ in the control group,and there was obvious difference between the values in the two groups (P＜0.05).The observation group had the decreased degree of carrying angle statistically lower than that in the control group (P＜0.05),while the reduced flexion-extension at the fractured side was not significantly different from that in the control group (P＞0.05).Conclusion Open reduction and external fixation with two parallel Kirschner wires and closed reduction both gain their advantages when used to treat Gartland type llⅢ fractures in children,the former has high clinical effect,significantly recovered elbow joint function and low incidence rate for cubitus varus,and the latter has slight invasion,small incision while relatively low clinical effect and high incidence rate for cubitus varus.

OBJECTIVETo investigate the efficacy and toxicity of methotrexate (MTX) give intravenously in the primary treatment of gestational trophoblastic tumor (GTT).

METHODSA total of 37 patients with low-risk GTT was primarily treated by single MTX in Women's Hospital, School of Medicine, Zhejiang University. Data on the patients' age, clinical stage, WHO classification criteria, antecedent pregnancy, presenting level of human chorionic gonadotropin, courses of chemotherapy required to achieve complete remission, and toxicity related to chemotherapy treatments were collected.

RESULTSThirty-seven patients with low-risk GTT totally received 137 cycles of MTX between Oct. 1999 and Sep. 2002, 34 patients (91.9%) achieved complete remission. Twenty-nine patients received multiple courses of MTX, complete remission was induced in 26 patients (89.7%). The complete response rates of I stage and III stage were 100.0% and 70.0% (P = 0.03) respectively in patients who were received multiple courses of MTX. However, eight patients received single course of chemotherapy, 7 patients achieved complete remission, and 1 achieved complete remission after another additional course of MTX was conducted. Grade III side effects (WHO criteria) only appeared in 7 courses (5.1%) during MTX treatment. Follow-up data showed that only one patient with single course of chemotherapy relapsed after 6 months.

CONCLUSIONSingle MTX chemotherapy may be effective and well tolerated for low-risk GTT.

Adolescent ; Adult ; Antimetabolites, Antineoplastic ; administration & dosage ; Choriocarcinoma ; drug therapy ; Drug Administration Schedule ; Female ; Gestational Trophoblastic Disease ; drug therapy ; Humans ; Methotrexate ; administration & dosage ; Pregnancy ; Uterine Neoplasms ; drug therapy

OBJECTIVE: To study the expression of vascular endothelial growth factor (VEGF) mRNA isoforms in ovarian carcinoma and to explore their role in tumorigenesis and development of ovarian carcinoma. METHODS: The types and levels of VEGF mRNA isoforms of surgical samples from 30 patients with ovarian carcinoma were determined by relatively quantative RT-PCR, nest PCR and sequence analysis. RESULTS: VEGF(121), VEGF(145), VEGF(165) and VEGF(189)mRNA were detected in normal ovaries and ovarian carcinoma tissues. The expression level of VEGF(121) was significantly higher than that of VEGF(145), VEGF(165) and VEGF(189) (P<0.001, respectively). The expression of all 4 isoforms in carcinoma tissues was increased significantly compared with that in normal ovaries (P<0.05). CONCLUSION: Overexpression of VEGF(121), VEGF(145), VEGF (165) and VEGF(189) mRNA, especially VEGF(121), was found in varian carcinoma tissues. This findings suggest that all 4 VEGF isoforms may be involved in the tumorigenesis and development of ovarian carcinoma and VEGF(121) may play a key role.

Objective To construct a new conditionally replicative adenovirus (CRAds) targeting carcinoembryonic antigen (CEA) positive colorectal cancer cells. Methods The DNA fragment of the CEA gene promoter was amplified through PCR and cloned into the vector carrying fusion reporter gene EGFP-Luc to construct expression plasmid pCEA-EGFPLuc. The constructed plasmid pCEA-EGFPLuc was transfected into CEA positive and negative cells by liposome. The activity of CEA gene promoter was evaluated by detecting the expression of EGFP and luciferase activity. The conditionally replicative adenovirus Ad.CEA-E1A/CMV-TK carrying suicide gene HSVtk was constructed, in which the E1A gene was controlled under CEA promoter. CEA positive(Lovo and SW620)and negative tumor cells(HeLa) were infected with Ad.CEA-E1A/ CMV-TK. The selective cytotoxicity of Ad.CEA-E1A/CMV-TK and the synergistic effect of the virus with GCV in CEA positive tumor cells were evaluated by the expression of E1A, cytopathic effect and cell survival rate. Results CEA promoter possesses a good specificity as well as high activity. The expression of E1A only presented in CEA positive tumor cells. After infection with Ad. CEA-E1A/CMV-TK, the cell survival rates of Lovo and SW620 were (36.72±2.49)% and (39.82±4.76)%, respectively, significantly lower than that of Hela[(87.44±2.76)%1 (P<0.01). When combined with GCV, Ad.CEA-E1A/CMV-TK had better oncolytic effect on Lovo and SW620 cells, with cell survival rates of (17.26±3.65) % and (23.93±5.40) %, respectively, significantly lower than those without GCV[(36.72±2.49) % and (39.82±4.76) %, respectively] (P<0.01). Conclusion Ad. CEA-E1A/CMV-TK under the control of CEA promoter has selective cytotoxic effect on CEA positive colorectal cancer cells, and the effect can be enhanced when combined with GCV.

Objective To investigate the factors associated with the occurrence of pulmonary complication in patients with cervical spine fiactures and concurrent cervical cord injury in Wenchuan earthquake. Methods A retrospective analysis was conducted among the 9 patients with cervical spine fractures and cervical cord injuries treated in our department between May 12 to August 6, 2008. Results All the patients received surgical treatment for cervical spine fractures and cervical cord injuries. Six of the patients developed pulmonary complications 5 days after the injury, including 3 patients with pneumonia, 2 with ventilation disorder, and 1 with lung edema and hemopnenmothorax. Aggressive respiratory management was administered in these patients, and the pulmonary complications were effectively controlled. Conclusion Patients with cervical spine fractures and concurrent cervical cord injury often experience severe pulmonary complications during the acute phase (<5 days), which can be more likely in patients with high level injury, chest trauma, old age, preexisting pulmonary illnesses or smoking history. Early detection of the complications results in better therapeutic effect with conservative therapy.