A new method of establishing orthotopic bladder transplantable tumor in mice.

OBJECTIVE: The present study aims to find a convenient, rapid, and stable method to establish bladder tumor in mice. METHODS: Female Balb/C-nu-nu nude mice (or female T739 mice) were narcotized by sodium pentobarbital at a dosage of 60 mg/kg. The stylet of the 24# venous retention needles was bent in a 5° to 7° angle at a distance of 15 mm from the needlepoint to form a circle with 2.61 mm to 3.66 mm radius when the stylet is rotated. The pipe casing was lubricated with liquid paraffin, and inserted into the bladder cavity. The drift angle stylet was inserted into the pipe casing slowly, rotated for five times, and then pulled out. A cell suspension (0.1 mL) of approximately 1×10(6) T24 cells (or BTT cells) was then injected immediately. RESULTS: A total of 60 T739 mice and 60 Balb/C-nu-nu nude mice were inoculated with BTT cells and T24 cells, respectively. The bladder tumor incidence and the average survival time of the tumor-bearing mice were 100% and (26.69±9.24) d and 100% and (34.59±9.8) d for the T739 mice and Balb/C-nu-nu nude mice, respectively. CONCLUSIONS: Using the drift angle stylet to injure the mucous membrane of the urinary bladder can establish a stable bladder transplantable tumor model in mice.

[Expression and clinical significance of nuclear matrix protein 22 and cytokeratin 18 in transitional cell carcinoma of the bladder].

OBJECTIVE: To evaluate the expression and clinical significance of urinary nuclear matrix protein (NMP22) and cytokeratin 18 (CK18) for transitional cell carcinoma of the bladder. METHODS: Urinary NMP22 and CK18 levels of 293 patients with transitional cell carcinoma of the bladder, 400 patients with non-transitional cell carcinoma of the bladder, and 105 bladder benign disease were analysed by enzyme-linked-immunosorbent assay (ELISA). RESULTS: The levels of urinary NMP22 and CK18 in the patients with transitional cell carcinoma of the bladder (M = 17.3 U/ml, M(CK18) = 484.2 U/L) were significantly higher than those in the non-transitional cell carcinoma of the bladder (M = 6.8 U/ml, M(CK18) = 156.0 U/L) and the benign disease group (M(NMP22) = 2.3 U/ml, M(CK18) = 66.6 U/L) (P < 0.001). The sensitivity and specificity of urinary NMP22 and CK18 were 79.2%, 88.6% and 78.2%, 82.9%, respectively, for transitional cell carcinoma of the bladder before any treatment. The joint sensitivity of the two markers was 91.7%. The NMP22 and CK18 levels were significantly lower in the recovered patients after surgical operation (P < 0.01), while in patients with recurrence or metastasis the levels of the markers were significantly higher (P < 0.01). There was a significant relationship between NMP22 and CK18, (r = 0.689, P < 0.01). The levels of urinary nmp22 and CK18 were significantly different among pathological grade G1, G2, G3, and stage Ta, T1, T2, T3 (P < 0.01). CONCLUSION: NMP22 and CK18 are useful tumor marker for diagnosis of transitional cell carcinoma of the bladder and for monitoring the state of illness. The joint use of the two markers can improve the sensitivity of cancer detection. NMP22 and CK18 may become a new class of tumor markers, and to be the basis for development of a new assay with an increased efficacy for the detection and treatment of bladder cancer.

[Testosterone induces different-featured prostate hyperplasia in castrated and uncastrated mice].

OBJECTIVE: To study the different features of hyperplasia in castrated and uncastrated mice after testosterone (T) treatment. METHODS: Forty-eight BALB/c mice were randomly divided into 6 groups of 8 in each: castrated (A), uncastrated (B) , castrated + low T (C), uncastrated + low T (D), castrated + high T (E), uncastrated + high T (F). Groups C and D were treated with testosterone solution at the dose of 12.5 mg/(kg d) and Groups E and F at 125 mg/(kg d) for 20 consecutive days, while Groups A and B received saline only. All the mice were sacrificed on the 21st day, their ventral and dorsal prostate glands weighed and their pathological features studied. RESULTS: Atrophic prostates were observed in Group A, but normal in Group B; prostatic hyperplasia was found in both Group C and D, but more obvious in the latter (P <0.05); and a slightly higher degree of hyperplasia was noted in Groups E and F than in C and D. There was an increase in serum T and vascular endothelial growth factor (VEGF) concentration and a decrease in serum estrogen (E2) concentration in the testosterone treated groups. CONCLUSION: Both castrated and uncastrated mice develop prostate hyperplasia after short-term testosterone treatment, although in different degrees and with different features, which may help further the studies on the association of castration and androgen with prostate diseases.

