Objective In this study,we tested different disinfectants and different action times to establish an efficient protocol for zebrafish embryo disinfection,which may in turn improve the management of zebrafish facilities.Methods We tested three reagents that can be used for embryo disinfection and are readily available in the domestic market:reagent-grade povidone-iodine,sodium hypochlorite,and chlorine dioxide.We evaluated the toxic effects of the reagents at three concentrations and two action times on zebrafish embryos at three stages,in term of survival,dechorioning,and malformation rates.The effects of the three disinfectants were also compared in terms of the amounts of embryos with surface bacteria after disinfection.Results Chlorine-containing disinfectants,i.e.sodium hypochlorite and chlorine dioxide,were better able to kill bacteria on the embryo surface,while povidone-iodine was not quite effective.Survival,dechorioning,and malformation rates were similar in embryos treated with 30 ppm sodium hypochlorite or chlorine dioxide to those in control embryos.Treatment with chlorine-containing disinfectants for 10 min were more effective in sterilizing than for 5 min.Conclusions The result of this study suggest treating 6～30 hpf embryos with 30 ppm sodium hypochlorite for 10 min as an operational method.

Depression and circadian rhythms exhibit bidirectional interactions,suggesting a close association with the biological clock system.The biological clock in the suprachiasmatic nucleus of the hypothalamus is regulated by circadian genes.Recent clinical and basic research has revealed multifaceted associations between depression and circadian genes.For instance,significant phase abnormalities in BMAL1,PER2,and PER3 were detected in brain tissue from depressed patients,while plasma levels of CRY1,ARNTL,and PER1 proteins showed marked reduction,demonstrating good diagnostic value.Mice with CLOCK and BMAL1 knockouts exhibited depression-like behaviors.Single nucleotide polymorphisms(SNPs)in genes such as PER1 and PER3 directly influence depression susceptibility.Methylation levels of BMAL1,PER3,and CLOCK genes correlate closely with depressive symptoms.Antidepressant mechanisms like ketamine exert their effects by downregulating PER2 and other genes.This review summarizes the differential expression patterns of circadian clock genes in depression and associated therapeutic approaches,aiming to provide new theoretical foundations for precision diagnosis and personalized treatment strategies for depression.

Objective In this study,we tested different disinfectants and different action times to establish an efficient protocol for zebrafish embryo disinfection,which may in turn improve the management of zebrafish facilities.Methods We tested three reagents that can be used for embryo disinfection and are readily available in the domestic market:reagent-grade povidone-iodine,sodium hypochlorite,and chlorine dioxide.We evaluated the toxic effects of the reagents at three concentrations and two action times on zebrafish embryos at three stages,in term of survival,dechorioning,and malformation rates.The effects of the three disinfectants were also compared in terms of the amounts of embryos with surface bacteria after disinfection.Results Chlorine-containing disinfectants,i.e.sodium hypochlorite and chlorine dioxide,were better able to kill bacteria on the embryo surface,while povidone-iodine was not quite effective.Survival,dechorioning,and malformation rates were similar in embryos treated with 30 ppm sodium hypochlorite or chlorine dioxide to those in control embryos.Treatment with chlorine-containing disinfectants for 10 min were more effective in sterilizing than for 5 min.Conclusions The result of this study suggest treating 6～30 hpf embryos with 30 ppm sodium hypochlorite for 10 min as an operational method.

Depression and circadian rhythms exhibit bidirectional interactions,suggesting a close association with the biological clock system.The biological clock in the suprachiasmatic nucleus of the hypothalamus is regulated by circadian genes.Recent clinical and basic research has revealed multifaceted associations between depression and circadian genes.For instance,significant phase abnormalities in BMAL1,PER2,and PER3 were detected in brain tissue from depressed patients,while plasma levels of CRY1,ARNTL,and PER1 proteins showed marked reduction,demonstrating good diagnostic value.Mice with CLOCK and BMAL1 knockouts exhibited depression-like behaviors.Single nucleotide polymorphisms(SNPs)in genes such as PER1 and PER3 directly influence depression susceptibility.Methylation levels of BMAL1,PER3,and CLOCK genes correlate closely with depressive symptoms.Antidepressant mechanisms like ketamine exert their effects by downregulating PER2 and other genes.This review summarizes the differential expression patterns of circadian clock genes in depression and associated therapeutic approaches,aiming to provide new theoretical foundations for precision diagnosis and personalized treatment strategies for depression.

