A robust, selective and highly sensitive chemiluminescent （CL） platform for protein assay was presented in this paper. This novel CL approach utilized rolling circle amplification （RCA） as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier. Typically, the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence. This aptamer-primer sequence was then employed as the primer of RCA. Based on this design, a number of the biotinylated probes and streptavidin-horseradish peroxidase （SA-HRP） were captured on the plate, and the CL signal was amplified. In summary, our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized, and devoid of light source. Therefore, this new technique .will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases, by taking advantages of high sensitivity and selectivity.

A robust,selective and highly sensitive chemiluminescent (CL) platform for protein assay was presented in this paper.This novel CL approach utilized rolling circle amplification (RCA) as a signal enhancement technique and the 96-well plate as the immobilization and separation carrier.Typically,the antibody immobilized on the surface of 96-well plate was sandwiched with the protein target and the aptamer-primer sequence.This aptamer-primer sequence was then employed as the primer of RCA.Based on this design,a number of the biotinylated probes and streptavidin-horseradish peroxidase (SA-HRP) were captured on the plate,and the CL signal was amplified.In summary,our results demonstrated a robust biosensor with a detection limit of 10 fM that is easy to be established and utilized,and devoid of light source.Therefore,this new technique will broaden the perspective for future development of DNA-based biosensors for the detection of other protein biomarkers related to clinical diseases,by taking advantages of high sensitivity and selectivity.

Cell culture plays an important role in virology. It provides a platform for the detection and isolation of viruses as well as for the biochemistry and molecular biology based studies of viruses. In the present work, a new system that could permits multiple (different) cell lines to be simultaneously cultured in one dish was developed. In the system, each cell line was cultured in an isolated zone in the same dish or well and the system is therefore called an isolated co-culture system. The usefulness of this novel approach for virus isolation was demonstrated using a model system based on adenovirus.

BACKGROUND: Hemorrhagic cystitis remains a common complication of hematopoietlc stem cell transplantation.Granulocyte-macrophage colony stimulating factor (GM-CSF) affects proliferation and differentiation of hematopoietic stem/progenitor cells, adjusts functions of monocytes, granulocytes, lymphocytes and endothelial cells.OBJECTIVE: To investigate the protective effects of GM-CSF bladder irrigation in hemorrhagic cystitis after allogeneic hematopoietic stem call transplantation.DESIGN: Case analysis.PARTICIPANTS: A total of 15 hematopathy patients undergoing allogenic hematopoietic stem cell transplantation at the Zhongshan Hospital of Sun Yat-sen University from January 2004 to August 2006 (routine treatment group). A total of 16 hematopathy patients undergoing allogenic hematopoietic stem cell transplantation from September 2006 to December 2008 (GM-CSF group).METHODS: In the routine treatment group, patients received mesna, hydration, alkalization and forced diuresis in the prevention of hemorrhagic cystitis. In the GM-CSF group, GM-CSF was infused into the bladder in addition to mesna,hydration, alkalization and forced diuresis in the prevention of hemorrhagic cystitis 24 hours before cyclophosphamide treatment. Catheter was extracted 3 days following cyclophosphamide withdraw. Following washing with saline, the bladder was emptied. 10 mL of saline and 5 mL of lidocaine were added into 300 μg of GM-CSF. The mixture was infused into the bladder for 60-120 minutes.MAIN OUTCOME MEASURES: The following parameters were measured: occurrence of hemorrhagic cystitis and its correlation to graft versus host disease, as well as the occurrence of cytomegalovirus infection and urinary system infection.RESULTS: Compared with routine treatment group, the occurrence rate of hemorrhagic cystitis was significantly decreased in the GM-CSF group (x2=4.39, P < 0.05), mean duration of hemorrhagic cystitis and duration of hospitalization were significantly shortened (t=3.97, P < 0.05; t=3.13, P < 0.05), and the occurrence rate of over grade HI hemorrhagic cystitis was significantly reduced (x2=5.04, P < 0.05). Cystitis degree was associated with degree and duration of graft-versus-host disease (r = 0.76).Compared with the routine treatment group, cytomegalovirus infection rate was slightly decreased in the GM-CSF group (x2=0.28, P> 0.05), and occurrence rate of over grade Ⅲ hemorrhagic cystitis was higher in patients with cytomegalovirus infection.Compared with the routine treatment group, the occurrence rate of urinary system infection was slightly reduced in the GM-CSF group (x2=0.28, P > 0.05).CONCLUSION: GM-CSF bladder irrigation is well tolerated and often effective, and should be considered as a preparative regimen of hemorrhagic cystitis after allogeneic hematopoietic stem call transplantation.

