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

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

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

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.

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.

OBJECTIVETo evaluate the correlation of epididymal protease inhibitor(Eppin) and Semenogelin(Sg) on human ejaculated spermatozoa.

METHODSThe experimental approaches include: (1) Immunoprecipitation of Eppin with anti-Eppin from semen; (2) Colocalization of Eppin and Sg by immunofluorescence; (3) Immunoprecipitation of rEppin and rSg;(4) Far-Western blotting of rEppin and rSg;(5) Competition of saturated 125I-rSg binding to rEppin with unlabeled Sg, and direct binding of 125I-rSg to rEppin on a blot; (6) Autoradiography of 125I-rSg with rEppin.

RESULTSEppin-Sg complex present on the surface of human ejaculated spermatozoa, Cys-239 is the only cystein for rEppin binding rSg. Reduction and carboxymethylation of Cys-239 blocks binding of 125I-rEppin to rSg.

CONCLUSIONOur study demonstrates that Eppin and Sg bind to each other on human ejaculated spermatozoa. A disulfide linkage occurs between Sg and Eppin, indicating the specificity of binding.

Humans ; Male ; Protein Binding ; Proteinase Inhibitory Proteins, Secretory ; Proteins ; chemistry ; metabolism ; Recombinant Proteins ; Seminal Vesicle Secretory Proteins ; chemistry ; metabolism ; Spermatozoa ; metabolism

AIMTo study the molecular mechanism of epididymal protease inhibitor (Eppin) modulating the process of prostate specific antigen (PSA) digesting semenogelin (Sg).

METHODSHuman Sg cDNA (nucleotides 82-849) and Eppin cDNA (nucleotides 70-723) were generated by polymerase chain reaction (PCR) and cloned into pET-100D/TOPO. Recombinant Eppin and Sg (rEppin and rSg) were produced by BL21 (DE3). The association of Eppin with Sg was studied by far-western immunoblot and radioautography. In vitro the digestion of rSg by PSA in the presence or absence of rEppin was studied. The effect of anti-Q20E (N-terminal) and C-terminal of Eppin on Eppin-Sg binding was monitored.

RESULTSEppin binds Sg on the surface of human spermatozoa with the C-terminal of Eppin (amino acids 75-133). rSg was digested with PSA and many low molecular weight fragments were produced. When rEppin is bound to rSg, then digested by PSA, incomplete digestion and a 15-kDa fragment results. Antibody binding to the N-terminal of rEppin did not affect rSg digestion. Addition of antibodies to the C-terminal of rEppin inhibited the modulating effect of rEppin.

CONCLUSIONEppin protects a 15-kDa fragment of rSg from hydrolysis by PSA.

Animals ; Antibodies ; pharmacology ; Autoradiography ; Humans ; Hydrolysis ; Male ; Prostate-Specific Antigen ; metabolism ; Proteinase Inhibitory Proteins, Secretory ; genetics ; immunology ; metabolism ; Rabbits ; Recombinant Proteins ; genetics ; metabolism ; Semen ; cytology ; metabolism ; Seminal Vesicle Secretory Proteins ; metabolism ; Spermatozoa ; metabolism

OBJECTIVETo study the etiopathogenesis, clinical manifestations, diagnosis and management of persistent Müllerian duct syndrome (PMDS).

METHODSTwo cases of PMDS were reported, one accompanied by transverse testicular ectopia and the other associated with cryptorchidism. Corporeal hysterectomy and orchidopexy were given to both the patients and cryptorchidectory the latter.

RESULTSVascular supply and texture of the testis were normal in both the 2 patients after 1.5-2 years' follow-up.

CONCLUSIONPMDS is male pseudohermaphroditism, for which means should be taken to preserve the blood supply and fertility function of the testis in surgical management, and attention should be paid to possible development of testis tumor in follow-up.

Adult ; Disorders of Sex Development ; pathology ; Follow-Up Studies ; Humans ; Male ; Mullerian Ducts ; abnormalities ; Syndrome