Objective As SELDI-TOF-MS (Surface Enhanced Laser Desorption/Ionization Time of Flight Mass Spectrometry) has been broadly used to screen biomarkers for a variety of diseases, the identification and validation of the revealed biomarkers requires more focused attention.Method In this paper, the serum samples from 60 cholangiocarcinoma, 146 lung cancer, 65 LGC and 58 LPC, 49 benign diseases of hepatobiliary and 53 normal individuals were analyzed by SELDI-TOF-MS. Results Among a set of proteins automatically selected as specific biomarkers by Biomarker Wizard software, three protein peaks, with molecular weights of 13. 71 × 103 , 13.83 × 103 and 13. 99 ×103 , were found significantly decreased in cholangiocarcinoma samples. The candidate biomarkers obtained from Tricine-SDS-PAGE gel bands by matching the molecular weight with peaks on CM10 chips were identified by Mass spectrometry as the native transthyretin(native TTR),cysTTR and glutTTR.These preliminary results were further proven by immunoprecipitation using commercial TTR antibodies. This allowed us to re-measure the TTR levels in all the groups more simply by ELISA assay. It showed a firm consistency between ELISA and SELDI analysis. In addition, while TTR levels in cholangiocarcinoma were found to be lower than those in normal healthy controls, TTR levels in benign diseases of the hepatobiliary system were found to be higher than those in healthy controls.Conclusion TTR could be a biomarker that better discriminates cholangiocarcinoma patients from the benign diseases compared to other biomarkers presently available.

Objective: To investigate whether acupoint penetration acupuncture (APA) could regulate chondrocyte autophagy and apoptosis via the PI3K/Akt-mTOR signaling pathway to reduce cartilage degeneration in knee osteoarthritis (KOA) rats. Methods: KOA was induced in rats via intra-articular injection of sodium iodoacetate resolution. Forty male Sprague-Dawley rats were randomly assigned to blank control, model, APA, electro-acupuncture (EA), and sham model groups (n = 8) and those in the APA and EA groups received their respective therapies. Following completion of the treatment course, histological examinations of cartilage and muscle were conducted. Levels of apoptosis- and autophagy-related factors, including Bax, Bcl-2, mTOR, ULK-1, and Beclin-1 protein, and mRNAs were assessed. Additionally, β-endorphin (β-EP) concentrations in the brain and serum were measured. Results: Histological analysis revealed that APA alleviated cartilage and muscle damage compared with the model group. APA inhibited cartilage degeneration by modulating the expression of apoptosis- and autophagy-related proteins and mRNA, thus preventing chondrocyte apoptosis. In the APA group, Bax and mTOR protein levels were significantly lower than those in the model group (both P = .024). Conversely, the Bcl-2 expression level was significantly higher than that in the EA group (P = .035). Additionally, ULK-1 expression was significantly lower than that in the EA group (P = .045). The mRNA level of Bax was significantly higher than that in the blank control group (P < .001). However, Beclin-1 levels were significantly higher than those in both the model and EA groups (both P < .001). ELISA results showed a significant decrease in the concentration of β-EP in the brains of the rats in the APA group compared with those in the model group (P = .032). Conclusions APA reduced osteoarthritis-related pain and alleviated cartilage damage by upregulating chondrocyte autophagy and down-regulating apoptosis via signaling pathways involving PI3K/Akt-mTOR in KOA rats.