Correlations of UGT1A1 gene polymorphisms with onset and prognosis of non-small cell lung cancer.

OBJECTIVE: The aim of this study was to investigate the correlations of UDP glucuronosyltransferase family 1 member A1 (UGT1A1) gene polymorphisms with the onset and prognosis of non-small cell lung cancer. PATIENTS AND METHODS: A total of 400 patients with non-small cell lung cancer (disease group) and healthy controls (control group) in our hospital were selected as research subjects. Genomic DNA was extracted from the peripheral blood. UGT1A1 gene polymorphisms rs8330, rs4148323 and rs35003977 were detected after Polymerase Chain Reaction (PCR) amplification. RT-qPCR was performed to measure the expression level of UGT1A1. The survival of patients was analyzed combined with their prognosis. Moreover, the expression of UGT1A1 gene in lung cancer patients from The Cancer Genome Atlas (TCGA) database was analyzed by bioinformatics, and the prognosis was analyzed. RESULTS: According to the expression level of UGT1A1 gene from TCGA and GTEx databases, UGT1A1 gene was highly expressed in lung cancer tissues but lowly expressed in normal lung tissues, and the difference was statistically significant (p<0.05). Combined with the expression level of UGT1A1 and the prognostic information of lung cancer patients from TCGA database, patients with higher expression level of UGT1A1 gene exhibited significantly better prognosis than those with lower level (p=0.0013), suggesting that UGT1A1 gene is an anti-oncogene. There were statistically differences in allele distribution of UGT1A1 gene polymorphism rs8330 between the disease group and control group (p=0.003), and the frequency of allele G was higher in disease group. Moreover, the distribution of genotypes of UGT1A1 gene polymorphisms rs8330 (p=0.006) and rs4148323 (p=0.003) in the disease group was significantly different from that in the control group, and the frequencies of GG genotype of polymorphisms rs8330 and rs4148323 were higher in the disease group. Statistically significant differences in the distribution of recessive models of UGT1A1 gene polymorphism rs8330 were observed between the disease group and control group (p=0.047), and the disease group exhibited a lower frequency of recessive model GC + CC than control group. There were evident differences in the distribution of haplotype GGT of UGT1A1 gene polymorphisms rs8330, rs4148323 and rs35003977 between the disease group and control group (p=0.004), and the frequencies of haplotype GGT were higher in the disease group than those in the control group. UGT1A1 gene polymorphism rs8330 was remarkably associated with gene expression (p<0.05). Meanwhile, the expression of UGT1A1 gene in patients carrying genotype CC declined notably compared with that in patients carrying genotypes GG and GC (p<0.05). Furthermore, the polymorphism rs8330 had a significant correlation with the survival of patients in disease group (p=0.0001), and patients with genotype CC had the worst prognosis. CONCLUSIONS: UGT1A1 gene polymorphisms are prominently correlated with the onset and prognosis of non-small cell lung cancer.

Phase-shifting differential interference contrast microscope with Savart shear prism and rotatable analyser.

A novel differential interference contrast microscope (DICM) is proposed in this research. It is constituted by inserting a Savart shear prism between the objective and sample of a polarising microscope having a rotatable analyser as the phase-shifter, and it is with the ability to enhance image contrast using the principle of shearing interferometry. This letter is to introduce the configuration, interpret the interference patterns and present the experimental setup of the DICM. In addition, this letter is to display the experimental results from the uses of the setup; the results demonstrate the validity and ability of the DICM.

Antimicrobial and antitumor activity and diversity of endophytic fungi from traditional Chinese medicinal plant Cephalotaxus hainanensis Li.

Endophytes from Cephalotaxus hainanensis Li, an important source of anti-leukemia drugs, have not been widely explored. In this study, 265 endophytic fungal isolates from C. hainanensis Li were screened for antimicrobial activities against tilapia, banana, rice, and rape and for antitumor activities against human leukemia cell lines (K562, NB4, and HL-60). Diversity was also analyzed. The results showed that 17.7% of the endophytic fungi had antimicrobial activities against at least three different test microbes, and activity against Fusarium oxysporum RKY102 was the highest at 15.8%. Cytotoxicity against at least one tumor cell line tested was observed in 18.5% of the endophytic fungi; with the highest value of 10.6% against K562. The endophytic fungal strains also showed relatively high activities against K562, NB4, and HL-60 while relatively fewer strains were cytotoxic against the human hepatic Hep-G2 and colon LoVo cancer cell lines. Thirty endophytic fungal strains showed both high antimicrobial and antitumor activities. Moreover, the analyses of the diversity of the 30 highly active strains showed they belonged to 20 species from 14 genera, and this is the first report of endophytic fungi Albonectria rigidiuscula, Colletotrichum magnisporum, and Nemania diffusa being isolated from Cephalotaxus plants. These findings suggest that natural antibacterial products for humans and tilapia; antifungal compounds for rice, rape, and banana; and antitumor compounds for leukemia therapy could be isolated from fungal strains derived from C. hainanensis Li.

