Objective： To discover HER2/neu tyrosine kinase inhibitor through computer-aided drug design approach. Methods： The three-dimensional(3D) structure of HER2/neu and EGFR tyrosine kinase domain was modeled using MODERLAR software. To search database and pick up candidate compounds based 3D structure of HER2/neu tyrosine kinase domain. Inhibition of HER2/neu tyrosine kinase phosphorylation by the compounds was detected using Western blot analysis. Inhibition of cell proliferation was determined by MTT assay. Results： The amino acid identity and similarity between HER2/neu and EGFR kinase domain, insulin receptor kinase domain, FGFR1 kinase domain, Src kinase domain were compared. The identity was 35％ -41％ and the similarity was 52％ -55％ . The 3D structure of HER2/neu and EGFR was obtained by MODERLAR software. Through searching, screening and optimization, the authors found that ST2325 had significantly inhibitory effect on HER2/neu tyrosine kinase phosphorylation with IC50 of 6.6 μ mol/L. The inhibition of HER2/neu phosphorylation was selective and reversible. ST2325 inhibited cell proliferation of HER2/neu-overexpressing MDA-MB-453m1 preferentially compared with EGFR-overexpressing MDA- MB-468, The IC50 values were 29.05 and 60.4 μ mol/L. Conclusion： ST2325 which was discovered through structure-based approach had notable inhibitory effect on HER2/neu tyrosine kinase.

AIMTo isolate and identify a glucuronide metabolite of SFZ-47 [3H-1,2-dihydro-2-(4-methyl-phenylamino)methyl-1-pyrrolizinone], which is difficult to synthesize because it undergoes hydrolysis and intramolecular acyl migration at physiological pH, in rabbit urine.

METHODSTwo rabbits were ig 200 mg doses of SFZ-47. Urine was collected for 24 h, adjusted to pH 4.0 with acetic acid and lyophilized. The residues were reconstituted in 25 mL methanol and centrifuged at 5,000 r.min-1 for 10 min. The supernatant was filtered (0.45 micron) and then isolated with semi-preparative reversed phase HPLC. The eluent collected from individual peaks was evaporated by rotary evaporation and freeze-drying. Compounds were then identified with electrospray ion trap mass spectrometry and 1HNMR spectroscopy.

RESULTSThe 1HNMR and ESI-MSn results indicate that the metabolite is the 1-O-acyl beta-D-glucuronide conjugate of 4-(3H-1,2-dihydro-1-pyrrolizinone-2-methylamino) benzoic acid.

CONCLUSIONThis method was shown to be rapid and simple and gave excellent resolution from endogenous constituents in urine, and it is suitable for preparation of the glucuronide metabolites of SFZ-47 and its analogues.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; metabolism ; urine ; Chromatography, High Pressure Liquid ; Male ; Molecular Structure ; Pyrroles ; chemistry ; metabolism ; urine ; Rabbits ; Spectrometry, Mass, Electrospray Ionization

Adenocarcinoma ; diagnosis ; Biomarkers, Tumor ; metabolism ; Carcinosarcoma ; diagnosis ; Gastrointestinal Stromal Tumors ; metabolism ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Neoplasm Metastasis ; Prostate ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; surgery ; Stromal Cells ; pathology ; Treatment Outcome

Objective To investigate the clinical diagnosis,treatment and prevention of fallopian tube prolapse(FTP)after hysterectomy.Methods A total of 7949 patients received hysterectomy from 1983 to Aug 2005 in Peking Union Medical College Hospital,including 6229 cases of trans-abdominal hysterectomy(TAH),780 cases of transvaginal hysterectomy(TVH),and 940 cases of laparoscopic assisted vaginal hysterectomy(LAVH).Nine cases(including 1 case from other hospital)of FTP after hysterectomy were analyzed retrospectively for their symptoms,diagnosis and treatment.All of them were diagnosed according to the results of histology and follow-up.Results The overall incidence of FTP after hysterectomy was 0.11%(9/7949).Incidence of FTP after trans-abdominal hysterectomy was 0.08% (5/6229),after vaginal hysterectomy 0.51%(4/780),and after laparoscopic assisted vaginal hysterectomy 0(0/940).There were no symptoms in 3 cases,but the other 6 cases had symptoms.The pelvic examination revealed the typical prolapsed fimbrial end of a fallopian tube in 3 cases and red granulation tissue in the other 6 cases.All of them were excised vaginally and cauterized.The results were confirmed by histological examination.No recurrent cases were reported in follow up.Conclusions FTP is a rare complication after hysterectomy.The prognosis is well after proper diagnosis and treatment.Salpingectomy or fixation of accessories into the pelvic wall and complete peritonealisation at the time of hysterectomy are important methods to prevent FTP after hysterectomy.

