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

BACKGROUNDSustained hypotension during hemodialysis (HD) is an important clinical issue. Plasma adrenomedullin (AM) is increased in HD patients with sustained hypotension, but little is known about whether removing AM can improve hypotension. The objective of this study was to investigate the beneficial effects of hemodialysis using a high-flux dialyzer on removal of increased plasma AM levels and improving low blood pressure in elderly HD patients with sustained hypotension.

METHODSForty-eight elderly patients (age 65 or older) who had undergone maintenance HD for more than one year were recruited and studied. We evaluated plasma levels of AM in sustained hypotension (SH; n = 28) and normotensive (NT; n = 20) patients. The patients with hypotension were further divided into two subgroups and treated with either high-flux dialyzer or low-flux dialyzer for 3 months. Plasma adrenomedullin levels and blood pressure were analyzed at days 0 and 181.

RESULTSPlasma levels of AM were significantly higher in SH than in NT patients ((24.92 ± 3.7) ng/L vs. (15.52 ± 6.01) ng/L, P < 0.05), and were inversely correlated with mean arterial blood pressure (MAP) at pre-HD. After 3 months, the level of plasma AM in high-flux group was decreased ((24.58 ± 4.36) ng/L vs. (16.18 ± 5.08) ng/L, P < 0.05), but MAP was increased ((67.37 ± 4.31) mmHg vs. (74.79 ± 3.59) mmHg, P < 0.05). There was no obvious change in low-flux group.

CONCLUSIONSPlasma AM levels were significantly elevated in elderly HD patients with SH. High-flux dialyzer therapy can decrease plasma AM level and improve hypotension.

Adrenomedullin ; blood ; Aged ; Female ; Humans ; Hypotension ; blood ; Male ; Renal Dialysis

OBJECTIVETo investigate the effect of reverse saphenous nerve neurocutaneous flaps for skin defects at forepart of feet.

METHODSFrom January 2004 to October 2008,15 cases of skin defects at forepart of feet were repaired with reverse saphenous nerve neurocutaneous flaps. The flap size ranged from 3.5 cm x 3.0 cm to 8 cm x 5 cm. The wounds at donor site were closed with skin graft.

RESULTSAll the flaps survived completely with no ulcer at the donor site. 10 patients were followed up for 1 to approximately 9 months. The skin color and texture were satisfactory. The patients could walk very well.

CONCLUSIONSIt is reliable to repair the skin defects at forepart of feet with reverse saphenous nerve neurocutaneous flaps. It is easily performed with less morbidity.

Adolescent ; Adult ; Child ; Female ; Foot Injuries ; surgery ; Humans ; Male ; Skin ; injuries ; Skin Transplantation ; Surgical Flaps ; blood supply ; innervation ; Young Adult

OBJECTIVETo explore the apoptosis and the express of proliferating cell nuclear antigen (PCNA) in spermatogenic cells, and study generation function of the rabbit after scrotal reconstruction with flaps.

METHODSThe 48 New Zealand white rabbits were randomly divided into three groups as experimental group (the scrotum reconstructed with flaps, n = 48), the control group (the sham operated group, n = 24) and the blank group (n = 18). The apoptosis and the expression of PCNA in the spermatogenic cells were detected with TUNEL and the immunohistochemistry from the 3rd to the 8th week after operation. 8 weeks later, 12 animals in each group were fed respectively with one female rabbit to observe the procreation.

RESULTSThe apoptotic index of the spermatogenic cells in blank group was 7.73 +/- 4.95. 3 weeks after operation, the apoptotic index of spermatogenic cells was 22.59 +/- 3.04 in the experimental group, and 21.13 +/- 1.68 in control group, showing no significant difference between the two groups (P > 0.05). 8 weeks after operation, the apoptotic index of spermatogenic cells was 71.85 +/- 2.69 in the experimental group, and 13.64 +/- 2.09 in control group, show a significant difference between them (P < 0.05). The apoptotic index in experimental group increased gradually from the 3rd to 8th week after scrotal reconstruction , which was markedly higher than that in the blank group (P < 0.05). The apoptotic index in control group was higher than that in the blank group at the 3rd week (P < 0.05), but not at the 8th week (P > 0.05). The proliferation index of spermatogenic cells was 9.32 +/- 9.30 and 12.52 +/- 3.87 in experimental group at the 3rd and 4th week, respectively, which was significantly lower than that in blank group (43.07 +/- 2.25) and control group (45.69 +/- 4.98) at the 3rd week (P < 0.05). The proliferation index of spermatogenic cells was 46.98 +/- 18.92 and 49.53 +/- 9.79 in experimental group at the 7th and 8th week, respectively, 39.90 +/- 5.10 in control group at the 8th week, showing no difference between the two groups (P > 0.05). The proliferation index of spermatogenic cells in control group at the 3rd and 8th week was not different from that in the blank group (P > 0.05). The female pairing rabbits in the blank and control group were all pregnant, and the average childbirths were 6.0 +/- 1.28 and 5.92 +/- 1.31 respectively, with no difference between the two groups (P > 0.05). All the female pairing rabbits in the experimental group were not pregnant, showing a significant difference from those in the blank and control group (P < 0.05).

CONCLUSIONSThe rabbit generation functional disturbance after scrotal reconstruction with flaps is due to the excessive apoptosis of spermatogenic cell. The spermatogenic cell proliferation is affected only in the early postoperative period, but can recover later.

Animals ; Apoptosis ; Cell Proliferation ; Male ; Proliferating Cell Nuclear Antigen ; metabolism ; Rabbits ; Scrotum ; surgery ; Seminiferous Epithelium ; cytology ; Skin Transplantation ; Surgical Flaps