Objective To investigate the role of negative-regulatory factors of toll-like receptors (TLRs)signal pathways in immunological pathogenesis of Kawasaki disease(KD).Methods Thirty-two chil- dren with Kawasaki disease and 16 age-matched healthy children were studied.Reverse-transcription PCR (RT-PCR)and real-time PCR were used to evaluate the mRNA expression levels of toll-like receptor 4(TLR4), MD-2,MyD88,IRAK-4,TRAF6,T1/ST2,IRAK-M,Triad 3A,and proinflammatory factors such as IL-1?, IL-6,IL-8 and TNF-?,in peripheral blood monocytes/macrophages(MC).The expression of TLR4 protein in MC was analyzed by flow cytometry.Results①Compared with the control group,the mRNA levels of TLR4, MD-2,MyD88,IRAK-4 and TRAF6 in KD group were up-regulated significantly(P＜0.01),and the expression level of TLR4 protein was also found to be up-regulated in KD group during acute phase.It was detected that expression levels of TLR4 protein in KD with coronary artery lesion(KD-CAL~+)was significantly higher than that of KD without coronary artery lesion(KD-CAL-)[flow cytometry:(6.5?1.7)% vs(11.9_+2.4)%,P＜0.01].②The expression level of negative-regulatory factors such as IRAK-M and Triad3A were significantly up-regulat- ed in acute phase of Kawasaki disease,while the mRNA levels of IRAK-M and Triad3A in KD-CAL~+ group was found to be significantly lower than those of KD-CAL~- group(P＜0.01).No difference of T1/ST2 mRNA expres sion level was detected among all groups(P＞0.05).③The expressions of proinflammatory eytokines such as IL-1?, IL-6,IL-8 and TNF-?in monoeytes/macrophages during acute phase of Kawasaki disease were higher than those of the control group(P＜0.01),and expression of proinflammatory cytokines in KD-CAL~+ group was significantly higher than that of KD-CAL~- group.Conclusion Relative insufficient expression of negative-regulatory factors, such as IRAK-M and Triad3A,maybe correlate with immunological pathogenesis of Kawasaki disease.

Objective To investigate the features of chronic prostatitis during puberty(CPP)and the effects of pelvic floor biofeedback therapy.Methods Totally,25 CPP children (mean age,16 years) and 15 children (mean age,16 years) with normal lower urinary tract as controls were included.In CPP group,NIH-CPSI scores were evaluated,expressed prostatic secretions (EPS) were examined,and bacterial culture was done;and CPP patients were categorized based on the definitions of NIH types.In both groups, urodynamic examination was performed,including evaluation of uroflow curve,maximum flow rate (Q_(max)), post-voiding residual urine (PVR),detrusor-sphincter dyssynergia (DSD),maximum detrusor pressure (P_(det,max))and maximum urethral closure pressure (MUCP).CPP patients underwent biofeedback therapy, and 10 weeks later the effects were assessed.Results In CPP group,NIH typing showedⅡ,ⅢA andⅢB in 1,3 and 21 cases,respectively.Before treatment in CPP and control groups,the incidence of staccato voiding (20 cases vs 1 case),DSD (22 cases vs 1 case),Q_(max)(10.7?3.7 vs 15.0?4.3ml/s),PVR (7.7?4.1vs 3.2?2.6ml),P_(det,max)(115.1?33.6vs 76.8?16.6cm H_2O)and MUCP(176.5?45.7 vs 86.2?28.5cm H_2O)all showed significant differences between the 2 groups(P＜0.05).In CPP group,the differences in pain(4.6?2.2 vs 2.1?1.6),urination (7.9?2.0vs 2.2?1.7),life impact (9.4?2.2vs 2.6?2.1)and total scores(22.0?5.2vs 7.0?4.2) of NIH-CPSI and Q_(max)(10.7?3.7 vs 14.9?5.6) between pre-and post-biofeedback were significant (P＜0.05).Conclusions The main type of CPP is categoryⅢB.The primary symptom is voiding disorder,which leads to greater psychological stress in patients.Children with CPP have pelvic floor dysfunctions and multiple abnormal urodynamic param- eters.The short-term effect of biofeedback strategies for CPP is satisfactory.

Comparison studies of recombinant prourokinase(pro-UK) in various host cells,and expression vectors were carried out. Expression levels of vectors constructed in this study in different cell lines were compared. Mammalian cells expressing pro-UK were established. The levels of pro-UK expression in recombinant Namalwa, Vero and Sp2/0 cells are 200,12.5 and 50 IU/( 106 cells 24h). pro-UK purities separated by immunoaffinity chromatograph are above 90 %. Immunoabsorbent assay showed the ratio of pro-UK in CHO cells is lower than that from Vero and Namalwa cells. This study provide new host cells for pro-UK production.

