Objective:To induce myeloma specific cytotoxic T cells response in vitro mediated by autologous dendritic cells generating from multiple myeloma(MM) patients.Methods:Monocytes isolated from the peripheral blood of MM patients were cultured in serum free medium with 800 U/ml GM CSF and 600 U/ml IFN? for 8 days to generate DCs.These DCs were pulsed by U 266 cells treated with mitomycin C and their lysates,and then incubated with autologus T lymphocytes for 5～7 days to induce antigen specific CTL.MTT assay was performed to examine the U 266 cell specific lysis.Results:DCs derived from MM patients carried a typical dendritic like morphology,they highly expressed CD86,CD54 and class II MHC molecule(HLA DR) on cell surface.Specific killing of U 266 cells mediated by DCs were tested with MTT assay.At an effector/target ratio of 20/1,specific cytotoxic activities against U 266 cells(28.0%?7.6% and 21.2%?5.4%,respectively),were observed,but without antigen stimulation,the killing rate was 11.7%?4.3%,without DC mediation,the killing rate was 15.6%?4.8% and 13.1%?5.5%(P

Objective To analyze the molecular biological characteristics of Streptococcus pyogenes (GAS) causing children tonsillitis in Yangfangdian community Haidian district,Beijing between 2011 and 2013.Methods From Apr.2011 to Apr.2013,there were 94 children with tonsillitis treated in outpatient of Beijing Shijitan Hospital Affiliated to Capital Medical University,and 17 strains GAS were isolated from these pediatrician outpatients.Their encoding mature M protein gene (emm gene),7 pairs of alleles housekeeping gene (gki,gtr,murI,mutS,recP,xpt and yqiL),superantigen speA and speC genes,macrolide-resistance genes ermB,mefA and ermRT were amplified by polymerase chain reaction (PCR).The PCR amplified products of emm gene and 7 pairs of alleles housekeeping gene were sequenced.The emm gene sequences were stored in the Centers for Disease Control and Prevention database.The emm types of GAS were identified.The 7 pairs of alleles housekeeping gene sequences of GAS were sent to the multilocus sequence typing (MLST) website for their iso-allelic sequences to identify their sequence types (ST).The PCR amplified products of superantigen gene speA and speC,macrolide-resistance genes ermB,mefA and ermRT were performed with agar gel electrophoresis.The corresponding qualitative results were obtained.Results Five emm-types were discovered:emm12.0 (76.4％),emm1.0 (5.9％),emm89.0 (5.9％),emm75.0 (5.9％) and emm22.0 (5.9％) ;emm12.0 and its subtype strains had the same allelic profiles which belonged to ST36 type; emm1.0 strain belonged to ST28 ; emm89.0 strain belonged to ST101 ; emm75.0 strain belonged to ST49 ; emm22.0 strain belonged to ST46 ; 94.1％ (16/17) strains carried macrolide-resistance gene ermB.None carried macrolide-resistance genes mefA and ermRT.None carried superantigen speA gene.100.0％ (17/17 cases) carried superantigen speC gene.Conclusions emm12.0 (ST36) strain was the most common type of GAS causing the children tonsillitis in Yangfangdian community Haidian District,Beijing between 2011 and 2013.These strains mainly carry macrolide-resistance ermB and superantigen speC gene.

