[The clinical features, survival analysis, and geriatric assessment of 85 patients with follicular lymphoma: a single-center study].

Objective: To retrospectively analyze the clinical characteristics and prognosis of 85 newly diagnosed patients with follicular lymphoma (FL), as well as the prognostic value of comprehensive geriatric assessment (CGA) in patients with FL aged ≥ 60 years old. Methods: The clinical data and prognosis of 85 newly diagnosed FL patients admitted from August 2011 to June 2022 were collected. The clinical features, laboratory indicators, therapeutic efficacy, survival and prognostic factors of patients were statistically analyzed, and the prognosis of patients was stratified using various geriatric assessment tools. Results: ① The patients with FL were mostly middle-aged and older, with a median age of 59 (20-87) years, including 41 patients (48.2%) aged ≥60 years. The ratio of male to female was 1∶1.36. Overall, 77.6% of the patients were diagnosed with Ann Arbor stage Ⅲ-Ⅳ, and 17 cases (20.0%) were accompanied by B symptoms. Bone marrow involvement was the most common (34.1%). ②Overall, 71 patients received immunochemotherapy. The overall response rate was 86.6%, and the complete recovery rate was 47.1% of 68 evaluated patients. Disease progression or relapse in the first 2 years was observed in 23.9% of the patient. Overall, 14.1% of the patients died during follow-up. ③Of the 56 patients receiving R-CHOP-like therapies, the 3-year and 5-year progression-free survival (PFS) rates were 85.2% and 72.8%, respectively, and the 3-year and 5-year overall survival (OS) rates were 95.9% and 88.8%, respectively. The univariate analysis showed that age ≥60 years old (HR=3.430, 95% CI 1.256-9.371, P=0.016), B symptoms (HR=5.030, 95% CI 1.903-13.294, P=0.016), Prognostic Nutritional Index (PNI) <45.25 (HR=3.478, 95% CI 1.299-9.310, P=0.013), Follicular Lymphoma International Prognostic Index (FLIPI) high-risk (HR=2.918, 95% CI 1.074-7.928, P=0.036), and PRIMA-prognostic index (PRIMA-PI) high-risk (HR=2.745, 95% CI 1.057-7.129, P=0.038) significantly predicted PFS. Moreover, age ≥60 years old and B symptoms were independent risk factors for PFS. Progression of disease within 24 months (POD24) significantly predicted OS in the univariate analysis. Conclusions: FL is more common among middle-aged and older women. Age, B symptoms, PNI score, FLIPI high-risk, PRIMA-PI high-risk, and POD24 influenced PFS and OS. The CGA can be used for treatment selection and risk prognostication in older patients with FL.

Sex identification of ducklings based on acoustic signals.

Sex identification of ducklings is a critical step in the poultry farming industry, and accurate sex identification is beneficial for precise breeding and cost savings. In this study, a method for identifying the sex of ducklings based on acoustic signals was proposed. In the first step, duckling vocalizations were collected and an improved spectral subtraction method and high-pass filtering were applied to reduce the influence of noise. Then, duckling vocalizations were automatically detected by using a double-threshold endpoint detection method with 3 parameters: short-time energy (STE), short-time zero-crossing rate (ZCR), and duration (D). Following the extraction of Mel-Spectrogram features from duckling vocalizations, an improved Res2Net deep learning algorithm was used for sex classification. This algorithm was introduced with the Squeeze-and-Excitation (SE) attention mechanism and Ghost module to improve the bottleneck of Res2Net, thereby improving the model accuracy and reducing the number of parameters. The ablative experimental results showed that the introduction of the SE attention mechanism improved the model accuracy by 2.01%, while the Ghost module reduced the number of model parameters by 7.26M and the FLOPs by 0.85G. Moreover, this algorithm was compared with 5 state-of-the-art (SOTA) algorithms, and the results showed that the proposed algorithm has the best cost-effectiveness, with accuracy, recall, specificity, number of parameters, and FLOPs of 94.80, 94.92, 94.69, 18.91M, and 3.46G, respectively. After that, the vocalization detection score and the average confidence strategy were used to predict the sex of individual ducklings, and the accuracy of the proposed model reached 96.67%. In conclusion, the method proposed in this study can effectively detect the sex of ducklings and serve as a reference for automated sex identification of ducklings.

