An extracellular yellow laccase from white rot fungus Trametes sp. F1635 and its mediator systems for dye decolorization.

A novel extracellular laccase was purified from fermentation broth of the white rot fungus Trametes sp. F1635 by a three-step protocol including two consecutive ion-exchange chromatography steps on DEAE-Sepharose and SP-Sepharose, and a final gel-filtration on Superdex 75. The purified laccase (TsL) was a monomeric protein with the molecular mass of 64.8 kDa. It demonstrated high oxidation activity of 4.00 × 104 U/mg towards ABTS. Its N-terminal amino acid sequence was AIGPVADLTIINNAV which was unique and sharing high similarity of other fungal laccases. TsL was a yellow laccase based on absorption spectrum analysis. It demonstrated an acidic pH optimum of 2.6 and temperature optimum of 50 °C towards ABTS. The Km and Vmax values towards ABTS were estimated to 18.58 µM and 1.35 µmol/min, respectively. TsL manifested effective decolorization activity towards eriochrome black T (EBT), remazol brilliant blue R (RBBR), malachite green (MG), and eriochrome black T (EBT) (over 60%). Violuric acid (VA) and acetosyringone (AS) were the optimal mediators for the laccase in dye decolorization. Results suggest that TsL demonstrates great potential for dye decolorization and water treatment.

Comparison of Curative Effects of HAG and CAG Regimens for Patients with AML and Medium/High-Risk MDS / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To study the curative effect of HAG and CAG regimens for patients with acute myelocytic leukemia (AML) and high/medium-risk myelodysplastic syndrome (MDS).</p><p><b>METHODS</b>Fifty two patients from January 2010 to January 2014 were enrolled in this study, 32 were diagnosed with AML and 20 with MDS. All the patients were divided into 2 groups: 26 in HAG group (26 cases) and another 26 in CAG group (26 cases). The bone marrow examination, remission rate, PFS, OS and side reaction rates were compared between 2 groups.</p><p><b>RESULTS</b>After treatment, the bone marrow hyperplasia and juvenile cells were decreased significantly. In HAG group, the remission rate was 57.69% and that was 76.92% in CAG group, the difference between these 2 groups was statistically significant (P<0.05), but the survival time was not statistically significant different between 2 groups (P>0.05). The incidence of side reaction in HAG group was 11.54%, that in CAG group was 7.69%, there was no statistically significant difference (P>0.05).</p><p><b>CONCLUSION</b>Both CAG and HAG regimens have shown significant curative effects for acute myelocytic leukemia and high/medium-risk myelodysplastic syndrome.</p>

Effect of jianpi liqi recipe on mRNA expressions of TP, DPD, and CYP3A4 in mice with human gastric cancer transplanted tumor / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of Jianpi Liqi Recipe (JLR) on 5-fluorouracil (5-FU) relevant metabolic enzymes and CYP3A4 (the same enzyme of many chemotherapeutics) of mice with human gastric cancer transplanted tumor.</p><p><b>METHODS</b>Totally 80 mice were randomly divided into the model group, the chemotherapy group, the JLR group, and the combination group (using chemotherapy combined JLR), 20 in each group. The human gastric cancer transplanted tumor mouse model was duplicated by hypodermic inoculating MKN-8 tumor cell suspension from the left armpit. Physiological saline or JLR was given to those in the model group or the JLR group at 0.25 mL each time, twice daily by gastrogavage from the 2nd day after transplantation. Mice in the chemotherapy group were given 0.25 mL physiological saline, twice daily by gastrogavage 2 days after transplantation, for 5 days in succession, and then they were peritoneal injected with 5-FU at the daily dose of 20 mg/kg, once daily for 5 days in succession from the 7th day of transplantation. Those in the combination were given 0.25 mL JLR, twice daily by gastrogavage, for 5 days in succession, and then they were peritoneal injected with 5-FU at the daily dose of 20 mg/kg, once daily for 5 days in succession from the 7th day of transplantation. The mRNA expressions of thymidine phosphorylase (TP), dihydropyrimidine dehydrogenase (DPD), and CYP3A4 were detected using RT-PCR.</p><p><b>RESULTS</b>Compared with the model group and the chemotherapy group, mRNA expressions of TP and CYP3A4 obviously increased, mRNA expression of DPD obviously decreased in the JLR group and the combination group (P < 0.01). There was no statistical difference in mRNA expressions of TP, DPD, and CYP3A4 between the JLR group and the combination group (P > 0.05).</p><p><b>CONCLUSION</b>JLR could promote the activation of 5-FU, suppress the decomposition and inactivation of 5-FU in the tumor tissue of mice, and improve the chemotherapeutic efficacy through up-regulating mRNA expressions of TP and CYP3A4, and suppressing the mRNA expression of DPD.</p>

