Objective:To explore the effect of siRNA targeting myeloid cell leukemia-1(Mcl-1)on the biological behavior of salivary adenoid cystic carcinoma cells.Methods:The chemically synthesized Mcl-1-siRNA was transfected into salivary adenoid cystic carci-noma SACC-2 cells.The expression levels of Mcl-1-mRNA and Mcl-1protein were examined by Real-time PCR and western blotting respectively.MTT assay,transwell chamber and flow cytometry were used to determine the effect of Mcl-1-siRNA on SACC-2 cell pro-liferation,migration and apoptosis.Results:Compared with the control group,liposome group and NC-siRNA group,SACC-2 cell proliferation rate of Mcl-1-siRNA group was obviously slowed down.48 h after transfection,the migration of SACC-2 cells in Mcl-1-siRNA group(39 ±9.0)were lower than that in control group(69 ±6.0).The apoptosis rate of Mcl-1-siRNA group(8.6%)was sig-nificantly higher than that in control group(1.9%).Conclusion:Silence Mcl-1 can inhibit cell proliferation and migration and pro-mote apoptosis of salivary adenoid cystic carcinoma cells.

Objective:To investigate the effect of c-fos on the proliferation and migration of oral squamous cell carcinoma and potential mechansism.Methods:The expression of c-fos,CyclinD1 and p16 in 60 oral squamous cell carcinoma samples and 60 oral mucosa tissue samples was examined by immunohistochemistry.HN6 and SCC9 cells were respectively transfected with siRNA-c-fos and siRNA-scramble,then were respectively divided into control group,siRNA-scramble group and siRNA-c-fos group.The mRNA and protein expressions of CyclinD1 and p16 were decteted,meanwhile cell proliferation and migration were tested.Results:Compared with the oral mucosa tissue samples,the expressions of CyclinD1 and c-fos were increased in the carcinoma samples,while the expression of p16 was reduced.Compared with control group,the expressions of CyclinD1 in siRNA-c-fos group were significantly reduced,while p16 enpression was increased,with the inhibition of cell proliferation and migration.Conclusion:c-fos may regulate pl6/CyclinD1 signaling pathways and promote the proliferation and migration of oral squamous cell carcinoma.

Objective:To explore the effects of cysteine-rich 6 1 (Cyr6 1 )on biological behavior of human adenoid cystic carcinoma ACC-LM and ACC2 cells.Methods:The chemically synthesized Cyr6 1-siRNA was transfected into ACC-LM and ACC2 cells.Cell proliferation was measured by the MTT method,the invasive ability was evaluated by Transwell chamber assay,and cell apoptosis was analyzed using flow cytometry by double staining with Annexin V and propidium iodide.Results:Cyr61-siRNA significantly down-regu-lated Cyr61 protein expression in ACC-LMand ACC2 cells.Cyr61-siRNA markedly inhibited the proliferation and invasion of the cells, however,there was no significant difference in cell apoptosis between Cyr6 1-siRNA and control groups.Conclusion:Cyr6 1 promote the proliferation and invasion of adenoid cystic cancer cells.

Objective:To investigate the effects of IGF-1 and IL-1β on the proliferation and apoptosis of cultured human condylar chondrocytes( CCs) of temporomandibular joint( TMJ) . Methods:Cultured CCs were derived from human TMJ condylar cartilage tis-sue, and identified by immunocytochemistry staining. The cultured cells were divided into 6 groups:control group, IL-1β(10 μg/L) group and IL-1 group (10 μg/L) + IGF-1 group (0, 1, 10, 50 and 100 μg/L, respectively). The cell proliferation ability was de-tected by MTT assay. The cell cycle and apoptosis were detected by flow cytometry. The expression of apoptosis-associated factors Bcl-2, Bax and p38 MAPK/NF-κB proteins were detected by Western blot. Results:Type II collagen was positively expressed in cultured CCs. IL-1β treatment decreased cell proliferation, increased cell apoptosis with concomitant increase of the percentage of early apopto-sis and late apoptotic cells, increased Caspase-3 expression, decreased Bcl-2/Bax ratio, and increased the expression of p38 MAPK/NF-κB proteins. Whereas, with 1-100 μg/L IGF-1 pretreatment, the proliferation ability and Bcl-2/Bax ratio of the cells were in-creased(P<0. 05), the apoptotic cells were decreased, the expression of Caspase-3 and p38 MAPK/NF-κB proteins was decreased ( P<0. 05) in a dose-dependent manner. Conclusion:IGF-1 may inhibit IL-1β-induced cell apoptosis and attenuate the activation of p38 MAPK/NF-κB of human condylar chondrocytes.