[Analysis of the common respiratory viruses in children with acute respiratory infection in a hospital in Lanzhou City from 2021 to 2022].

To explore the situation of 8 common respiratory pathogens in children with acute respiratory infection (ARI) from 2021 to 2022.The retrospective study selected 8 710 ARI patients from September 2021 to August 2022 in the Maternal and Child Health Hospital of Gansu Province as the study object, patients aged 0 to 17 years old, including 5 048 male children and 3 662 female children. Indirect immunofluorescence was used to detect 8 common respiratory pathogens, including influenza virus A (FluA), influenza virus B (FluB), parainfluenza virus (PIV), respiratory syncytial virus (RSV), adenovirus (ADV), Mycoplasma pneumoniae (MP), Chlamydia pneumoniae (CP), and Coxsackie virus group B (CoxB) IgM antibodies. χ2 test was used to analyze the results. The results showed that 1 497 of 8 710 children with ARI were positive, with a positive rate of 17.19%. The detection rate of MP among 8 common respiratory pathogens was 11.34%, accounting for 66.0%, followed by FluB, CoxB, PIV, RSV, ADV, FluA and CP, accounting for 13.83%, 9.55%, 6.01%, 2.61%, 1.47%, 0.40% and 0.13%, respectively. Respiratory tract viruses (FluA, FluB, RSV, ADV, PIV, CoxB) accounted for 33.86%.There were significant differences in the detection rates of PIV, ADV and MP among children of different genders (χ2=6.814, 5.154 and 17.784, P<0.05). The detection rate of school-age children (6-17 years old) was the highest, accounting for 33.27% (184/553). The detection rates of 8 common respiratory pathogens in patients with ARI were higher in spring and winter and lower in summer and autumn. To sum up, from 2021 to 2022, MP and FluB infection were dominant in ARI patients in our hospital. The peak period of 8 common respiratory pathogens was in spring and winter. The physical examination rate of 8 common respiratory pathogens in ARI patients aged 6-17 years old was the highest.

RTKN2 Enhances Radioresistance in Gastric Cancer through Regulating the Wnt/ß-Catenin Signalling Pathway.

Adjuvant therapy and radiotherapy improves the survival of patients with metastatic and locally advanced gastric cancer (GC). However, the resistance to radiotherapy limits its clinical usage. Rhotekin 2 (RTKN2) functions as an oncogene and confers resistance to ultraviolet B-radiation and apoptosis- inducing agents. Here, the role of RTKN2 in radiosensitivity of GC cell lines was investigated. RTKN2 was found to be elevated in GC tissues and cells. A series of functional assays revealed that overexpression of RTKN2 induced GC cell proliferation, promoted GC cell migration and invasion, while inhibiting GC cell apoptosis. However, silence of RTKN2 promoted GC cell apoptosis, while repressing GC cell proliferation, invasion and migration. GC cells were exposed to irradiation, and data from cell survival and apoptotic assays showed that knock-down of RTKN2 enhanced radiosensitivity of GC through up-regulation of apoptosis and down-regulation of proliferation in irradiation-exposed GC cells. Moreover, the protein expression of ß-catenin and c-Myc in GC cells was enhanced by RTKN2 over-expression, but reduced by RTKN2 silence. Interference of RTKN2 down-regulated nuclear ß-catenin expression, while up-regulating cytoplasmic ß-catenin in GC. In conclusion, RTKN2 contributed to cell growth and radioresistance in GC through activation of Wnt/ß-catenin signalling.

[Dose-effect relationship between serum polycyclic aromatic hydrocarbon adducts and serum complements among children in a city of East China].

