OBJECTIVETo produce recombinant human prostate-specific antigen (PSA) by molecular cloning technology and to identify its activity.

METHODSThe human PSA cDNA and PET-12a vector were digested by NdeI and BamH1 before ligated by T4 ligase. The correct sequence was verified and transformed into high competent E. coli BL21 (DE3). Recombinant PSA was expressed and purified by hydrophobic interaction phenyl Sepharose column and activated by trypsin digestion. Enzymatic activation assay was done by hydrolysis of the substrate S-2586 and semenogelin.

RESULTSNon-active recombinant PSA was digested by trypsin and demonstrated enzyme activity. The activated PSA hydrolyzed S-2586 and its physiological substrate semenogelin (Sg).

CONCLUSIONRecombinant pro-PSA can be an active serine protease by trypsin digestion and demonstrate native PSA enzymatic activity.

Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; genetics ; Escherichia coli ; genetics ; Gene Expression ; Humans ; Hydrolysis ; Male ; Oligopeptides ; metabolism ; Prostate-Specific Antigen ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; isolation & purification ; metabolism ; Seminal Vesicle Secretory Proteins ; metabolism ; Trypsin ; metabolism

OBJECTIVETo establish a mouse model of hypospadias induced by benzoate estradiol to further the studies on the molecular mechanisms of hypospadias.

METHODSA total of pregnant mice were randomly divided into 5 groups, Group A, B, C, D and E, and injected subcutaneously (sc) with estradiol benzoate at the dose of 0, 0.2, 1, 5 and 25 mg x kg(-1) d(-1) respectively from the 12th to the 16th gestational day. The mortality of the newborn mice was recorded and the male neonates of 2 pregnant mice from each group were anatomized to observe the testis position and prostate agenesis on the delivery day. Examinations were made for urethra and cryptorchidism on the 28th postnatal day.

RESULTSThe death rates of the neonates in Group A, B, C, D and E were 21.6%, 21.5%, 41.4%, 56. 6% and 75.0%, respectively. Hypospadias was detected in Group C (3.3%, 1/30), D (20.0%, 4/20) and E (23.0%, 3/13), with significant difference between Group D and A (P < 0.05) and E and A (P < 0.05), but not between Group D and E (P > 0.05). Cryptorchidism was found in Group C (6.6%, 2/30) , D (30.0%, 6/20) and E (61.6%, 8/13), with significant difference between Group D and A (P < 0.05) and E and A (P < 0.05) , but not between Group D and E (P >0.05).

CONCLUSIONExposure of pregnant mice to large dose of estradiol benzoate can induce hypospadias and cryptorchidism in their neonates. And the right dose of estradiol benzoate for the establishment of the mouse model of hypospadias should be 5 mg x kg(-1) x d(-1).

Animals ; Animals, Newborn ; Disease Models, Animal ; Dose-Response Relationship, Drug ; Estradiol ; analogs & derivatives ; toxicity ; Female ; Hypospadias ; chemically induced ; Male ; Mice ; Mice, Inbred ICR ; Pregnancy ; Random Allocation

OBJECTIVETo study the effect of combined androgen block therapy on hemoglobin (Hb) and hematocrit value (Ht) in patients with prostate cancer.

METHODSOne hundred and thirty-six patients with adenocarcinoma of the prostate were treated with combined androgen block (orchiectomy and flutamide 250 mg, Tid). Complete blood counts were detected before initiation and after 1, 2, 3, 6, 9 and 12 months of therapy.

RESULTSHb level declined significantly in all patients from a mean baseline of (136 +/- 14) g/L to (126 +/- 16) g/L, (121 +/- 14) g/L, (120 +/- 15) g/L, (113 +/- 12) g/L, (121 +/- 13) g/L and (123 +/- 15) g/L at 1, 2, 3, 6, 9 and 12 months. Ht decreased from a mean baseline of 0.424 +/- 0.041 to 0.390 +/- 0.038, 0.381 +/- 0.042, 0.378 +/- 0.038, 0.366 +/- 0.041, 0.384 +/- 0.039 and 0.387 +/- 0.040. The differences between Hb, Ht before and after treatment were significant (P < 0.05).

