Prognostic significance of USP22 as an oncogene in papillary thyroid carcinoma.

Ubiquitin-specific protease 22 (USP22), a novel deubiquitinating enzyme, has been associated with metastasis, therapy resistance, and cell cycle progression. The purpose of this study was to investigate the expression level of USP22 in papillary thyroid carcinoma (PTC) samples and to evaluate its clinical significance in PTC patients. USP22 expression was examined in 30 fresh PTC tissues and paired adjacent noncancerous tissues by real-time quantitative RT-PCR. Immunohistochemistry for USP22 was performed on additional 156 PTC tissues. The clinical significance of USP22 expression was analyzed. We found that the expression levels of USP22 mRNA and protein in PTC tissues were both significantly higher than those in noncancerous tissues. Clinicopathological analysis showed that USP22 expression was significantly correlated with tumor size (p=0.036), extracapsular invasion (p=0.012), multifocality (p=0.014), lymph node metastasis (p=0.022), distant metastasis (p=0.005), and TNM stage (p=0.002). The Kaplan-Meier survival curves revealed that USP22 expression was associated with poor prognosis in PTC patients. USP22 expression was an independent prognostic marker of overall patient survival in a multivariate analysis. Our findings suggest that USP22 is an independent predictor of poor prognosis of PTC patients.

A subanesthetic dose of isoflurane during postconditioning ameliorates zymosan-induced neutrophil inflammation lung injury and mortality in mice.

Anesthetic isoflurane (ISO) has immunomodulatory effects. In the present study, we investigated whether a subanesthetic dose of ISO (0.7%) protected against zymosan (ZY) induced inflammatory responses in the murine lung and isolated neutrophils. At 1 and 6 hrs after ZY administration intraperitoneally, ISO was inhaled for 1 hr, and 24 hrs later, lung inflammation and injury were assessed. We found that ISO improved the survival rate of mice and mitigated lung injury as characterized by the histopathology, wet-to-dry weight ratio, protein leakage, and lung function index. ISO significantly attenuated ZY-induced lung neutrophil recruitment and inflammation. This was suggested by the downregulation of (a) endothelial adhesion molecule expression and myeloperoxidase (MPO) activity in lung tissue and polymorphonuclear neutrophils (b) chemokines, and (c) proinflammatory cytokines in BALF. Furthermore, ZY-induced nuclear translocation and DNA-binding activity of NF- κ B p65 were also reduced by ISO. ISO treatment inhibited iNOS expression and activity, as well as subsequent nitric oxide generation. Consistent with these in vivo observations, in vitro studies confirmed that ISO blocked NF- κ B and iNOS activation in primary mouse neutrophils challenged by ZY. These results provide evidence that 0.7% ISO ameliorates inflammatory responses in ZY-treated mouse lung and primary neutrophils.

4,4'-Dimethoxybenzhydryl substituent augments performance of bis(imino)pyridine cobalt-based catalysts in ethylene polymerization.

A series of cobalt complexes with bis(imino)pyridine derivatives featuring unsymmetrical substitution with bulky groups has been synthesized and characterized. The molecular structures of two representatives have been determined by the single-crystal X-ray diffraction study, revealing distorted tetrahedral geometry with different degrees of steric hindrance imparted by the two inequivalent aryl groups attached to the imine nitrogen atoms. On activation with either MAO or MMAO, these complexes display high activity toward ethylene polymerization, reaching 8.71 × 106 g of PE (mol of Co)-1 h-1 at 60 °C and produce polyethylene of high molecular weight (M w = 5.27 × 105 g mol-1) and low dispersity. The presence of the methoxy-substituent noticeably enhances the activity of the cobalt catalyst and increases the molecular weight of the resultant polyethylene.

Application of colour Doppler in encircling constriction of the superficial femoral vein in primary deep venous insufficiency of the lower limbs.

1. The aim of the present study was to evaluate the clinical value of colour Doppler application in encircling constriction of the superficial femoral vein in deep vein insufficiency. 2. A total of 87 patients with primary deep venous insufficiency (PDVI) using ascending venography were randomly divided into group A (44 patients) and group B (43 patients). All patients underwent encircling constriction of the superficial femoral vein, high ligation and ablation of the great saphenous vein and perforator vein. The duration of venous reflux at operation was monitored with colour Doppler in group A (but not group B) to evaluate the immediate effects. Clinical grading and scoring of the clinical, etiological, anatomical, pathophysiological (CEAP) classification system were used to evaluate the follow-up curative effect. 3. In four cases from group A, completely destroyed valves were identified at the time of operation and autografting of the vein segment with a valve was carried out. The intraoperative examination of colour Doppler in group A showed a much shorter duration of vein reflux after the encircling constriction procedure than the presurgery condition. According to the results of CEAP grading, the success rate of group A (95.0%, 38/40) was significantly higher than that of group B (76.7%, 33/43). Postoperative clinical scores were markedly lower than preoperative scores in both groups A and B. 4. In conclusion, our data suggest that application of colour Doppler in encircling constriction of superficial femoral vein might enhance surgical pertinence and improve surgical effect for PDVI.