Objective: To investigate the dose-response relationship between serum polycyclic aromatic hydrocarbon adducts and serum complement C3 and C4 levels among children from a city in East China. Methods: In September 2016, two boarding schools in the air pollution exposure area and the control area (beyond the upwind of 30 km in the air pollution exposure area) in a city in East China were selected as the research site, and the eligible school-age children were recruited as the research objects. A total of 273 children were included, including 163 in the exposure group and 110 in the control group. The annual air pollutant data (PM2.5, PM10 and NO2) of the two regions during the study period were collected. The exposure level of tobacco was evaluated by cotinine in urine. The levels of serum complement C3 and C4 were determined by automatic biochemical analyzer. The serum anti-7, 8, -dihydrodiol-9, 10-epoxide benzo[a]pyrene (BPDE)-albumin adduct levels were detected by ELISA. Linear regression model was used to explore the dose-response relationship between BPDE-albumin adducts and serum complement C3 and C4. Results: The age of 273 subjects was (13.67±0.37) years old, including 165 boys (60.4%). The average annual exposure levels of PM2.5, PM10 and NO2 and the level of serum BPDE-albumin adducts in the exposure group were higher than those in the control group (P<0.05). The results of linear regression model analysis showed that after adjusting age, sex, BMI z-score and urinary cotinine level, when the serum BPDE-albumin adduct level increased by 10%, the serum complement C4 level decreased by 1.2% (P=0.017). After adjusting age, BMI z-score and urinary cotinine level, for every 10% increase in serum BPDE-albumin adduct level in boys, the serum complement C4 level decreased by 1.68% (P=0.024). After adjusting age, sex and BMI z-score, the levels of serum complement C3 and C4 decreased by 1.31% and 3.57% respectively for every 10% increase in serum BPDE-albumin adducts among children in the urinary cotinine detection group (P<0.05). Conclusion: There is a significant dose-response relationship between serum BPDE-albumin adducts and the complement C4 among children.

[Effect of different mechanical ventilation modes on patient-ventilator synchrony and diaphragm function in rabbit model of acute respiratory distress syndrome].

Objective: To observe the effect of different modes of mechanical ventilation on patient-ventilator synchrony and diaphragm function in rabbits with acute respiratory distress syndrome(ARDS). Methods: Eighteen New Zealand rabbit models of ARDS were induced by intratracheal infusion hydrochloric acid until the oxygenation index (PaO(2)/FiO(2)) was less than 200 mmHg, and then divided into three groups with random number: assisted-controlled mechanical ventilation (A/C) group, pressure support ventilation (PSV) group and neurally adjusted ventilatory assist (NAVA) group. All of them were ventilated for four hours with the targeted tidal volume (V(T)) (6 ml/kg) and the positive end-expiratory pressure (PEEP) titrated with the maximum oxygenation method. Gas exchange, pulmonary mechanics and patient-ventilator synchrony were determined during 4 h of ventilation and the concentrations of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) in diaphragm were measured after 4 h of ventilation. The q test was used for the multiple comparison of the sample mean. Results: There were no significant differences in PaO(2)/FiO(2) between three groups during ventilation 1-4 h (F=1.029, P>0.05). The V(T) in NAVA group was obviously lower than that in PSV group and the respiratory rate (RR) and the electrical activity of diaphragm(EAdi) were higher than those in A/C group(all P<0.05).The trigger delay and off cycle delay the in NAVA group were markedly lower than those in A/C and PSV group during ventilation 1-4 h(F=14.312, 9.342, both P<0.05). Asynchrony index in NAVA group (3.1%±1.0%) was obviously lower than those in A/C group (22.3%±5.2%) and PSV group(8.4%±2.3%) (F=7.192, P<0.05). In NAVA group, peak EAdi (EAdi(peak)) and peak airway pressure (Ppeak) were markedly correlated (r=0.97±0.16, P<0.05), while Ppeak delivery in A/C and PSV group was not correlated to EAdi(peak) (r=0.38±0.13,0.46±0.15, both P>0.05).Compared with A/C group, the concentration of MDA in the diaphragm in NAVA group was obviously lower(P<0.05). SOD and GSH level inthe diaphragm in NAVA group were both obviously higher than those in A/C group (both P<0.05). Conclusions: It is helpful to avoid eccentric contraction of diaphragm, lessen oxidative stress and alleviate ventilator-related diaphragm dysfunction by keeping spontaneous breathing as far as possible and subject-ventilator synchrony when ventilation in ARDS with NAVA.

