Objective To observe the dynamic serological test results of 28 patients with acute brucellosis,and to investigate the relationship between serological test results and diagnosis,curative effect and prognosis of brucellosis.Methods Twenty eight patients(infected with sheep brucellosis) with acute brucellosis in the Department of Brucellosis in Heilongjiang Provincial Land Reclamation Headquarters General Hospital were selected as research subjects,and their serological changes were tested by means of tube agglutination test (SAT)and clinical outcomes were compared before and after each stage of treatment.In addition,symptoms of fever,weakness,sweating,joint pain,swollen lymph nodes and biochemical parameters [alanine aminotransferase (ALT),lactate dehydrogenase (LDH)] were also tested 3,6 and 9 weeks after the treatment.Results Antibody titer reached the peak at the third week,1 ∶ 400(++),which accounted for 39.29％ (11/28); 2857％(8/28) of the patients became negative at the sixth week; 50.00％ (14/28) became negative at the ninth week.Before the treatment,20(71.43％) patients had the symptom of fever,8 (28.57％) patients had the symptom of hyperhidrosis,28 (100.00％) patients had the symptom of joint pain,7(25.00％) patients had the symptom of lymph node enlargement,28 (100.00％) patients' ALT was elevated,and 26(92.86％) patients' LDH was elevated.After three weeks of treatment,except the three patients (10.71％) who occasionally had fever,the rest of the patient's temperature was returned to normal.Also the numbers of patients with the symptoms of fatigue,sweating and joint pain were significantly reduced,and specifically,the conesponding number was 13(46.43％),2(7.14％),and 21 (75.00％)patients,respectively.ALT and LDH returned to normal(only one patient's ALT was out of the range).At the sixth week,all the patients' symptoms of fever and hyperhidrosis disappeared.The number of patients with the symptoms of joint pain and lymph node enlargement reduced to 12(42.86％) and 3(10.71％),respectively.The results of biochemical tests(ALT and LDT) returned to normal.At the ninth week,most patients' clinical symptoms disappeared.A few patients still had the symptoms of weakness[2(7.14％)] and joint piin[6(21.43％)].Conclusions After effective treatment,antibody titer of patients decreases rapidly,at the same time,the clinical symptoms improve quickly.There is a parallel relationship between the change of antibody titer and clinical symptoms.It is demonstrated that the appearing time of brucellosis specific antibodies,the ampfitude and speed of change of antibody titers can be used in diagnosis,therapeutic evaluation and prognosis of the disease.

Objective To observe the clinical features of brucellosis spondylitis and analyse the reasons for its misdiagnosis,and improve the level of diagnosis and differential diagnosis.Methods Forty-two clinically diagnosed patients with brucellosis spondylitis were studied retrospectively,and these patients were diagnosed and hospitalized in the General Hospital of Heilongjiang Land Reclamation Bureau.Their medical records were analyzed,which included the general information,medical history,clinical symptoms,results of magnetic resonance imaging(MRI) and serum tube agglutination test(SAT) and so on.Results Main clinical symptoms and signs were severe persistent neck,back and leg pain.They also had plate shape low back but without kyphosis.In addition,patients had to keep in one posture because their spinal activity was limited.Also,scoliosis or pelvic tilt and lameness may occur when standing,which were typical symptoms of nerve root compression.Thirteen cases were diagnosed as tuberculosis,accounting for 30.95％(13/42); 6 cases were diagnosed as lumbar disc herniation,accounting for 14.28％ (6/42); 2 cases were diagnosed as ankylosing spondylitis,accounting for 4.76％ (2/42).Therefore,the total rate of misdiagnosis was 50％ (21/42).Abnormal MRI signal intensity can be seen in the pathological vertebrae.Specifically,T1-weighted images (T1WI) showed low signal,T2-weighted images (T2WI) showed high signal,or mixed high and low signal intensity was observed.Vertebral showed wedge deformation without collapse and sequestrum; strip and sheet abnormal signal can also be found within the intervertebral disc.Normal structure disappeared and disc space became narrow.Accordingly,the plane dural sac was compressed to form visible abscess near the spine,but psoas abscess was not found.Patients with positive SAT result accounted for 92.85％ (39/42).Conclusion Reasons for misdiagnosis include lack of detailed medical records,atypical clinical symptoms and similar imaging changes and so on.

OBJECTIVETo develop a human ovarian carcinoma SKOV3 model in severe combined immunodeficiency (SCID) mouse and to study its biologic characteristics.

METHODSHuman ovarian carcinoma SKOV3 cells were injected intraperitoneally into female SCID mouse to establish a transplantation model of human ovarian carcinoma. The biological characteristics, metastasis and morphology of transplanted tumors were studied.

RESULTAll tumors grew progressively with no sign of regression. The tumor cells spread around the peritoneal cavity and mainly on the diaphragm, mesentery, peritoneum and around the liver, which was confirmed by histopathology. The morphology, growth pattern and CA125 secretion of primary culture of transplanted cells remained as same as those of ovarian carcinoma cell line SKOV3.

CONCLUSIONAn intraperitoneal transplantation model of human ovarian carcinoma SKOV3 in SCID mice has been developed successfully, which can simulate the biological behavior of peritoneal metastasis of human ovarian carcinoma.

Animals ; Disease Models, Animal ; Female ; Humans ; Mice ; Mice, SCID ; Neoplasm Transplantation ; Ovarian Neoplasms ; pathology ; ultrastructure ; Peritoneal Neoplasms ; secondary ; Transplantation, Heterologous

OBJECTIVETo investigate the effects of estrogen receptor alpha (ERa) and insulin-like growth factor 1 (IGF1) on the proliferation of prostatic smooth muscle cells (PSMCs) in vitro.