CONCLUSIONPatients with prostate cancer being treated with combined androgen block would develop a significant degree of anemia. Hemoglobin and hematocrit level should be monitored periodically. This kind of anemia can be treated by recombinant human erythropoietin.

Aged ; Aged, 80 and over ; Androgen Antagonists ; adverse effects ; Anemia ; chemically induced ; Hematocrit ; Hemoglobins ; analysis ; Humans ; Male ; Middle Aged ; Prostatic Neoplasms ; blood ; drug therapy

OBJECTIVETo further study gene expression and characterization of voltage-dependent anion channels (VDACs) on human spermatozoa.

METHODSVDACs were cloned by PCR from the testis cDNA library. Recombinant human sperm VDACs were produced in E. coli system by molecular cloning technology. Sperm membrane protein was extracted by 1% Triton X-100 and separated by chloroform/methanol.

RESULTSThe gene expression of VDACs was found in the human testis cDNA library and VDAC protein was detected located on the sperm membrane by alpha-helix.

CONCLUSIONVDAC proteins, abundant on the human sperm membrane and responsible for anion transportation, play an important role in sperm signaling transduction and fertility.

Blotting, Western ; Gene Expression ; Humans ; Male ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Signal Transduction ; physiology ; Spermatozoa ; metabolism ; Testis ; metabolism ; Voltage-Dependent Anion Channels ; biosynthesis ; physiology

OBJECTIVESTo study the effect of acute urinary retention on the serum prostate-specific antigen (PSA) concentration.

METHODSBlood samples from 34 benign prostatic hyperplasia (BPH) patients with acute urinary retention were drawn immediately before suprapubic cystomy and 48 hours after relief of urinary retention. Serum PSA concentrations were measured with radioimmunoassay.

RESULTSThe mean serum PSA levels of BPH patients with acute urinary retention was (24.6 +/- 16.1) micrograms/L (range from 2.6 micrograms/L to 45.8 micrograms/L). Forty-eight hours after relief of urinary retention, the mean serum PSA levels declined to (9.4 +/- 6.3) micrograms/L (range from 1.7 micrograms/L to 16.6 micrograms/L). The difference was significant (P < 0.01).

CONCLUSIONSAcute urinary retention could dramatically increase the serum PSA value of patients with BPH. After relief of the urinary retention, the patients had a great than 50% decreased of PSA values.

Acute Disease ; Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostate-Specific Antigen ; blood ; Prostatic Hyperplasia ; blood ; Urinary Retention ; blood

