ABSTRACT
OBJECTIVE@#To observe the effect of the small needle knife through the Zusanli(ST 36) on behavior and hippocampal expression of NLRP3 and IL-1β in myalgia comorbid depressed rats.@*METHODS@#The rat models of myalgia comorbid depression were prepared by intraperitoneal injection of acute reserpine. Twenty-four SD male rats were randomly divided into control group, model group, small needle knife group and amitriptyline group, 6 rats in each group. The open field behavior and mechanical pain threshold of each group were detected. The thermal pain threshold was detected by intelligent hot plate test. The expression of NLRP3 and IL-1β in hippocampus of rats was detected by Western blotting.@*RESULTS@#Compared with the model group, the mechanical pain threshold of the foot was significantly improved in the small needle knife group (0.05). The expressions of NLRP3 and IL-1β in the hippocampus of the model group were significantly increased(0.05).@*CONCLUSIONS@#Small needle knife can improve the pathological state of myalgia comorbid depression caused by reserpine in rats. The mechanism may be related to the inhibition of NLRP3 inflammasome and IL-1β expression in central hippocampus.
Subject(s)
Animals , Male , Rats , Hippocampus , Inflammasomes , Interleukin-1beta , Myalgia , NLR Family, Pyrin Domain-Containing 3 Protein , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To study the effect of PC-1 gene knockdown on the biological action of prostate cancer cell line C4-2.</p><p><b>METHODS</b>Recombinant plasmids of expressing short hairpin RNA targeting PC-1 mRNA were constructed using DNA recombinant technology and transfected into C4-2 cells via liposome. The positive cell clones were selected by G418. The expression of PC-1 gene was analyzed by RT-PCR and Western blotting technology. MTT and soft agar cloning formation were applied to observe the changes of the growth rate and independent anchor ability of C4-2 cells.</p><p><b>RESULTS</b>PC-1 RNA interference severely affected the expression of PC-1 gene and reduced the growth and colony formation ability of C4-2 cells.</p><p><b>CONCLUSION</b>RNA interference-mediated PC-1 gene knockdown can decrease the growth and cloning formation ability of C4-2 cells.</p>
Subject(s)
Humans , Male , Cell Line, Tumor , Down-Regulation , Gene Expression , Phosphoric Diester Hydrolases , Genetics , Prostatic Neoplasms , Genetics , Metabolism , Pathology , Pyrophosphatases , Genetics , RNA Interference , RNA, Messenger , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVES</b>To identify and clone human genes transactivated by HCV F protein by constructing a cDNA subtractive library using the suppression subtractive hybridization technique.</p><p><b>METHODS</b>Suppression subtractive hybridization (SSH) and bioinformatics techniques were used for screening and cloning of the target genes transactivated by HCV F protein. The mRNA was isolated from HepG2 cells transfected with pcDNA3.1 (-)-F or with pcDNA3.1(-) empty vector as a control, and SSH method was employed to analyze the differentially expressed DNA sequence between the two groups. After restriction enzyme Rsa I digestion, small sized cDNAs were obtained. Then tester cDNA was divided into two groups and ligated to the specific adaptor 1 or adaptor 2. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, it was then subcloned into T/A plasmid vectors to set up the subtractive library. Amplification of the library was carried out with E. coli strain DH5 alpha. The cDNA was sequenced and analyzed in GenBank with blast search after PCR.</p><p><b>RESULTS</b>The subtractive library of genes transactivated by HCV F protein was constructed successfully. The amplified library contains 71 positive clones. Colony PCR shows that 56 clones contain 200-1000 bp inserts. Sequence analysis was performed on 28 clones randomly, and the full length sequences were obtained with using the bioinformatics method. Altogether 19 coding sequences were obtained, consisting of 17 known and 2 unknown.</p><p><b>CONCLUSIONS</b>The obtained sequences may be target genes transactivated by HCV F protein, and some gene coding proteins are those involved in cell cycle regulation, metabolism, and cell apoptosis.</p>