Detection of Prostate Cancer Antigen 3 and Prostate Cancer Susceptibility Candidate in Non-DRE Urine Improves Diagnosis of Prostate Cancer in Chinese Population.

Although prostate biopsy is the gold standard for the diagnosis of prostate cancer, it also leads to high incidence of negative biopsies and the diagnosis of clinically low-risk prostate cancer and the subsequent overtreatment. It remains an unmet need to discover new biomarkers in order to defer the unnecessary biopsies in clinical practice. In this study, we described a new method, LBXexo score, to measure the urine exosomal PCA3/PRAC expression from non-DRE urine as a noninvasive diagnosis to improve the detection rate in Chinese population with a low serum PSA level. First-voided urine samples were collected to isolate exosomes, and exosomal RNAs of PCA3 and PRAC were measured by quantitative reverse transcription PCR. A significant increase in exoPCA3/PRAC was observed in both any-grade and high-grade prostate cancer groups when compared with the biopsy-negative group. Receiver-operating characteristic curve analyses showed that the LBXexo score significantly improved diagnostic performance in predicting biopsy results, with AUCs of 0.723 (p=0.017) and 0.736 (p=0.038) for any-grade and high-grade (GS ≥ 7) prostate cancer, respectively. For high-grade cancer, LBXexo had the negative and positive predictive values of 100% and 27.59%, respectively, and could potentially avoid unnecessary biopsy. This is the first report in Chinese population that demonstrates the predictive value of the exosomal expression of PCA3 and PRAC derived from non-DRE urine in predicting prostate biopsy outcomes. It could be used in clinical practice to make a better informed biopsy decision and avoid unnecessary biopsies in Chinese population.

Successful second allogeneic stem-cell transplantation from the same sibling donor for a patient with recurrent hepatosplenic gamma-delta (γ/δ) T-cell lymphoma: A case report.

RATIONALE: Hepatosplenic T-cell lymphoma (HSTCL) is a rare but aggressive type of peripheral T-cell lymphoma (PTCL). There is an urgent need for effective treatment due to the poor prognosis of HSTCL. Here, for the 1st time we describe the rare successful case of HSTCL who relapsed after a previous allogeneic stem-cell transplantation (allo-SCT), achieved remission with the second allo-SCT from the same donor. PATIENT CONCERNS: A 24-year-old male, presented with a 2-week history of fever, drenching night sweats and nonquantified weight loss. DIAGNOSES: Laboratory studies, flow cytometry of immunophenotyped, and physical examination results strongly suggested hepatosplenic γ/δ T-cell lymphoma, stage IVB. INTERVENTIONS: We proceeded to an allo-SCT with a human leukocyte antigen (HLA) identical sibling donor. The bone marrow examination and fluorescent in situ hybridization were observed for complete donor chimerism of bone marrow cells on day 34. On day 157 after the initial allo-SCT, the bone marrow examination revealed the relapse of the sinusoidal infiltration with lymphoma cells. Considering the disease persistence, we conducted the second allo-SCT from the same HLA-identical sibling donor immediately. OUTCOMES: Bone marrow examination indicated hematologic recovery without residual lymphoma cells. LESSONS: Our encouraging outcome suggests that the latter allo-SCT needs to be considered early for patients with disease recurrence, and it also demonstrates that graft-vs-lymphoma conferred by allo-SCT may play an essential role on HSTCL treatment. Furthermore, detecting related genes at diagnosis may have prognostic implications and guidance value for personal chemotherapy program.

Co-administration of tissue plasminogen activator and hyperbaric oxygen in ischemic stroke: a continued promise for neuroprotection.

Intravenous recombinant tissue-type plasminogen activator (r-tPA, alteplase) remains the recommended therapy for acute ischemic stroke. However, several factors are limiting its practical use. It makes it urgent for us to search more efficient strategies that can save the ischemic neurons, and safely extend the time window, while in the mean time reducing the detrimental effects for stroke thrombolysis. Hyperbaric oxygen therapy (HBOT) is considered to be potentially neuroprotective. Co-administration of r-tPA and HBOT has already been proved to be effective, safe and feasible in myocardial infarction. In this article, we would like to review whether HBOT has any beneficial effects on r-tPA thrombolysis. If there is, what is the underlying possible mechanisms and how to optimize for maximal effects?

SPARC silencing inhibits the growth of acute myeloid leukemia transformed from myelodysplastic syndrome via induction of cell cycle arrest and apoptosis.

