Stem cells are characterized by self-renewing, multipotent differentiation, and high proliferation and receiving more and more attention for their roles in the development and management of various diseases. There are epithelial stem cells and mesenchymal stem cells in the prostate. The markers of the epithelial stem cells include cytokeratin, stem cell antigen-1, and integrins alpha2beta1, CD49f, CD133, CD117, and CD44. The markers of the mesenchymal stem cells include CD30, CD44, CD133, neuron-specific enolase, and vascular endothelial growth factor receptor-1. Prostate stem cells are involved in the development and treatment of prostatic diseases. This review focuses on the latest progress in the studies of prostate stem cells.
Antigens, CD ; Biomarkers ; Cell Differentiation ; Humans ; Integrin alpha2beta1 ; Male ; Prostate ; cytology ; Stem Cells ; chemistry ; cytology ; Vascular Endothelial Growth Factor A

OBJECTIVETo culture rat prostate glandular epithelial cells and study their barrier functions in vitro.

METHODSRat prostate glandular epithelial cells were cultured in vitro. The expression of the tight junction protein claudin-1 was determined by immunohistochemistry, the structure and composition of the epithelial cells observed under the inverted microscope and transmission electron microscope. The transepithelial electrical resistances (TEERs) were monitored with the Millicell system. The permeability of the prostate glandular epithelial cells was assessed by the phenol red leakage test.

RESULTSCompact monolayer cell structures were formed in the prostate glandular epithelial cells cultured in vitro. Immunohistochemistry showed the expression of the tight junction protein claudin-1 and transmission electron microscopy confirmed the formation of tight junctions between the adjacent glandular epithelial cells. The TEERs in the cultured prostate glandular epithelial cells reached the peak of about (201.3 ± 3.5) Ω/cm2 on the 8th day. The phenol red leakage test manifested a decreased permeability of the cell layers with the increase of TEERs.

CONCLUSIONThe structure and function of rat prostate glandular epithelial cells are similar to those of brain capillary endothelial cells, retinal capillary endothelial cells, and intestinal epithelial cells. In vitro cultured prostate glandular epithelial cells have the barrier function and can be used as a model for the study of blood prostate barrier in vitro.

Animals ; Cell Membrane Permeability ; Cells, Cultured ; Claudin-1 ; metabolism ; Electric Impedance ; Epithelial Cells ; pathology ; physiology ; ultrastructure ; In Vitro Techniques ; Male ; Microscopy, Electron, Transmission ; Phenolsulfonphthalein ; pharmacokinetics ; Prostate ; metabolism ; pathology ; Rats ; Tight Junctions

OBJECTIVETo investigate the inhibitory effect of bone marrow mesenchymal stem cells (BMSCs) on E coliinduced prostatitis in rats.

METHODSBMSCs were isolated, cultured and amplified by the attached choice method. Fifty SD rats were randomized into five groups of equal number: normal control, acute bacterial prostatitis (ABP) , chronic bacterial prostatitis (CBP), ABP + BMSCs, and CBP + BMSCs, and the animals in the latter four groups were injected with E. coli into both sides of the prostate under ultrasound guidance for 1 - 14 days to induce ABP and for 4 - 12 weeks to induce CBP. The control rats were injected with the same amount of PBS. Two weeks after injection of BMSCs into the prostates, pathomorphological changes in the prostate were observed under the light microscope and the mRNA and protein levels of IL-1β and TNF-α determined by RT-PCR and ELISA, respectively, followed by statistical analysis with SPSS 18.0.