Rectal carcinoma metastatic to the male breast after 7 years: case report.

BACKGROUND: Colorectal cancer metastasis to a mammary location is very rare. CASE REPORT: A 38-year-old male, who had undergone anterior resection of an advanced rectal carcinoma 7 years earlier, presented with a right mammary mass. Core needle biopsy of the mass indicated cytology consistent with breast adenocarcinoma. After neoadjuvant chemotherapy and modified radical mastectomy, pathology identified the mass as rectal carcinoma. CONCLUSION: The authors highlight the difficulty of making an accurate diagnosis of rectal cancer metastasis to the breast of a male.

Achieving polydispersive HDPE by N,N,N-Co precatalysts appended with N-2,4-bis(di(4-methoxyphenyl)methyl)-6-methylphenyl.

A family of unsymmetrical 2-(2,4-bis(di(4-methoxyphenyl)methyl)-6-MeC6H2N)-6-(1-(arylimino)ethyl)pyridine-cobalt dichloride complexes has been synthesized and characterized by NMR spectroscopy, FT-IR spectroscopy and elemental analysis as well as single crystal X-ray diffraction for Co2 and Co4. Activated with either MAO or MMAO, all the cobalt precatalysts displayed high activities toward ethylene polymerization and produced highly linear polyethylenes with high molecular weights as well as wide polydispersities; for example, the performance using Co1/MAO at 50 °C reached 9.17 × 106 g PE (mol of Co)-1 h-1 with the production polyethylene of molecular weight as high as M w = 3.14 × 105 g mol-1, T m = 134.3 °C besides its wide polydispersity of M w/M n of 54.6. Besides the terminal vinyl group of the resultant polyethylenes, it is rare for a late-transition metal catalyst to achieve highly linear polyethylenes with not only wide polydispersity but also high molecular weights, being similar to high-density polyethylenes produced using Phillips catalyst.

Expression of MUC1 and its significance in hepatocellular and cholangiocarcinoma tissue.

AIM: To investigate the relation between MUC1 expression, distribution, and prognosis in hepatocellular and cholangiocarcinoma (HCC and CC) and cirrhotic liver tissues, and their significance in HCC and CC diagnosis. METHODS: Expression and distribution of MUC1 were examined by immunohistochemical assay with anti-MUC1 mAb in 59 samples of HCC and 37 samples of CC, 20 samples of cirrhotic liver tissues, and 10 samples of normal liver tissues, seeking possible associations between MUC1 positive expression, distribution in HCC and CC (primary liver cancer, PLC) cases and the studied clinical data. RESULTS: Immunohistochemical analysis of MUC1 expression showed that in the 96 PLC samples, 68 (70.8%) were strong positive, and 6 (6.2%) were weak positive. Only 4 in the 20 cirrhotic liver tissues were found to be weak positive, while no expression of MUC1 was detected in normal liver tissues. Apparently, the high expression rate of MUC1 in PLC tissues was statistically significant in comparison to that in cirrhotic and normal liver tissues. The expressed MUC1 protein, stained in dark brownish or brownish-yellow particles, chiefly localized on the cancer cell membranes or in cytoplasm. In the 68 strong positive samples, 40 were detected on cell membrane and the other 28 were in cytoplasm. In addition, follow-up studies of those PLC cases demonstrated that MUC1 expression on cell membrane or in cytoplasm was closely associated with PLC prognosis. The expression of MUC1 in PLC had little statistical significance in respect of the pathological types and sizes of the tumors, but a strong relationship regarding histological differentiation, metastasis of lymph nodes, portal canal emboli, and post-operational recurrence of the carcinomas. After 3 years of tumor excision, the metastasis rate in MUC1 positive expression group (67.6%) was much higher than that in MUC1 weak expression group (33.3%) and negative expression group (31.8%), and thus the survival rate in MUC1-positive expression group was significantly different from that in weak and negative expression groups. CONCLUSION: Expression and localization of MUC1 proteins in primary liver carcinomas (PLCs) may act as prognostic markers, and MUC1 molecules might be helpful in differential diagnosis.

Patchouli alcohol dampens lipopolysaccharide induced mastitis in mice.