Salmonella induce autophagy in melanoma by the downregulation of AKT/mTOR pathway.

Salmonella have been demonstrated to inhibit tumor growth. However, the mechanism of Salmonella-induced tumor cell death is less defined. Autophagy is a cellular process that mediates the degradation of long-lived proteins and unwanted organelles in the cytosol. Tumor cells frequently display lower levels of basal autophagic activity than their normal counterparts and fail to increase autophagic activity in response to stresses. Autophagy is involved in the cell defense elimination of bacteria. The signaling pathways leading to activation of Salmonella-induced autophagy in tumor cells remain to be elucidated. We used autophagy inhibitor (3-Methyladenine) and apoptosis inhibitor (Z-VAD-FMK) to demonstrate that Salmonella may induce cell death via apoptosis and autophagic pathway. Meanwhile, we suggested that Salmonella induce autophagy in a dose- and time-dependent manner. The autophagic markers were increased after tumor cell infected with Salmonella. In addition, the protein express levels of phosph-protein kinase B (P-AKT), phosph-mammalian targets of rapamycin (P-mTOR), phosph-p70 ribosomal s6 kinase (P-p70s6K) in tumor cells were decreased by western analysis after Salmonella infection. In conclusion, our results point out that Salmonella induce the autophagic signaling pathway via downregulation of AKT/mTOR pathway. Herein, our findings that Salmonella in controlling tumor growth may induce autophagic signal pathway.

Suppressor of cytokine signaling 6 (SOCS6) promotes mitochondrial fission via regulating DRP1 translocation.

Mitochondria are highly motile organelles that constantly undergo fission and fusion. Impairment of mitochondrial dynamics is associated with mitochondrial dysfunction and is frequently linked to the pathogenesis of neurodegenerative diseases and cancer. We have previously shown that biallelic inactivation of the suppressor of cytokine signaling 6 (SOCS6) gene is a frequent event in human gastric cancer. In this study, we recapitulated the event of SOCS6 loss using a Lentivirus-based knockdown approach, and demonstrated the linkage between SOCS6 depletion and the suppression of programmed cell death. SOCS6 promotes intrinsic apoptosis, with increased Bax conformational change, mitochondrial targeting, and oligomerization. Most importantly, SOCS6 is targeted to mitochondria and induces mitochondrial fragmentation mediated through an increase in DRP1 fission activity. Here, we show that SOCS6 forms complex with DRP1 and the mitochondrial phosphatase PGAM5, attenuates DRP1 phosphorylation, and promotes DRP1 mitochondrial translocation. Based on mutation analyses, SOCS6-mediated apoptosis is tightly coupled to its ability to induce mitochondrial fission. This study demonstrates an important role for SOCS6 in modulating mitochondrial dynamics and apoptosis.

Flexible optitrode for localized light delivery and electrical recording.

We present optitrode, a miniaturized flexible probe for integrated, localized light delivery and electrical recording. This device features an annular light guide with transparent polymer and fused silica layers surrounding a twisted-wire tetrode. We have developed a novel fabrication process, V-groove guided capillary assembly, to achieve high-precision, coaxial alignment of the various layers of the device. Optitrode with a length-to-diameter ratio ∼500 (5 cm long, 100 µm diameter) has been fabricated, and both the electrical and optical functions have been characterized. The prototype can deliver 11% (110 mW) of the total laser power under abrupt bending angle ∼25°.

A novel RING finger protein, Znf179, modulates cell cycle exit and neuronal differentiation of P19 embryonal carcinoma cells.

Znf179 is a member of the RING finger protein family. During embryogenesis, Znf179 is expressed in a restricted manner in the brain, suggesting a potential role in nervous system development. In this report, we show that the expression of Znf179 is upregulated during P19 cell neuronal differentiation. Inhibition of Znf179 expression by RNA interference significantly attenuated neuronal differentiation of P19 cells and a primary culture of cerebellar granule cells. Using a microarray approach and subsequent functional annotation analysis, we identified differentially expressed genes in Znf179-knockdown cells and found that several genes are involved in development, cellular growth, and cell cycle control. Flow cytometric analyses revealed that the population of G0/G1 cells decreased in Znf179-knockdown cells. In agreement with the flow cytometric data, the number of BrdU-incorporated cells significantly increased in Znf179-knockdown cells. Moreover, in Znf179-knockdown cells, p35, a neuronal-specific Cdk5 activator that is known to activate Cdk5 and may affect the cell cycle, and p27, a cell cycle inhibitor, also decreased. Collectively, these results show that induction of the Znf179 gene may be associated with p35 expression and p27 protein accumulation, which lead to cell cycle arrest in the G0/G1 phase, and is critical for neuronal differentiation of P19 cells.