METHODSThe ERalpha shRNA expression frame was subcloned to the pGSadeno adenovirus vector by homologous recombination technology to construct the pGSaaeno-ERalpha vector. After the mouse PSMCs were transfected in vitro by pGSaaeno-ERalpha, the mRNA and protein expression levels of ERalpha were detected by RT-PCR and Western blot respectively. The expression of IGF1 in the ERa-reduced cells was determined by Western blot 6 hours after treatment with 17beta-estradiol (E2) at 10(-8) mol/L. The post-transfection activity of estrogen or exogenous IGF1 in the proliferation of PSMCs was evaluated by MTT chlormetric analysis.

RESULTSAfter treatment with E2, the proliferation of PSMCs and the expression of the IGF1 gene were significantly increased in the normal control group (P <0.05), but not obviously changed in the ERalpha-siRNA group (P> 0.05). And exogenous IGF1 failed to induce the proliferation of the ERalpha-reduced PSMCs.

CONCLUSIONE2 induces the expression of IGF1 via ERalpha, and IGFl, with the interaction of ERalpha, promotes the proliferation of PSMCs.

Animals ; Cell Proliferation ; Cells, Cultured ; Estradiol ; pharmacology ; Estrogen Receptor alpha ; metabolism ; Insulin-Like Growth Factor I ; metabolism ; Male ; Mice ; Myocytes, Smooth Muscle ; cytology ; Prostate ; cytology ; RNA, Messenger ; genetics

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

OBJECTIVETo investigate the changes in cell cycle induced by cisplatin (DDP) and the effect of antisense oligonucleotide (AsODN) targeting Chk1/2 on DDP-induced apoptosis in lung cancer cell line A549 cells.

METHODSThe characteristics of cell cycle and apoptosis induced by DDP were detected by flow cytometry using SubG1 method. Chk1/2 mRNA and protein expression were assayed by RT-PCR and Western blot under best condition of transfection of AsODN targeting Chk1/2 by lipofection. Apoptosis of A549 cells induced by DDP was determined by flow cytometry using AnnexinV-FITC staining after transfection of Chk1/2 AsODN.

RESULTSAsynchronized A549 cells were treated with 10 micromol/L DDP, and significant S-phase arrest was observed at 12 h later. Transfection with antisense oligonucleotide targeting Chk1/2 inhibited the Chk1/2 expression at both mRNA and protein levels. Either Chk1 or Chk2-specific AsODN consistently enhanced DNA damage-induced apoptosis by 100% - 200%, compared with that in the sODN control (P < 0.05), but combined use of Chk1- and Chk2-specific AsODN did not show synergistic effects as compared with that induced by treatment with Chk1- or Chk2-specific AsODN alone (P > 0.05).

CONCLUSIONChk1 and Chk2 may be regarded as effective targets of chemotherapy for lung cancer. Silencing the key effector Chk1 and Chk2 genes may significantly increase the chemosensitivity of lung cancer cells.

Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Checkpoint Kinase 1 ; Checkpoint Kinase 2 ; Cisplatin ; pharmacology ; Gene Silencing ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Oligonucleotides, Antisense ; genetics ; Protein Kinases ; genetics ; metabolism ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Transfection

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

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.

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

OBJECTIVESTo investigate the role of hMLH1 promoter hypermethylation and microsatellite instability (MSI) in the development of ovarian mucinous tumors.

METHODSOne hundred and seven of paraffin-embedded specimens of ovarian mucinous tumors (malignant 49, borderline 35, and benign 23) were collected from Women's Hospital, School of Medicine, Zhejiang University from 1995 to 2001. The assessment of MSI was based on the use of a panel of six microsatellite markers (BAT-25, BAT-26, BAT-40, D5S346, D17S250, and D2S123) by polymerase chain reaction (PCR). Hypermethylation of hMLH1 promoter region was detected using restriction cut analysis.

RESULTS4.3% (1/23), 14.3% (5/35), and 36.7% (18/49) of benign tumors, borderline tumors, and malignant tumors respectively displayed hypermethylation of the hMLH1 promoter. The hMLH1 promoter hypermethylation rate of malignant group was significantly higher than that of borderline and benign group (P = 0.023, 0.004), but no significant difference between the borderline group and the benign group (P = 0.438); 4.3% (1/23), 8.6% (3/35), and 16.3% (8.49) of benign tumors, borderline tumors, and malignant tumors showed MSI positive phenotype. But there were no significant differences each other in the MSI positive phenotype rate; 75% (9/12) MSI positive phenotype ovarian mucinous tumors were hypermethylated at hMLH1 promoter, while the MSI-phenotype tumors were unmethylated in 84.2% (80.95) of cases. There was significant correlation between MSI positive phenotype and hMLH1 promoter hypermethylation (P = 0.000).

CONCLUSIONSIn ovarian mucinous tumors, malignant, borderline, and benign tumors exist hMLH1 promoter hypermethylation. Hypermethylation of hMLH1 promoter results MSI in ovarian mucinous tumors. Methylation of hMLH1 promoter and MSI may be involved in the carcinogenesis of ovarian mucinous cancer.

Adaptor Proteins, Signal Transducing ; Base Pair Mismatch ; Carrier Proteins ; Chromosomal Instability ; Cystadenocarcinoma, Mucinous ; genetics ; DNA Methylation ; DNA Repair ; DNA, Neoplasm ; genetics ; DNA, Satellite ; Female ; Genes, Neoplasm ; Humans ; Microsatellite Repeats ; genetics ; MutL Protein Homolog 1 ; Neoplasm Proteins ; genetics ; Nuclear Proteins ; Ovarian Neoplasms ; genetics ; Promoter Regions, Genetic ; genetics