Objective To improve the diagnosis accuracy and treatment quality of juxtaglomerular cell tumor.Methods The clinical data of 2 female patients(20 and 36 years,respectively)with juxtaglo- merular cell tumor were presented and discussed in combination with review of the literature,including the onset characteristics,imaging features,treatment,pathology and prognosis.Case 1 presented with hyperten- sion of 180/110 mm Hg.The laboratory examinations showed that in decubitus and standing position,the plasma rennin activity(PRA)was 3.2?g - L~(-1)?h~(-1)and 36.5?g?L~(-1)?h~(-1);angiotensinⅡwas 54.3 pg/ml and 183.5 pg/ml;aldosterone was 193.5 prnol/L and 489.4 pmol/L,respectively;serum kalium was 2.6 mmol/L.Case 2 presented with hypertension of 210/120 mm Hg.The laboratory examination results were as follows:in decubitus and standing position,PRA was 4.3?g?L~(-1)?h~(-1)and 37.0?g?L~(-1)?h~(-1);angio- teusinⅡwas 55.6 pg/ml and 200.4 pg/ml;aldosterone was 162.4 pmol/L and 506.3 pmol/L,respectively; serum kalium was 3.0 mmol/L.On CT scan,both cases had renal tumor,with the diameter of 3.0 cm and 3. 5 cm,respectively.Results Case 1 underwent laparoscopic partial nephrectomy.Case 2 who had artery stricture and severe functional injure of the right kidney underwent laparoscopic right nephrectomy.The oper- ative time was 3.0 h and 2.0 h,and the blood loss was 175 ml and 112 ml,respectively.There was no mor- tality or postoperative complication.In 1 or 2 postoperative weeks,Case 1 had blood pressure(BP)of 120/70 mm Hg;in deeubitus and standing position,PRA was 1.5?g ~ L~(-1)?b~(-1)and 12.8?g?L~(-1)?h~(-1);angio- tensinⅡwas 30.6 pg/ml and 97.5 pg/ml;aldosterone was 78.5 pmol/L and 192.2 pmol/L,respectively ;se- rum kalium was 4.2 mmol/L.Case 2 had BP of 125/75 mm Hg;in decubitus and standing position,PRA was 1.6?g.L~(-1)?h~(-1)and 12.3?g.L~(-1)?h(-1);angiotensinⅡwas 34.3 pg/ml and 83.5 pg/ml;aldoste- rone was 62.6 pmol/L and 292.5 pmol/L,respectively;serum kalium was 4.8 mmol/L.Pathology showed that the juxtaglomerular cell tumor had intact envelop.Light microscopically,the tumor was very much like a hemangiopericytoma,showing active proliferation and nuclear atypia.The immunohistochemical staining showed positive Vimentin,CD_(34)expression,and negative MSA,EMA,Bcl-2,?-SmA,AG/AG3,34?En, CD_(117),CD_(31),Iv glue,Ki-G~-(＜2%)expression.Ultrastructural changes of the nuclei and some organelles in the cytoplasm were observed under electron microscope.The conspicuous ultrastructural feature was the pres- ence of secretion granules and rhomboid-shaped,crystal-like structures in the dilated cisternae of rough endo- plasmic reticulum and vesicles of Golgi complex.The follow-up was 14 and 6 months,respectively;the renal function was normal and no tumor recurrence was found.Conelusions Juxtaglomerular cell tumor is a rare tumor which can produce renin.It is characterized by severe hypertension and low serum potassium.La- boratory examination results are helpful for the diagnosis:PRA and angiotensinⅡincrease obviously ;aldoste- rone is 1-10 times more than normal;serum kalium is commonly between 2.1-3.5 mmol/L.The definite diagnosis depends on clinical presentations,immunohistochemistry,light and electron microscopic examina- tions.Laparoscopie operation is the first choice of surgical treatment.

Objective To discuss the treatment of ureteropelvic junction obstruction by laparoscopic pyeloplasty. Methods A retrospective review of consecutive laparoscopic pyeloplasty in 102 patients between September 2001 and December 2007 was performed. The ureterpelvic junction was dissected and the obstruction portion was excised. Anastomosis was then performed through the ureter and the renal pelvis walls with a stent. Results The mean operating time was 120 min and the average blood loss was 80ml. No major complication occurred intraoperative. The drainage was removed in 3-10 days. The average hospital stay was 8.5 days. The stent was kept for 30-60 days. IVU and B ultrasound examination revealed that the hydronephrosis alleviated during the follow-up and no anastomosis stricture occurred. Conclusions Laparoscopic dismembered pyeloplasty could provide lower morbidity, shorter hospital stay, and faster convalescence. It could be an effective treatment for ureteropelvic junction obstruction.

BACKGROUNDWith potent suppressive effect on responder T cells, CD(4)(+)CD(25)(+) regulatory T (Treg) cells have become the focus of attention only recently and they may play an important role in transplantation tolerance. However, the mechanism of action is not clear. This study was designed to assess the possibility of using CD(4)(+)CD(25)(+) Treg cells to induce transplantation tolerance and to investigate their mechanism of action.

