[Clinicopathological features and prognostic analysis of 151 patients with primary extra-nodal non-Hodgkin's lymphoma].

OBJECTIVE: To evaluate the epidermiology, clinicopathological features and prognosis of primary extra-nodal non-Hodgkin's lymphoma (PE-NHL). METHODS: The clinicopathological data of 151 patients diagnosed as PE-NHL in the First Affiliated Hospital of Jilin University between January 2007 and December 2011 were reviewed and analyzed. RESULTS: The proportion of PE-NHL patients was 58.8% among all the non-Hodgkin's lymphoma cases, with the average age of 52 years, and the male/female ratio was 1.16:1. The most frequently involved site was the stomach (30.5%) followed by nose and sinuses (19.9%) and Waldeyer's ring (17.2%). The most common subtype was diffuse large B-cell lymphoma (DLBCL) (55.0%), followed by NK/T (18.5%) and extra-nodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT) ( 13.2%). The distribution of clinical stages was as follows: stage I 20.5%, II 29.8%, III 29.1%, and IV 20.5%. Most nasal PE-NHL is in early stages, with a proportion of 76.7% in stages I & II. The 3-year overall survival rate was 73.2% and 3-year progression free survival rate was 46.6% in the PE-DLBCL patients. The International Prognosis Index (IPI) could be used to estimate the prognosis of PE-DLBCL well. Multivariate analysis showed that ESR and curative effect of the first treatment were independent prognostic factors for PE-DLBCL patients. CONCLUSIONS: The incidence of PE-NHL is quite high, and the most common primary extra-nodal site is the gastro-intestinal tract and the most common subtype is diffuse large B-cell lymphoma. Risk groups based on IPI can indicate the prognosis of PE-DLBCL to some extent, but only the ESR and curative effect of the first treatment are confirmed to be independent risk factors.

Influence of the mixture of Epimedii Herba and Ginkgo Folium extracts on the coronary flow of isolated hearts in rats.

BACKGROUND: In clinical practice, Epimedii Herba and Ginkgo Folium preparations are widely used in treatment of diseases such as coronary heart disease (angina) in China. However, there are no studies on the two-drug combination. OBJECTIVE: To explore the effect of the mixture of the Epimedii Herba extract (EE) and Ginkgo Folium extract (GE) on coronary flow of isolated hearts in rats. MATERIALS AND METHODS: EE and GE were prepared by reflux in alcohol, and processed with HPD-100 macro-reticular resins; icariin from EE and total bilobalides from GE were determined by high performance liquid chromatography (HPLC). Fifty male Sprague-Dawley (SD) mice were subdivided into five groups (10 rats each): Normal control group (NC), EE - 10 mg group, GE - 10 mg group, EE - 5 mg + GE - 5 mg group, and EE - 10 mg + GE - 10 mg group. Isolated hearts uniform pressure perfusion was proceeded with Langendorff system. RESULTS: The content of icariin in EE was 20.8%. The total content including four kinds of bilobalides (ginkolide A-C and bilobalide) in GE was 8.6%. The coronary flow in the NC group remained stable before and after treatment, and the coronoray flow in the EE, GE, EE + GE groups was increased and the relative magnitude of heightening was 25.0-33.3%, and the coronary flow in EE + GE was significantly different from that in the single EE or GE group. CONCLUSION: EE or GE itself can heighten coronary flow of isolated hearts in rats. The activity of the mixture including EE and GE is better than that of single EE or GE, and the activity becomes larger when the dosage is doubled, and is related with dosage.

Differential expression of filamin A and its clinical significance in breast cancer.