Objective To establish the chick embryo chorioallantioc membrane(CAM)as a model for in vivo research on endometriosis.The model was used to investigate the mechanism of anti-vascular endothelial growth factor(VEGF)antibody for treatment of endometriosis.Methods Human endometrial fragments were explanted onto the CAM.Then anti-VEGF antibody was used for the endometriosis-like lesions after transplantation of human endometrial fragments.The CAM models were treated respectively as control groups and experimental groups.The terminal deoxynucleotidyl transferase-mediated biotin- deoxyuridine triphosphate(dUTP)nick end labeling(TUNEL),proliferating cell nuclear antigen(PCNA) and microvessel density(MVD)were used in vivo for analysis of anti-angiogenesis.Results The apoptosis intensity of anti-VEGF antibody treated groups(6.7?0,9,6.9?0.8)was significantly higher than that of the control groups(5.0?0.9,5.4?1.1;P

OBJECTIVETo explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on the vitality and toxicity of E. coli ATCC 25922, and to analyze the potential mechanism.

METHODS(1) In vitro experiments. Standard strains of E. coli ATCC 25922 were divided into groups A (without addition), B, C, D, and E according to the random number table, and then the latter 4 groups were respectively cultured with 1.2 mmol/L CORM-2, 1.6 mmol/L CORM-2, 1.2 mmol/L inactive CORM-2 (iCORM-2), 1.6 mmol/L iCORM-2, with six samples in each group. After being cultured for 0, 3, 5, 8, 10, 12, 16, 20, 24, 27, 30, 48 hours, proliferative vitality of E. coli was examined (denoted as absorption value under 600 nm wavelength), and bacteria number was counted. Other standard strains of E. coli ATCC 25922 were divided into groups F (without addition) and G (cultured with 0.8 mmol/L CORM-2), the expressions of genes fliA, dnaK, marA, and waaQ related to E. coli were detected by quantitative real-time (qRT)-PCR. (2) In vivo experiments. Other standard strains of E. coli ATCC 25922 were grouped as A', B', C', D', and E' and treated with the same method as that in groups A, B, C, D, and E, and 0.5 mL bacterial liquid of each group were collected when the absorption value of bacterial liquid in group A' was equal to 0.4 (under 600 nm wavelength). Seventy-two C57BL/6 mice were divided into groups, namely blank control (without treatment), H, I, J, K, and L according to the random number table, with 12 mice in each group. The mice in the latter 5 groups were intraperitoneally injected with 0.5 mL bacterial suspension of groups A', B', C', D', and E' respectively. After injection, general condition of mice in groups H, I, J, K, and L was observed. The serum levels of TNF-α and IL-6 were determined at post injection hour (PIH) 6, 12. The liver and lung samples were harvested for determination of myeloperoxidase (MPO) activity at PIH 12. The same process was carried out in blank control group. Data were processed with repeated measure analysis of variance (ANOVA), factorial design ANOVA, one-way ANOVA, and t test.

