OBJECTIVETo investigate the expression of the Pim-1 gene in the LNCaP cells of the animal model of orthotopically implanted prostate cancer by surgical castration simulating androgen-deprivation therapy.

METHODSWe equally allocated 32 male BALBc-nu mice into 4 groups, androgen-dependent prostate cancer (ADPC), androgen-deprivation therapy (ADT) , castration-resistant prostate cancer (CRPC) and blank control, and established the models of orthotopically implanted tumor using human prostate cancer LNCaP cells. We detected and ,compared the expressions of Pim-1, PSA, and androgen receptor (AR) in the tumor tissues of different groups by RT-PCR. qRT-PCR, ELSIA and immunohistochemistry.

RESULTSThe relative gray scales in the ADPC and CRPC groups were 0.59 ± 0.01 and 1.14 ± 0.02, with statistically significant differences from 0.62 ± 0.03 in the ADT group (P < 0.05), and the Δ Ct values of Pim-1 were 6.15 ± 0.34 and 4.56 ± 0.23 in the former two groups, also with significant differences from 5.11 ± 0.21 in the latter (P < 0.05). The results of 2-ΔΔ Ct relative quantification analysis showed that the amplification products of Pim-1 in the ADT and CRPC groups increased 2.05 and 3.01 times respectively that of the ADPC group. The concentration of PSA was significantly higher in the ADPC ([480 ± 25] pg/ml) and CRPC ([870 ± 23] pg/ml) than in the ADT ([170 ± 32] pg/ml) and blank control groups (0 µg/L) (P < 0.01). The mean optical densities of Pim-1 and AR proteins were 0.017 ± 0.002 and 0.032 ± 0.009 in the ADPC group and 0.024 ± 0.002 and 0.040 ± 0.011 in the CRPC group, both with significant differences from those in the ADT group (0.018 ± 0.001 and 0.019 ± 0.006) (P < 0.01).

CONCLUSIONPim-1 is highly expressed in nude mice with prostate cancer receiving androgen-deprivation therapy and plays an important role in the progression and metastasis of prostate cancer.

Androgen Antagonists ; therapeutic use ; Animals ; Disease Progression ; Gene Expression ; Heterografts ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasms, Hormone-Dependent ; metabolism ; Prostate-Specific Antigen ; metabolism ; Prostatic Neoplasms, Castration-Resistant ; genetics ; metabolism ; therapy ; Proto-Oncogene Proteins c-pim-1 ; metabolism ; Receptors, Androgen ; metabolism

OBJECTIVETo explore the role and function of stromal cell-derived factor-1 (SDF-1) in stem cells migrating into injured brain area.

METHODSRat-derived nerve stem cells (NSCs) were isolated and cultured routinely. Transwell system was used to observe the migration ability of NSCs into injured nerve cells. Immunocytochemistry was used to explore the expression of chemotactic factor receptor-4 (CXCR-4) in NSCs. In vivo, we applied immunofluorescence technique to observe the migration of NSCs into injured brain area. Immunofluorescence technique and Western blotting were used to test expression level of SDF-1. After AMD3100 (a special chemical blocker) blocking CXCR-4, the migration ability of NSCs was tested in vivo and in vitro, respectively.

RESULTSNSCs displayed specific tropism for injured nerve cells or traumatic brain area in vivo and in vitro. The expression level of SDF-1 in traumatic brain area increased remarkably and the expression level of CXCR-4 in the NSCs increased simultaneously. After AMD3100 blocking the expression of CXCR-4, the migration ability of NSCs decreased significantly both in vivo and in vitro.

CONCLUSIONSSDF-1 may play a key role in stem cells migrating into injured brain area through specially combining with CXCR-4.