Patchouli alcohol (PA), a tricyclic sesquiterpene isolated from Pogostemonis Herba, has been known to exhibit antioxidant, anti-inflammatory, and other important therapeutic activities. The aim of this study was to investigate the effects of PA on LPS-induced mastitis in vivo and the possible mechanism. The mouse model of mastitis was induced by injection of LPS through the duct of mammary gland. Mice were pretreated with dexamethasone or PA 1 h before and 12 h after induction of LPS. The myeloperoxidase activity and inflammatory cytokines production in mammary tissues were determined. The effects of PA on NF-κB signal pathways were analyzed by Western blotting. The results showed that PA inhibited the LPS-induced TNF-α, IL-6, and IL-1ß production in a dose manner. It was also observed that PA attenuated mammary histopathologic changes. Furthermore, Western blot analysis showed that PA could inhibit the phosphorylation of NF-κB and IκB induced by LPS. These results indicate that PA inhibits NF-κB signaling pathways to attenuate inflammatory injury induced by LPS. PA may be a potent therapeutic reagent for the prevention of mastitis.

Subanesthetic isoflurane reduces zymosan-induced inflammation in murine Kupffer cells by inhibiting ROS-activated p38 MAPK/NF-κB signaling.

Volatile anesthetic isoflurane (ISO) has immunomodulatory effects. The fungal component zymosan (ZY) induces inflammation through toll-like receptor 2 or dectin-1 signaling. We investigated the molecular actions of subanesthetic (0.7%) ISO against ZY-induced inflammatory activation in murine Kupffer cells (KCs), which are known as the resident macrophages within the liver. We observed that ISO reduced ZY-induced cyclooxygenase 2 upregulation and prostaglandin E2 release, as determined by western blot and radioimmunoassay, respectively. ISO also reduced the production of tumor necrosis factor-α, interleukin-1ß, IL-6, high-mobility group box-1, macrophage inflammatory protein-1α, macrophage inflammatory protein-2, and monocyte chemoattractant protein-1 as assessed by enzyme-linked immunosorbent assays. ISO blocked the ZY-induced nuclear translocation and DNA-binding activity of nuclear factor- (NF)-κB p65. Moreover, ISO attenuated ZY-induced p38 mitogen-activated protein kinase (MAPK) activation partly by scavenging reactive oxygen species (ROS); the interregulation that ROS activated p38 MAPK followed by NF-κB activation was crucial for the ZY-induced inflammatory responses in KCs. An in vivo study by peritoneal injection of ZY into BALB/C mice confirmed the anti-inflammatory properties of 0.7% ISO against ZY in KCs. These results suggest that ISO ameliorates ZY-induced inflammatory responses in murine KCs by inhibiting the interconnected ROS/p38 MAPK/NF-κB signaling pathways.

Palliative surgery for primary sarcoma in the abdominal aorta: A case report and review of the literature.

Primary sarcoma of the aorta is extremely rare and accounts for <1% of all sarcomas. The present study describes the case of a 45-year-old male who presented with lower limb and abdominal pain. Abdominal computed tomography (CT) and magnetic resonance (MR) arteriography revealed a tumor that extended from the infrarenal aorta to the aortic bifurcation. The external and internal iliac arteries were occluded by the tumor incursion. Palliative surgery was performed for the sarcoma since the patient refused a radical resection. To improve the blood supply to the lower limbs, an axillary bifemoral bypass was established. Following the surgery, the pain was significantly reduced. However, the patient succumbed due to extensive metastasis 6 months after this surgery. Aortic sarcoma is an extremely rare disease with a poor prognosis. A diagnosis at a relatively early stage is necessary for a longer survival time. Radical surgery is the most significant treatment. Patients at advanced stages should consider palliative surgery in order to improve their quality of life.

Metal (Mg, Fe, Co, Zr and Ti) complexes derived from aminosilyl substituted aminopyridinato ligand: synthesis, structures and ethylene polymerization behaviors of the group 4 complexes.