RESULTS(1) In vitro experiments. Compared with those of groups A and D, the proliferative vitality and bacteria number of E. coli in group B were all decreased (with F values respectively 1170.80, 217.52, P values all below 0.01). Compared with those of groups A and E, the proliferative vitality and bacteria number of E. coli in group C were also obviously decreased (with F values respectively 7948.34, 14 432.85, P values all below 0.01). Compared with those in group F, the expression of fliA was downregulated, while the expressions of dnaK, marA, and waaQ were upregulated in group G (with t values 30.28, -165.54, -168.88, -187.28, P values all below 0.01). (2) In vivo experiments. Symptoms including listlessness and tachypnea were observed in mice in groups H, K, and L, and they were ameliorated or not obvious in groups I and J. At PIH 6, the serum levels of TNF-α and IL-6 in groups H and K were respectively (647.3 ± 3.8) pg/mL, (3.44 ± 0.22) ng/mL and (639.3 ± 0.8) pg/mL, (2.47 ± 0.32) ng/mL, which were obviously higher than those in group I [(124.6 ± 10.7) pg/mL, (1.03 ± 0.16) ng/mL, with t values from 15.22 to 84.03, P values all below 0.01]. The serum levels of TNF-α and IL-6 in group J at PIH 6, 12 were also obviously decreased as compared with those in groups H and L (with t values from 19.27 to 245.34, P values all below 0.01). MPO activity of liver and lung tissues were significantly attenuated in group I at PIH 12 as compared with those in groups H and K, and it was also attenuated in group J when compared with those in groups H and L (with t values respectively from 17.36 to 18.92 and 2.35 to 3.61, P values all below 0.01).

CONCLUSIONSCORM-2 can obviously inhibit the vitality and toxicity of E. coli, which might be attributable to regulation of expressions of genes fliA, dnaK, marA, and waaQ of E. coli.

Animals ; Carbon Monoxide ; metabolism ; DNA-Binding Proteins ; metabolism ; Escherichia coli ; drug effects ; metabolism ; physiology ; Escherichia coli Proteins ; metabolism ; Glycosyltransferases ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Interleukin-6 ; blood ; Liver ; enzymology ; Lung ; enzymology ; Mice ; Mice, Inbred C57BL ; Organometallic Compounds ; pharmacology ; Peroxidase ; metabolism ; Sigma Factor ; metabolism ; Tumor Necrosis Factor-alpha ; blood

Objective To study the clinical value of serum cytokeratin 19 fragment (CYFRA21-1) as a biomarker in evaluating its prognosis,monitoring and follow-up of the postoperative patients with non- small cell lung cancer (NSCLC).Methods Serum CYFRA21-1 was determined by radioimmunoassay for 207 patients with NSCLC before and after surgical operation at Tongji Hospital,Shanghai.Relationship between serum CYFRA21-1 and the prognosis,recurrence and metastasis of lung cancer was analyzed retrospectively.Results The patients were followed-up for 37 months in average.Preoperative serum CYFRA21-1 was positive in 42.0% (87/207) of all the cases,48.8% (60/123) of squamous cell earcinama and 34.6% (27/78) of adenocarcinoma,with statistical significance (X~2=3.901,P

OBJECTIVETo explore a method of extracting tumor interstitial fluid (TIF) which is similar to muddy phlegm in Chinese medicine (CM), interleukin-8 (IL-8) in concentration was taken as the representative of the content of TIF, analyzed in the extracted TIF and the original tumor tissue, and examined to see whether TIF has an interfering effect on tumor recurrence.

METHODSTumor tissue was ground, centrifuged, and filtered for intercellular substances. Tumor-bearing Kunming S180 mice were raised for 21 days and then the tumors were removed to observe the influence of intervention with TIF, normal saline (NS) and a blank control on tumor recurrence.

RESULTSThe content of IL-8 in the filtered and unfiltered tumor tissue was not significantly different (P>0.05). Postoperative tumor recurrence in TIF intervention group was significantly higher than that in the NS intervention and control groups (60%, 12/20 vs. 20%, 4/20. vs. 15%, 3/20, χ(2) =11.058, P<0.01). Tumor cells grew vigorously and infiltrated to muscular tissue in TIF intervention group. Large numbers of tumor cells were seen necrotic in the NS intervention group, and small numbers of tumor cells were seen necrotic in the blank control group.

CONCLUSIONSTIF can be effectively extracted by the means described. It does not contain tumor cells, but its contents such as IL-8 may stimulate tumor cell growth and promote postoperative tumor recurrence, which provided preliminary experimental basis for hypothesis of "tumor-phlegm microenvironment".