Clinicopathological significance of PTEN and Caspase-3 expressions in breast cancer / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the expressions of PTEN and Caspase-3 proteins in human breast carcinoma, and to evaluate their clinicopathological implications during the tumorigenesis and progression of breast cancer.</p><p><b>METHODS</b>The expressions of PTEN and Caspase-3 proteins in 95 cases of breast cancer and 15 cases of benign breast diseases were investigated immunohistochemically. Correlations between the expression of PTEN protein, Caspase-3 protein, and clinicopathological features of breast cancers were analyzed.</p><p><b>RESULTS</b>The loss expression rate of PTEN protein in tumor tissues was significantly higher than that in benign breast diseases (33.7% vs. 0, P < 0.01). Analysis of the clinicopathological features showed that PTEN expression level was negatively correlated with TNM stage, histological grade, axillary lymph node status, recurrence, and metastasis (P < 0.05). The positive expression level of Caspase-3 was negatively correlated with TNM stage (P < 0.01), but not related with histological grade, axillary lymph node status, recurrence, or metastasis (P > 0.05). In addition, the expression of PTEN protein had significantly positive correlation with the expression of Caspase-3 protein in breast cancer (P < 0.01).</p><p><b>CONCLUSION</b>The combination detection of PTEN and Caspase-3 may serve as an important index to estimate the pathobiological behavior and prognosis of breast cancer.</p>

Mutation of mitochondrial 12S rRNA in gastric carcinomas and its significance / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect the alterations of mitochondrial 12S rRNA in patients with gastric cancer, and further evaluate their effects on development of gastric carcinomas.</p><p><b>METHODS</b>Mitochondrial 12S rRNA of 22 samples of gastric cancer tissues and 22 corresponding normal gastric mucosa taken from the distal portion of surgical specimens were PCR amplified, followed by direct DNA sequencing. Laser capture microdissection technique (LCM) was used to isolate cancerous cells and dysplastic cells from patients with specific mutations. Denaturing high-performance liquid chromatography (DHPLC) plus allele-specific PCR (AS-PCR), nest-PCR and polyacrylamide gel electrophoresis (PAGE) were applied to further evaluate this mutant property and quantitative difference of mutant type between cancerous and dysplastic cells. Finally, RNAdraw bio-soft was used to analyze the RNA secondary structure of mutant type 12S rRNA.</p><p><b>RESULTS</b>Compared with mitomap database, some variations were firstly found, among which np652 G insertion and np716 T-G transversion were only found in cancers. There existed statistically significant difference in variant frequency of 12S rRNA between intestinal type and diffuse type of gastric carcinoma, 5/17 (29.4%) and 12/17 (70.6%) respectively (P < 0.05). DHPLC analysis showed that 12S rRNA np652 G insertion and np716 T-G transversion were heteroplasmic mutation. Variant frequency of 12S rRNA in cancer was higher than that in dysplasia (P < 0.01). 12S rRNA 652G insertion had more adverse effect on secondary structure stability of 12S rRNA than T-G transversion did.</p><p><b>CONCLUSION</b>Highly variant frequency of mitochondrial 12S rRNA may be associated with intestinal type of gastric cancer. Most parts of variations exist in both cancer and normal tissues and may not be characteristic of tumor specificity. However np652 G insertion and np716 T-G transversion may possess some molecular significance on gastric cancerogenesis. During the process of progression from normality through dysplasia to cancer, 12S rRNA tended to transit from homoplasmy (wild type) and heteroplasmy to homoplasmy (mutant type, np717 T-G).</p>

The use of bioluminescent image in monitoring islet graft in vivo after islet transplantation / 中华器官移植杂志