Changes in filamin A (FLNa) expression contribute to the development and progression of numerous malignancies. However, in vitro studies of breast cancer have shown conflicting results. Thus, the present study aimed to detect the expression of FLNa in breast cancer tissue samples and the association with clinicopathological data, in order to provide insightful ex vivo data. A total of 96 breast cancer and distant normal breast tissues and 20 benign tumor tissue specimens were subjected to immunohistochemistry or reverse transcription polymerase chain reaction (RT-PCR) analysis of FLNa expression. Clinicopathological data were collected to analyze the association with FLNa expression. The FLNa protein was overexpressed in breast cancer tissues compared with distant normal mammary gland and benign breast tissues. The FLNa protein was expressed in 63.5% of breast cancer, with positive rates of 36, 66.7 and 84.6%, respectively, in stage I, II and III breast cancer patients (P<0.05). Overexpression of the FLNa protein was associated with advanced stage, lymph node metastasis, vascular or neural invasion, menstruation state and other risk stratifications for breast cancer. The overexpression of FLNa in breast cancer was validated by RT-PCR, indicating transcriptional regulation of FLNa overexpression in breast cancer. FLNa mRNA and protein were overexpressed in breast cancer tissues, which was associated with advanced stage, lymph node metastasis and vascular or neural invasion of breast cancer, suggesting that FLNa contributes to breast cancer development and progression.

In vivo and in vitro inhibition of human liver cancer progress by downregulation of the µ-opioid receptor and relevant mechanisms.

Opiates have long been used as analgesics to relieve pain associated with various medical conditions. µ-opioid receptor (MOR) is the main member of the opioid receptor super-family and the excitation or overexpression of MOR promotes the proliferation of many kinds of tumor cells. It was found in our previous studies that MOR was highly expressed in the tissue and cells of human liver cancer. However, the impact of MOR on the progress of human liver cancer remains unknown. The purpose of this study is to investigate the impact of MOR downregulation on the progress of human liver cancer and the mechanisms involved. RNA interfering or specific inhibitor was administered to downregulate the MOR in human hepato-cellular carcinoma cells and it was found that the proliferation of hepatocellular carcinoma cells was significantly inhibited with the increase of the apoptotic rate, while the cell cycle was blocked in G0/G1 phase and the tumor growth in the mice was retarded. In addition, downregulation of MOR resulted in the increase of phosphorylation of the MKK7 expression and JNK activation. On the contrary, blockade of MKK7 pathway can reverse the antitumor role of MOR. In summary, downregulation of MOR is able to inhibit both in vivo and in vitro human liver cancer progress and it shows potential to be used in cancer therapy.

Shockwaves induce osteogenic differentiation of human mesenchymal stem cells through ATP release and activation of P2X7 receptors.

Shockwave treatment promotes bone healing of nonunion fractures. In this study, we investigated whether this effect could be due to adenosine 5'-triphosphate (ATP) release-induced differentiation of human mesenchymal stem cells (hMSCs) into osteoprogenitor cells. Cultured bone marrow-derived hMSCs were subjected to shockwave treatment and ATP release was assessed. Osteogenic differentiation and mineralization of hMSCs were evaluated by examining alkaline phosphatase activity, osteocalcin production, and calcium nodule formation. Expression of P2X7 receptors and c-fos and c-jun mRNA was determined with real-time reverse transcription polymerase chain reaction and Western blotting. P2X7-siRNA, apyrase, P2 receptor antagonists, and p38 MAPK inhibitors were used to evaluate the roles of ATP release, P2X7 receptors, and p38 MAPK signaling in shockwave-induced osteogenic hMSCs differentiation. Shockwave treatment released significant amounts (≈ 7 µM) of ATP from hMSCs. Shockwaves and exogenous ATP induced c-fos and c-jun mRNA transcription, p38 MAPK activation, and hMSC differentiation. Removal of ATP with apyrase, targeting of P2X7 receptors with P2X7-siRNA or selective antagonists, or blockade of p38 MAPK with SB203580 prevented osteogenic differentiation of hMSCs. Our findings indicate that shockwaves release cellular ATP that activates P2X7 receptors and downstream signaling events that caused osteogenic differentiation of hMSCs. We conclude that shockwave therapy promotes bone healing through P2X7 receptor signaling, which contributes to hMSC differentiation.

Inhibition of filamin-A reduces cancer metastatic potential.