Secreted protein acidic and rich in cysteine (SPARC) plays key roles in erythropoiesis; haploinsufficiency of SPARC is implicated in the progression of the 5q- syndrome. However, the role of SPARC in other subtypes of myelodysplastic syndrome (MDS) is not fully understood, particularly in the del(5q) type with a complex karyotype, which has a high risk to transform into acute myeloid leukemia (AML). In the present study, we investigated the role of SPARC in the proliferation and apoptosis of SKM-1 cells, an acute myeloid leukemia cell line transformed from an MDS cell line. SKM-1 cells were infected with SPARC-RNAi-LV or NC-GFP-LV lentivirus. Apoptosis and cell cycle profiling were assessed by flow cytometry, and cell proliferation was evaluated by MTS assay. The mRNA and protein expression levels of SPARC, p53, caspase-3, caspase-9 and Fas were detected by RT-PCR, real-time PCR and western blot assay. The SPARC shRNA constructed by us led to a significant reduction in SPARC expression in SKM-1 cells. SPARC knockdown inhibited the proliferation of SKM-1 cells by inducing cell cycle arrest at the G1/G0 phase and apoptosis. SPARC knockdown elevated the expression of p53, caspase-9, caspase-3 and Fas at both the mRNA and protein levels. SPARC silencing inhibited the growth of AML transformed from MDS by activating p53-induced apoptosis and cell cycle arrest. These data indicate that SPARC acts as an oncogene in transformed MDS/AML and is a potential therapeutic target in MDS/AML.

Epigenetic inactivation of PLCD1 in chronic myeloid leukemia.

Phospholipase C Î´1 (PLCD1), is located at the important tumor suppressor locus 3p22. It encodes an enzyme that mediates regulatory signaling of energy metabolism, calcium homeostasis and intracellular movements. PLCD1 has been studied in some human solid tumors relating to the CpG island methylation of the gene promoter as a functional tumor suppressor. However, no such information is available in chronic myeloid leukemia (CML). In this study, we investigated PLCD1 expression in the CML K562 cell line (0/1) and 15% (2/13) of bone marrow mononuclear cells with CML by using semi-quantitative PCR. The CpG island (CGI) methylation status of the PLCD1 promoter was detected in K562 (0/1) and 56% (23/41) of CML patients by methylation-specific PCR (MSP), but not in the normal adult bone marrow mononuclear cells. Furthermore, the DNA demethylation agent 5'-aza-2'deoxycytidine restored the expression of PLCD1 in K562 cells. Functional studies showed that ectopic expression of PLCD1 in K562 cells was able to dramatically inhibit their colony formation and induce cell cycle G1 arrest, suggesting that PLCD1 acts as a functional tumor suppressor and may serve as a biomarker for possible early detection and prognosis of CML.

[Construction and identification of SPARC shRNA lentiviral vector and its expression in human SKM-1 cells].

AIM: To construct a lentiviral vector expressing small-hairpin RNA(shRNA) targeting SPARC gene and investigate its silenced effect on SPARC in human myelodysplastic syndromes(MDS) cell line SKM-1. METHODS: The targeting sequence of SPARC gene which can be effectively silenced in RNA interference was confirmed in our previous study. The designed and synthesized single-stranded primers were annealed to double-stranded oligo sequences and subcloned into linear pGCSIL-GFP lentiviral plasmid digested by enzyme Age I and EcoR I to produce GC-shSPARC lentiviraL vector. After being identified by PCR and sequencing, plasmids GC-shSPARC with pHelper 1.0 and pHelper 2.0 were cotransfected into 293T cells to package lentiviral particles. The recombinant lentiviral vector was transfected into human SKM-1 cells, transfection efficiency was evaluated with expression of green fluorescent protein(GFP) determined by fluorescent microscope. Expression of SPARC in SKM-1 cells was detected using RT-PCR and Western blotting. RESULTS: A recombinant lentiviral vector, GC-shSPARC, expressing shRNAs targeting SPARC gene was constructed and confirmed by DNA sequencing. The recombinant lentivirus was harvested from 293T cells with a viral titer of 1×10(9); TU/mL. GFP was observed in the 70% of SKM-1 cells after transfection. Expression of SPARC mRNA and protein was significantly reduced in the GC-shSPARC transfected group than that in the control group (P<0.05). CONCLUSION: The lentivirus RNAi vector targeting SPARC has been successfully constructed, and can effectively inhibit the expression of SPARC in SKM-1 cell line, which shed light on the foundation for researching the inhibition of SPARC siRNA target against human MDS cells proliferation, induction apoptosis and gene therapy.

Protocadherin-10 is involved in angiogenesis and methylation correlated with multiple myeloma.

