[A multicenter study of neonatal stroke in Shenzhen, China]. / 深圳市新生儿脑卒中多中心现况调查.

OBJECTIVES: To investigate the incidence rate, clinical characteristics, and prognosis of neonatal stroke in Shenzhen, China. METHODS: Led by Shenzhen Children's Hospital, the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022. The incidence, clinical characteristics, treatment, and prognosis of neonatal stroke in Shenzhen were analyzed. RESULTS: The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137, 1/6 060, and 1/7 704, respectively. Ischemic stroke accounted for 75% (27/36); boys accounted for 64% (23/36). Among the 36 neonates, 31 (86%) had disease onset within 3 days after birth, and 19 (53%) had convulsion as the initial presentation. Cerebral MRI showed that 22 neonates (61%) had left cerebral infarction and 13 (36%) had basal ganglia infarction. Magnetic resonance angiography was performed for 12 neonates, among whom 9 (75%) had involvement of the middle cerebral artery. Electroencephalography was performed for 29 neonates, with sharp waves in 21 neonates (72%) and seizures in 10 neonates (34%). Symptomatic/supportive treatment varied across different hospitals. Neonatal Behavioral Neurological Assessment was performed for 12 neonates (33%, 12/36), with a mean score of (32±4) points. The prognosis of 27 neonates was followed up to around 12 months of age, with 44% (12/27) of the neonates having a good prognosis. CONCLUSIONS: Ischemic stroke is the main type of neonatal stroke, often with convulsions as the initial presentation, involvement of the middle cerebral artery, sharp waves on electroencephalography, and a relatively low neurodevelopment score. Symptomatic/supportive treatment is the main treatment method, and some neonates tend to have a poor prognosis.

[Efficacy and safety of the 177Lu-PSMA-617 therapy in the treatment of metastatic castration-resistant prostate cancer: A meta-analysis].

OBJECTIVE: To evaluate the clinical efficacy and safety of the 177Lu-PSMA-617 therapy in the treatment of metastatic castration-resistant prostate cancer (mCRPC). METHODS: We searched PubMed, Medline, Embase, Cochrane Library, CNKI, VIP, CBM and Wanfang Database from inception to July 2019 for published retrospective studies on the treatment of mCRPC. We performed literature screening, data extraction and quality evaluation according to the inclusion and exclusion criteria, and conducted a meta-analysis of the data obtained using the STATA15.1 software. RESULTS: A total of 12 studies involving 508 cases of mCRPC were included in this analysis. After the first cycle of treatment, the pooled rate of PSA decline was 69.30% (95% CI: 65.40%－73.30%), and that of >50% PSA decline was 35.90% (95% CI: 31.80%－40.00%). No significant adverse events were reported in any of the studies. CONCLUSIONS: The177Lu-PSMA-617 therapy is a safe and effective option for the treatment of metastatic castration-resistant prostate cancer.

[Effects of long non-coding RNA RP1-90L14.1 on the biological behaviors of cancer prostate LNCaP cells and its regulating mechanisms].