RESULTSHistopathological evaluation showed typical pathological inflammatory changes in the prostates of the rats in the ABP and CBP groups, including glandular structural changes, interstitial edema, inflammatory cell infiltration, and fibrous hyperplasia, which were all remarkably relieved after treated with BMSCs. The mRNA and protein levels of IL-β ([0.829 ± 0.121] and [271.75 ± 90.59] pg/ml) and TNF-α ([0.913 ± 0. 094] and [105.78 ± 19. 05] pg/ml) in the ABP and those of IL-1β ([0. 975 ± 0. 114] and [265. 31 ± 71. 34] pg/ml) and TNF-α ([0. 886 ± 0. 084] and [107. 45 ± 26. 11 ] pg/ml) in the CBP groups were significantly higher than those in the control rats ([0. 342 ± 0.087] and [45.76 17. 99] pg/ml, P <0. 05); ([0.247 ± 0.054] and ([19.42 ± 7. 75] pg/ml, P <0. 01) as well as than those in the ABP + BMSCs ([0. 433 ± 0. 072] and [51. 34 ± 22. 13] pg/ml, P < 0. 05 ) ; ( [0. 313 ± 0. 076] and [28. 38 ± 8. 78] pg/ml, P < 0. 01) and the CBP + BMSCs group ([0.396 ± 0.064] and [56.37 ± 21.22] pg/ml, P <0.05); ([0.417 ± 0.068] and [29.21 ± 10.22] pg/ml, P <0.01).

CONCLUSIONInjection of BMSCs can reduce E coli-induced prostatic inflammation reaction, which.may be associated with its reduction of inflammatory cell infiltration and the expressions of IL-1β and TNF-α in the prostate tissue.

Acute Disease ; Animals ; Bone Marrow Cells ; physiology ; Chronic Disease ; Escherichia coli Infections ; therapy ; Humans ; Interleukin-1beta ; genetics ; Male ; Mesenchymal Stromal Cells ; physiology ; Prostate ; metabolism ; Prostatitis ; metabolism ; microbiology ; therapy ; RNA, Messenger ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; genetics ; metabolism

Icaritin, a prenylflavonoid derivative from Epimedium Genus, has been shown to exhibit many pharmacological and biological activities. However, the function and the underlying mechanisms of icaritin in human non-small cell lung cancer have not been fully elucidated. The purpose of this study was to investigate the anticancer effects of icaritin on A549 cells and explore the underlying molecular mechanism. The cell viability after icaritin treatment was tested by MTT assay. The cell cycle distribution, apoptosis and reactive oxygen species (ROS) levels were analyzed by flow cytometry. The mRNA and protein expression levels of the genes involved in proliferation and apoptosis were respectively detected by RT-PCR and Western blotting. The results demonstrated that icaritin induced cell cycle arrest at S phase, and down-regulated the expression levels of S regulatory proteins such as Cyclin A and CDK2. Icaritin also induced cell apoptosis characterized by positive Hoechst 33258 staining, accumulation of the Annexin V-positive cells, increased ROS level and alteration in Bcl-2 family proteins expression. Moreover, icaritin induced sustained phosphorylation of ERK and p38 MAPK. These findings suggested that icaritin might be a new potent inhibitor by inducing S phase arrest and apoptosis in human lung carcinoma A549 cells.

Xanthomonas campestris pv. campestris (Xcc), the pathogenic agent of black rot disease in cruciferous plants, produces large amount of extracellular polysaccharide (EPS), which has found wide applications in industry. For the great commercial value of the xanthan gum, many of the genes involved in EPS biosynthesis have been cloned and the mechanism of EPS biosynthesis also has been studied. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5 gusA5, and a number of EPS-defective mutants were isolated in our previous work. The Tn5 gusA5 inserted sites of these mutants were located by using thermal asymmetric interlaced PCR, and results showed that two EPS-defective mutants were insertion mutants of the gene wxcA which involved in lipopolysaccharide (LPS) biosynthesis. The gene wxcA involved in lipopolysaccharide biosynthesis but dose not extracellular polysaccharide in others' report. wxcA::Tn5 gusA5 mutant 021C12, the polar mutant, was complemented with recombinant plasmid pLATC8570 harboring an intact wxcA gene in this work, but the yield of EPS of the wxcA::Tn5 gusA5 mutant was not restored. In order to identify the function of wxcA gene of Xcc 8004 strain, the gene wxcA was deleted by gene replacement strategy, and the no-polar mutant of wxcA was obtained. DeltawxcA mutant strain, named Xcc 8570, was confirmed by using both PCR and southern analysis. Beside the LPS biosynthesis of deltawxcA mutant was affected, The EPS yield of deltawxcA mutant strain reduced by 50% as compared with the wild-type strain 8004. DeltawxcA mutant could be complemented in trans with the intact wxcA gene, and the EPS yield of the mutant was restored. The combined data showed that wxcA gene not only involved in LPS biosynthesis but also EPS yield in Xcc 8004 strain.
Cell Proliferation ; Genes, Bacterial ; physiology ; Lipopolysaccharides ; biosynthesis ; Mutation ; Polysaccharides, Bacterial ; biosynthesis ; Xanthomonas campestris ; genetics