MicroRNA-23a inhibits the growth of papillary thyroid carcinoma via regulating cyclin G1.

OBJECTIVE: The aim of this study was to investigate the regulatory mechanism of miR-23a on biological behaviors of papillary thyroid carcinoma (PTC) cells, such as cell proliferation, cell cycle and apoptosis. PATIENTS AND METHODS: The expression of miR-23a in 28 paired of PTC tissue samples and matched adjacent tissues was detected by quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR). Meanwhile, miR-23a expression in PTC cell lines was also detected by qRT-PCR. Subsequently, miR-23a mimics and inhibitor were transfected into PTC cells. The effects of gain or loss of miR-23a on cell proliferation, cell cycle and apoptosis were analyzed. Bioinformatics analysis, Dual-Luciferase activity assay and Western blot were recruited to validate the potential target gene of miR-23a. RESULTS: The expression of miR-23a was significantly decreased in PTC tissue samples and cell lines. Upregulation of miR-23a in PTC cells markedly decreased cell proliferation, induced cell cycle arrest at G0/G1 phase and promoted cell apoptosis. However, decreased miR-23a exerted the opposite effects. Dual-Luciferase, qRT-PCR and Western blot showed that CCNG1 was a target gene of miR-23a. Furthermore, the silence of CCNG1 intensified the suppressive effect of miR-23a on cell growth. CONCLUSIONS: MiR-23a was involved in the development of PTC via targeting CCNG1, which might provide a new prospect for PTC diagnosis and therapy.

Inhibition of the androgen receptor induces a novel tumor promoter, ZBTB46, for prostate cancer metastasis.

Current therapeutic regimens for prostate cancer focus on targeting androgen receptor (AR) signaling. However, the AR is a key factor in luminal epithelium differentiation and was shown to have a role as a tumor suppressor. Thus, its inhibition may activate oncogenic pathways that contribute to metastatic castration-resistant prostate cancer (CRPC). Herein, we report a novel tumor promoter, ZBTB46, which is negatively regulated by AR signaling via microRNA (miR)-1-mediated downregulation. ZBTB46 is associated with malignant prostate cancer and is essential for metastasis. Its overexpression can overcome the antitumor effects of miR-1 and promote androgen-independent proliferation. We demonstrated that ZBTB46 can transcriptionally regulate SNAI1, a key epithelial-to-mesenchymal transition (EMT) driver, which could contribute to induction of the EMT after androgen-deprivation therapy and metastasis. Our findings are supportive of the model that disruption of AR's function may predispose prostate cancer to progress to metastatic CRPC.

TCF7 is suppressed by the androgen receptor via microRNA-1-mediated downregulation and is involved in the development of resistance to androgen deprivation in prostate cancer.