Animals ; Brain Injuries ; pathology ; Cell Movement ; Cells, Cultured ; Chemokine CXCL12 ; analysis ; physiology ; Neurons ; cytology ; Rats ; Receptors, CXCR4 ; analysis ; physiology ; Stem Cells ; physiology ; Tropism

Streptomyces hygroscopicus 17997 produces the antiviral and antitumor ansamycin antibiotic, geldanamycin. Studies on geldanamycin biosynthetic pathway will provide good tools for genetic manipulation of the antibiotic-producing strain to improve the productivity or to facilitate making novel geldanamycin analogs. The structural similarities between geldanamycin and ansamycins such as rifamycin or ansatrienin suggest that both geldanamycin and ansamycins has a closely related pathways of biosynthesis and that biosynthetic system for geldanamycin is similar to the one of type I polyketide synthase (PKS) enzyme system. To explore the possible PKS genes involved in geldanamycin biosynthesis, the degenerate primers were designed according to the conserved sequence of KS-AT region from erythromycin and oleandomycin type I PKS genes. Cosmids containing multiple PKS genes (pCGBK2,4,6,10,11,18) were obtained by hybridization with the PCR products, which were amplified from S. hygroscopisus 17997 genomic DNA. The designed primers above were used for PCR. Development of a Streptomyces temperate phage phiC31-derivative KC515( tsrR) transduction system was carried out for identification of cosmids containing the PKS gene related to biosynthesis of geldanamycin. Several factors, mainly the Ca2+ and Mg + concentrations in different culture media affecting the frequency of gene transfection, were optimized .Transfection efficiency could reach up to 10(3) /microg DNA on YMG medium supplemented with 10mmol/L MgSO4. Reversely, the transfection efficiency decreased when YMG medium was supplemented with 30mmol/L MgSO4. Gene transfection system based on the integration-defective phage KC515 had been established for S. hygroscopicus17997. Recombinant phages (ph111, 258, 287, 116, 105) were constructed by insertion of the homologous to PKS gene fragments into the KC515 phage vector. Gene disruption experiments were performed by transduction of recombinant phages into S. hygroscopicus 17997 genome, and disruption of geldanamycin production was observed as a result of homologous recombination between the cloned insert in recombinant phage and the S. hygroscopicus 17997 genome by integration. Thiostrepton resistant transductants were selected and integration event was analyzed by Southern hybridization. The fermentation broth extracts from five resistant transductants were analyzed by TLC and HPLC. The results showed that only G16 mutant failed to produce geldanamycin. This result showed that the integration of the insert DNA fragment in recombinant phage phl6 into the chromosome of S. hygroscopicus disrupted the expression of the geldanamycin biosynthetic genes. The original cosmid pCGBK10 containing this cloned insert was predicted to encode PKS genes in the geldanamycin biosynthesis. This study laid the foundation for cloning the PKS genes involved in geldanamycin biosynthetic gene cluster from S. hygroscopicus 17997.
Alcohol Oxidoreductases ; genetics ; metabolism ; Bacterial Proteins ; genetics ; metabolism ; Bacteriophages ; genetics ; Benzoquinones ; metabolism ; Blotting, Southern ; Chromatography, High Pressure Liquid ; Chromatography, Thin Layer ; Genetic Vectors ; genetics ; Lactams, Macrocyclic ; metabolism ; Multigene Family ; genetics ; physiology ; Polymerase Chain Reaction ; Streptomyces ; genetics ; metabolism