Objective To discuss the technique and value of preoperative reformatting with three-dimensional computed tomography technique for C_2 pedicle screw track.Methods GE Light Speed 16 Pro spinal CT scans of 15 adult dry vertebrae were loaded into an imaging station (software ADW4.2).Two methods of C_2 pedicle screw techniques were analyzed through virtual screw trajectory by VR (volume rendering) and MPR (multiple planar reformatting) techniques,in method A,screw entry point was the intersection between the media-vertical and the cranial line of C_2 inferior facet joint,in method B,the screw track was from the cranial and medial quadrant of the dorsal part of C_2 inferior facet joint.Results The screw track could be observed dynamically from any plane.Two vertebrae were ob- served with smaller height in isthmus and the medial edge of the transverse foramen since no space was a- vailable for the screw.The screw trajectories data were compared between method A and method B,which showed that the angles towards the cephalad (in sagittal plane) and midline (in transverse plane) were bigger in method A than in method B (P＜0.05,0.01),but the safe screw diameter was smaller in method A than in method B (P＜0.05),and there was no difference of the screw length between the two methods(P＜0.05 ).Conclusion In this research,the individual C_2 pedicle screw entry points, screw diameter and security screw angle can be simulated,and the screw track can be observed dynami- cally to make sure if it transits the bone structure completely.Preoperative three-dimensional computed tomography reformatting for pedicle screw track is of great value in clinical and basic researches.

To study the expressions of CD34 and CD117 in the tissues of hepatocelluar carcinoma (HCC) and to explore the relationship with clinical pathology and it's evaluation on the prognosis of HCC patients. The expressions of CD34 and CD117 were examined by two-step methods of PV-9000 of immunohistochemistry in 55 HCC cases, 10 liver cirrhotic specimens and 6 normal liver specimens. Clinical-pathological data, tumor recurrent rate and survival rate after hepatectomy were recorded and analyzed with Fisher's Exact Test, Pearson X2 Test, Kaplan-Meier, Log-Rank Test and Cox Regression. The positive expression of CD34 was found in 65.4% of HCC, 20% of cirrhostic liver specimens and 16.7% of normal liver specimens, respectively. Significant differences found among the three groups, and the CD34 expression was significantly associated with vessel embolus (X2 = 4.000, P = 0.046) and the histological grades (X2 = 11.008, P = 0.001). The positive expression of CD117 was 47.3%, 10% and 0% in HCC, cirrhotic liver specimens and normal liver tissues, respectively, and statistical differences esxisted among the three groups. The CD117 expression was dramatically related to the histological grades (X2 = 5.115, P = 0.024) and clinical stages (X2 = 15.459, P = 0.000). Median disease free survival time after hepatectomy was significantly shorter in the group with positive-expression of CD34 (X2 = 4.105, P = 0.043) and CD117 (X2 = 28.023, P = 0.000) than the negative-expressed groups, respectively. Multivariate analysis showed that CD117 expression status, serum AFP levels and the size of tumor were independently prognostic factors for HCC patients. Tthe results demonstrated that CD34 and CD117 might play an important role in liver carcinogenesis and the progression of HCC, and they might potentially serve as markers for HCC prognosis.
Carcinoma, Hepatocellular ; Hepatectomy ; Humans ; Kaplan-Meier Estimate ; Liver Neoplasms ; Neoplasm Recurrence, Local