BACKGROUND: Resistance to androgen deprivation therapy (ADT) represents a key step in the malignant progression of prostate cancer, and mutation to androgen receptor (AR) is one major driver to an androgen-independent phenotype. However, alternative oncogenic pathways that bypass AR signaling have emerged as an important mechanism promoting resistance to ADT. It is known that AR activation can prevent the interaction between ß-catenin and T cell factor/lymphoid enhancer-binding factor (TCF/LEF) family, inhibiting the Wnt signaling pathway. The aim of this study was to determine the role of transcription factor 7 (TCF7), a transcription factor best known as a Wnt effector that forms a complex with ß-catenin, in the development of advanced prostate cancer. We further investigated the molecular mechanisms by which TCF7 is induced when AR signaling is inactivated. METHODS: A novel AR signaling pathway that induces microRNA-1 (miR-1) to suppress metastatic prostate cancer was recently demonstrated (AR-miR-1 signaling axis), and its regulation of Wnt signaling was explored in the current study. Clinical data sets were analyzed for potential targets of AR-miR-1 signaling in the TCF/LEF family, and tissue samples were utilized to validate the relationship. The molecular mechanism and biological functions were demonstrated in prostate cancer cell lines and a mouse xenograft model. RESULTS: We demonstrated a molecular mechanism of AR signaling suppressing TCF7 partly through miR-1-mediated downregulation. TCF7 exhibited oncogenic properties and compromised the tumor-suppressive effects of miR-1. Our results also showed that overexpression of TCF7 or disruption of miR-1 function promoted androgen-independent proliferation. CONCLUSIONS: We demonstrated that the AR-miR-1 axis negatively regulates the novel oncogenic factor, TCF7. Dysregulation of TCF7 promoted a survival advantage and resistance to androgen deprivation, suggesting its therapeutic potential for castration-resistant prostate cancer.

Androgen deprivation leads to increased carbohydrate metabolism and hexokinase 2-mediated survival in Pten/Tp53-deficient prostate cancer.

Prostate cancer is characterized by a dependence upon androgen receptor (AR) signaling, and androgen deprivation therapy (ADT) is the accepted treatment for progressive prostate cancer. Although ADT is usually initially effective, acquired resistance termed castrate-resistant prostate cancer (CRPC) develops. PTEN and TP53 are two of the most commonly deleted or mutated genes in prostate cancer, the compound loss of which is enriched in CRPC. To interrogate the metabolic alterations associated with survival following ADT, we used an orthotopic model of Pten/Tp53 null prostate cancer. Metabolite profiles and associated regulators were compared in tumors from androgen-intact mice and in tumors surviving castration. AR inhibition led to changes in the levels of glycolysis and tricarboxylic acid (TCA) cycle pathway intermediates. As anticipated for inhibitory reciprocal feedback between AR and PI3K/AKT signaling pathways, pAKT levels were increased in androgen-deprived tumors. Elevated mitochondrial hexokinase 2 (HK2) levels and enzyme activities also were observed in androgen-deprived tumors, consistent with pAKT-dependent HK2 protein induction and mitochondrial association. Competitive inhibition of HK2-mitochondrial binding in prostate cancer cells led to decreased viability. These data argue for AKT-associated HK2-mediated metabolic reprogramming and mitochondrial association in PI3K-driven prostate cancer as one survival mechanism downstream of AR inhibition.

[Effect of miR- 155 on immune- factors and its mechanism in mesenchymal stem cells under hypoxic environment].

OBJECTIVE: To investigate the effect of miR-155 on immune-factors and its mechanism in mesenchymal stem cells under hypoxia. METHODS: The microRNA sequences targeting miR-155 mimic and mimic NC gene was designed and transfected into MSC by lipofectamineTM 2000. Lipopolysaccharide was used to stimulate the immunity of MSC under hypoxic environment. Transfection efficiency of miR- 155 and immune-related genes (IL-6, IL-8, iNOS, TGF-ß, HIF-1α) were detected by real-time RT-PCR. The cell surface antigens (CD29, CD73, CD90, CD105, CD31, CD45) and supernatant cytokines (IL- 6, IL- 8, TGF- ß, SDF- 1α) were analyzed by flow cytometry and ELISA, respectively. Western Blot was applied to evaluate related proteins, iNOS and HIF- 1α. RESULT: miR- 155 was transfected into MSC effectively (53.447±8.361 vs 1.070±0.174, P<0.01). In miR-155 high-expressed groups, the expressions of IL-6 and IL-8 were up-regulated [(24.201±1.536) vs (1.802±0.058), P<0.01; (24.406±4.611) vs (7.407± 1.553), P<0.01] and iNOS was markedly suppressed [(0.151 ±0.035) vs (32.925±1.632), P<0.01]. Hypoxia up-regulated expressions of HIF-1α [(45.093±3.371) vs (2.210±0.498), P<0.01] and promoted the regulation of miR-155. MiR-155 and hypoxia had effect on mRNA expression of SDF-1α and TGF-ß [(5.690±1.655) vs (0.841±0.194), P<0.01; (6.982±1.353) vs (0.632±0.184), P<0.01]. However, there was no influence on cytokines of SDF- 1α and TGF- ß [(24.609±2.584) vs (25.359±2.455), P=0.760;(0.568±0.019) vs (0.345±0.037), P=0.002]. CONCLUSION: Hypoxia environment may promote miR-155 to positively modulate immune factors of MSC through up-regulating the expression of HIF-1α, which down-regulated iNOS protein.