Lithium N-[(N,N-dimethylamino)dimethylsilyl]-2-pyridylamidate (2) was prepared from the reaction of new compound N-[(N,N-dimethylamino)dimethylsilyl]-2-pyridylamine (1) and LiBu(n). Treatment of the lithium salt (2) with an equal equivalent of MgBr(2)(THF)(2), FeCl(2) and CoCl(2) afforded the corresponding dinuclear complexes , , and , in which metal atoms possess similar trigonal bipyramidal geometries and each ligand functions as a bimetallic bridging binding aminopyridinato moiety with N-donation from the dimethylamino group. While the stoichiometric reaction of with ZrCl(4) gave the mononuclear zirconium complex (6); the seven coordinated zirconium atom adopts a distorted pentagonal bipyramid geometry and the ligand acts as monoanionic η(2)-aminopyridinato moiety with the pendant arm coordinated via N(CH(3))(2). The reaction of with one equivalent of TiCl(4)(THF)(2) produced the interesting dinuclear titanium complex (7) owing to the elimination of a (N,N-dimethylamino)dimethylsilyl group from the original ligand, and the two titanium centers present different coordination geometries. The molecular structures of the crystalline metal complexes have been confirmed by X-ray single crystal diffraction analysis. Upon activation with methylaluminoxane (MAO), both complexes and exhibited moderate catalytic activities toward ethylene polymerization and produced high molecular weight polyethylenes with broad molecular weight distributions.

A bulky silyl shift-directed synthesis of a silyl-linked amidinate-amidine and its Zr(IV) complex.

A novel silyl-linked amidinate-amidine monoanionic ligand 3 was synthesized by double additions of PhCN starting from silyl-linked bis(amino) monoanion 2, which underwent an intramolecular Li/H metathesis and double silyl shift. The related Zr(IV) complex 4 was prepared and confirmed by X-ray diffraction revealing a structural rearrangement from its precursor. The mechanisms for the reaction processes have been proposed. Each of compounds 2, 3 and 4 was characterized by NMR spectroscopy, elemental analysis and X-ray diffraction. Complex 4 exhibited moderate activity for ethylene polymerization in the presence of methylalumoxane (MAO).

Mononuclear titanium compounds based on the silyl-linked bis(amidinate) ligand: synthesis, characterization and ethylene polymerization.

The mononuclear silyl-linked bis(amidinate) titanium compound 2 was prepared by salt metathesis of 1 and TiCl(4)(thf)(2). Alkylation of 2 with methyllithium gave analogue 3. Both 2 and 3 exhibited a configuration similar to ansa-metallocene and showed good activity towards ethylene polymerization after activation with MAO.

T-vector and in vivo recombination as tools to construct a large antibody library of breast cancer.

The emergence of phage antibody libraries is an important advance in the field of antibody engineering. It provides a useful methodology to produce human antibodies and has the potential to replace traditional hybridoma technology. In our research, we used T-vector and in vivo recombination to construct a large antibody library from breast cancer patients. The use of T-vector considerably increased the cloning efficiency, and the diversity of the library could be increased easily using in vivo recombination. Taken together, a combination of these two techniques might be valuable in constructing a large antibody library.

[Construction and expression of eukaryotic coexpression plasmid containing human MUC1 gene and GM-CSF gene].

AIM: To construct an eukaryotic coexpression plasmid containing the coding region of human MUC1 tandem repeats gene and GM-CSF gene and to identify its expression in COS-7 cell. METHODS: MUC1 tandem repeats gene was obtained by synthesizing the segments. After identified by restriction endonuclease digestion analysis and DNA sequencing, MUC1 tandem repeats gene and GM-CSF gene were cloned into eukaryotic expression vector pcDNA3.1(+) to construct recombinant plasmid pcDNA3.1(+)-MUC1-GM-CSF. Then the recombinant plasmid was transfected into COS-7 cell by electroporation and the expression of target gene was detected by immunoflourescence and ELISA. RESULTS: Restriction analysis and DNA sequencing showed that the recombinant plasmid contained the coding region of human MUC1 tandem repeats gene and GM-CSF gene. The expression of MUC1 and GM-CSF was detected. CONCLUSION: The suuessful construction and expression of recombinant plasmid pcDNA3.1(+)-MUC1-GM-CSF lay a foundation for further development of DNA vaccine against breast cancer.

[Enhanced inhibitory effect of MUC1 gene vaccine on breast cancer growth by GM-CSF].