OBJECTIVE: To investigate the effects of long non-coding RNA RP1-90L14.1 on the proliferation, migration and invasion of prostate cancer LNCaP cells and the expressions of GRIN2A and BACE2. METHODS: Using RT-PCR, we detected the expression of RP1-90L14.1 in LNCaP and LNCaP-AI cells, transiently transfected the RP1-90L14.1 overexpression plasmid (the RP1-90L14.1 group) and vector plasmid (the LNCaP-NC group) into the LNCaP cells, and cultured the two groups of cells with ordinary medium and phenol red-free activated carbon adsorption medium (PRF-ACA). Then we examined the proliferation, migration and invasiveness of the cells by CCK-8 and Transwell, and determined the mRNA and protein expressions of GRIN2A and BACE2 by RT-PCR and Western blot. RESULTS: The expression of RP1-90L14.1 was significantly higher in the LNCaP-AI than in the LNCaP cells (8.49 ± 0.43 vs 2.53 ± 0.95, P < 0.05), and so was that of LNCaP-RP1-90L14.1 in the RP1-90L14.1 than in the LNCaP-NC group after transfection (0.71 ± 0.22 vs 0.02 ± 0.01, P < 0.05). The optical densities (OD) of the cells were 51.95% and 50.69% higher in the RP1-90L14.1 than in the LNCaP-NC group after 72 hours of culture with ordinary medium and phenol red-free ACA (1.22 ± 0.08 vs 0.08 ± 0.05, P < 0.05; 0.79 ± 0.02 vs 0.53 ± 0.05, P < 0.05), and 51.72% and 60.23% higher in the former than in the latter after 96 hours (1.72 ± 0.07 vs 1.13 ± 0.05, P < 0.05; 1.18 ± 0.05 vs 0.73 ± 0.08, P < 0.05). The numbers of the migrating cells cultured with common medium and PRF-ACA were markedly higher in the RP1-90L14.1 than in the LNCaP-NC group after transfection (682.0 ± 42.7 vs 422.0 ± 37.1, P < 0.05; 419.0 ± 42.9 vs 251.0 ± 25.9, P < 0.05), and so were those of the invading cells (507.0 ± 22.2 vs 274.0 ± 19.6, P < 0.05; 352.0 ± 14.1 vs 216.0 ± 14.3, P < 0.05). Statistically significant differences were observed between the RP1-90L14.1 and LNCaP-NC groups in the mRNA and protein expressions of GRIN2A (5.13 ± 0.89 vs 2.09 ± 0.54, P < 0.05; 5.88 ± 0.29 vs 2.03 ± 0.22, P < 0.05) and BACE2 (5.82 ± 0.50 vs 2.53 ± 0.30, P < 0.05; 4.89 ± 0.19 vs 3.37 ± 0.13, P < 0.05). CONCLUSIONS: 　lncRNA RP1-90L14.1 may play important roles in the proliferation, migration and invasiveness of prostate cancer cells. RP1-90L14.1 can promote the expressions of GRIN2A and BACE2 and may have an endogenous competitive relation with GRIN2A and BACE2.

Preparation and characterization of WSe2 nano-films by magnetron sputtering and vacuum selenization.

Graphene-structure WSe2 nano-films (NFMs) were prepared by magnetron sputtering and vacuum selenization. The as synthesized WSe2 NFMs were characterized using various techniques such as SEM, XRD and EDS. Our work provides a simple and effective method to prepare WSe2 NFMs. We have observed the high photo responsivity of WSe2 NFMs up to 10 A W-1, which is 40% higher than the previously reported data. Our photoelectrochemical tests demonstrate that WSe2 NFMs have excellent photoelectric properties, indicating that NFMs have a wide application potentialsin solar cells and optoelectronic devices.

[Long non-coding RNAs in prostate cancer: An update].

In the human genome, there is a group of RNAs, called long non-coding RNA (lncRNA) with do not have the function of encoding proteins and whose transcript length is greater than 200 nucleotides. The disorders of lncRNAs are often involved in the occurrence and progression of malignant tumors. A large number of studies have indicated the aberrant expression of lncRNAs in prostate cancer (PCa) can regulate gene expressions at epigenetic, transcriptional and post-transcriptional levels and cause changes in the biological behaviors of PCa cells. Some lncRNAs have been shown to be closely related to the castration resistance of PCa. In recent years, a variety of lncRNAs have been detected in the PCa tissue, prostatic fluid, serum, and urine, and somehow influenced radiotherapy and chemotherapy of tumors. The expressions of some lncRNAs are also associated with disease prognosis. Thus, lncRNAs are expected to become new diagnostic markers and a therapeutic target for PCa. This review focuses on the roles and action modes and mechanisms of some lncRNAs as well as their potential value of clinical application in the diagnosis, treatment and prognosis of PCa.

The role of fascin in migration and invasion of urothelial carcinoma of the bladder.

