OBJECTIVETo explore the protective effect and mechanism of total flavones of Bidens pilosa L. (TFB) on IgA1 induced injury of venous endothelial cells in Henoch-Schönlein purpura (HSP) children patients. METHODS Human umbilical venous endothelial cells (HUVECs) were taken as subject. They were intervened by normal IgA1 and HSP children patients' serum IgA1, and added with different concentrations TFB at the same time. Then they were divided into the blank control group, the normal control group, the HSP IgA1 group, and HSP IgA1 plus TFB (1.0, 0.5, 0.25 mg/mL) groups. Levels of TNF-α and IL-8 in supernate were detected by ELISA. The NO level was detected by nitrate reductase method. mRNA and protein expressions of NF-κB and ICAM-1 in HUVECs were detected by fluorescent quantitative PCR and Western blot respectively.

RESULTSCompared with the normal control group and the blank control group, levels of IL-8, TNF-α, and NO all significantly increased in the HSP group (P < 0.05). Compared with the HSP group, levels of IL-8, TNF-α, and NO significantly decreased after intervention of TFB (1.0 and 0.5 mg/mL; P < 0.05, P < 0.01). Results of fluorescent quantitative PCR and Western blot showed, as compared with the blank control group and the normal control group, mRNA and protein expressions of NF-κB and ICAM-1 in HSP children patients' serum IgA1 induced venous endothelial cells significantly increased with statistical difference (P < 0.05, P < 0.01). Compared with the HSP group, mRNA and protein expressions of NF-KB and ICAM-1 were obviously down-regulated after intervention of TFB (1.0, 0.5, 0.25 mg/mL), with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONTFB could protect vascular damage by inhibiting in vivo high expression of NF-κB, reducing the production of IL-8, TNF-α, and NO in vascular endothelial cells of HSP children patients.

Bidens ; chemistry ; Child ; Flavones ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Immunoglobulin A ; blood ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-8 ; metabolism ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Purpura, Schoenlein-Henoch ; blood ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the mid-term outcome of operation for thoracolumbar tuberculosis. METHODS : Twenty-eight patiens with thoracolumbar tuberculosis underwent one stage anterior debridement,interbody fusion with bone graft and posterior pedicle screw internal fixation treatment from July 2006 to July 2011. There were 17 males and 11 females. Total 17 patients had nerve injuries ,including 6 cases of grade B, 5 cases of grade C, 6 cases of grade D according to Frankel classification. The poisoning symptoms of tuberculosis and recovery of spinal function were observed. The bone fusion and recovery of [umbar function were evaluated.

RESULTSAll the patients were followed up ,and the duration ranged from 39 to 85 months (mean 57 months). The clinical symptoms were controlled gradually, and the thoracolumbar back pain was alleviated after operation. Among the 17 patients with complications of nerve injuries, 3 patients were improved from preoperative grade B to postoperative grade D, 3 patients were improved from preoperative grade B to postopertive E, 5 patients with preoperative grade C and 6 patients with preoperative D were almostly recovered to normal after operation. According to JOA scoring system for curative effect evaluation, the excellent and good rate at the 3rd month, the 1st year, the 3rd year and the 5th year after operation were 67.86% ,82.14% ,85.71% ,89.29% and 91.30% respectively. The results at the 6th month and the 1st year had no statistical differences compared to the results at the 3rd month (P > 0.05); but the results at the 3rd year and the 5th year were better than that at 3 months after operation (P < 0.05); and the results between 3 yesrs and 5 years after operation had no statistical differences (P < O.05). The degeneration of adjacent segments were evaluated according to the California University (Universith of California at Los Angeles , UCLA) score. The degeneration rate was 53.57% (15/28) at the 3rd year after surgery, which was better than that before surgery. Twenty-three patients were followed up for 5 years ,and the degeneration rate was 86.96% (20/23) ,which was better than those of before surgery and 3 years after surgery.

CONCLUSIONThe surgical treatment for thoracolumbar spinal tuberculosis can achieve the thorough debridement, reconstruction of spinal stability, recovery of lumbar function and promote the functional recovery of the spinal cord, which is an effective method of treatment. However, the mid term follow-up showed that more severe degenerative changes were found in the postoperative adjacent segment.

Adult ; Aged ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Spinal Fusion ; methods ; Thoracic Vertebrae ; surgery ; Tuberculosis, Spinal ; physiopathology ; surgery

OBJECTIVETo explore clinical outcomes and advantages of anterior small-incision focus debridement with posterior internal fixation through muscle spa ring in treating patients with lumbar spinal tuberculosis.