Clinical significance of CTHRC1 protein expression in human cancers: a meta-analysis.

The aim of this meta-analysis was to investigate the overall diagnostic and prognostic values of CTHRC1 expression in human cancer development. Based on the inclusion and exclusion criteria, 8 cohort studies were included in the meta-analysis. The data were extracted, and analyses were performed using a random-effects model. Summary odds ratios (ORs) and effect sizes (ESs) with 95% confidence intervals (CIs) were calculated to assess the strength of the associations. A total of 1065 cancer patients from the 8 studies were included in the meta-analysis. The results revealed a positive correlation of CTHRC1 protein expression in tumors with tumor-node-metastasis (TNM) stage and with lymph node (LN) metastasis (TNM: OR = 2.98, 95%CI = 1.48-6.00, P = 0.002; LN: OR = 4.26, 95%CI = 1.88-9.67, P = 0.001). CTHRC1 expression was higher in tumors with sizes ≥5 cm than in tumors with sizes <5 cm (OR = 2.39, 95%CI = 1.12-5.09, P = 0.024). Patients with higher CTHRC1 expression had decreased overall survival (OS) (ES = 1.78, 95%CI = 1.23-2.33, P < 0.001) and poorer disease-free survival (DFS) (ES = 1.71, 95%CI = 1.11-2.31, P < 0.001). Disease-stratified analyses yielded significantly different estimates of CTHRC1 levels in the majority of the subgroups (all P < 0.05). In conclusion, increased CTHRC1 expression is associated with advanced TNM stage, increased LN metastasis and tumor size, and decreased OS and DFS, indicating that CTHRC1 may be a biomarker for prognosis of cancer patients.

Hepatic phosphoenolpyruvate carboxykinase expression after gastric bypass surgery in rats with type 2 diabetes mellitus.

The objective of this study was to investigate the mRNA expression of hepatic phosphoenolpyruvate carboxykinase (PEPCK) after gastric bypass surgery (GBS) in rats with type 2 diabetic mellitus (T2DM). Thirty-six male Goto-Kakizaki rats, aged 12 weeks, were randomly divided into the GBS, sham operation with diet restriction (SO), and sham operation alone (control) groups (N = 12 per group). Liver specimens from all rats were obtained during the operation and 8 weeks after operation. Blood lipid levels were measured before and 8 weeks after operation. Fasting blood glucose (FBG), food intake, and body weight were recorded at weekly time points after operation. The blood glucose area under the curve (AUC) was calculated, and insulin sensitivity indices (ISI) were assessed. The expression PEPCK mRNA and protein were measured by real-time polymerase chain reaction and western blot. Compared with those of the SO and control groups, the blood lipid levels and the FBG in the GBS group was significantly decreased (P < 0.05), as was the AUC (P < 0.05), whereas the ISI was significantly increased (P < 0.05). PEPCK mRNA and protein levels in the GBS group were lower than those in the control group, whereas those in the SO group were significantly higher than those in controls (P < 0.05). In conclusion, GBS can reduce blood glucose in T2DM rats while improving glucose tolerance and hyperglycemia, and the mechanism appears to be associated with a decrease of hepatic PEPCK mRNA and protein expression.

Transforming growth factor-ß promotes prostate bone metastasis through induction of microRNA-96 and activation of the mTOR pathway.