BACKGROUND:Donor selection for high-risk acute leukemia is still controversial.OBJECTIVE:To compare the therapeutic efficacy of the unrelated donor peripheral blood and matched sibling allogeneic hematopoietic stem cell transplantation for high-risk acute leukemia.METHODS:Total 65 patients with high-risk acute leukemia treated during January 2008 to January 2016 were included,in which 30 patients chose the unrelated donor peripheral blood stem cell transplantation (UD), and other 35 chose the matched sibling allogeneic hematopoietic stem cell transplantation (MS) according to the wishes of patients and their own situation. After treatment, the chi-square test, Kaplan-Meier survival analysis method, and other methods were used to compare the implanted and hematopoietic reconstitution, the occurrence of graft-versus-host disease, relapse mortality and long-term survival between the two groups.RESULTS AND CONCLUSION:The implantation rate, platelet hematopoietic reconstitution time, the incidence of acute and chronic graft-versus-host disease, and its type exhibited no significant differences between the two groups (P > 0.05).The relapse rate, total death rate, and transplant-related mortality rates were 10.0%, 50.0%, and 40.0% in the UD group and 20.0%, 48.6%, and 25.7% in the MS groups, respectively, and the intergroup difference was insignificant (P > 0.05).The expected 2-year cumulative disease-free free survival and overall survival rates were (49.4±9.2)% and (52.6±9.2)% in the UD group and (53.9±8.5)% and (53.9±8.5)% in the MS group, respectively, and the intergroup difference was also insignificant (P > 0.05). Our experimental findings show that unrelated donor peripheral blood stem cell transplantation can be used as an effective alternative in the absence of sibling donors.

Objective To explore the impact of IL10-592 (rs1800872) single nucleic acid polymorphism (SNP) on the prognosis of HLA matched unrelated hematopoietic stem cell transplantation (HSCT).Methods The polymorphism of IL10-592 in 104 recipient-donor pairs and 100 healthy volunteers was analyzed with sequence based typing (SBT).Results When the genotype of IL1 0-592 in donors and recipients matched,AA/AA genotype had higher incidence of Ⅲ-Ⅳ aGVHD than AC/AC or CC/CC genotype (47.1％,3.7％,0,P=0.002).When the genotype of IL10-592 in donors and recipients mismatched,recipients with AC genotype or donors with AA genotype,there was significant different incidence of Ⅲ-Ⅳ aGVHD among donors or recipients with different genotype (P=0.046,P=0.041).The recipients with AA genotype had higher incidence of Ⅲ-Ⅳ aGVHD than AC or CC genotype (27.8％ vs 10.2％,11.1％;P=0.072),and higher incidence of intestinal aGVHD (22.2％vs 5.1％,11.1％;P=0.040),lower incidence of 2-year overall survival (OS:48.2％ vs 75.1％,85.7％;P=0.002),lower incidence of 2 year disease free survival (DFS:48.5％ vs 66.3％,76.2％;P=0.045).Patients had higher incidence of Ⅲ-Ⅳ aGVHD with donors of AA genotype than with donors of AC or CC genotype (26.5％ vs 8.9％,0;P=0.024),and higher incidence of intestinal aGVHD (20.4％ vs 4.4％,0;P=0.026).In multivariate analysis,the genotype of IL10-592AA in recipients and donors had increased risk of Ⅲ-Ⅳ aGVHD (OR=3.3,P=0.049;OR=3.9,P=0.043).There were no statistical differences on the incidence of cGVHD and relapse.Conclusion In HLA-10/10 matched unrelated HSCT,the presence of IL10-592 AA genotype in recipients and/or donors is an adverse factor for Ⅲ-ⅣaGVHD,worse OS and 2-year DFS.