METHODSFrom February 2010 to February 2014, totally 82 patients with lumbar spinal tuberculosis were treated by posterior individual fixation with small-incision focus debridement,including 50 males and 32 females with an average of 50.5 years old. All patients were divided into two groups according to different procedures. Forty-nine patients in group A were treated with anterior small-incision focus debridement with posterior internal fixation through muscle spa ring at stage I ; and 33 patients in group B were treated with focus debridement with posterior internal fixation by extraperitoneal approach at stage I . Postoperative mechanical ventilation time, preoperative and postoperative Cobb angle, visual analogue scale (VAS), erythrocyte sedimentation rate (ESR) and Frankel grading were observed and compared. Postoperative complications, stability of internal fixation and bone union were compared.

RESULTSAll patients were followed-up from 15 to 36 months with an average of 23.7 months. Psoas abscess of three patients in group A and 1 patient in group B on the opposite side increased and were healed by the secondary apocenosis. The other 78 cases were healed at stage I, and no sinus tract formation, incisional hernia, leakage of cerebrospinal and occurrence of spinal tuberculosis were occurred. Fracture healing time ranged from 3 to 7 months with an average of 4.6 months. Postoperative mechanical ventilation time and VAS score in group A was better than group B. There were no statistical differences in Cobb angle, ESR and Frankel grading at the final following-up between two groups.

CONCLUSIONAnterior small-incision focus debridement with posterior internal fixation through muscle spa ring in treating patients with lumbar spinal according to degree of damage is a safe and effective method.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Debridement ; methods ; Female ; Fracture Fixation, Internal ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Treatment Outcome ; Tuberculosis, Spinal ; surgery ; Young Adult

P300/CBP-associated factor(PCAF),an important member of histone acetyltransferase family(HATs) within eukaryotic cells,is capable of inducing the acetylation of histone,promoting the transcription of specific genes and involving in many biological effects.In the present study,full-length cDNA of PCAF was inserted into plasmid pGEX-5x-1,then the soluble protein GST-PCAF was expressed in E.coli BL21(DE3) after the optimization of inducing conditions.The recombinant protein was further purified with affinity chromatography and tested the activity by in vitro acetylation assays.High efficient PCAF protein produced by this method could serve for the study on the role of PCAF in gene regulation and the interaction between PCAF and other proteins.

Objective To screen differentially expressed genes in placentas with hepatitis B virus (HBV)infection and to discuss the molecular mechanism of HBV intrauterine infection.Methods Thirty placenta tissue specimens from HBsAg and HBV DNA positive pregnant women were used as the study group and 30 placenta tissue specimens from normal pregnant women with HBsAg and HBV DNA negativity were served as the control group.The suppression subtractive hybridization(SSH)technique was used.Total RNAs of placenta tissue of the study group were mixed as the tester,and total RNAs of placenta tissue of the control group were mixed as the driver.A subtractive cDNA library was constructed by PCR-selective cDNA subtraction systems.Amplifications of the library were carried out with E.coil strain DH5? by reverse spot hybridization.RT-PCR confirmed that phosphatidylinositol 3-kinase(PI3K)was up-regulated in placenta tissue with HBV infection.Results Colony PCR showed that the clones contained 200-1000 bp inserts. Thirty five clones were confirmed by reverse spot hybridization and analyzed by sequencing and bioinformatics.Thirty three known genes and 2 genes with unknown function were obtained.RT-PCR preliminarily confirmed that PI3K gene was up-regulated in HBV infected placenta.Conclusions The differentially expressed genes in placentas with hepatitis B virus(HBV)infection using SSH technique has been screened out successfully.These differentially expressed genes encoding proteins participating in cell vital metabolism and malformation,and signal conduction-antiapoptosis pathway.This finding brings some new clues for studying the mechanisms of HBV intrauterine infection.

OBJECTIVETo explore the dynamics of hepatitis B virus covalently closed circular DNA (cccDNA) and optimal duration of treatment after serum virology response.

METHODSHBV cccDNA in liver biopsies and the serum HBV DNA were quantified by real time PCR, the serum makers were detected by enzyme-linked immunosorbent assay.

RESULTS(1) The cccDNA in biopsy samples continued to decrease after serum virology responded. (2) The longer the treatment after serum virology response, the lower the cccDNA level in liver tissue. (3) Anti-HBe positive patients had lower cccDNA in liver tissue than anti-HBe negative patients. (4) cccDNA in liver tissue was undetectable in 12 out of the 18 case anti-HBe(+) patients. Serum virology response lasted 35 months and anti-HBe(+) lasted 30 months.