Transforming growth factor-ß (TGFß) is enriched in the bone matrix and serves as a key factor in promoting bone metastasis in cancer. In addition, TGFß signaling activates mammalian target of rapamycin (mTOR) functions, which is important for the malignant progression. Here, we demonstrate that TGFß regulates the level of microRNA-96 (miR-96) through Smad-dependent transcription and that miR-96 promotes the bone metastasis in prostate cancer. The enhanced effects in cellular growth and invasiveness suggest that miR-96 functions as an oncomir/and metastamir. Supporting this idea, we identified a downstream target of the TGFß-miR-96 signaling pathway to be AKT1S1 mRNA, whose translated protein is a negative regulator of mTOR kinase. Our findings provide a novel mechanism accounting for the TGFß signaling and bone metastasis.

MiR-1 and miR-200 inhibit EMT via Slug-dependent and tumorigenesis via Slug-independent mechanisms.

Epithelial-mesenchymal transition (EMT) is a developmental program of signaling pathways that determine commitment to epithelial and mesenchymal phenotypes. In the prostate, EMT processes have been implicated in benign prostatic hyperplasia and prostate cancer progression. In a model of Pten- and TP53-null prostate adenocarcinoma that progresses via transforming growth factor ß-induced EMT, mesenchymal transformation is characterized by plasticity, leading to various mesenchymal lineages and the production of bone. Here we show that SLUG is a major regulator of mesenchymal differentiation. As microRNAs (miRs) are pleiotropic regulators of differentiation and tumorigenesis, we evaluated miR expression associated with tumorigenesis and EMT. Mir-1 and miR-200 were reduced with progression of prostate adenocarcinoma, and we identify Slug as one of the phylogenetically conserved targets of these miRs. We demonstrate that SLUG is a direct repressor of miR-1 and miR-200 transcription. Thus, SLUG and miR-1/miR-200 act in a self-reinforcing regulatory loop, leading to amplification of EMT. Depletion of Slug inhibited EMT during tumorigenesis, whereas forced expression of miR-1 or miR-200 inhibited both EMT and tumorigenesis in human and mouse model systems. Various miR targets were analyzed, and our findings suggest that miR-1 has roles in regulating EMT and mesenchymal differentiation through Slug and functions in tumor-suppressive programs by regulating additional targets.

Immunolocalization of alpha-synuclein in the rat spinal cord by two novel monoclonal antibodies.

This study provides the first immunohistochemical evidence of the presence and distribution patterns in the rat spinal cord of alpha-synuclein (alpha-Syn), a soluble acidic protein, widely expressed in the CNS and closely associated to the pathogenesis of neurodegenerative conditions such as Parkinson's and Alzheimer's diseases. We used two novel homemade monoclonal antibodies (2E3 and 3D5) recognizing two different epitopes of alpha-Syn. Both antibodies localized alpha-Syn within the nerve terminals, whereas 3D5 alone also localized it within the neuronal nuclei. alpha-Syn-immunoreactive nervous elements were widely recognized throughout rat spinal cord and in almost all the gray matter laminae. However, they appeared particularly concentrated within laminae I, II, VII and X and more scattered in the others. Double immunofluorescent labeling showed that alpha-Syn colocalized with synaptophysin in the presynaptic nerve terminals, with neuropeptide Y (NPY) in lamina I, II, IX and X, and had close relationships with tyrosine hydroxylase (TH) immunoreactive neurons in laminae VII and X. Interestingly, the alpha-Syn-immunoreactive nerve elements, in lamina X, contained little of calbindin-28KD and calretinin-31KD. Our findings could help in understanding the genesis of some early clinical symptoms of Parkinson's disease (PD), such as pain and dysautonomic disorders, and indicate the spinal cord as their probable starting point, according to the ascending theory of PD, proposed by Braak.

UVA photoirradiation of anhydroretinol--formation of singlet oxygen and superoxide.