INTRODUCTION: To investigate the roles of fascin in migration and invasiveness in bladder urothelial carcinoma. MATERIALS AND METHODS: Immunohistochemical detection of fascin in urothelial carcinoma samples and inhibition the expression of fascin in the urothelial carcinoma cell line were performed, then the differences in cell behaviors before and after silencing of the fascin gene were tested. RESULTS: In our study, we found that overexpression of fascin was more frequent in urothelial carcinoma tissues (p < 0.001). Fascin expression was positively correlated with histological grade (p = 0.024) and pT stage (p < 0.001). After transfection of fascin shRNA, the expressions of fascin in 5637 cells and BIU87 cells were efficiently decreased according to real-time RT-PCR and Western blot analysis. When fascin was inhibited, a significant decrease in migration and invasion, and increase in adhesion were observed in 5637 cells and BIU87 cells. However, there was no significant change in the proliferation of 5637 cells or BIU87 cells with or without inhibition of the fascin gene. CONCLUSIONS: Fascin expression can be used as a predictor for transformation and progression of urothelial carcinoma, and reduction of fascin levels may represent a novel therapeutic strategy for urothelial carcinoma of the bladder.

[Clinical efficacy of daily low-dose tadalafil on erectile dysfunction-no sexual life].

OBJECTIVE: Erectile dysfunction-no sexual life (ED-NS) is defined as the inability to have enough penile erection hardness and duration so as to have enough confidence in attempting sexual intercourse for more than six months. This study was to investigate the effect of daily low-dose tadalafil on ED-NS. METHODS: We treated 35 ED-NS patients aged 17-35 (25.9 +/- 3.9) years with oral tadalafil at 5 mg qd for 3 months and followed them up for another 3 months after drug withdrawal. We obtained the scores of the patients on Self-estimation Index of Erectile Function-No Sexual Life (SIEF-NS) and compared them before and after medication and at 3 months after drug withdrawal. RESULTS: The patients' SIEF-NS scores were 43.2 +/- 7.1 after medication and 42.1 +/- 7.4 at 3 months after drug withdrawal, both significantly higher than 21.2 +/- 5.9 before treatment (P < 0.05), though there was no significant difference between the former two scores (P > 0.05). CONCLUSION: Daily medication of low-dose tadalafil can significantly improve the erectile function of the patients with ED-NS.

[Hydrochemical and Isotopic Evidence for Groundwater Conversion of Surface Water in Alpine Arid Areas: A Case Study of the Datong River Basin].

Global warming and regional extreme climates will lead to unbalanced rainfall, melting glaciers, and permafrost degradation in alpine and arid mountain plains, thereby changing the regional hydrological cycle. The relationship between surface water and groundwater conversion is one of the important scientific issues of hydrological cycle climate response in alpine arid areas. Taking the Datong River Basin at the southern foot of the Qilian Mountains as the study area, based on 119 sets of basic hydrochemical parameters and deuterium-oxygen isotope data, using multivariate statistical analysis and isotopic techniques, the hydrochemical characteristics of surface water and groundwater in the basin and their mutual transformation process were studied. The results showed that the surface water was HCO3-Mg·Ca type, which was mainly controlled by rock weathering, whereas the groundwater was HCO3-Mg·Ca type and Cl·SO4-Na type, which was controlled by rock weathering and evaporation concentration. There was a small amount of calcium and magnesium feldspar dissolved in the upstream groundwater, and the chemical components of the midstream groundwater were mainly the weathering and dissolution of carbonate rocks. The contribution rates of weathering filtration, anthropogenic activities, native sedimentary environment, alternating adsorption of cations, and other factors to the chemical components of surface water and groundwater in the study area were 39.1%, 15.0%, 12.6%, 13.8%, and 19.5%, respectively. The deuterium and oxygen isotope contents of Datong River water showed a trend of enrichment to depletion along the groundwater flow direction. The δD and δ18O isotope test results showed that the deuterium and oxygen isotope content in the Datong River along the groundwater flow showed a trend of enrichment to depletion. The upper and middle reaches of the Datong River were mainly recharged by atmospheric precipitation, whereas the lower reaches were affected by geological structure and influenced by hydrogeological conditions, which was mainly due to diving and spring water overflow to supply river water, as the discharge area of groundwater.

Cloning, characterization and expression analysis of a cold shock domain family member YB-1 in turbot Scophthalmus maximus.