Protocadherin-10 (PCDH10) which is located at 4q28.3, is a member of the cadherin superfamily of cell adhesion molecules. PCDH10 is broadly expressed in normal adult, but nearly undetectable in multiple myeloma (ΜΜ) tissues and cell lines. Its promoter methylation was detected in virtually all the silenced or downregulated cell lines. The silencing of PCDH10 could be reversed by pharmacological demethylation, indicating a methylation-mediated mechanism. In the current study, we investigated 44 patients (23 females, 21 males), 77.27% (34/44) of whom presented high methylation of PCDH10. We found no associations between promoter hypermethylation and gender or age at the time of initial diagnosis. We also examined the role of PCDH10 as a mediator of MM cell proliferation, cell cycle progression, and its involvement in angiogenesis. Our results demonstrate that the PCDH10 gene is a target for epigenetic silencing in MM and provide a link between the dysregulation of angiogenesis and DNA methylation.

[Effect and mechanism of matrine on apoptosis of Raji cells].

OBJECTIVE: To invest the effect and mechanism of matrine on apoptosis of human Burkitt's lymphoma Raji cells. METHODS: Raji cells were cultured in vitro and treated by different final concentrations (0.4, 0.8, 1.6 mg/mL) of matrine or combined with SB203580 (p38 MAPK inhibitor) before matrine was added, then cocultured for 48 h, cell apoptosis rate was detected by Annexin V-FITC/PI double staining method and the P-p38 MAPK, Fas, FasL protein expresssion of Raji cells were evaluated by Western blot. RESULTS: After cells were treated by matrine (0.4, 0.8, 1.6 mg/mL), the corresponding total apoptosis rate (15.77 +/- 0.53)%, (27.88 +/- 1.52)%, (48.08 +/- 2.87)%, had statistical significance compared with SB203580 groups (11. 48 +/- 0.64)%, (19.34 +/- 0.91)%, (33.98 +/- 1.26)% (P < 0.05 or P < 0.01), and control group (8.78 +/- 0.66)% (P < 0.05 or P < 0.01). As the concentration of matrine gradually increased,the protein expresssion levels of P-p38MAPK, Fas, FasL increased, and decreased after SB203580 were added, the correlation of P-p38MAPK and Fas, FasL was obvious. CONCLUSION: Matrine can upregulation of Fas and FasL to promote the apoptosis of Raji cells, it may be related to p38MAPK Activation.

Proteomic analysis of a rat model of depression.

Major depression is one of the most disabling disorders, yet the pathogenesis of this mental disorder is poorly understood. The techniques of proteomics provide us with powerful tools, while the animal models of depression enable research that cannot be performed on humans due to practical difficulties or ethical reasons. In this review, we summarize the characteristics of some most commonly applied rat models of depression, and explore what could be done with novel proteomic approaches to offer an insight to the pathogenesis of major depression, biomarker establishment and drug development.

1H magnetic resonance spectroscopy study of thalamus in treatment resistant depressive patients.

Treatment-resistant depression (TRD) is a common clinical problem, and represents a considerable challenge to treatment, however, the pathogenesis of this disease is poorly understood. Thalamus is generally believed to have a role in the pathophysiology of depression. In this study, we adopted 1.5T (1)H magnetic resonance spectroscopy ((1)H MRS) to examine possible alterations of thalamus metabolism in 20 adult TRD patients. Our results suggested there might be damage and loss of neurons, as well as membrane phospholipids associated metabolism abnormality in the TRD thalamus.

Neurogenesis and major depression: implications from proteomic analyses of hippocampal proteins in a rat depression model.

Major depression is one of the most disabling disorders. Yet, the pathogenesis of this mental disorder is poorly understood. Hippocampus is generally believed to be associated with pathogenesis of depression. In this study, we adopted a proteomic approach to examine possible alterations of protein expression in the hippocampus of a rat depression model. Our results suggest that neurogenesis in hippocampus may play an important role in the pathogenesis of major depression.

[Functional magnetic resonance image study on the brain areas involved in reading Chinese, English, and Nepali in Nepalese].

OBJECTIVE: To explore the different brain areas activated by Chinese, English, and Nepali word tasks in Nepalese by using the functional magnetic resonance image (fMRI). METHODS: To determine the neuroanatomic functional brain areas responsible for Chinese, English and Nepali reading as well as sentence-formation, blood oxygenation level dependent (BOLD) block design by fMRI was performed in 6 healthy Nepalese volunteers. RESULTS: During Chinese reading, the activated areas included bilateral motor area, subfrontal gyri, superior temporal gyri, and superior parietal lobule; during English reading, the activated areas were left motor area, left subfrontal gyrus, left supra temporal gyrus, left insula and bilateral cerebellum; and the Nepali task demonstrated the activation of left anterior central gyrus, superior frontal gyrus, superior temporal gyrus. Aside from the bilateral occipital lobes, both English and Nepali activated areas were the left cerebral hemisphere dominant. CONCLUSION: The more familiar with the language, the fewer areas are activated. Superior temporal gyrus might be involved in sentence-formation.