To investigate the technological parameters of the isolation and purification of 10-hydroxycamptothecin and vincoside-lactam from Camptotheca acuminata seed by polyamide. The static arid dynamic adsorption characteristics of 10-hydroxycamptothecin and vincoside-lactam on polyamide were studied, and the contents were determined by HPLC. The optimum parameters for adsorption were as follows: the contents of 10-hydroxycamptothecin and vincoside-lactam in the extracts were 0.189 g x L(-1) and 0.334 g x L(-1), respectively, pH 6, flow rate was 1.0 mL x min(-1), processing volume was 3 BV; for desorption: ethanol-water (60:40), flow rate was 1.0 mL x min(-1), 5 BV as an eluent. After treated with polyamide, the contents of 10-hydroxycamptothecin and vincoside-lactam were 17.52% and 32.87%, respectively, the recovery yields were 66.05% and 75.86%, respectively. Results showed that polyamide revealed a good ability to separate 10-hydroxycamptothecin and vincoside-lactam. Therefore, we concluded that results in this study may provide scientific references for the large-scale production of 10-hydroxycamptothecin and vincoside-lactam extracted from C. acuminata seed.
Camptotheca ; chemistry ; Camptothecin ; analogs & derivatives ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Hydrogen-Ion Concentration ; Nylons ; chemistry ; Seeds ; chemistry

OBJECTIVETo summarize the experience in one-stage urethroplasty with pedicled scrotal skin flap for hypospadias, and improve its therapeutic effect.

METHODSWe retrospectively analyzed the clinical data of 310 cases of hypospadias (except coronal hypospadias) treated by one-stage urethroplasty with pedicled scrotal skin flap. All the patients were followed up for 6 to 24 months.

RESULTSNo postoperative complications were observed except urinary fistula, which occurred in 12.6% of the patients. Postoperative fistula formation was associated with the type of hypospadias, the length of the urethral defect and postoperative comprehensive medication, but not with the stent indwelling time after surgery. Most of the fistulae were located at the base of the penis.

CONCLUSIONOne-stage urethroplasty with pedicled scrotal skin flap is a simple and effective option for all types of hypospadias except the coronal type, and postoperative treatment is very important.

Adolescent ; Adult ; Child ; Child, Preschool ; Humans ; Hypospadias ; surgery ; Infant ; Male ; Reconstructive Surgical Procedures ; methods ; Retrospective Studies ; Scrotum ; surgery ; Skin Transplantation ; Surgical Flaps ; Urethra ; surgery ; Young Adult

OBJECTIVETo investigate effect of the serum concentration of epidermal growth factor receptor (EGFR) in the pre- and postoperative peripheral blood in patients with meningiomas.

METHODSUsing ELISA, the EGFR concentration were measured in the pre- and postoperative serum in 53 patients with meningiomas. In this study, the patients were divided into preoperative group, postoperative group (including 42 total resection, 11 subtotal resection), and 28 healthy control.

RESULTSIn 53 patients with meningiomas, concentration of preoperative serum EGFR was (352.93 +/- 66.18) fmol/ml, to show higher than control group (159.11 +/- 40.50) fmol/ml (P < 0.0001); Concentration of postoperative serum EGFR was (220.74 +/- 70.63) fmol/ml, to show lower than preoperative group (P < 0.001). Including in 42 patients with meningomas by total resection, serum EGFRs of the 38 patients were decreased (191.20 +/- 32.13) fmol/ml, the 4 patients with peritumoral edema were decreased (248.75 +/- 10.31) fmol/ml, to show lower than preoperative group (P < 0.001); the 11 patients with subtotal resection were decreased (322.14 +/- 89.53) fmol/ml, not to show different than preoperative group (P < 0.05).

CONCLUSIONSIn 92.16% patients with meningiomas, the more poison of the tumor is resected, the less the serum EGFR concentration is detected postoperatively, while the total resection of mangionmas resulted in a lowest level of EGFR. These results support the concept that human meningiomas may be autocrine EGFRs. The measurement of serum EGFR can be useful to patients with meningiomas for follow-up after surgery.