The Y-box proteins are a family of highly conserved nucleic acid binding proteins. In this report we have identified a new member, YB-1 from turbot (Scophthalmus maximus) spleen cDNA library. The full-length cDNA sequence of turbot YB-1 was obtained and then the expression at transcriptional level was researched by qRT-PCR. In normal organs, the expression of YB-1 was higher in liver, brain, gill and heart, respectively. YB-1 had the highest expression level at gastrula stage during the early stages of embryo development. In the liver, kidney and spleen, the turbot YB-1 expression level was the highest at 72 h after challenge with lymphocystis disease virus (LCDV) and the highest at 12 h after challenge with Vibrio anguillarum (V. anguillarum). Furthermore, the expression of turbot YB-1 also distinctly increased in turbot kidney cells (TK) at 24 h after challenge with V. anguillarum and LCDV. These results indicated that the turbot YB-1 protein may play a significant role in the immune response of turbot.

Bioaccumulation of differently-sized polystyrene nanoplastics by human lung and intestine cells.

The bioaccumulation of nanoplastics (NPs) has been intensively examined using the fluorescence-labeling technique. As the fluorescence intensity per particle is different for NPs with different physicochemical properties, it's hard to directly compare their bioaccumulation based on fluorescence. Therefore, how physicochemical properties may affect NPs' bioaccumulation remains unclear. In the present study, we chose polystyrene NPs (PSNPs) with the primary particle size of 70 nm (PS70), 200 nm (PS200), and 500 nm (PS500), and examined their uptake by human lung and intestine cells. We found that PSNPs had low cytotoxicity, but could be taken up by both cell lines. The particle-mass-, particle-number-, and particle-surface-area-based accumulation of the differently-sized PSNPs were then compared. Smaller PSNPs showed lower particle-mass-based but higher particle-number-based uptake rate than the larger ones. Nevertheless, much less difference was observed when the unit of uptake rate was based on particle surface area, suggesting the critical role of surface area during PSNPs' interaction with the cell membrane. Additionally, all three PSNPs could enter the cells by phagocytosis and PS70 could also be internalized by clathrin- and caveolae-mediated endocytosis. Overall, the effects of size on the bioaccumulation of NPs need to be considered when evaluating their environmental and health risks.

Graphene-supported polyoxometalate entrapped in a MIL-88A network with highly efficient conversion of polysulfides in Li-S batteries.

It has been demonstrated that polyoxometalates (POMs) have strong anchoring abilities with efficient catalysis of lithium polysulfides (LiPSs). However, the severe aggregation that buries the effective active sites of POMs along with poor electrical conductivity limits the practical application of POMs in lithium sulfur batteries (LSBs). In our strategy, we utilized reduced graphene oxide (rGO) to support a POM catalyst entrapped in a MIL-88A(FeCo) network with a hollow shell skeleton as the sulfur host material. H4PW11VO40 (PW11V) with optimal vanadium atom implantation ensures the ruggedness and integrity of the hollow structure, which is conducive to achieving high sulfur loading as well as accommodating the volume change of the sulfur cathode during the charging and discharging process. Importantly, PW11V can capture polysulfides through firm chemical adsorption and accelerate redox reactions of LiPS conversion by effective electrochemical catalysis. Furthermore, the satisfactory electrical conductivity of rGO provides access for electrons to reach the interface of PW11V and polysulfides and trigger Li-S conversion reactions. Thus, the constructed PW11V-based sulfur cathode exhibited a superior specific capacity of 905 mA h g-1 after 100 cycles under 0.2 C and long cycling life with a capacity recession rate of 0.046% for each cycle upon 500 cycles under 3 C. This research reveals the effect of vanadium atom substitution of POMs on the cycling performance of a sulfur cathode and affords insight for developing high-performance LSBs.

Manipulation of the MoO2/MoSe2 Heterointerface Boosting High Rate and Durability for Sodium/Potassium Storage.