Filamin-A cross-links actin filaments into dynamic orthogonal networks, and interacts with an array of proteins of diverse cellular functions. Because several filamin-A interaction partners are implicated in signaling of cell mobility regulation, we tested the hypothesis that filamin-A plays a role in cancer metastasis. Using four pairs of filamin-A proficient and deficient isogenic cell lines, we found that filamin-A deficiency in cancer cells significantly reduces their migration and invasion. Using a xenograft tumor model with subcutaneous and intracardiac injections of tumor cells, we found that the filamin-A deficiency causes significant reduction of lung, splenic and systemic metastasis in nude mice. We evaluated the expression of filamin-A in breast cancer tissues by immunohistochemical staining, and found that low levels of filamin-A expression in cancer cells of the tumor tissues are associated with a better distant metastasis-free survival than those with normal levels of filamin-A. These data not only validate filamin-A as a prognostic marker for cancer metastasis, but also suggest that inhibition of filamin-A in cancer cells may reduce metastasis and that filamin-A can be used as a therapeutic target for filamin-A positive cancer.

Prognostic values of filamin-A status for topoisomerase II poison chemotherapy.

Filamin-A, also called Actin Binding Protein-280, is not only an essential component of the cytoskeleton networks, but also serves as the scaffold in various signaling networks. It has been shown that filamin-A facilitates DNA repair and filamin-A proficient cells are more resistant to ionizing radiation, bleomycin, and cisplatin. In this study, we assessed the role of filamin-A in modulating cancer cell sensitivity to Topo II poisons, including etoposide and doxorubicin. Intriguingly, we found that cells with filamin-A expression are more sensitive to Topo II poisons than those with defective filamin-A, and filamin-A proficient xenograft melanomas have better response to etoposide treatment than the filamin-A deficient tumors. This is associated with more potent induction of DNA double strand breaks (DSBs) by Topo II poisons in filamin-A proficient cells than the deficient cells. Although the expression of filamin-A enables cells a slightly stronger capability to repair DSB, the net outcome is that filamin-A proficient cells bear more DSBs due to the significantly enhanced DSB induction by Topo II poisons in these cells. We further found that filamin-A proficient cells have increased drug influx and decreased drug efflux, suggesting that filamin-A modulates the intra-cellular drug kinetics of Topo II poisons to facilitate the generation of DSB after Topo II poison exposure. These data suggest a novel function of filamin-A in regulating the pharmacokinetics of Topo II poisons, and that the status of filamin-A may be used as a prognostic marker for Topo II poisons based cancer treatments.

K-Ras mutation-mediated IGF-1-induced feedback ERK activation contributes to the rapalog resistance in pancreatic ductal adenocarcinomas.

Mammalian target of rapamycin complex 1 (mTORC1) is frequently activated in human cancers; however, clinical trials of rapalog (the mTORC1 inhibitors) have shown that pancreatic ductal adenocarcinomas (PDACs) resist to the treatment. Rapalog treatment activated the extracellular signal-regulated kinase (ERK) pathway in K-Ras mt PDAC cells. K-Ras knockdown abolished the insulin-like growth factor-1 (IGF-1)-induced ERK pathway in the K-Ras mt PDAC cells and enhanced the therapeutic efficacy of everolimus in treating K-Ras mt PDAC cells-derived mouse xenografts. The results indicate that targeting of K-Ras mutation may lead to the development of therapies that overcome rapalog resistance in PDAC.

Cisplatin restores TRAIL apoptotic pathway in glioblastoma-derived stem cells through up-regulation of DR5 and down-regulation of c-FLIP.

Glioblastoma-derived stem cells (GSCs) are responsible for the cancer resistance to therapies. We show here that GSC-enriched neurospheres are resistant to the treatment of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) due to the insufficient expression of the death receptor DR4 and DR5 and the overexpression of cellular Fas-associated death domain-like interleukin-1ß-converting enzyme-inhibitory protein (c-FLIP). However, treatment with cisplatin leads to the upregulation of DR5 and downregulation of c-FLIP and restores TRAIL apoptotic pathway in the neurospheres. This study suggests that the combined treatment of TRAIL and cisplatin can induce apoptosis in GSCs and thus provide an effective treatment of glioblastomas.

The effect of green tea extract and EGCG on the signaling network in squamous cell carcinoma.

