Allogeneic stem cell transplantation is the only treatment which may cure multiple myeloma (MM).The high transplantation related mortality (TRM) limits the wide clinical application of myeloablative allogeneic stem cell transplantation.Non-myeloablative allogeneic stem cell transplantation can reduce the TRM,but the risk of relapse and progress of the disease may get increased. Autononmyeloablative allogeneic stem cell tandem transplantation can reduce both the TRM and the relapse and progress of the disease. At present,there exist many new methods which may improve the effect of allogeneic stem cell transplantation for MM in clinical.

【Objective】 To study the effects of different activators and doses on the concentration of growth factor in platelet rich plasma(PRP). 【Methods】 15 healthy volunteers, recruited to prepare PRP by COM.TEC blood cell separator, were divided into 5 groups: PRP group with no activator, Calcium gluconate PRP group, Thrombin 100U/ mL-PRP group, Thrombin 50U/ mL-PRP group, and Thrombin 100U/ml-calcium gluconate -PRP group. The white blood cell count and platelet count in PRP and whole blood were detected. The concentrations of PDGF-AA, TGF-βand VEGF in PRP inactive group and 4 different activators 1 hour after activation were determined by ELISA. 【Results】 The mean concentration of platelets in PRP was 1 462.86×10⁹/L, which was 5.77 times that in the whole blood. The mean concentrations (pg/mL) of PDGF-AA(174 348.00±132 872.39 vs 217 909.67±182 517.96 vs 221 020.38±153 321.51 vs 208 550.35±177 100.47), TGF-β(12 573.14±3 173.20 vs 14 678.45±5 880.96 vs 14 694.39±5 083.90 vs 12 675.65±4 981.83) and VEGF (653.45±489.82 vs 671.61±506.68 vs 690.05±416.13 vs 678.93±501.07) in 4 different activator groups were significantly higher than those in inactivated group (P<0.05). There was no significant difference in the concentrations of PDGF-AA, TGF-βand VEGF among different activators. Platelet concentration(1 462.86±628.41×10⁹/L) in PRP had a strong positive correlation with PDGF concentration (221 020.38±153 321.51 pg/mL)in thrombin 50 U/ mL-PRP group (P<0.05) and TGF-βconcentration(pg/mL) (12 573.14±3 173.20 vs 14 678.45±5 880.96 vs 14 694.39±5 083.90 vs 12 675.65±4 981.83, respectively) in four different activator groups (P<0.05), but had no correlation with VEGF in each group. 【Conclusion】 PRP was prepared by blood cell separator with high purity. There was no difference in the concentrations of PDGF-AA, TGF-βand VEGF among different activators and different thrombin doses. The correlation between platelet concentration and growth factor concentration in PRP was related to the type of activator and growth factor.

Objective: To explore radiosensitivity-associated genes in esophageal squamous cell carcinoma by targeted sequencing panel. Methods: The peripheral blood from 22 esophageal squamous cell carcinoma (ESCC) patients received radiotherapy alone were collected, respectively. The genomic DNA (gDNA) of peripheral blood was extracted and used to create a library of gDNA restriction fragments. The gDNA restriction fragments were hybridized to the HaloPlex probe capture library, which comprises 356 cancer genes selected from the Catalogue of Somatic Mutations in Cancer (Cosmic) database of 2011 updated edition. The sequencing data were aligned by the Genome Analysis Toolkit GATK (version 3.0) and Picar. The single nucleotide polymorphism and inserted-deletion (SNP/InDel) variations were annotated by online database. The pathway enrichment was analyzed by Ingenuity Pathway analysis (IPA). Moreover, according to the short-period curative effect, 22 patients were divided into two groups: the radiation- sensitivity group (CR+ PR) and the radiation-resistant group (PD+ SD). The nonsynonymous mutation sites were statistically analyzed and the genes associated with radiosensitivity of ESCC were screened. Results: More than 97% sequencing reads were aligned to human genome reference sequence and more than 90% sequencing reads were the target sequences. SNP/InDel database annotation results showed that the mutations of 22 cases mainly distributed in exons, and the mutant types were mainly missense and synonymous single nucleotide variant (SNV). There were 23 genes of high-frequency mutation associated with esophageal cancer. Pathway enrichment by IPA showed that 3 pathways were associated with the development of esophageal cancer, which were roles of BRCA1 in DNA damage response pathway, DNA double-strand break repair by non-homologous end joining pathway and ATM signaling pathway. According to the curative effect, five genes including mismatch repair system component (PMS1), fibronectin 1(FN1), mutL homolog 1 (MLH1), B-Raf proto-oncogene, serine/threonine kinase (BRAF), patched 1 (PTCH1) and cytochrome P450 family 2 subfamily C member 19 (CYP2C19) were associated with radiosensitivity of ESCC patients.Moreover, the PTCH1 was mutated in all of 22 ESCC patients, while the variations of rs199476092 and rs202111971 sites of PTCH1 were only identified in the radiation-resistant group. Conclusions We find that the variations of rs199476092 and rs202111971 in the encoding region of PTCH1 gene are significantly associated with radiosensitivity of ESCC patients.