The main challenge for sodium/potassium ion storage is to find the suitable host materials to accommodate the larger-sized Na+/K+ and conquer the sluggish chemical kinetics. Herein, by selenation of polyoxometalate in electrospinning fiber, a novel MoO2/MoSe2 heterostructure embedded in one-dimensional (1D) N,P-doped carbon nanofiber (MoO2/MoSe2@NPC) is rationally constructed to show distinct enhancement of rate performance and cycle life for sodium ion batteries (SIBs) and potassium ion batteries (PIBs). The 1D skeleton of MoO2/MoSe2@NPC decreases the diffusion pathway of Na+/K+, and the doping of N/P heteroatoms in carbon fiber creates abundant active sites and provides good reachability for Na+/K+ transportation. MoSe2 nanosheets grow in the bulk phase of MoO2 via in situ local phase transformation to achieve effective and firm heterointerfaces. Especially, the exposure extent of heterointerfaces can be controlled by treatment temperature during the preparation process, and the optimized heterointerfaces result in an ideal synergic effect between MoO2 and MoSe2. DFT calculations confirm that the internal electric field in the heterogeneous interface guides the electron transfer from MoO2 to MoSe2, combined with strong adsorption capacity toward sodium/potassium, facilitating ion/electron transfer kinetics. It is confirmed that the MoO2/MoSe2@NPC anode for SIBs delivers 382 mA h g-1 under 0.1 A g-1 upon 200 cycles; meanwhile, a reversible capacity of 266 mA h g-1 is maintained even under 2 A g-1 after 2000 cycles. For PIBs, it can reach up to 216 mA h g-1 in the 200th cycle and still retain 125 mA h g-1 after 2000 cycles under 1 A g-1. This study opens up a new interface manipulation strategy for the design of anode materials to boost fast Na+/K+ storage kinetics.

Melatonin inhibits ESCC tumor growth by mitigating the HDAC7/ß-catenin/c-Myc positive feedback loop and suppressing the USP10-maintained HDAC7 protein stability.

BACKGROUND: Melatonin, a natural hormone secreted by the pineal gland, has been reported to exhibit antitumor properties through diverse mechanisms of action. However, the oncostatic function of melatonin on esophageal squamous cell carcinoma (ESCC) remains elusive. This study was conducted to investigate the potential effect and underlying molecular mechanism of melatonin as single anticancer agent against ESCC cells. METHODS: ESCC cell lines treated with or without melatonin were used in this study. In vitro colony formation and EdU incorporation assays, and nude mice tumor xenograft model were used to confirm the proliferative capacities of ESCC cells. RNA-seq, qPCR, Western blotting, recombinant lentivirus-mediated target gene overexpression or knockdown, plasmids transfection and co-IP were applied to investigate the underlying molecular mechanism by which melatonin inhibited ESCC cell growth. IHC staining on ESCC tissue microarray and further survival analyses were performed to explore the relationship between target genes' expression and prognosis of ESCC. RESULTS: Melatonin treatment dose-dependently inhibited the proliferative ability and the expression of histone deacetylase 7 (HDAC7), c-Myc and ubiquitin-specific peptidase 10 (USP10) in ESCC cells (P < 0.05). The expressions of HDAC7, c-Myc and USP10 in tumors were detected significantly higher than the paired normal tissues from 148 ESCC patients (P < 0.001). Then, the Kaplan-Meier survival analyses suggested that ESCC patients with high HDAC7, c-Myc or USP10 levels predicted worse overall survival (Log-rank P < 0.001). Co-IP and Western blotting analyses further revealed that HDAC7 physically deacetylated and activated ß-catenin thus promoting downstream target c-Myc gene transcription. Notably, our mechanistic study validated that HDAC7/ß-catenin/c-Myc could form the positive feedback loop to enhance ESCC cell growth, and USP10 could deubiquitinate and stabilize HDAC7 protein in the ESCC cells. Additionally, we verified that inhibition of the HDAC7/ß-catenin/c-Myc axis and USP10/HDAC7 pathway mediated the anti-proliferative action of melatonin on ESCC cells. CONCLUSIONS: Our findings elucidate that melatonin mitigates the HDAC7/ß-catenin/c-Myc positive feedback loop and inhibits the USP10-maintained HDAC7 protein stability thus suppressing ESCC cell growth, and provides the reference for identifying biomarkers and therapeutic targets for ESCC.

Efficacy and safety of Fuzheng Huayu tablet on persistent advanced liver fibrosis following 2 years entecavir treatment: A single arm clinical objective performance criteria trial.