Long-term results of Taiwan Pediatric Oncology Group studies 1997 and 2002 for childhood acute lymphoblastic leukemia.

The long-term outcome of 1390 children with acute lymphoblastic leukemia (ALL), treated in two successive clinical trials (Taiwan Pediatric Oncology Group (TPOG)-ALL-97 and TPOG-ALL-2002) between 1997 and 2007, is reported. The event-free survival improved significantly (P=0.0004) over this period, 69.3+/-1.9% in 1997-2001 to 77.4+/-1.7% in 2002-2007. A randomized trial in TPOG-97 testing L-asparaginase versus epidoxorubicin in combination with vincristine and prednisolone for remission induction in standard-risk (SR; low-risk) patients yielded similar outcomes. Another randomized trial, in TPOG-2002, showed that for SR patients, two reinduction courses did not improve long-term outcome over one course. Decreasing use of prophylactic cranial irradiation in the period 1997-2008 was not associated with increased rates of CNS relapse, prompting complete omission of prophylactic cranial irradiation from TPOG protocols, beginning in 2009. Decreased use of etoposide and cranial irradiation likely contributed to the low incidence of second cancers. High-risk B-lineage ALL, T-cell, CD10 negativity, t(9;22), infant, and higher leukocyte count were consistently adverse factors, whereas hyperdiploidy >50 was a consistently favorable factor. Higher leukocyte count and t(9;22) retained prognostic significance in both TPOG-97 and TPOG-2002 by multivariate analysis. Although long-term outcome in TPOG clinical trials is comparable with results being reported worldwide, the persistent strength of certain prognostic variables and the lower frequencies of favorable outcome predictors, such as ETV6-RUNX1 and hyperdiploidy >50, in Taiwanese children warrant renewed effort to cure a higher proportion of patients while preserving their quality of life.

ErbB4 (JM-b/CYT-1)-induced expression and phosphorylation of c-Jun is abrogated by human papillomavirus type 16 E5 protein.

Human papillomavirus type 16 E5 (HPV-16 E5) is a highly hydrophobic membrane protein with weak-transforming activity, which is associated with ErbB4 receptor in HPV-16-infected cervical lesions. Presently, we investigated the transforming mechanisms of E5 involving ErbB4 signaling. Firstly, we report a role for ErbB4 (JM-b/CYT-1) receptor that activates c-jun gene expression and phosphorylating at Ser63 and Ser73 of the c-Jun protein in ligand-independent and Ras-c-jun NH(2)-terminal kinase-dependent pathway. Secondly, we show that HPV-16 E5 protein can form a complex with ErbB4 via binding to the extracellular and transmembrane domains of ErbB4 (JM-b/CYT-1). When co-expressing HPV-16 E5 and ErbB4 in cells, E5 can abrogate ErbB4-induced c-Jun protein expression and phosphorylation resulted in increasing cell proliferation compared to ErbB4-expressing cells. The interaction between of HPV-16 E5 and ErbB4 provides more insight into the mechanisms of HPV-16 E5 transformation induction.

Renoprotective effects of combination of angiotensin converting enzyme inhibitor with mycophenolate mofetil in diabetic rats.