Anhydroretinol is a metabolite of vitamin A (retinol) and a major photodecomposition product of retinyl palmitate and retinyl acetate. Anhydroretinol is biologically active, inducing cell death in lymphoblastoid cells, prevention of N-methyl-N-nitrosourea-induced mammary cancer, and inhibition of cell growth in lymphocytes. We have previously determined that photoirradiation of anhydroretinol in the presence of a lipid, methyl linoleate, with UVA light-induced lipid peroxidation. In the present study, electron spin resonance (ESR) spin-trap techniques were employed to explore the mechanism of lipid peroxidation initiation. Irradiation of anhydroretinol by UVA in the presence of 2,2,6,6-tetramethylpiperidine (TEMP), a specific probe for singlet oxygen, resulted in the formation of TEMPO, indicating that singlet oxygen was generated. During photoirradiation in the presence of 5,5-dimethyl N-oxide pyrroline (DMPO), a specific probe for superoxide, ESR signals for DMPO-OOH were formed, and these signals were quenched by superoxide dismutase. The involvement of singlet oxygen on the induction of lipid peroxidation was also evidenced by the observation that lipid peroxidation was inhibited by sodium azide and enhanced by deuterium oxide. Our overall results provide evidence that photoirradiation of anhydroretinol with UVA light generates reactive oxygen species, e.g. singlet oxygen and superoxide, which mediate the induction of lipid peroxidation.

Ganoderma lucidum extract inhibits proliferation of SW 480 human colorectal cancer cells.

AIM: Ganoderma lucidum is a commonly used Chinese herb and an important ingredient in traditional Chinese medicine herbal formulations for immune dysfunction related illnesses. The effects of this medicinal mushroom on human colorectal cancer cells have not yet been evaluated. In this study, we investigated the effects of Ganoderma lucidum extract using SW 480 human colorectal cancer cell line. MATERIALS AND METHODS: Two different fractions of Ganoderma lucidum extract, i.e., a fraction containing mainly polysaccharides (GLE-1), and a triterpenoid fraction without polysaccharides (GLE-2) were analyzed. Their antiproliferative activity was evaluated by cell proliferation assay and 3H-thymidine incorporation assay. Scavenging effects of DPPH radical were assessed using ESR-spectroscopy. RESULTS: Our data showed that both GLE-1 and GLE-2 significantly inhibited the proliferation of SW 480 cells. The inhibitory effect of GLE-2 was much stronger than that of GLE-1. GLE-1 inhibited DNA synthesis in the cells and reduced the formation of DPPH radicals. CONCLUSION: Ganoderma lucidum extract inhibits proliferation of human colorectal cancer cells and possesses antioxidant properties.

Chromosome 18 suppresses prostate cancer metastases.

Loss of heterozygosity and allelic imbalance data has shown that there are two distinct regions of loss on chromosome 18q associated with the progression of prostate cancer (CaP). To investigate the functional significance of chromosome 18q loci in CaP, we utilized the technique of microcell-mediated chromosome transfer to introduce an intact chromosome 18 into the human prostate cancer cell line, PC-3. Three of the resulting hybrid lines were compared to the PC-3 cells in vitro and in vivo. The hybrid cell lines, containing an intact copy of the introduced chromosome 18, exhibited a substantial reduction in anchorage-dependent and independent growth in vitro. These hybrid cell lines also made smaller tumors in nude mice following subcutaneous injection compared to PC-3 cells. Because tumor growth was not completely eliminated by introduction of chromosome 18, we assessed the ability of the hybrids to metastasize to bone after intra-cardiac inoculation in a nude mouse model. Mice inoculated with PC-3 hybrids containing intact copies of chromosome 18 had significantly fewer bone metastases and dramatically improved survival compared to PC-3 cells. In addition, the introduction of chromosome 18 significantly reduced tumor burden in extraskeletal sites. This was not because of differences in growth rates because mice bearing hybrids were monitored for metastases over twice as long as mice bearing PC-3 cells. Taken together, these data suggest that chromosome 18 has a functional role in CaP to suppress growth and metastases. Identification of the responsible gene(s) may lead to molecular targets for drug discovery.