Objective To explore the application value of 320-slice volume CT in detecting the cerebral transient ischemic attack (TIA). Methods One-stop CT scan was performed at intermission of onset on 11 patients with cerebral TIA; the data of plain CT scans, CT angiography (CTA) and whole brain perfusion on these patients were collected. Analysis by synthesis was performed on these imaging data. Results No lesions responsible for the attack were found in the brains of 11 patients with TIA under plain CT scan; 3 of them performed MRI did not have positive results. CTA indicated that the vascular networks in 10 patients were obviously sparse in the region of lesions responsible for the attack;the corresponding vessels in 8 of them became thin; both vascular thrombosis and compensatory angiogenesis occurred in 2 patients. The cerebral perfusion indicated that perfusion areas corresponding to clinical symptoms displayed delayed mean transit time and time to peak level in these 11 patients.Decreased regional cerebral blood flow was noted in 5 patients and regional cerebral volume increased mildly in 3 patients. Conclusion The 320-slice-volume CT perfusion imaging can obtain the information about whole brain perfusion, routine CT scan and CTA by injecting a kind of contrast medicament for only one time, thus can assess the ischemic cerebrovascular disease comprehensively.

The infection status of anisakid larvae was examined in 290 marine fish of 25 species and in 108 cephalopods of 3 species purchased in Bayuquan region, Yingko city nearby the coast of the Bohai Sea from may to August 1992. A total of 7,327 larvae were collected from 156 fish of 19 species and 8 squids of one species. The 3rd-stage larvae of Anisakis simplex were collected from 121 fish (63.4%) of 15 species (N = 191) and from 8 squids (14.8%) of one species (N = 54), and they were total, 5,992 (81.8%). Out of remaining 1,335 larvae, 154 (2.1%) were classified as Thynnascaris type B from 23 fish of 4 species, 1,013 (13.8%) as Thynnascaris type C from 79 fish of 13 species. 164 (2.2%) as Hysterothylacium China type V from 20 fish of 4 species, 3 (0.04%) as Raphidascaris from 3 fish of 2 species and one was Pseudoterranova decipiens larva.
Animal ; Anisakiasis/veterinary* ; Anisakiasis/parasitology ; Anisakiasis/epidemiology ; Anisakis/isolation & purification ; Anisakis/classification* ; China ; Fish Diseases/parasitology* ; Fish Diseases/epidemiology ; Fishes ; Larva ; Seawater ; Squid/parasitology*

This study was aimed to investigate the T cell (helper T cells) immune status in ITP patients and its relation with therapeutic response. 20 de novo ITP patients were enrolled (8 males, 12 females) with a median age of 41 (20 to 81). Real-time RT-PCR method was used to measure the gene expression of Th cells including T-bet, IFN-γ, GATA-3, TGF-β, Foxp3, IL-2, IL-4 in PBMNC of ITP patients before and after conventional dose of prednisone therapy [1 mg/(kg·d)] and in PBMNC of 20 normal controls. The results showed that T-bet, IFN-γ and IL-2 were significantly over-expressed in PBMNC of ITP patients before treatment compared with that in normal controls (p < 0.01), and compared with that before treatment, T-bet, IFN-γ, and IL-2 were markedly down-regulated in ITP patients after treatment. Before treatment, the expressions of Foxp3, TGF-β, GATA3 and IL-4 in ITP patients did not show difference from normal controls, while after treatment Foxp3 were more up-regulated than that before treatment (p < 0.05). After treatment, TGF-β expression showed a different pattern between old and young patients. TGF-β expression was down-regulated (p < 0.05) among ITP patients younger than 60, while up-regulated in older patients. It is concluded that there is an imbalance of Th1/Th2/Treg cytokines in ITP patients, which can be reversed by glucocorticoid treatment. The conventional dose of glucocorticoid may be regarded as effective therapy for de novo ITP patients, it may correlate with improvement of imbalance between Th1/The2/Treg cytokines.
Adult ; Aged ; Aged, 80 and over ; Case-Control Studies ; Female ; Forkhead Transcription Factors ; metabolism ; Gene Expression ; Gene Expression Regulation ; Glucocorticoids ; therapeutic use ; Humans ; Interferon-gamma ; metabolism ; Interleukin-2 ; metabolism ; Male ; Middle Aged ; Purpura, Thrombocytopenic, Idiopathic ; drug therapy ; metabolism ; pathology ; T-Box Domain Proteins ; metabolism ; T-Lymphocytes, Helper-Inducer ; metabolism ; Transforming Growth Factor beta ; metabolism ; Young Adult