Objective:To evaluate the psychological pain of patients with head and neck cancer aged ≥60 years old before and after intensity-modulated radiotherapy (IMRT).Methods:Distress Thermometer (DT)(Chinese version) was used to investigate the degree and problems of psychological pain before and after IMRT for 85 elderly patients with head and neck cancer. The results before and after IMRT were compared by paired t-test. Relevant factors were identified by Logistic regression analysis. Results:The median age in the cohort was 66 years old (60-85 years old). The incidence rates of psychological pain were 73% and 87% before and after IMRT ( P<0.001). The corresponding incidence rates of severe distress were 6% and 34%( P<0.001). The main distress problems before IMRT were memory loss/attention deficit, worry, oral pain, economic problems, stress, sleep problems, and dry nose. The significantly-increased distress problems after IMRT were oral pain, constipation, eating, nausea, and dry nose. Logistic regression analysis showed gender ( OR=5.520, 95% CI 1.437-21.212, P=0.013), pre-treatment PG-SGA score ( OR=1.220, 95% CI 1.048-1.421, P=0.010) and medical insurance ( OR=0.230, 95% CI 0.053-0.995, P=0.049) were the relevant factors of the severe psychological distress before IMRT. Occupation ( OR=2.286, 95% CI 1.291-4.050, P=0.005) and medical insurance ( OR=0.089, 95% CI 0.029-0.276, P<0.001) were the relevant factors of severe psychological distress after IMRT. Conclusion:The incidence rate of distress is high in elderly patients with head and neck cancer before IMRT, which can be aggravated after IMRT, primarily the treatment-related physical pain problems.

Objective To analyze gene mutations and the etiology of six fetuses with osteogenesis imperfecta detected by prenatal ultrasonography.Methods From March 2016 to May 2017,six gravidas of singleton pregnancy and their fetuses that were diagnosed with osteogenesis imperfecta by prenatal ultrasonography were enrolled in this study.Gravida 1 came to the Center of Prenatal Diagnosis of the First Affiliated Hospital of Zhengzhou University for prenatal diagnosis,while the other five were referred to that center after termination to identify genetic defects with their fetal tissues.Next-generation sequencing technology was carried out for exome sequencing in the genomes of six fetuses.Suspected mutations were confirmed by polymerase chain reaction and Sanger sequencing.Two hundred unrelated healthy individuals were analyzed with Sanger sequencing for validation of novel mutations.Results Fetus 1 carried a heterozygous mutation in collagen,type Ⅰ,alpha-1 (COL1A1) gene,c.724G＞C(p.Gly242Arg),which was found in the mother and brother but not in the father.Fetus 2 carried a known heterozygous mutation in COL1A 1 gene,c.2461G＞A(p.Gly821Ser),which was found in the mother but not in the father.Four heterozygous mutations,c.2282G＞A(p.Gly761Asp),c.1002+5G＞A in COL1A1 gene,c.1774G＞A(p.Gly592Ser) and c.3277G＞T(p.Gly1093Cys) in collagen,type Ⅰ,alpha-2 (COL1A2) gene,were respectively carried by fetuses 3 to 6,but not by their parents.Mutations of c.724G＞C(p.Gly242Arg),c.2282G＞A (p.Gly761Asp) and c.1002+5G＞A in COL1A1 gene and c.3277G＞T (p.Gly1093Cys) in COL1A2 gene were four novel mutations,which were not found in the 200 unrelated healthy individuals.The mother of fetus 1 who was highly suspected with osteogenesis imperfecta selected to continue the pregnancy because the family members had mild symptoms.After delivery,cord blood was collected for genetic test and the result was consistent with that of prenatal genetic diagnosis.Fetus 1 had no fractures during a six-month follow-up after birth.Conclusions Mutations in the COL1A1 and COL1A2 genes may be the etiology of osteogenesis imperfecta in these six fetuses.Results of this study could enrich the data on COL1A1 and COL1A2 mutations relating to osteogenesis imperfecta,and provide a basis for genetic counseling.

Objective:To evaluate the efficacy and safety of hypofractionated radiotherapy for lung metastases (LMs).Methods:From March 2007 to April 2019, 193 patients with 317 LMs including 124 male and 69 female admitted to our hospital were enrolled. The median age was 58 years old and the median KPS was 80. The primary tumors were mainly distributed in the lung (33.7%), colorectum (21.2%), head and neck (13.5%) and breast (10.9%), respectively. The clinical efficacy and side effects of hypofractionated radiotherapy for LMs were evaluated.Results:The median follow-up time was 59.9 months (95% CI: 55.1-64.6 months). Among 193 patients with 317 LMs, 90.7% of them were treated with 4D-CT, 69.4% for intensity-modulated radiation therapy (IMRT), 28.0% for volumetric-modulated arc therapy (VMAT) and 2.6% for tomotherapy (TOMO), respectively. The median gross tumor volume (GTV) and planning target volume (PTV) were 5.0 cm 3(0.2-142.3 cm 3) and 12.0 cm 3(1.0-200.1 cm 3). The prescription dose regimen was 60 Gy in 4 to 15 fractions. The median dose for PTV was 60 Gy (45-70 Gy) and biological effective dose was 96 Gy (60-150 Gy), respectively. The 1-, 3-and 5-year local control rates (LCR) were 95.7%, 91.3% and 89.9%, respectively. The median time from primary cancer diagnosis to lung metastases was a prognostic factor for LCR ( P=0.027). The overall survival (OS) and progression-free survival (PFS) rates were 90.1%, 60.8%, 46.2%, and 54.3%, 30.3%, 19.9%, respectively. The median time from primary cancer diagnosis to lung metastases and extrapulmonary metastases was the prognostic factor for OS and PFS. No Grade 3 toxicities were seen. Conclusion:Image-guided hypofractionated precision radiotherapy is an efficacious and safe treatment for LMs.