Adult ; Biomarkers, Tumor ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Male ; Meningeal Neoplasms ; blood ; Meningioma ; blood ; Middle Aged ; Postoperative Period ; Receptor, Epidermal Growth Factor ; blood

OBJECTIVETo discuss whether vascular endothelial growth factor (VEGF) in the peripheral blood can reflect the biological activities of pituitary adenomas.

METHODSThe concentrations of VEGF in peripheral blood were measured with ELISA in 203 patients with pituitary adenomas, 22 patients with pituitary hyperplasia, 7 patients with pituitary Rathke' pouch and 3 patients with pituitary abscess.

RESULTSThe serum VEGF levels were (366.8 +/- 211.1) pg/ml and (286.8 +/- 107.6) pg/ml in patients with pituitary adenomas and pituitary hyperhasia, respectively, which were higher than those in patients with pituitary Rathke' pouch [(180.5 +/- 61.7) pg/ml], patients with pituitary abscess [(147.5 +/- 46.3) pg/ml] and the health control [(180.8 +/- 56.2) pg/ml] (P < 0.05). In patients with pituitary adenomas, the VEGF levels were (380.0 +/- 234.5) pg/ml in macroadenomas and (380.1 +/- 2870.3) pg/ml in giant adenomas, higher than those in microadenomas [(294.6 +/- 111.6) pg/ml] and in pituitary hyperhasia respectively (P < 0.05). The serum VEGF levels were not significantly different in pituitary adenoma in terms of invasive growth, apoplexy, cyst and hormone secretory functions (P > 0.05).

CONCLUSIONSThe upregulation of serum VEGF expression may reflect the biological activities of pituitary adenoma. However, it may not be associated with pituitary Rathke' pouch, pituitary abscess, adenoma with invasiveness, apoplexy, cyst and hormone secretory function. The serum VEGF levels could be helpful in differentiating pituitary adenoma from pituitary Rathke' pouch and pituitary abscess.

Adenoma ; blood ; diagnosis ; Adult ; Diagnosis, Differential ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Hyperplasia ; blood ; diagnosis ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Pituitary Diseases ; blood ; diagnosis ; Pituitary Neoplasms ; blood ; diagnosis ; Vascular Endothelial Growth Factor A ; blood

OBJECTIVEUsing HPLC To determine hypoxanthine in co-hirudo injection for establishing its HPLC fingerprint, and evaluating its internal quality.

METHODThe chromatographic separation was performed on a Kromasil C18 column (4.6 mm x 250 mm,5 microm). A linear gradient elution with A (0.01 mol x L(-1) x KH2PO4) and B (50% methanol) was used, the flow rate was 0.8 mL x min(-1), the detection wavelength was set at 254 nm, and the column temperature was at normal.

RESULTHypoxanthine was used as the reference substance in the fingerprint of co-hirudo injection, it showed 15 common peaks and theirs similarity threshod was 0.97.

CONCLUSIONThis method was accurate, repeatable and useful for the quality control of co-hirudo injection.

Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Hypoxanthine ; chemistry ; Leeches ; chemistry ; Reproducibility of Results

Adult ; Chylous Ascites ; therapy ; Female ; Fibrin Tissue Adhesive ; therapeutic use ; Humans ; Middle Aged ; Parenteral Nutrition, Total ; Somatostatin ; therapeutic use

AIM:To investigate the synergistic effect of imperatorin on enhancing the anti -tumor effect of TNF-related apoptosis-inducing ligand(TRAIL)on breast cancer and the mechanisms.METHODS:T-47D and MCF-7 breast cancer cells were divided into control group ,imperatorin group,TRAIL group,imperatorin+TRAIL group and imperatorin+TRAIL+death receptor 5(DR5)siRNA group.The viability of T-47D and MCF-7 cells was measured by MTT assay. The apoptosis and mitochondrial membrane potential in T-47D cells were analyzed by flow cytometry.Western blot and flow cytometry analysis were performed to evaluate the expression of DR 5 on T-47D cell surface and the activation of caspase-8 and caspase-3.RESULTS:Imperatorin significantly enhanced the inhibition of cell viability induced by TRAIL of T -47D and MCF-7 cells,and significantly increased the apoptosis of T-47D cells induced by TRAIL.Imperatorin treatment ob-viously induced upregulation of DR5 expression and production of reactive oxygen species in the T-47D cells.In addition,imperatorin enhanced the TRAIL-induced damage of mitochondrial membrane potential and activation of caspase -8 and caspase-3.CONCLUSION:Imperatorin enhances the anti-tumor effect of TRAIL on breast cancer via upregulating the ex-pression of DR5.