METHODSCD(4)(+)CD(25)(+) Treg cells were isolated using magnetic cell separation techniques. Mixed lymphocyte reactions were used to assess the ability of Treg cells to suppress effector T cells. Before skin transplantation, various numbers of CD(4)(+)CD(25)(+) Treg cells, which have been induced using complex skin antigens from the donor, were injected into the host mice either intraperitoneally [0.5 x 10(5), 1 x 10(5), 2 x 10(5), 3 x 10(5), 4 x 10(5), or 5 x 10(5)] or by injection through the tail vein [5 x 10(3), 1 x 10(4), 2 x 10(4), 5 x 10(4), 1 x 10(5), 2 x 10(5)]. Skin grafts from two different donor types were used to assess whether the induced Treg cells were antigen-specific. The survival time of the allografts were observed. Single photon emission computed tomography was also used to determine the distribution of Treg cells before and after transplantation.

RESULTSTreg cells have suppressive effect on mixed lymphocyte reactions. Grafts survived longer in mice receiving CD(4)(+)CD(25)(+) Treg cell injections than in control mice. There was a significant difference between groups receiving intraperitoneal injection of either 2 x 10(5) or 3 x 10(5) CD(4)(+)CD(25)(+) Treg cells and the control group (P < 0.05, respectively). Better results were achieved when Treg cells were injected via the tail vein than when injected intraperitoneally. The transplantation tolerance induced by CD(4)(+)CD(25)(+) Treg cells was donor-specific. Analysis of the localization of Treg cells revealed that Treg cells mainly migrated from the liver to the allografts and the spleen.

CONCLUSIONSCD(4)(+)CD(25)(+)Treg cells can induce donor-specific transplantation tolerance. Cell-to-cell contact may be the primary mechanism by which Treg cells act on effector T cells.

Animals ; Graft Rejection ; Immune Tolerance ; Lymphocyte Culture Test, Mixed ; Mice ; Mice, Inbred BALB C ; Skin Transplantation ; immunology ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo evaluate the clinical efficacy of alpha-1 A adrenoceptor antagonist (tamsulosin) in the treatment of benign prostatic hyperplasia (BPH) patients with acute urinary retention.

METHODSSeventy-two BPH patients with acute retention of urine were randomly divided into treatment group and control group of 36 patients each. All the patients were treated with indwelling catheter, oral antibiotics and the patients in treatment group tamsulosin 0.4 mg once a day for 3 days. The catheter was removed after 72 hours of treatment.

RESULTSAfter removal of the catheter, 44% (32/72) of patients voided successfully. The effect rates were 61% (22/36) in the tamsulosin treatment group and 28% (10/36) in the control group(P < 0.01).

CONCLUSIONSTreatment with tamsulosin was effective in raising the success rate of voiding without catheter after an episode of acute urinary retention. The efficacy of treatment was not influenced by the volume of prostate.

Acute Disease ; Adrenergic alpha-Antagonists ; administration & dosage ; therapeutic use ; Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Prostatic Hyperplasia ; complications ; drug therapy ; Sulfonamides ; administration & dosage ; therapeutic use ; Treatment Outcome ; Urinary Catheterization ; Urinary Retention ; complications ; drug therapy

AIMTo study the effect of combined androgen block therapy on hemoglobin and hematocrit values in patients with prostate cancer.

METHODSOne hundred and thirty-six patients with adenocarcinoma of prostate were treated with combined androgen block (orchiectomy and flutamide 250 mg, tid). Complete blood counts were determined before and after 1, 2, 3, 6, 9 and 12 months of therapy.

RESULTSThe hemoglobin and hematocrit levels declined significantly in all patients and at all the time points after treatment (P<0.05).

CONCLUSIONProstate cancer patients treated with combined androgen block would develop obvious anemia. Recombinant human erythropoietin can be used to treat patients with severe anemia.

Adenocarcinoma ; complications ; drug therapy ; therapy ; Adult ; Androgen Antagonists ; adverse effects ; therapeutic use ; Anemia ; chemically induced ; Antineoplastic Agents, Hormonal ; therapeutic use ; Combined Modality Therapy ; Flutamide ; therapeutic use ; Hematocrit ; Hemoglobins ; metabolism ; Humans ; Male ; Orchiectomy ; Prostatic Neoplasms ; complications ; drug therapy ; therapy ; Prostatic Secretory Proteins ; analysis