Green tea and its major active component, epigallocatechin-3-gallate (EGCG), have been reported to have anticancer activity on various cancers. However, the exact molecular mechanism of its anticancer activity is still not well understood. We investigated the anticancer activity of green tea extract (GTE) and EGCG on 3 human squamous carcinoma cell lines (CAL-27, SCC-25, and KB) in vitro. We also examined the effects of GTE and EGCG on cell signaling networks using our newly developed Pathway Array technology, which is an innovative proteomic assay to globally screen changes in protein expression and phosphorylation. Our results demonstrated that GTE and EGCG inhibited all 3 squamous carcinoma cells' growth via S and G(2)/M phase arrest, but different sensitivities to GTE and EGCG in different cell lines were observed: CAL-27 cells were more sensitive to the both agents than SCC-25 and KB cells, and GTE at an EGCG equivalent concentration displayed a stronger inhibition than EGCG alone. The Pathway Array assessment of 107 proteins indicated that different signaling pathways were activated in different cell lines, suggesting heterogeneity at the signaling network level. After treatment with GTE or EGCG, a total of 21 proteins and phosphorylations altered significantly in all 3 cell lines based on analysis of variance (ANOVA) (P < 0.05). The major signaling pathways affected by GTE and EGCG were EGFR and Notch pathways, which, in turn, affected cell cycle-related networks. These results suggested that GTE and EGCG target multiple pathways or global networks in cancer cells, which resulted in collective inhibition of cancer cell growth. The finding pointed out the future direction to study the underlying mechanism of the chemotherapeutic and chemopreventive activities of EGCG and GTE.

Construction, expression and characterization of a chimeric multi-domain protein mediating specific DNA transfer.

The delivery of plasmid DNA to target cells using a simple, defined, non-viral system is an area of intense research in gene therapy. Here, we describe a novel DNA carrier protein termed TG, consisting of the DNA-binding domain of the yeast transcriptional activator GAL4 and human immunodeficiency virus type 1 Tat protein, which can transfer modified naked plasmid DNA into target cells to express foreign genes of interest. The TG protein was expressed in Escherichia coli (E. coli), refolded and purified on an immobilized Ni(2+) affinity chromatography column. SDS-PAGE and Western blotting revealed that the fusion protein was highly expressed with a yield of approximately 275 mg/L. We also constructed the pIRES-UAS-EGFP DNA vector, consisting of upstream activating sequences (UASs) for the specific binding of the DNA-binding protein and the enhanced green fluorescent protein (EGFP) gene. The TG protein could bind specifically to pIRES-UAS-EGFP, forming a complex which could efficiently transfect target cells and result in detectable EGFP protein expression. Thus, these results provide a basis for development of efficient non-viral DNA transfer vectors for further improvements of gene therapy strategies.

Shockwaves increase T-cell proliferation and IL-2 expression through ATP release, P2X7 receptors, and FAK activation.

Shockwaves elicited by transient pressure disturbances are used to treat musculoskeletal disorders. Previous research has shown that shockwave treatment affects T-cell function, enhancing T-cell proliferation and IL-2 expression by activating p38 mitogen-activated protein kinase (MAPK) signaling. Here we investigated the signaling pathway by which shockwaves mediate p38 MAPK phosphorylation. We found that shockwaves at an intensity of 0.18 mJ/mm(2) induce the release of extracellular ATP from human Jurkat T-cells at least in part by affecting cell viability. ATP released into the extracellular space stimulates P2X7-type purinergic receptors that induce the activation of p38 MAPK and of focal adhesion kinase (FAK) by phosphorylation on residues Tyr397 and Tyr576/577. Elimination of released ATP with apyrase or inhibition of P2X7 receptors with the antagonists KN-62 or suramin significantly weakens FAK phosphorylation, p38 MAPK activation, IL-2 expression, and T-cell proliferation. Conversely, addition of exogenous ATP causes phosphorylation of FAK and p38 MAPK. Silencing of FAK expression also reduces these cell responses to shockwave treatment. We conclude that shockwaves enhance p38 MAPK activation, IL-2 expression, and T-cell proliferation via the release of cellular ATP and feedback mechanisms that involve P2X7 receptor activation and FAK phosphorylation.