ETHNOPHARMACOLOGICAL RELEVANCE: Antiviral therapy can alleviate liver fibrosis in chronic hepatitis B, but it has a limited effect on advanced liver fibrosis/cirrhosis. Traditional Chinese medicine (TCM), particularly FuZheng HuaYu (FZHY) tablet, appears to have an antifibrotic effect, but its improving resolution of hepatitis b virus (HBV) -associated advanced fibrosis and experienced anti-viral treatment has not been investigated. AIM OF THE STUDY: To observe the safety and efficacy of adjunctive FZHY on the HBV-associated cirrhosis patients who received 2 years of entecavir but still with advanced fibrosis. METHODS: An open-label, multicentre, single arm trial. 251 patients were included and treated with TCM consisted of FZHY tablets 1.6 g and granules, three times a day in addition to entecavir 0.5 mg daily for an additional 48 weeks. Primary outcome was regression of fibrosis (the proportion of patients with a 1-point decrease in the Ishak liver fibrosis score from baseline to week 48). RESULTS: Fibrosis regression occurred in 94 of 184 patients with paired liver biopsy (51.09%, 95% CI: 43.9～58.0). In 132 compensated cirrhosis patients (Ishak score ≥5), 56.06% (74/132, 95% CI: 47.5～64.2) showed fibrosis regression and reached the goal of 54% (15% more than entecavir mono-therapy). 10 patients occurred adverse reaction, most of them were mild, and all recovered or achieved remission. CONCLUSIONS: The combination therapy of FZHY, TCM granules and ETV could regress the liver fibrosis in the patients with HBV cirrhosis, who experienced 2 years of ETV treatment, and it is safe and well tolerated.

[Geochemical Characteristics and Driving Factors of NO3-Type Groundwater in the Rapidly Urbanizing Pearl River Delta].

In response to rapid economic development, nitrate pollution of groundwater is becoming a serious issue in many parts of China. Urbanization and industrialization are the main drivers of NO3-type groundwater expansion. Focusing on the Pearl River Delta, the occurrence and driving factors of shallow nitrate groundwater are discussed. Overall, groundwater nitrate concentrations are generally high in this region. Of 1538 groundwater samples, 5.7% had nitrate concentrations higher than the groundwater quality standard(88.6 mg·L-1) and 18.5% were classified as NO3-type waters, which are mainly distributed in the hilly and piedmont areas. Guangzhou, Dongguan, Foshan, Zhuhai and other areas show high total dissolved solid(TDS)-concentration NO3-type waters, which are affected by urbanization and industrialization. In comparison, low-TDS NO3-type waters are distributed in the hilly and valley areas. In the Xijiang and Dongjiang plains, the TDS concentrations on groundwater increased significantly due to inputs of industrial wastewater and saline seawater. The NO3- concentration in the groundwater in this area exceeded the class III water standard but did not change the hydrochemical type classification. However, industrialization has led to the frequent appearance of SO4-type water in this area. The NO3-type water occurs in acidic or weakly acidic environments, typically characterized by low TDS and total hardness concentrations, and high Cl-, SO42-, and K+ concentrations. The formation of NO3-type water is mainly affected by domestic sewage, industrial wastewater, agricultural nitrogen fertilizer, septic tank outflows, and landfill leachate leakage. Generally, the pollution loads of high-TDS NO3-type waters are higher than low-TDS NO3-type waters. The delineation of NO3-type waters, especially the low-TDS type, is helpful for identifying groundwaters posing greater risks for human activities, and those with low nitrate concentrations but potential pollution risk, which is of great significance in the prevention and control of groundwater pollution.

The complete mitochondrial genome of an Asian warty newt, Paramesotriton aurantius (Caudata: Salamandridae).

Paramesotriton aurantius (Caudata: Salamandridae) is a new species that found in southeastern China. Its complete mitochondrial genome (mitogenome) sequence was 16,313 bp in length with with A + T contents of 60.9%, and contained 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, one control region (D-loop), and one non-coding region. Our molecular tree showed that P. aurantius was positioned near P. hongkongensis, and formed a clade with other Paramesotriton species. The first complete mitogenome sequence of P. aurantius could provided fundamental data for resolving phylogenetic and genetic problems related to genus Paramesotriton.