CONCLUSIONAfter serum virology responded, the longer the treatment, the lower the liver cccDNA. The cccDNA is undetectable in about 2/3 of the patients if the serum virological clearance lasts more than 35 months and anti-HBe lasts more than 30 months.

Adult ; Aged ; Antiviral Agents ; therapeutic use ; Biopsy ; DNA, Circular ; analysis ; DNA, Viral ; analysis ; blood ; Female ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; Hepatitis B, Chronic ; blood ; drug therapy ; virology ; Humans ; Liver ; pathology ; virology ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Time Factors ; Viral Load ; Young Adult

UNLABELLEDOBJECTIVE To study the clinical and biological characteristics and prognosis of t(8;21)/AML1-ETO-positive childhood acute myeloid leukemia (AML).

METHODSThe clinical data of 55 children who were diagnosed as t (8; 21)/AML1-ETO-positive AML were retrospectively studied. Event-free survival (EFS), disease-free survival (DFS), and overall survival (OS) rates were estimated by the Kaplan-Meier method. Prognostic factors were evaluated by COX regression analysis software.

RESULTSOf the 55 patients, 4 patients gave up treatment after the diagnosis was confirmed and 4 patients were lost to follow-up after the first chemotherapy course. The remaining 47 patients received a double-induction therapy. The total complete remission (CR) rate was 71% and 94% after the first and second chemotherapy course, respectively. The disease was relapsed in 10 patients (21%). The 5-year EFS, DFS and OS rates were (56.1 ± 7.9)%, (59.8 ± 8.1)%, and (72.0 ± 8.1)%, respectively. Multivariate analysis showed that age was an independent risk factor for the long-term prognosis. The older children had a greater risk of experiencing an accident or death (P<0.05). The 5-year OS rate in 27 patients with regular consolidation chemotherapy was significantly higher than 13 patients with irregular chemotherapy after CRz [(47.5 ± 17.1)% vs (38.9 ± 17.3)%; P<0.01].

CONCLUSIONSChildhood t(8;21)/AML1-ETO-positive AML is a highly heterogeneous disease, with a high CR rate and a good long-term prognosis. Age is one of the important factors affecting the long-term therapeutic effect. Regular consolidation chemotherapy applied after CR usually is helpful.

Adolescent ; Bone Marrow Examination ; Child ; Child, Preschool ; Chromosomes, Human, Pair 21 ; Chromosomes, Human, Pair 8 ; Core Binding Factor Alpha 2 Subunit ; analysis ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; mortality ; Male ; Oncogene Proteins, Fusion ; analysis ; Prognosis ; RUNX1 Translocation Partner 1 Protein ; Translocation, Genetic

OBJECTIVESome recent studies revealed that phenthiazine might be able to reverse tumor cell drug-resistance. Chlorderazin belongs to the phenthiazine compounds. The study aimed to investigate the reversing effect and mechanism of chlorderazin on multidrug resistance of leukemic cell line K562/AO2.

METHODS(1) The cytotoxicities of chlorderazin were assayed with the tetrazolium dye, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. (2) The reverse effect of chlorderazin on K562/AO2 cells was analyzed with MTT method. The multidrug resistance reversal index (RI) was equal to the ratio of control group IC(50)/test group half inhibition concentration IC(50). (3) The intracellular daunorubicin (DNR) concentrations were measured by the flow cytometry. (4) Mdr1 mRNA expression was detected by reverse transcription-polymerase chain reaction (RT-PCR). The ratio of mdr-1/beta-actin density was calculated.

RESULTS(1) Chlorderazin 3 micro g/ml showed little toxicity to K562/AO2 cells and the suppression rate was less than 5%, so the concentration of 3 micro g/ml chlorderazin was selected as the experiment concentration. (2) The cytotoxicities of DNR to K562/AO2 were enhanced by 3 micro g/ml of chlorderazin (P < 0.05) and RI was 1.901. (3) Chlorderazin of 3 micro g/ml could increase the intracellular DNR accumulation significantly (P < 0.05), and the fluorescence staining by the flow cytometry was higher (250.95 +/- 18.96) than the control group (112.75 +/- 15.78) and shift right in K562/AO2 cells treated with chlorderazin, and the difference was significant (P < 0.05). (4) Chlorderazin has no significant influence to the expression level of mdr-1 mRNA. Both test group and control group showed a clear mdr-1 mRNA band located at the position of 157 kb. The ratios of mdr-1/beta-actin density were 0.414 +/- 0.012 in the test group and 0.447 +/- 0.027 in the control group, respectively, and the difference was not significant (P > 0.05).