OBJECTIVE AND DESIGN: Previously it was shown that blocking of the renin-angiotensin system (RAS) by angiotensin converting enzyme (ACE) inhibitors, or suppression of inflammatory responses by immunosuppressive drugs such as mycophenolate mofetil (MMF) could attenuate renal injury in diabetic rats. Whether RAS blockade combined with an immunosuppressive drug provides superior renoprotection against diabetic nephropathy has not been clearly delineated. MATERIALS: Diabetes was induced by injection of streptozotocin after uninephrectomy. TREATMENT: Rats were randomly separated into five groups: control, diabetes, diabetes treated with enalapril (an ACE inhibitor, 10 mg/kg/d by gastric gavage), diabetes treated with MMF (10 mg/kg/d by gastric gavage), or diabetes treated with a combination of both agents and were followed for 8 weeks. METHODS: 24 h urinary albumin excretion rate (AER) was determined, renal injury was evaluated, immunohistochemistry for ED-1 macrophage marker, intercellular adhesion molecule-1 (ICAM-1) and monocyte chemoattractant protein-1 (MCP-1) were performed, and expression of transforming growth factor (TGF)-beta1 protein was determined by Western blotting analysis. RESULTS: Diabetes was associated with a considerable increase in AER. Both enalapril and MMF retarded the increase in albuminuria, which was nearly completely abrogated by combination therapy. Increased glomerular volume and tubulointerstitial injury index in diabetic rats was attenuated by treatment with either enalapril or MMF and further reduced by the combination of the two. Elevated malondialdehyde levels in renal tissue were reduced by enalapril or MMF and, more effectively, by combined enalapril with MMF. Renal overexpression of ICAM-1 was not retarded by enalapril and attenuated by MMF or combined enalapril with MMF. Combination therapy was associated with a superior suppression of diabetes-induced macrophage recruitment and overexpression of MCP-1 and TGFbeta1 compared to either monotherapy in renal tissue. CONCLUSION: The combination of enalapril and MMF confers superiority over monotherapy in renoprotection, a mechanism which may be at least partly correlated with synergistic suppression of increased macrophage recruitment and overexpression of MCP-1 and TGF-beta1 in renal tissue in diabetic rats.

MPTP treatment of common marmosets impairs proteasomal enzyme activity and decreases expression of structural and regulatory elements of the 26S proteasome.

Dysfunction of the ubiquitin-proteasome system occurs in the substantia nigra (SN) in Parkinson's disease (PD). However, it is unknown whether this is a primary cause or a secondary consequence of other components of the pathogenic process. We have investigated in nonhuman primates whether initiating cell death through mitochondrial complex I inhibition using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP) altered proteasomal activity or the proteasomal components in the SN. Chymotrypsin-like, trypsin-like and peptidylglutamyl-peptide hydrolase (PGPH) activating of 20S proteasome were decreased in SN homogenates of MPTP-treated marmosets compared to naïve animals. Western blotting revealed a marked decrease in the expression of 20S-alpha subunits, but no change in 20S-beta subunits in the SN of MPTP-treated marmoset compared to naïve animals. There was a marked decrease in the expression of the proteasome activator 700 (PA700) and proteasome activator 28 (PA28) regulatory complexes. The 20S-alpha4 subunit immunoreactivity was decreased in the nucleus of colocalized tyrosine hydroxylase (TH)-positive cells of MPTP-treated animals compared to naïve animals but no difference in the intensity of 20S-beta1i subunit staining. Immunoreactivity for PA700-Rpt5 and PA28-alpha subunits within surviving TH-positive cells of MPTP-treated marmoset was reduced compared to naïve controls. Overall, the changes in proteasomal function and structure occurring follow MPTP-induced destruction of the SN in common marmosets were very similar to those found in PD. This suggests that altered proteasomal function in PD could be a consequence of other pathogenic processes occurring in SN as opposed to initiating cell death as previously suggested.

Paper electrophoresis of serum from patients with Philippine h-fever

The serum protein fractions from Philippine Hemorrhagic Fever cases were studied by means of paper electrophoresis as a preliminary step in the clarification of tbe biochemical nature of this disease. Variations in the electrophoretic pattern might indicate the presence of an aberrant chemical component or simply a quantitative increase or decrease in a well characterize constituent. Also, when abnormal patterns exist, these if consistent, may serve as a diagnostic tool or great clinical value. The limited clinical material available for study precludes the present work from becoming a definitive study. Analysis of the serum protein fractions of the Philippine Hemorrhagic Fever cases studied when compared to those of the control group exhibited certain characteristics of interest. Among these was the observation that the averages for each separated serum protein fraction of the Philippine Hemorrhagic Fever cases when compared to those of the control group were found to be significantly different with the exception of the alpha, globulin fraction. A study of the percentage of serum protein fractioin values for Philippine Hemorrhagic Fever cases that fall outside the standard deviation from the mean of the control group values showed 65 percent of the albumin fraction to be below this mean value, while none were above this values. Ten percent of the alpha cases were below while 30 percent were above the mean value. No alpha fractions were below but 55 percent were found to be above the control group mean value. Beta globulin was below the control mean value in 15 percent of the cases and above in 40 percent. Lastly the gamma globulin fraction was above the control group mean value in 60 percent, and below in only 5 percent of cases. No reversal of the albumin-globulin ratio was observed at any time. Total protein values were found to be within the normal range of the determination employed. A group of children studied separately showed no difference from the Philippine Hemorrhagic Fever case group as a whole. Examination of clinical records revealed a below average platelet count and positive tourniquet test producing petechial hemorrhages to be a consistent characteristic among Philippine Hemorrhagic Fever cases.(Summary)