[Human chorionic gonadotrophin regulates epidermal growth factor in the phallus of hypospadias mice].

OBJECTIVE: To investigate the etiopathogenesis of congenital hypospadias and the regulating effect of exogenous human chorionic gonadotrophin (hCG) on the epidermal growth factor (EGF) in the phallus of hypospadiac mice. METHODS: Mouse models of congenital hypospadias were established. Fifty healthy male mice randomly selected as normal controls received intraperitoneal injection of normal saline, and another 50 with hypospadias were equally divided into an experimental control group, intraperitoneally injected with 1 ml normal saline, and 4 hCG dose groups treated by hCG intraperitoneal injection at 50 IU, 100 IU, 150 IU and 200 IU respectively for 7 consecutive days. The concentrations of EGF in the phallus and serum were detected in different groups by ELISA. RESULTS: The concentrations of EGF in the phallus were significantly lower in the hypospadias rats than in the normal controls (P < 0.05), the 150 IU and 200 IU hCG groups showing significant differences from the 50 IU, 100 IU and experimental control groups (P < 0.05), as the 50 IU, 100 IU and experimental control groups from the normal control (P < 0.05). But no obvious difference was found in EGF concentration in the serum among different groups (P > 0.05). CONCLUSION: Nonsteroidal antiandrogen and decreased concentration of EGF in the mouse phallus may be associated with the etiology of hypospadias. And exogenous hCG at 150 IU or 200 IU can increase the concentration of EGF in the phallus of mice with congenital hypospadias.

[The diagnosis and monitoring of transitional cell cancer of the urinary tract through nuclear matrix protein 22].

OBJECTIVE: To evaluate the urinary nuclear matrix protein (NMP22) as an adjuvant diagnostic index for transitional cell carcinoma of urinary tract and monitoring the state of disease. METHODS: Urinary samples were collected from 262 patients with transitional cell carcinoma, 198 non-transitional cell carcinoma of the urinary tract and 65 patients with benign diseases. Urinary NMP22 concentration was determined through enzyme linked immunosorbent assay (ELISA). RESULTS: The urinary NMP22 concentration had significant difference among the three groups (Kruskal Wallis, chi(2) = 197.17 P < 0.001). The detection sensitivity and specificity of urinary NMP22 to transitional cell carcinoma were 71.37% and 87.69% respectively. The NMP22 concentration showed significant difference among three groups divided according to the pathological grade (Kruskal-Wallis test, chi(2) = 34.06 P < 0.01). The NMP22 concentration was significant lower in the recovery patients after the operation than the peoples of pre-operation and recurrence (Kruskal-Wallis test, chi(2) = 37.53, P < 0.001). CONCLUSION: MP22 is a helpful tumor marker for the diagnosis of transitional cell carcinoma and monitoring the state of illness with increased efficacy.

[Nifedipine modifies plasma membrane functional integrity and inhibits osmosensitive calcium influx in human sperm].

OBJECTIVE: To investigate the effect of nifedipine of therapeutic dosage on the plasma membrane functional integrity and osmosensitive calcium influx in human sperm in vitro. METHODS: Sperm samples were aseptically obtained from 10 healthy fertile men by masturbation and prepared by swim-up technique to produce a spermatozoal solution of high motility. The solution was then incubated with nifedipine of 20, 100 and 20 x 10(3) ng/ml respectively at 37 degrees C in vitro. The hypo-osmotic swelling (HOS) test was done to assess the sperm function. Intracellular calcium concentration was measured by fluorescent probe fura-2/AM before and after sperm medium dilution in distilled water. RESULTS: The 20 x 103 ng/ml group showed significantly lower HOS scores than the control (P < 0.01). The 20, 100 and 20 x 10(3) ng/ml groups all showed significantly lower Ca2+ fluorescence D-value than the control (P < 0.01). CONCLUSION: Nifedipine can modify plasma membrane functional integrity and inhibit osmosensitive calcium influx in human sperm and affect male fertility in vitro in therapeutic dose.