CONCLUSIONChlorderazin could reverse the multidrug resistance by increasing the intracellular DNR accumulation in K562/AO2 cells. The effects had no correlation to the mdr-1 gene. Further study is needed.

ATP-Binding Cassette, Sub-Family B, Member 1 ; genetics ; Antiemetics ; pharmacology ; Cell Division ; drug effects ; Chlorpromazine ; pharmacology ; Drug Resistance, Multiple ; drug effects ; genetics ; Drug Resistance, Neoplasm ; drug effects ; Flow Cytometry ; Humans ; K562 Cells ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the expression of the Wilms' tumor 1 (WT1) mRNA in childhood myelodysplastic syndrome (MDS), and to evaluate WT1 as a tool to differentiate MDS from aplastic anemia(AA).

METHODSThe quantitative expression of WT1 transcript by using real-time quantitative polymerase chain reaction (RQ-PCR) was performed in the bone marrow samples of 36 childhood MDS and 49 childhood AA, the samples were collected from September 2008 to December 2011.

RESULTS(1) The positive rate of WT1 in severe AA (SAA) was 0, 14.3% in chronic AA (CAA), 58.6% in refractory cytopenia (RC), 100% in refractory anemia with excessive blast (RAEB) and 97.5% in acute myeloid leukemia (AML). The mean level of WT1 in SAA, CAA, RC, RAEB and AML was 0.041%, 0.357%, 7.037%, 12.680% and 24.210%, respectively. The positive rate of WT1 in RC patients was higher than that of SAA (P = 0.000) and CAA (P = 0.001). (2) The positive rate of WT1 in patients with hypoplastic MDS was 66.7% and was higher than that of SAA (P = 0.000) and CAA (P = 0.001). The mean level of WT1 in patients with hypoplastic MDS was (3.022 ± 5.040)% and higher than that of SAA \[(0.041 ± 0.047)%, P = 0.000\] and CAA\[(0.351 ± 0.479)%, P = 0.002\].

CONCLUSIONSThe level of WT1 in childhood MDS was higher than that of childhood AA. The degree of WT1 expression in MDS increased during disease progression. WT1 is a useful tool for differentiating the childhood hypoplastic MDS from AA.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Male ; Myelodysplastic Syndromes ; genetics ; metabolism ; pathology ; WT1 Proteins ; genetics ; metabolism

The tumor multidrug resistance reversal effect of NPB304, a novel taxane, was studied. MTT assay was used to determine the IC50 of chemotherapy drugs. Western blotting assay was applied to analyze the expression of P-glycoprotein (P-gp). The effect of compounds on the P-gp function and P-gp ATPase activity was determined by rhodamine 123 (Rh123) accumulation assay and analysis kit, respectively. Molecular docking was employed to predict the binding force between compounds and P-gp. Transmembrane transport of NPB304 was analyzed using MDCK II and MDR1-MDCK II cell model. NPB304 displayed multidrug resistance reversal effect on KBV cells and MCF-7/paclitaxel cells, NPB304 collaborative with P-glycoprotein (P-gp) inhibitors verapamil enhanced the reversal activity, specifically, 10 μmol x L(-1) verapamil in combination with paclitaxel reversed resistance by 56.5-fold, while combined with NPB304 increased the reversal fold; NPB304 synergistically increased Rh123 accumulation in the resistant cells when combined with verapamil, and NPB304 at 0-1 μmol x L(-1) enhanced the ATPase activity activated by verapamil was observed. NPB304 existed the hydrophobic interactions with the TM regions of P-gp, and the binding force between NPB304 and the A chain of the TM region was stronger. P-gp ATPase activity assay demonstrated NPB304 at lower concentrations (0-1.5 μmol x L(-1)) could activate the P-gp ATPase, playing a role on inhibition of P-gp function. However, NPB304 did not have an obvious feature of P-gp substrate. NPB304 exerted itself and synergy with verapamil activity on reversing tumor resistance via inhibiting the P-gp function.
ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antineoplastic Agents ; pharmacology ; Biological Transport ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Drug Synergism ; Humans ; MCF-7 Cells ; Rhodamine 123 ; Taxoids ; pharmacology ; Verapamil ; pharmacology