Dichlorido{2-[2-(piperidin-1-yl)ethyl-imino-meth-yl]phenolato}zinc(II) monohydrate.

In the title mononuclear zinc(II) complex, [ZnCl(2)(C(14)H(20)N(2)O)]·H(2)O, the Zn(II) atom is four-coordinated by the phenolate O and imine N atoms of the Schiff base ligand and by two Cl atoms in a tetra-hedral geometry. In the crystal structure, O-H⋯Cl, O-H⋯O and N-H⋯O hydrogen bonds involving the water mol-ecules bridge adjacent complexes into a ladder-like structure running along the c axis.

[Therapeutic effects of combination of arsenic trioxide with low-dose all-trans retinoic acid on induction of remission acute promyeloeytic leukemia].

OBJECTIVE: To observe the therapeutic efficacy and side effects of arsenic trioxide (As2O3)combined with low-dose all-trans retinoic acid (ATRA) on remission induction in newly-diagnosed and relapsed patients with acute promyeloeytic leukemia (APL). METHODS: 224 patients of APL, 156 newly diagnosed patients, aged 34 (13 approximately 62), with a male/female ratio of 1.56, and 28 relapsed patients, aged, aged 34 (12 approximately 63), with a male/female ratio of 1.89, underwent As2O3 + ATRA therapy. The therapeutic effects was compared with that of As2O3 alone treatment on 40 newly diagnosed patients and 25 relapsed patients and that of ATRA alone treatment on 36 newly diagnosed patients and 15 relapsed patients. The treatment protocol for the combination group was as following: As2O3 was administered intravenously at a dose of 10 mg/day and ATRA was given orally three times per day at a dose of 10 mg. The complete remission (CR) rate, period to CR, incidence of early death and side effects were observed in the three groups. RESULTS: In the newly-diagnosed patients, there was no significant difference in CR rate among the three groups (92.5% for the As2O3/LD-ATRA group, 83.8% for the ATRA group, and 90% for the As2O3 group respectively). In comparison with As2O3 alone, administration of LD-ATRA to the patients in the As2O3/LD-ATRA group significantly shortened the period to CR (the medium time to CR was 28 days for the As2O3/LD-ATRA group and 39 days for the As2O3 group respectively). As compared to ATRA alone, treatment with As2O3 with low-dose ATRA showed a significantly lower incidence of early death (2.5% for the As2O3/LD-ATRA group and 13.9% for the ATRA group respectively). In the relapsed patients, the CR rate was significantly higher in the group treated with As2O3/LD-ATRA (71.4% for the As2O3/LD-ATRA group, 32.0% for the ATRA group, and 43.0% for the As2O3 group respectively). The combined use of LD-ATRA with As2O3 did not further enhance toxic side effects as compared to As2O3 alone or ATRA alone. CONCLUSION: As2O3/LD-ATRA regimen is superior to either regimen given alone to patients with APL. It is an efficient therapeutic approach to APL patients using a combination of As2O3 with low-dose ATRA.

An efficient therapeutic approach to patients with acute promyelocytic leukemia using a combination of arsenic trioxide with low-dose all-trans retinoic acid.

The use of arsenic trioxide (As2O3, ATO) combined with all-trans retinoic acid (ATRA) has recently been reported to induce remission in patients with acute promyelocytic leukemia (APL). However, its efficiency remains inconclusive mainly due to the small number of the available cases. In this study, therefore, we present a clinical study using a combination of ATO with low-dose ATRA (LD-ATRA) to treat 108 APL patients (80 newly diagnosed patients, 28 relapsed patients). Therapeutic outcomes using the ATO/LD-ATRA approach were compared with those of APL patients treated either with ATO alone (65 patients) or ATRA alone (51 patients). The results showed that the ATO/LD-ATRA approach provided significantly better therapeutic outcomes as compared to either ATO or ATRA alone, as evidenced by lower mortality, a higher CR rate and a reduced period to CR. In addition, the toxic side-effects have been no worse with the combined ATO/LD-ATRA treatment than with either ATO or ATRO alone and in some cases have been reduced. These data suggest that the ATO/LD-ATRA regimen is superior to either regimen given alone to patients with APL.