AIM: To investigate if GM-CSF adjuvant could enhance the inhibitory effect of the MUC1 gene vaccine on EMT6 breast cancer growth. METHODS: Female BALB/c mice were immunized intramuscularly with 100 microgram MUC1 gene vaccine 3 times at 3 week intervals. On 1, 3 and 5 days after intramuscular immunization, GM-CSF 100 microL(1 microg/100 microL) was given s.c. respectively. Three weeks after the last immunization, tumor challenge experiments were performed by using MUC1 expressing tumor cell line EMT6. Tumor growth inhibition was observed two weeks later. After 43 d of challenge experiments, all mice were killed and tumors were weighted. CTL-specific cytotoxicity was detected by (51)Cr release assay. RESULTS: After 43 d of challenge experiments, the size of EMT6 tumor in MUC1 DNA+GM-CSF, MUC1 DNA, pcDNA3.1(+)+GM-CSF and pcDNA3.1(+) group were (135+/-33.8) mm(3), (250+/-34.3) mm(3), (568+/-43.6) mm(3) and (596+/-48.2) mm(3), respectively. The average weight (g) of EMT6 tumors in those four groups were 0.81+/-0.42, 1.23+/-0.41, 2.30+/-0.48 and 2.28+/-0.58, respectively. EMT6 breast cancer growth in mice of MUC1 gene vaccine group was suppressed significantly(P<0.05), compared with that in pcDNA3.1(+) control group. Co-delivery of GM-CSF adjuvant and MUC1 DNA immunization enhanced the antitumour effects(P<0.05). The cytotoxicity of MUC1-specific CTLs to EMT6 target cells was different at various ratios of effector cells to target cells. At ratios of 100:1, 50:1, 25:1 and 12.5:1, the specific lysis for MUC1 DNA+GM-CSF group reached 68.5%, 53.4%, 35.9% and 28.5%, MUC1 cDNA group 54.1%, 39.8%, 26.4% and 20.1%, while for control groups 13.2%, 10%, 8.2%, 7.2% and 11.7%, 9.8%, 7.7%, 7.0%, respectively. GM-CSF enhanced the cytotoxicity of CTL induced by MUC1 DNA immunization(P<0.05). CONCLUSION: GM-CSF adjuvant significantly enhanced the inhibitory effect of the MUC1 gene vaccine on EMT6 breast cancer growth.

[Induction of specific cytotoxic T lymphocytes and humoral immune response by MUC1 DNA vaccine in mice].

AIM: To observe the specific cytotoxic T lymphocyte and humoral immune response induced by MUC1 DNA inmunization in mice. METHODS: Female BALB/c mice were immunized intramuscularly with 100 microg pcDNA3.1-MUC1 3 times at intervals of 3 weeks. Three weeks after the last immunization, tumor challenge experiments were performed by inoculation of MUC1 positive breast cancer cell line EMT6. Both humoral immune response and CTL-specific cytotoxicity were detected by immunohistochemical staining and (51)Cr release assay, respectively. RESULTS: The cytotoxicity of MUC1-specific CTLs to EMT6 target cells showed that the cytotoxic effect was different at various ratio of effector cells to target cells. At ratios of 100:1, 50:1, 25:1 and 12.5:1, the specific lysis in MUC1cDNA group reached 54.1%, 39.8%, 26.4% and 20.1%, while two control groups 13.2%, 10%, 8.2%, 7.2% and 11.7%, 9.8%, 7.7%, 7.0%, respectively. The former was markedly higher than the two latters(P<0.01). MUC1 expression was exhibited to be positive in breast cancer tissue by immunohistochemical staining with BALB/c mouse sera against MUC1 cDNA. There was only 40% tumorigenic rate in MUC1 cDNA immunized group, while there 100% in pcDNA3.1(+) and NS control groups. There was a significant difference between MUC1 cDNA group and control groups (P<0.05). CONCLUSION: Specific CTLs and antibody prodution are elicited by MUC1 DNA inmunization. MUC1 cDNA inmunization partly inhibit the growth of implanted tumor in mice.

[Expression and activation of MAPK pathway signaling molecules in human breast cancer cell lines].

AIM: To detect the expression and status of extracellular regulatory kinase 1 and 2 (ERK1/2) and its upstream kinase MEK1/2 proteins in four breast cancer cell lines MCF-7, Bcap-37, SK-BR-3 and T47D and study the effects of cyclophosphamide and epirubicin on the growth of the cell lines and on the expression and status of the signaling molecules. METHODS: Western blot was used to examine the expression and status of MEK1/2 and ERK1/2 proteins in these cells and the effects of these two drugs on them. The effects of the two drugs on the proliferation of these breast cancer cell lines were detected by MTT colorimetry. RESULTS: The levels of expression and phosphorylation of MEK1/2 and ERK1/2 proteins in four breast cancer cell lines increased notably as compared with those in MCF-10 cells. Both drugs could inhibit the proliferation of breast cancer cells. And the levels of expression and phosphorylation of MEK1/2 and ERK1/2 proteins in breast cancer cell lines treated with the drugs were markedly lower than those in untreated breast cancer cells. CONCLUSION: Overexpression and phosphorylation of MEK and ERK may play an important role in the generation and development of human breast cancer. The inhibitory effect of cyclophosphamide and epirubicin on proliferation of the breast cancer cells may be by means of inhibiting expression and phosphorylation of MEK and ERK.