Animals ; Enzyme-Linked Immunosorbent Assay ; Extracellular Fluid ; Interleukin-8 ; analysis ; Mice ; Neoplasms, Experimental ; pathology ; Postoperative Period ; Recurrence

OBJECTIVETo study the changes of IL-12 produced by dendritic cells in peripheral blood in children with Henoch-Schonlein purpura (HSP), and to explore its influence on TH1/TH2 balance in order to elucidate its significance in the pathogenesis of HSP.

METHODSThe levels of interferon-gamma (IFN-gamma), interleukin-4 (IL-4) and interleukin-12 (IL-12) in plasma were determined by ELISA in 60 HSP children (HSP group) and 21 healthy children (Control group). Peripheral blood mononuclear cells (PBMC) of 22 HSP patients and 21 healthy children were cultured in vitro and then were transformed into dendritic cells. The levels of IL-12 in the supernatant were detected by ELISA and the positive expression rate of CD1a(+) was detected by indirect immunofluorescence procedure.

RESULTS1) The levels of IFN-gamma and the ratio of IFN-gamma/IL-4 in plasma of the HSP group were lower than those of the Control group (IFN-gamma 30.59 +/- 11.27 pg/mL vs 43.38 +/- 19.19 pg/mL; IFN-gamma/IL-4 ratio 0.70 +/- 0.28 vs 1.33 +/- 0.57) (P < 0.01). The levels of IL-12 in the HSP group were also lower than those of the Control group (153.95 +/- 91.88 pg/mL vs 323.06 +/- 162.34 pg/mL; P < 0.01). In contrast, the levels of IL-4 were higher than those of the Control group (45.08 +/- 9.19 pg/mL vs 32.95 +/- 7.10 pg/mL; P < 0.01). The plasma levels of IL-12 positively correlated with the IFN-gamma levels (r=0.52, P < 0.01) and the ratio of IFN-gamma/IL-4 (r=0.43, P < 0.01) in the HSP group. 2) The IL-12 levels in the supernatant of the HSP group were lower than those of the Control group (357.06 +/- 153.56 pg/mL vs 489.80 +/- 213.45 pg/mL; P < 0.05), and had a positive correlation with the plasma IL-12 levels (r=0.74, P < 0.01). 3) The positive expression rate of CD1a(+) of the HSP group was lower than that of the Control group [(27.42 +/- 10.75)% vs (35.68 +/- 12.18)%; P < 0.05], and positively correlated with the IL-12 levels in the supernatants (r=0.57, P < 0.01) and in plasma (r=0.68, P < 0.01).

CONCLUSIONSThere was an imbalance of TH1/TH2 in HSP children. The decrease of TH1 function had a positive correlation with the low levels of IL-12 in plasma, while the latter correlated closely with decreased number and / or function of dendritic cells, suggesting that the decreased number and / or function of dendritic cells in peripheral blood resulted in the imbalance of TH1/TH2 indirectly.

Adolescent ; Antigens, CD1 ; analysis ; Cells, Cultured ; Child ; Child, Preschool ; Dendritic Cells ; immunology ; Female ; Humans ; Interferon-gamma ; blood ; Interleukin-12 ; biosynthesis ; blood ; Interleukin-4 ; blood ; Male ; Purpura, Schoenlein-Henoch ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

OBJECTIVESThe investigation of the testicular volume, the penis length and the T, FSH, LH, PRL levels in serum were taken in 289 adolescent males to provide the valuable data for andrology.

METHODSThe adolescent males were grouped according to their age. The testicular volume was measured with testicular model and the T, FSH, LH, PRL levels in serum were determined by immunoenzymetric assay.

RESULTSThe male sexual development was rapid from age 11 to 16 and close to that of adult at age 18. Serum PRL of adolescent males was higher than that of adult males.

CONCLUSIONSThe age 11 to 16 is a period of rapid growth in sexual maturation. PRL may play an important role in sexual maturation.

Adolescent ; Body Height ; Body Weight ; Gonadal Steroid Hormones ; blood ; Humans ; Male ; Penis ; physiology ; Testis ; physiology