Ursolic acid overcomes Bcl-2-mediated resistance to apoptosis in prostate cancer cells involving activation of JNK-induced Bcl-2 phosphorylation and degradation.

Androgen-independent prostate cancers express high levels of Bcl-2, and this over-expression of Bcl-2 protects prostate cancer cells from undergoing apoptosis. Ursolic acid (UA) has demonstrated an anti-proliferative effect in various tumor types. The aim of this study is to evaluate the difference between UA-induced apoptosis in androgen-dependent prostate cancer cell line LNCaP cells and androgen-independent prostate cancer cell line LNCaP-AI cells and to reveal the molecular mechanisms underlying the apoptosis. We found that UA treatment in vitro can effectively induce apoptosis in LNCaP and LNCaP-AI cells. UA can overcome Bcl-2-mediated resistance to apoptosis in LNCaP-AI cells. Intrinsic apoptotic pathways can be triggered by UA treatment because c-Jun N-terminal kinase (JNK) is activated and subsequently provokes Bcl-2 phosphorylation and degradation, inducing activation of caspase-9. Although further evaluation is clearly needed, the present results suggest the potential utility of UA as a novel therapeutic agent in advanced prostate cancer.

Differential phosphoprotein levels and pathway analysis identify the transition mechanism of LNCaP cells into androgen-independent cells.

BACKGROUND: Androgen withdrawal can prolong life in men with advanced prostate cancer, but these remissions are temporary because the surviving cells progress as hormone-refractory cancer. The mechanisms that are involved in the transition of androgen-dependent prostate cancer into androgen-independent prostate cancer (AIPC) are not fully understood. OBJECTIVE: To identify globally differentially expressed phosphoproteins in the androgen-independent prostate, to elucidate the molecular mechanisms that underlie the formation of AIPC and to identify new molecular targets that can be used to develop treatments for the disease. METHODS: An androgen-independent LNCaP cell line, LNCaP-AI, was established using androgen ablation. Differentially expressed phosphoproteins in LNCaP cells and LNCaP-AI cells were enriched by immunoprecipitation, analyzed by 2D-PAGE and identified by MALDI-TOF MS. Total protein expression levels for two regulated proteins were confirmed by Western blot. Association network analysis was carried out using the STRING database. RESULTS: The phosphorylation statuses of 17 proteins were significantly (P < 0.05) different between LNCaP-AI cells and LNCaP cells. Most proteins that were identified are known to be involved in tumor progression, and several of these proteins could be constructed into an association network. A further analysis by bioinformatics indicated that P53, HSP27, and the MAPK pathway may contribute to the transition from androgen-dependence to androgen-independence. CONCLUSION: Blocking the MAPK signaling pathway may be useful in the treatment of AIPC.

p53 and its isoforms in DNA double-stranded break repair.

DNA double-stranded break (DSB) is one of the most catastrophic damages of genotoxic insult. Inappropriate repair of DNA DSBs results in the loss of genetic information, mutation, and the generation of harmful genomic rearrangements, which predisposes an organism to immunodeficiency, neurological damage, and cancer. The tumor repressor p53 plays a key role in DNA damage response, and has been found to be mutated in 50% of human cancer. p53, p63, and p73 are three members of the p53 gene family. Recent discoveries have shown that human p53 gene encodes at least 12 isoforms. Different p53 members and isoforms play various roles in orchestrating DNA damage response to maintain genomic integrity. This review briefly explores the functions of p53 and its isoforms in DNA DSB repair.

[The role of mitogen-activated protein kinase cascades in inhibition of proliferation in human prostate carcinoma cells by raloxifene: an in vitro experiment].