Objective To demonstrate the superiority and feasibility of using bioluminescent image to monitor the islet graft after islet transplantation.Methods Diabetic models were established by intraperitoneal injection of streptomycin into mature male C57BL/6 mice.Islets were harvested from the pancreas of C57BL/6 and Bclb/c mice by digestion and purification,and transfected with Lueiferase gene.The mouse diabetic models were divided into iso-transplantaion group (n=20) and allo-transplantation group (n=7).The islets of C57BL/6 were transplanted into iso-transplantaion mice with different doses (10,50,110 and 200,n=5 in every dose),and Bclb/c mouse islets were transplanted into allo-transplantation group.The islets were transplanted into the subcutaneous fat tis- sue near left scapula.The receptor mice were scanned with CCD camera to get bioluminescent images at different scheduled time points,and the changes in random blood glucose of allo-transplantation group were observed.Results On day 6 after transplantation,the scanning image showed that the pi- xel intensity from the region of interest (ROI) was increased with the increased number of islet grafts and they had a positive correlation.The random blood glucose was reduced to the normal level in the first 2 days,and then increased again to the diabetic level on 11 days averagely,while pixel intensity from the ROI reached the peak on day 6-7,and then reduced rapidly after islet transplantation in allo- transplantation group.The beginning of pixel intensity reduction occurred on day (6.14?0.90), while that of the random blood glucose raise occurred on day (10.00?0.82) after transplantation,and the former alteration occurred obviously earlier than the latter (P

Altered expression of PTEN gene and LOH of its epigenetic microsatellite in gastric carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the expression of PTEN and loss of heterozygosity (LOH) of its epigenetic microsatellite in gastric carcinoma and explore their roles in progression of gastric carcinoma.</p><p><b>METHODS</b>LOH of epigenetic microsatellites of PTEN (D10S541, D10S583 and D10S1687) in advanced gastric cancer was detected by PCR-SSCP. Expression of PTEN mRNA and protein in normal gastric mucosa and gastric cancer was evaluated by RT-PCR and SABC immunohistochemistry, respectively. The relationship between expression of PTEN mRNA and protein and lymph node metastasis or LOH of microsatellites was discussed.</p><p><b>RESULTS</b>LOH of D10S541, D10S583 and D10S1687 was found in 37.5% (21/56) of advanced gastric cancers. The positive rates of PTEN mRNA expression were 80.4% (45/56), 45.5% (5/11) and 32.1% (18/56) in normal mucosa, early and advanced gastric carcinomas, respectively, while 78.6% (44/56), 44.5% (5/11) and 28.6% (16/56) at the protein level. PTEN mRNA and protein were less frequently expressed in early and advanced gastric carcinomas than that in normal gastric mucosa (P < 0.05). There was positive correlation between PTEN mRNA expression and LOH of microsatellites in advanced gastric carcinomas. PTEN protein expression paralleled with its mRNA expression (P < 0.05). The expression of PTEN mRNA and protein was negatively correlated with lymph node metastasis of advanced gastric carcinomas (P < 0.05).</p><p><b>CONCLUSION</b>Down-regulated expression of PTEN gene is found in different stages of gastric carcinoma, and is closely correlated with LOH of its epigenetic microsatellites, which probably is its underlying molecular mechanisms. It suggests that altered PTEN gene contributes to tumorigenesis and progression of gastric carcinomas.</p>

Expression of maspin and kai1 and their clinicopathological significance in carcinogenesis and progression of gastric cancer / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the roles of maspin and kai1 expression in tumorigenesis and progression of gastric cancer.</p><p><b>METHODS</b>Maspin and kai1 expressions were detected in normal gastric mucosa (n = 182), gastric dysplasia (n = 69), and gastric cancer (n = 113) by immunohisto-chemistry. Their expressions were compared with clinicopathological parameters of tumors. Relationship between maspin and kai1 expression was also concerned in gastric cancer.</p><p><b>RESULTS</b>The positive rates of maspin expression were 79.8% (145/182), 75.4% (52/69), and 50.4% (57/113) in normal gastric mucosa, gastric dysplasia, and gastric cancer, while those of kai1 expression were 81.9% (149/182), 65.2% (49/69), and 58.4% (66/113) in corresponding tissues respectively. Gastric cancer less frequently expressed maspin than the normal gastric mucosa and gastric dysplasia (P < 0.05), while dysplasia and cancer showed less frequent expression of kai1 than normal mucosa (P < 0.05). Maspin expression showed negative association with invasive depth, metastasis, Lauren's and histological classifications (P < 0.05), but not with tumor size, Borrmann's classification, growth pattern or TNM staging (P > 0.05). Kai1 expression was negatively correlated with invasive depth, metastasis, growth pattern, Lauren's and histological classifications (P < 0.05), but not with tumor size, Borrmann's classification or TNM staging (P > 0.05). Maspin and kai1 were collaboratively expressed in gastric cancer (P < 0.05).</p><p><b>CONCLUSIONS</b>Down-regulated expressions of maspin and kai1 play an important role in gastric carcinogenesis. Abnormal expression of maspin and kai1 might have inhibitory effects on invasion and metastasis of gastric cancer and act as an effective and objective marker to indicate the pathobiological behaviors of gastric cancer.</p>