Variant pulmonary vein anatomy (PVA) has been reported to influence the recurrence of atrial fibrillation (AF) after radiofrequency ablation.However,the effects of PVA on AF in patients undergoing cryoballoon ablation (CBA) remain unknown.The present study aimed to examine the impact of PVA on the long-term outcome of CBA for AF.A total of 78 patients (mean age 60.7±10.9 years,64.1％ males) with symptomatic and drug-refractory paroxysmal AF were enrolled in the study.Left atrium (LA) and PVA acquired at computed tomography angiography (CTA) were reconstructed with CARTO(R) 3 SYSTEM.Patients were routinely evaluated by 24-hour Holter monitoring following CBA.Cox regression was used to detect the predictors of AF recurrence after CBA.The results showed abnormal PVA in 30 patients (38.5％) and 18 patients (23.1％) had left common PV (LCPV).Electrical pulmonary vein isolation was achieved in all patients.After a mean follow-up of 689.5±103.8 days,it was found that patients with abnormal PVA had similar AF recurrence rate to those with normal PVA (26.7％ vs.25.0％,P=0.54),and there was no significant difference in AF recurrence rate between LCPV patients and non-LCPV patients (33.7％ vs.23.3％,P=0.29).Cox regression analysis showed that AF duration (72.9±9.0 vs.42.3±43.2 months,HR 1.001;95％CI 1.003-1.014;P＜0.001) and cryo-applications of right-side PVs (3.0±1.6 vs.4.7±1.7,HR 0.661;95％ CI 0.473-0.925;P=0.016) were independent predictors of freedom from AF,but PVA was not identified as a predictor of long-term success.In conclusion,the variant PVA cannot significantly influence the long-term outcome of AF patients undergoing CBA;longer AF duration and less cryo-applications of right-side PVs are associated with higher AF recurrent rate.

Purpose: The experience with laparoscopic nephrectomy or nephroureterectomy was reported.Methods: Since Feb. 2000, 17 patients (Including 3 cases of pevic or upper-segment transitional cell tumour, 7cases of renal cell carcinoma, 3 cases of renal atrophy-induced hypertensin, 2 cases of non-functioning calculuskidney and 2 cases of renal tuberculosis) underwent peritoneoscopic or retroperitoneoscopic nephrectomy,radicalnephrectomy or nephroureterectomy with the latter in combination with cystoscope. Results:All operations weresuccessful except the first case. Operation time for nephrectomy was 90 to 180 min with an average of 117 min.All patients did not receive blood transfusion and had no obvious complication. Conclusions: Laparoscopicnephrectomy had the characteristics of minimal morbidity,minimal postoperative discomfort and a short hospitalstay. It may be used well in the future.

OBJECTIVESTo elucidate the treatment of Ureaplasma urealyticum (UU) and Chlamydia trachomatis (CT) infectious chronic prostatitis.

METHODSForty-eight cases of chronic prostatitis patients with UU and CT infections were treated with minocycline, Chinese medicine "Qianlieshulekeli" and alpha 1A adrenoceptorblocker (tamsulosin) for 6 weeks. The change of symptoms, expressed prostatic secretion (EPS) routine, and UU and CT detection results were observed before and after the treatment. The efficacy of treatment were evaluated by CPSI score.

RESULTSRemarkably effective-41 cases (85.4%), effective-5 cases (10.4%), noneffective-2 cases (4.2%). CPSI score reduced from (22 +/- 8) before treatment to (7 +/- 3) after treatment (P < 0.01). UU in 20 of 24 cases (83%) and CT in 25 of 28 cases (89%) turned negative after treatment.

CONCLUSIONSThe therapy combining Chinese medicine with western medicine for the treatment of UU and CT infectious chronic prostatitis is successful.

Adolescent ; Adult ; Chlamydia Infections ; drug therapy ; Chlamydia trachomatis ; Chronic Disease ; Drug Therapy, Combination ; Humans ; Male ; Medicine, Chinese Traditional ; Minocycline ; therapeutic use ; Prostatitis ; drug therapy ; Ureaplasma Infections ; drug therapy ; Ureaplasma urealyticum

OBJECTIVETo study the distribution of aldehyde dehydrogenase-2 (ALDH2) polymorphisms between healthy Zhuang and Han ethnic individuals in Guangxi Autonomous Region and its influence to the behaviors of alcohol consumption.

METHODSPolymerase chain reaction with confronting two-pair primers (PCR-CTPP) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) techniques were used to genotype ALDH2, respectively, and alcohol consumption was recorded in a constructed questionnaire.

