ObjectiveTo investigate the regulatory effect of simvastatin on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) at middle/late stages by p38MAPK pathway under condition of osteoinductive environment. MethodsThe bone marrow of bilateral femur and tibia were harvested from 20 4-week-old female Sprague Dawley rats. BMSCs were isolated and cultured with whole bone marrow culture method; the second generation of cells were randomly divided into 5 groups: control group (complete medium, CM), simvastatin group (simvastatin medium, SIM), osteogenic induction group (osteogenic induction medium, OM), simvastatin and osteogenic induction group (simvastatin+osteogenic induction medium, OM+SIM), and blocker group (SB203580+simvastatin+osteogenic induction medium, OM+SIM+SB). MTT assay was used to detect the cell activity in CM group and SIM group at 2, 3, 4, 5, and 6 days, ELISA method to measure the content of alkaline phosphatase (ALP) in OM group and OM+SIM group at 7 and 14 days. The mRNA and protein expressions of osteocalcin (OCN) were detected by real-time quatitative PCR and Western blot after 1, 12, and 24 hours of osteogenic induction at 21 and 28 days. The protein expressions of phospho-p38 (p-p38) and p38 in OM group, OM+SIM group, and OM+SIM+SB group were detected by Western blot at the best induction time of simvastatin. ResultsMTT assay showed that no significant difference was found in absorbance (A) value between CM group and SIM group at each time point (P > 0.05), indicating no effect of 1×10-7 mol/L simvastatin on cell viability. ELISA results showed that ALP content significantly increased in OM+SIM group when compared with OM group at 7 and 14 days; the ALP content was significantly higher at 7 days than 14 days in OM group and OM+SIM group (P < 0.05). OCN mRNA and protein expressions at 12 hours were significantly higher than those at other time points in each group (P < 0.05), and the expressions of OM+SIM group was significantly higher than those of OM group (P < 0.05). The best induction time of simvastatin was 12 hours. At 12 hours after blocking intervention, the p-p38/p38 in OM+SIM+SB group was significantly lower than that in OM group and OM+SIM group (P < 0.05), and the p-p38/p38 in OM+SIM group was significantly higher than that in OM group (P < 0.05). ConclusionSimvastatin can increase the mRNA and protein expression levels of OCN and the protein of p-p38 in osteogenic differentiation of BMSCs at middle/ late stages, and its best induction time is 12 hours.
ObjectiveTo detect the frequency of anaplastic lymphoma kinase (ALK), ROS1 and RET fusion genes in non-small cell lung cancer (NSCLC) patients in Sichuan, and analyze their correlation with clinical features of NSCLC. MethodsReverse transcription-polymerase chain reaction (RT-PCR) was performed to examine gene rearrangement of ALK, ROS1 and RET fusion genes in 310 NSCLC patients who were admitted in Department of Pulmonary Neoplasm of Sichuan Cancer Hospital from March 2009 to March 2012. There were 234 male and 76 female patients with their median age of 60 years (range, 29 to 77 years). There were 164 patients with a smoking history. Histological types included adeno-carcinoma (AC) in 142 patients, squamous cell carcinoma (SCC) in 138 patients, adenosquamous carcinoma in 10 patients, and other types in 20 patients. Patients, gender, age, smoking history, histological types and TNM staging were also collected. Correlations between fusion genes and clinical features were analyzed. ResultsAmong the 310 patient:15 patients with ALK fusion genes were identified (EML4-ALK) with a positive rate of 4.84%, including 14 patients with AC and 1 patient with SCC. ALK fusion genes were more common in patients under 60 years, without a smoking history, and with AC (P < 0.05). ALK fusion genes were not significantly correlated with gender or histodifferentiation. One patient with ROS1 fusion genes (CD74-ROS1) was identified with a positive rate of 0.32%, who was AC patients. Two patients with RET fusion genes (KIF5B-RET) were identified with a positive rate of 0.64%, both of whom were AC patients. ConclusionsGene rearran-gement rates of ALK, ROS1 and RET in NSCLC patients in Sichuan are 4.84%, 0.32% and 0.64% respectively. Patients with negative gene mutation of epithelial growth factor receptor (EGFR), AC, younger age, without a smoking history or with a light smoking history are more common to have ALK gene rearrangement. Gene rearrangement rates of ROS1 and RET are low, and their clinical significance needs more research.
ObjectiveTo summarize the research progress of the effects and mechanisms of Hedgehog signaling pathway in regulating bone formation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). MethodsThe related literature concerning the regulations and mechanism of Hedgehog signaling pathway in osteogenic differentiation of BMSCs and bone formation in vivo, in vitro, and ex vivo studies in recent years was analyzed and summarized. ResultsThe in vitro studies indicate that Hedgehog signaling pathway can promote osteogenic differentiation of BMSCs via activation of key molecules Smoothened (Smo) and Gli1 which are downstream of Hedgehog signaling, and Hedgehog signaling can activate mTORC2-Akt signaling by upregulation of insulin-like growth factor which has similar effects. Hedgehog signaling regulates osteoblast differentiation via activation of Hh-Smo-Ptch1-Gli signaling pathway and inhibition of Hh-Gαi-RhoA stress fibre signaling. Hedgehog signaling can regulate key molecules of osteogenesis Runx2 for promoting osteogenic differentiation and matrix mineralization by synergism of bone morphogenetic protein and Wnt signaling, and promotes bone formation and repair and healing for bone defect and bone graft model in vivo. ConclusionHedgehog signaling can regulate bone formation and osteogenic differentiation of BMSCs via activation of Hedgehog signaling and other signaling pathways. Hedgehog signaling pathway may be a potential target for developing treatment for bone related diseases of osteoporosis and fracture healing disorders.
ObjectiveTo compare the long-term survival of elderly patients with esophageal squamous cell carcinoma (ESCC) treated with surgical versus non-surgical treatment. MethodsA retrospective analysis was conducted on the clinical data of elderly patients aged ≥70 years with ESCC who underwent esophagectomy or radiotherapy/chemotherapy at Sichuan Cancer Hospital from January 2009 to September 2017. Patients were divided into a surgical group (S group) and a non-surgical group (NS group) according to the treatment method. The propensity score matching method was used to match the two groups of patients at a ratio of 1:1, and the survival of the two groups before and after matching was analyzed. ResultsA total of 726 elderly patients with ESCC were included, including 552 males and 174 females, with 651 patients aged 70-79 years and 75 patients aged≥80 years. There were 515 patients in the S group and 211 patients in the NS group. The median follow-up time was 60.8 months, and the median overall survival of the S group was 41.9 months [95%CI (35.2, 48.5)], while that of the NS group was only 24.0 months [95%CI (19.8, 28.3)]. The 1-, 3-, and 5-year overall survival rates of the S group were 84%, 54%, and 40%, respectively, while those of the NS group were 72%, 40%, and 30%, respectively [HR=0.689, 95%CI (0.559, 0.849), P<0.001]. After matching, 138 patients were included in each group, and there was no statistical difference in the overall survival between the two groups [HR=0.871, 95%CI (0.649, 1.167), P=0.352]. ConclusionCompared with conservative treatment, there is no significant difference in the long-term survival of elderly patients aged≥70 years who undergo esophagectomy for ESCC. Neoadjuvant therapy combined with surgery is still an important choice to potentially improve the survival of elderly patients with ESCC.