Objective To investigate and analyze the level of homocysteine(Hcy)in Tibet and to analyze the differences of Hcy level in different altitude regions,genders and ages,and thus to provide the prevalence profile of hyperhomocysteine and the differences in relevant tests between HHcy(hyperhomocysteinemia)and non-HHcy pop-ulations.Methods Totally 1 615(male n＝585)subjects were selected from Ngari,Lhasa,Shigatse and Nyingchi plat-eau areas of Tibet by stratified cluster sampling.Serum Hcy level was analyzed and the difference of Hcy level in pop-ulations located at different altitude plateau areas,gender groups were found.The prevalence of hyperhomocysteine and related test were analyzed.Kruskal Wallis test was used to compare Hcy levels in different altitudes,genders and age groups,and Pearson Chi-square test was used to compare HHcy prevalence.Variance analysis was used for the differences of different test indicators between non-HHcy and HHcy populations.Results The level of Hcy in differ-ent regions and different genders were statistically significant,which was higher in males than that in females,and higher in Lhasa and Shigatse than in Nyingchi and Ngari.There was difference in serum HHcy prevalence among dif-ferent genders,regions and age groups.Males showed a higher level than females,people from Lhasa and Shigatse showed a higher level than those from Nyingchi and Ngari.Conclusions The incidence of hyperhomocysteinemia in Tibet is statistically significant in different areas,different genders and different age groups.So this study provides a scientific basis for the rational use of Hcy as an indicator in clinical practice of prevention and treatment of related diseases in plateau areas.

Objective:To explore the clinical features and survival of newly diagnosed follicular lymphoma (FL) patients with diffuse large B-cell lymphoma (DLBCL) component.Methods:1845 newly diagnosed FL patients aged ≥ 18 years with grades 1-3a in 11 medical centers in China from 2000 to 2020 were included, and patients with DLBCL component were screened. The clinical data and survival data of the patients were retrospectively analyzed, and the prognostic factors were screened by univariate and multivariate analysis.Results:146 patients (7.9% ) with newly diagnosed FL had DLBCL component. The median age was 56 (25-83) years, 79 males (54.1% ) . The pathology of 127 patients showed the proportion of DLBCL component. Patients were divided into two groups according to whether the proportion of DLBCL component was ≥ 50% . The study found that patients with DLBCL component ≥ 50% had higher grade 3 ratio (94.3% vs 91.9% , P=0.010) , Ki-67 index ≥ 70% ratio (58.5% vs 32.9% , P=0.013) and PET-CT SUVmax ≥ 13 ratio (72.4% vs 46.3% , P=0.030) than patients with DLBCL component<50% . All patients received CHOP or CHOP like ± rituximab chemotherapy. The overall response rate (ORR) was 88.2% , and the complete response (CR) rate was 76.4% . In the groups with different proportions of DLBCL component, there was no significant difference in the remission rate after induction treatment and the incidence of disease progression within 2 years after initiation of treatment (POD24) ( P<0.05) . The overall estimated 5-year progression free survival (PFS) rate was 58.9% , and the 5-year overall survival (OS) rate was 90.4% . The 5-year OS rate of POD24 patients was lower than that of non POD24 patients (70.3% vs 98.5% , P<0.001) . Compared with non maintenance treatment of rituximab, maintenance treatment of rituximab could not benefit the 5-year PFS rate (57.7% vs 58.8% , P=0.543) , and the 5-year OS rate had a benefit trend, but the difference was not statistically significant (100% vs 87.8% , P=0.082) . Multivariate analysis showed that failure to reach CR after induction treatment was an independent risk factor for PFS ( P=0.006) , while LDH higher than normal was an independent risk factor for OS ( P=0.031) . Conclusion:FL patients with DLBCL component ≥50% have more invasive clinical and pathological features. CHOP/CHOP like ± rituximab regimen can improve the clinical efficacy of patients. Rituximab maintenance therapy can not benefit the PFS and OS of patients. Failure to reach CR after induction therapy was the independent unfavorable factor for PFS.

Humans ; Hematopoietic Stem Cell Transplantation ; Multiple Myeloma/therapy* ; Transplantation, Autologous ; Retrospective Studies ; Combined Modality Therapy ; Stem Cell Transplantation/adverse effects*