OBJECTIVE: To investigate the role of mitogen-activated protein kinase (MAPK) in the apoptosis and cell cycle arrest of human prostate carcinoma cells induced by raloxifene (RAL). METHODS: Human prostate carcinoma cells of the line PC3 were cultured. RAL of the concentrations of 10(-4), 10(-5), 10(-6), and 10(-7) mol/L were added into the culture fluid. MTT method was used to detect the inhibitory rate of the PC3 proliferation. RAL of the concentrations of 10(-6) mol/L or 10(-6) mol/L +10 micromol/L PD98059, a MEK1/2 inhibitor, and 10(-6) mol/L RAL +10 micromol/L SB203580, JNK inhibitor, and RAL + SB203580, a p38 inhibitor were added respectively for 48 h, and the flow cytometry (FCM) was used to detect the cell cycle. The cell apoptosis percentage was measured by TUNEL staining. The activation of extra cellular regulated protein kinases (ERK1/2), c-Jun N-terminal protein kinase (JNK), and p38 mitogen-activated protein kinase (p38), and the expressions of Bcl-2, Bax, phospho-Bcl-2 (p-Bcl-2), and caspase-3 were determined by Western blotting. The expressions of estrogen receptor (ER) alpha, ERbeta, cyclin dependent kinase inhibitor (P21(WAF1)), and cyclinD1 mRNA were determined by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: A dose-dependent proliferation inhibition of RAL was demonstrated in the PC3 cells. A G(1) cell cycle arrest and apoptosis were induced in the PC3 cells exposed to 10(-6) mol/L RAL. 10(-6) mol/L RAL induced the activation of ERK1/2 and p38 with different time courses, but it did not induce the activation of JNK. Suppression ERK1/2 activation by treatment with PD98059 or p38 activation by treatment with SB203580 attenuated the cell-cycle arrest at the G(1) phase respectively. 48 h after the treatment of 10(-6) mol/L RAL the PC3 cells was arrested at G(1) stage, however, 48 h after the treatment of 10(-6) mol/L RAL +10 micromol/L PD98059 and 10(-6) mol/L RAL +10 micromol/L SB203580 the degree of PC3 cell arrest at the G(1) stage was lower. 18 h after the treatment of 10(-6) mol/L RAL, the expressions of cyclinD1 and P21(WAF1) gene were (0.50 +/- 0.02) and (4.48 +/- 0.12) times that of the control group, and the expressions of cyclinD1 and P21(WAF1) gene of the SB203580 pretreatment group and SB203580 pretreatment groups were (2.36 +/- 0.08) and (4.50 +/- 0.03) times, and (0.49 +/- 0.02) and (1.77 +/- 0.06) times respectively. 48 h after treatment with 10(-6) mol/L RAL, the apoptosis rates were 22.9% +/- 1.5%, significantly higher than those of the 10(-6) mol/L RAL +10 micromol/L PD98059 and 10(-6) mol/L RAL +10 micromol/L SB203580 groups (15.2% +/- 1.8% and 9.7% +/- 0.6% respectively, both P < 0.05). The expression of Bcl-2, Bax and caspase-3 (control, 10(-6) mol/L raloxifene, 10(-6) mol/L raloxifene +10 micromol/L PD98059 and 10(-6) mol/L raloxifene +10 micromol/L SB203580) was 1, 0.33 +/- 0.02, 0.34 +/- 0.01, 0.81 +/- 0.05; 1, 3.14 +/- 0.02, 1.67 +/- 0.11, 3.15 +/- 0.03; 1, 4.16 +/- 0.02, 2.66 +/- 0.03, 1.80 +/- 0.06, respectively. Western blotting showed that 48 h after the treatment of 10(-6) mol/L RAL the expression of Bcl-2 decreased, and the expression of caspace-3 increased. Pretreatment with PD98059 inhibited the increase of caspase-3 and Bax expression induced by RAL. 1.5 h after the treatment with RAL the p- Bcl-2 expression increased remarkable, pretreatment with SB203580 inhibited the p- Bcl-2 expression induced by RAL, and however, PD98059 did not show such effect. CONCLUSION: RAL inhibits the proliferation and induces the apoptosis and G(1) cell cycle arrest via MAPK cascades in human prostate carcinoma cells. Up-regulation of P21(WAF1) mRNA expression by activating ERK1/2 and down-regulation of cyclinD1 by activating p38 induces G(1) cell cycle arrest in the human prostate carcinoma cells. The ERK1/2 or p38 cascade is involved in the induction of apoptosis by RAL. The activation of ERK1/2 increases the expression of Bax. The activation of p38 phosphorylates Bcl-2 then inactivates Bcl-2.