Objective:To explore CT and MRI features of the endolymphatic sac tumor (ELST).Methods:The CT and MRI morphology confirmed by surgical pathology for 19 patients with ELST were retrospectively analyzed from June 2011 to May 2019 in Eye & ENT Hospital of Fudan University. The features of CT and MRI included location, size, adjacent structures invasion, CT values, bone destruction, features of T 1WI and T 2WI, enhancement distribution characteristics, dynamic enhancement curve morphology, DWI signal characteristics. The ADC values of the lesions and ipsilateral medial pterygoid muscles were compared using a paired t test. Results:Nineteen ELST patients (one with bilateral diseases) were included. Totally 20 ears (right 9 and left 11) of 13 females and 6 males were studied. The masses with slightly high-density and obscure boundary were located around the vestibular aqueduct at the posterior edge of the petrosal bone. Bone destruction involved mastoid process of the middle ear (16 ears), jugular foramen (11 ears), semicircular canal (10 ears), facial nerve canal (7 ears) and internal auditory canal (9 ears). A large amount of residual bone could be found in the interior of nineteen masses. The CT value was (78.6±21.9) HU. The lesion showed central iso-intensity and peripheral hyperintensity on T 1WI and T 2WI in 16 ears, while no obvious hyperintensity on T 1WI in the other 4 ears. The hyperintensity on T 1WI was around the margin of the lesion in 10 ears, situated at lateral side in 5 ears and all over the lesion in 1 ear. Flow voids signals could be seen in 9 ears as well. Liquid-liquid plane was seen on T 2WI in 2 ears. The solid mass portion which showed iso-intensity on both T 1WI and T 2WI presented marked enhancement on contrast-enhanced T 1WI, while other part of the mass no enhancement. DWI of 14 ears illustrates no evidence of restricted diffusion, and the ADC value [(1.25±0.08)×10 -3 mm 2/s] was slightly higher than that of the medial pterygoid muscles ( t=4.437, P=0.001). The style of time-signal intensity curves of the dynamic contrast-enhanced MRI was rapidly ascending followed by descending curves in 2 ears. Conclusion:Imaging findings of ELST have some characteristics, including located around the vestibular aqueduct at the posterior edge of the petrosal bone, bone destruction, peripheral hyperintensity on T 1WI and no restricted diffusion, which is helpful for its diagnosis.

AIM: To obtain the high expression of recombinant human stem cell factor - thrombopoitin (SCF-TPO) fusion gene and predict its structure property. METHODS: Tour primers were designed according to known sequence of TPO and SCF to amplify the functional amino acid domain of TPO and SCF by RT- PCR, respectively from fetus hepatocytes. The expression plasmid pET32a/SCF- TPO was constructed by VOE gene fusion technique and expressed in BL21(DE3)plysS. The fusion protein property, such as second structure, flexibility, and hydrophilicity were predicted by DS Gene and Protscale software. RESULTS: The expression vector, pET32a/SCF - TPO was constructed and the high expression of the SCF/TPO fusion protein was obtained, with the expression amount of up to 40% of the total cellular protein. DS Genel .5 and Protscale predict no new antigenicity in fusion protein, and the second structure and ioelectric point have no changes except four amino acids change in first structure. There are high flexibility and low hydrophilieity in the linker peptide. CONCLUSION: High expression of SCF- TPO fusion protein has been obtained and protein prediction shows that the fusion protein design is reasonable, which lay foundation for further study of biological fundation of SCF - TPO fusion protein.

Objective:Identification of some biochemical and physical properties for a new recombinant B7-2-PE40KDEL exotoxin fusion protein.Methods:12%SDS-PAGE separating and gel imaging analyzing,peptide mass fingerprinting,Western blot and MTT assasying were used respectively for identification of the protein.Results:Molecular weight of the recombinant B7-2-PE40KDEL was 72 628,5% of the difference to its theoretical value 69 561.The result of Western blot indicated that the purified recombinant B7-2-PE40KDEL could specifically bind with mAb anti-human B7-2 and the antibody against PEA,while the negative control did not.The recombinant B7-2-PE40KDEL digested with trypsin and then detected by MOLTI-TOF-MS.It was shown that the detected 15 peptides lied in the extracellular part of B7-2 and the truncated Pseudomonas extoxin PE40KDEL.Searching in the peptident data bank of Expasy website,we did not find any known proteins which was accordant with the above terms.The cytotoxic activity of the recombinant toxin with MTT method showed that the B7-2-PE40KDEL selectively killed Jurkat cell line which expressesed CD28 receptor well and had no killing effect on the Raji cell line unexpressing CD28 receptor.Conclusion:Recombinant B7-2-PE40KDEL exotoxin fusion protein we construct proves to be a new one with targeted killing bioactivity to B7:CD28 system.

Objective To investigate the treatment value of intensified chemotherapy combined with intrathe-cal Methotrexate(ITMTX)in the treatment of children with relapsed medulloblastoma.Methods From February 2011 to February 2014,40 relapsed patients in Beijing Shijitan Hospital,Capital Medical University,aged 10.6 years(2.7 to 17.7 years),were received 4 cycles of carboplatin and Etoposide continuously infusion for 96 h,and sequentially fo-llowed with Ifosfamide,Etoposide and Nedaplatin for 3 cycles,then administrated oral Temozolomide(TMZ)and Etopo-side for 12 cycles.All patients were divided into ITMTX group,which were performed ITMTX during every chemothera-py or every 3 weeks,and no intrathecal Methotrexate(no-ITMTX)group,which were not performed with ITMTX.Re-sponses were evaluated,and the objective response time,time to tumor progress(TTP),time to treatment failure (TTF),and recurrence-free interval(RFI)of the two groups were contrasted.The side-effects of chemotherapy and ITMTX were analyzed,overall survival was calculated using Kaplan-Meier method,and chi-square test or t test was used to compare the difference between two groups.Results Until 30thDecember 2016,the follow-up time was 29.4 months(ranging from 2 to 57 months). The 3-year overall survival(OS)rate was(53.5 ± 8.1)%,and 45.0% pa-tients were objective response,and the mean objective response time was 2 months,and the RFI was 18 months.For IT-MTX patients,the 3-year OS rate was(66.6 ± 11.5)%,the objective response ratio was 70.0%,and mean objective response time was 1.5 months,24 months free-event of TTP and TTF was 70.0% and 90.0%,respectively,and the RFI was 22.5 months.For no-ITMTX patients,the 3-year OS rate was(40.0 ± 11.0)%,objective response ratio was 20.0%,and mean objective response time was 2.5 months(1.5-4.0 months),24 months free-event of TTP and TTF was 35.0% and 50.0%,respectively,and the RFI was 5 months(0-40 months).Compared with those of no-ITMTX patients,the RFI,24 months TTP and TTF of ITMTX patients were much better,there were statistical signi-ficances(all P<0.05).The main side-effects of chemotherapy was gastrointestinal hematologic side-effects,and re-covered quickly with supportive treatment.No obvious Methotrexate induced neurotoxicity was found.Conclusion Based on high-dose chemotherapy,ITMTX can further improve the long-term survival of patients with relapsed medulloblas-toma,and it is more feasible and suitable for relapsed MB children in China.

Falls among the elderly are a serious public health problem. This paper reviews the application status of machine learning in fall risk identification, fall prevention training intervention, fall behavior detection and warning, analyzes the advantages and disadvantages of machine learning in the field of fall risk management for the elderly, and aims to provide ideas for personalized and accurate fall prevention practice for the elderly in the future.

Objective To summarize the radiological features of follicular dendritic cell tumor of spleen (FDCS).Methods The clinical, radiological and pathological data of 8 patients from November 2011 to November 2017 in 5 hospitals with FDCS confirmed by pathology were retrospectively analyzed. All patients underwent CT examinations including plan and enhanced CT. Three patients underwent additional MRI and two patients underwent PET‐CT examinations simultaneously. The imaging features such as location, number, shape, boundary, size, internal structure, density (or signal, 18F‐fluorodeoxyglucose uptake), enhancement model and the relationship with surrounding structures were observed and compared with pathological results. Results Of the 8 patients with FDCS, 7 were located in the spleen and 1 was located in the spleen of the ectopic spleen of the pancreas. Seven patients with splenic FDCS underwent splenectomy and 1 patient with pancreatic ectopic spleen FDCS underwent resection of the pancreas. Multiple lesions were detected in 1 case, while single in the others. Tumor was round or oval. The tumors were well‐circumscribed and presented as expansive growth. On unenhanced CT, the tumors showed a slightly lower density, and hemorrhage and necrosis could be detected in 6 lesions. Calcification was seen in 1 case, significant necrosis, and cystic change was presented in the pancreatic ectopic spleen FDCS. The solid part presented isointensity or slightly hypointensity on T1WI, and hyperointensity on T2WI. Cystic necrosis areas were hypointensitive on T1WI, and hyperointensitive on T2WI. Spoke‐like areas with hypointensity on T1WI and hyperointensity on T2WI were detected in the center of the solid part with the distribution among the substantial degenerative and necrotic regions. PET‐CT showed that the 18F‐fluorodeoxyglucose was uptaked obviously. The enhancement CT showed that at the arterial phase, the tumors were markedly enhanced and continuously enhanced at portal vein phase and balance phase. Multiple liver metastases were detected in 1 case with huge FDCS. One patient was followed up for 6 years, and gastric lymphoma was detected. The others were followed up for 6 to 53 months, there remained no transfer or recurrence.Conclusions The features of FDCS of spleen mainly manifest as solid or cystic mass with clear solitary sphenoma accompanied by scarring, calcification and hemorrhage. The enhancement mode is persistent enhancement. MRI and PET‐CT help to further reflect the tumor pathological basis and biological characteristics.

BACKGROUND: Bone tissue engineering based on pluripotent stem cells (PSCs) is a new approach to deal with bone defects. Protocols have been developed to generate osteoblasts from PSCs. However, the low efficiency of this process is still an important issue that needs to be resolved. Many studies have aimed to improve efficiency, but developing accurate methods to determine efficacy is also critical. Studies using pluripotency to estimate efficacy are rare. Telomerase is highly associated with pluripotency. METHODS: We have described a quantitative method to measure telomerase activity, telomeric repeat elongation assay based on quartz crystal microbalance (QCM). To investigate whether this method could be used to determine the efficiency of in vitro osteogenic differentiation based on pluripotency, we measured the pluripotency pattern of cultures through stemness gene expression, proliferation ability and telomerase activity, measured by QCM. RESULTS: We showed that the pluripotency pattern determined by QCM was similar to the patterns of proliferation ability and gene expression, which showed a slight upregulation at the late stages, within the context of the general downregulation tendency during differentiation. Additionally, a comprehensive gene expression pattern covering nearly every stage of differentiation was identified. CONCLUSION: Therefore, this assay may be powerful tools for determining the efficiency of differentiation systems based on pluripotency. In this study, we not only introduce a new method for determining efficiency based on pluripotency, but also provide more information about the characteristics of osteogenic differentiation which help facilitate future development of more efficient protocols.
Bone and Bones ; Down-Regulation ; Gene Expression ; In Vitro Techniques ; Methods ; Mouse Embryonic Stem Cells ; Osteoblasts ; Pluripotent Stem Cells* ; Quartz Crystal Microbalance Techniques ; Telomerase* ; Up-Regulation

Objective:Summarizing the clinical characteristics of extraneural metastasis in childhood medulloblastoma.Methods:A total of 616 cases with medulloblastoma treated in Beijing Shijitan Hospital from April 2010 to April 2019 were analyzed retrospectively, among which 11 cases developed extraneural metastasis.The age of onset, location and time of extraneural metastasis, pathological and molecular typing, treatment and prognosis were descriptively analyzed.The differences of blood biochemical indexes between medulloblastoma cases with and without extraneural metastasis were statistically analyzed by t test. Results:As of February 2020, the median follow-up period was 16 months (ranging from 3 to 69 months). Eleven cases, including 8 males and 3 females, were diagnosed with extraneural metastasis, with the incidence being about 1.8%.The median age of medulloblastoma was 6 years (2-10 years), and the median age at presentation of extraneural metastasis was 7 years (2-12 years). Extraneural metastasis occurred from 0.5 months to 38.0 months after the operation, and the affected location includes bone (6 cases), bone marrow (3 cases), lung (3 cases), pelvis (2 cases) and abdominal cavity (1 case). In these patients, the range of lactic dehydrogenase (LDH) was (2 298.00±1 570.70) U/L and neuron-specific enolase (NSE) was (201.00±68.34) μg/L, which were significantly higher than those in patients without extraneural metastasis [(249.50±46.28) U/L and (22.80±7.12) μg/L, all P<0.05]. Partial patients were treated with chemotherapy, while the majority of them were treated with palliative treatment in the terminal stage, with the survival period mostly less than 10 months. Conclusions:Although there is a low incidence of extraneural metastasis in medulloblastoma pediatric patients, the prognosis of these patients with extraneural metastasis is poor and most of them would die within one year.The most common sites include bone, followed by bone marrow and lungs, which may be related to the spread of cerebrospinal fluid and the increased levels of LDH and NSE.