RESULTSThe frequencies of ALDH2 alleles (ALDH2(1)/ALDH2(2)) among Zhuang and Han ethnics were 0.511, 0.489 and 0.508, 0.492 respectively (chi2 = 0.001, P > 0.05). The ALDH2 genotypes were verified with PCR-RFLP method. The frequencies of ALDH2(1) genotype in alcoholics (> or = 3 times drinking per week) were 35.59% and 15.67% in Zhuang and Han groups respectively (chi2 = 5.800, P = 0.016).

CONCLUSIONThere was no significant different distribution of ALDH2 genotype among healthy Zhuang and Han ethnic people. The genotype of ALDH2 in different ethnicity might influence individual behavior of alcohol consumption.

Adult ; Aldehyde Dehydrogenase ; genetics ; Aldehyde Dehydrogenase, Mitochondrial ; Alleles ; China ; DNA Primers ; genetics ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; methods ; Polymorphism, Restriction Fragment Length

OBJECTIVETo investigate the clinicopathological features, diagnosis, treatment and prognosis of primary clear cell carcinoma of the liver (PCCCL).

METHODSThe clinicopathological data of 24 cases with pathologically proven PCCCL in the First Affiliated Hospital of Guangxi Medical University from May 1996 to December 2003 were collected and analyzed.

RESULTSThere were 21 males and 3 females in this group, with an average age of 46 years (range: 30 approximately 78 years). HBV infection was detected in 83.3%, and AFP expression was found in 75.0% of them. Of the 24 cases, 28 tumors were found with an average size of (6.64 +/- 5.54) cm. Liver cirrhosis was found in 75.0% of the patients. Macroscopic and microscopic tumor thrombi were found in 20.8% and 29.2%, respectively. Lymph node metastasis was found in 4.2% of the patents. The 1-, 3-, and 5-year overall survival rates of the 24 cases were 75.0%, 41.7% and 27.8%, respectively, with a median survival time of 29 months.

CONCLUSIONThe clinical characteristics of primary clear cell carcinoma of the liver are similar to that of common hepatocellular carcinoma. It is difficult to be diagnosed preoperatively and final diagnosis depends on pathological examination. Surgical resection is an effective way to achieve favorable treatment outcome and even long-term survival.

Adenocarcinoma, Clear Cell ; blood ; pathology ; surgery ; virology ; Adult ; Aged ; Female ; Follow-Up Studies ; Hepatectomy ; methods ; Hepatitis B ; Humans ; Liver Neoplasms ; blood ; pathology ; surgery ; virology ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Survival Rate ; alpha-Fetoproteins ; analysis

BACKGROUND AND OBJECTIVEMultinodular hepatocellular carcinoma(HCC) might originate from multicentric occurrence (MO) or intrahepatic metastasis(IM). This study was to find out proteins which play important roles in clonal origin of multinodular hepatocellular carcinoma bt screening the differentially expressed proteins between the MO and IM tissues using comparative proteomic analysis.

METHODSTotal protein extracted was separated by two-dimensional gel electrophoresis. Comparative analyses of the 2-DE protein patterns between the two groups were carried out using computerized imaging techniques. Proteins exhibiting significant alternations were subsequently isolated and identified by mass spectrometry.

RESULTSA total 1025+/-52 and 900+/-98 spots were detected in the protein profile in IM and MO, respectively. Twenty-five protein spots were statistically different at expression levels between the two groups. Twenty of them were identified by MALDI-TOF-MS and bioinformatics.

CONCLUSIONSThe protein profile of MO HCC tissues is different from that in IM HCC tissues. The twenty differentially expressed proteins might play a key role in the carcinogenesis and progression of multinodular HCC. These newly identified proteins might be potential and valuable biomarkers for identifying the multinodular HCC of clonal origin.

Adult ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Electrophoresis, Gel, Two-Dimensional ; Female ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasms, Multiple Primary ; metabolism ; pathology ; Protein Array Analysis ; Proteomics ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization