[Effects of "Tiaoyi Sanjiao" acupuncture and moxibustion on cancer-induced fatigue and immune function in patients with advanced non-small cell lung cancer].

OBJECTIVE: To observe the therapeutic effect of "Tiaoyi Sanjiao" （regulating and tonifying the triple energizer）acupuncture and moxibustion in the treatment of cancer-related fatigue of advanced non-small cell lung cancer and observe its influence on lymphocyte count. METHODS: The cancer-related fatigue patients with advanced non-small cell lung cancer who met the inclusive criteria were randomly divided into 2 groups, with 77 cases in each group. Patients in the control group were treated with conventional Chinese and Western medicine. In the treatment group, on the base of the treatment as the control group, "Tiaoyi Sanjiao" acupuncture and moxibustion was applied（mild moxibustion at Danzhong ［CV17］, Zhongwan［CV12］, Qihai［CV6］, bilateral Zusanli［ST36］, and acupuncture at bilateral Xuehai［SP10］, Waiguan［SJ5］, Taichong［LR3］）. KPS score, Piper scale, percentage and absolute count of lymphocyte subsets of the two groups before and after treatment were observed. RESULTS: Compared with pre-treatment, the KPS scores of both groups were significantly increased after treatment (P<0.05); the behavioral dimension score of the Piper scale, and the percentage of B cells of the control group decreased significantly(P<0.05); the behavioral dimension, emotional dimension, sensory dimension scores, and total score of the Piper scale in the treatment group were significantly reduced(P<0.05), while the percentage of CD3+T cells and absolute counts of CD3+T cells, CD4+T cells and CD8+T cells of the treatment group were significantly increased (P<0.05). After treatment, in comparison with the control group, behavioral dimension score of the Piper scale in the treatment group decreased significantly(P<0.05); the percentage and absolute counts of B cells and CD4+T cells, the absolute count of CD3+T cells in the treatment group were up-regulated (P<0.05). CONCLUSION: "Tiaoyi Sanjiao" acupuncture and moxibustion can significantly alleviate the fatigue state and improve the quality of life in patients with advanced non-small cell lung cancer, which may be achieved by regulating the number of lymphocytes and improving immune function.

A novel tumor suppressor SPINK5 targets Wnt/ß-catenin signaling pathway in esophageal cancer.

Esophageal cancer is one of the most common tumor in the world, and the morbidity rate is as high as 100/100 000 in some parts of China. Therefore, it is important and urgent to explore the pathogenesis of esophageal cancer and find new therapeutic targets for esophageal cancer. In this study, we found that a novel tumor suppressor SPINK5 is significantly reduced in the development of esophageal cancer, and is closely related to the pathological differentiation and lymph node metastasis of esophageal cancer via bioinformatics analysis and esophageal cancer tissue array. Further studies have found that SPINK5 is closely related to Wnt/ß-catenin signaling pathway by bioinformatics analysis and western blot. In esophageal cancer cells, SPINK5 overexpression can inhibit Wnt/ß-catenin signaling pathway. Combined with LiCl or MG-132 treatment, SPINK5 can inhibit GSK3ß phosphorylation and promote ß-catenin protein degradation, thus inhibit Wnt/ß-catenin signaling pathway. In vivo study, SPINK5 overexpression can significantly inhibit the growth of esophageal cancer cells. Our study shows that SPINK5 can inhibit the proliferation, migration, and invasion of esophageal cancer cells by inhibiting Wnt/ß-catenin signaling pathway, and thus plays an important role in the development of esophageal cancer, and may serve as a treatment target of esophageal cancer.

[Value of DAPK Gene Methylation Patterns in the Diagnosis of Leukemia].

OBJECTIVE: To screen the differential methylation patterns of tumor suppressor gene DAPK and evaluate its value as a biomarker for the diagnosis of leukemia. METHODS: The methylation status of DAPK gene promoter's CpG island was analyzed in the genomes of normal human white blood cells and HL-60, U937 and Jurkat cell lines by bisulfite sequencing PCR (BSP). The effectiveness of differential methylation patterns of DAPK gene for diagnosis of leukemia was verified in the leukemia cell lines and peripheral blood samples by methylation specific PCR (MSP). RESULTS: The methylation pattern of DAPK gene in different cell genomes displayed that the degree of unmethylation in normal cell genome was higher than that of leukemia cell lines. The differential CpG sites were found and could be used to differentiate HL-60 and the other 3 cell lines by MSP. Meanwhile, the differential methylation patterns in clinical specimens could distinguish acute non-lymphocytic leukemia (ANLL) and other types of leukemia by MSP. The diagnostic sensitivity, specificity and accuracy were 59.1%, 100% and 82.7% respectively. No relationship was found between MSP diagnosis results and clinical pathological typing. CONCLUSION: The differential methylation patterns of DAPK gene as potential tumor biomarker for diagnosis of leukemia can enrich the means of diagnosis of leukemia, provide idea and basis for finding all kinds of tumor's DNA methylation biomarkers in the future.

[Improvement and Evaluation on Bisulfite Modification of DNA for DNA Methylation Detection].

OBJECTIVE: To establish a rapid and convenient method of DNA modification by bisulfite sodium for the detection of DNA methylation. METHODS: Through increasing the bisulfite sodium concentration and the temperature of treatment, cutting down the modification time, besides using glassmilk to adsorb the DNA in the purification and recovery, to improve the methods of DNA modification. Efficiency of cytosine converted to thymine in MAGE-A3 gene and DAP-K gene fragments were analyzed by bisulfite sequencing PCR in order to evaluate the DNA modification effect among the improved method, traditional method and kit method. RESULTS: The operating time of test was shortened to about 3 hours by the improved method; conversion rate of unmethylated cytosine to thymine was over 99%; compared with the traditional method and kit method, there was no significant difference (χ(2) = 0.0564, P > 0.05); the improved method was only for the unmethylated cytosine conversion modification, and there was no significant difference in process of methylated cytosine converted to thymine comparing with the traditional method (χ(2) = 0.0149, P > 0.05). CONCLUSION: The improved method has high efficiency of DNA modification and has no significant effect on excessive modification;meanwhile, it has many advantages such as time-saving and easy to operate etc.