Objective To explore the clinical outcome of HLA haploidentical vs HLA-matcbed peripheral blood hematopoietic stem cell transplantation (PBSCT) without in vitro T-cell depletion for malignant hematological diseases. Methods 111 patients with malignant hematological diseases underwent PBSCT without in vitro T-cell depletion between May 2004 and February 2009, including 51 patients with HLA-haploidentical and 60 patients with HLA-matched. All patients have received myeloablative conditioning regimen. A two-agent based graft-versus-host disease (GVHD) prophylaxis was used as cyclosporine A and a short course of methotrexate. Mycophenolate mofetile was added for the patients with one locus mismatch. Mycophenolate mofetile, antithymocyte globulin and CD25 monoclonal antibody were added for the patients with 2-3 loci mismatch. The grafts were granulocyte colony-stimulating factor-mobilized peripheral blood stem cells without in vitro T-cell depletion. Results 111 patients achieved sustained and full donor-type engraftment. The median time to reach an absolute neutrophil count above 0.5×10~9/L was 14 days and that to a platelet count exceeding 20×10~9/L was 15 days in 51 HLA-haploidentical patients, and that was 12 days and 13 days in 60 HLA-matched patients, respectively. In 51 HLA-haploidentical patients, 25 patients developed aGVHD, including 20 cases of grade Ⅰ aGVHD, and 5 cases of grade Ⅱ. Thirty-three patients developed cGVHD with limited in 30 and extensive in 3. The 4-year cumulative incidence of cGVHD was 70.4 %. The 3-year probabilities of leukemia-free survival (LFS) were 74.5% (77.3 % for standard risk patients and 68.2 % for high risk patients respectively). Seven patients had recurrence. In 60 HLA-matched patients, 14 patients developed aGVHD, including 10 cases of grade Ⅰ, 2 cases of grade Ⅱ and 2 cases of grade Ⅲ. Thirty-seven patients developed cGVHD with limited in 32 and extensive in 5. The 4-year cumulative incidence of cGVHD was 58.1%. The 3-year probabilities of LFS were 72.1% (77.6 % for standard risk patients and 52.7 % for high risk patients respectively). Ten patients had recurrence. The incidence of aGVHD in HLA-haploidentical cohort was significantly higher than in HLA-matched cohort (P<0.05). There was no significant difference in incidence of cGVHD, incidence of relapse and LFS between HLA-haploidentical and HLA-matched cohorts (P>0.05). Conclusion Haploidentical PBSCT is feasible and safe for malignant hematological diseases to use myeloablative conditioning regimen in combination with intensive immunosuppressants without in vitro T cell depletion.

BACKGROUND: Lack of human leucocyte antigen-matched family donors has restricted the application of hematopoietic cell transplantation. Due to immunological disorder of humam leucocyte antigen misfit, common way for haploidentical transplantation is associated with poor engraftment and severe graft-versus-host disease. Because not every patient has HLA-Jdentical family member, a substantial proportion of patients will receive haploidentical transplantation. OBJECTIVE: To explore the curative effect on malignant hematological diseases of haploidentical peripheral blood stem cell transplantation (PBSCT) using myeloablative conditioning regimen in combination of proper immunosuppressants without in vitro T-cell depletion. DESIGN, TIME AND SETTING: A case observation was performed at the Department of Hematology in the First Affiliated Hospital of Xinjiang Medical University from July 2002 to June 2008. PARTICIPANTS: Forty-two patients with malignant hematological diseases, including 29 standard-risk patients and 13 high-risk patients, age from 10 to 48 years, were transplanted with cells from a haploidentical family donor with 1-3 mismatched loci of HLA antigens. Seven patients had 1 locus mismatched donors and thirty-five patients had 2-3 loci mismatched donors. METHODS: The patients have received myeloablative conditioning regimen. A two-agent based graft-versus-host disease (GVHD) prophylaxis was used as cyclospodne A and a short course of methotrexate. Mycophenolate mofetile was added for 1 locus mismatched patients. Mycophenolate mofetile, antithymocyte globulin and CD25 mono-colonal antibody were added for 2-3 loci mismatched patients. The grafts were granulocyte colony-stimulating factor-mobilized peripheral blood stem cells without in vitro T-cell depletion. MAIN OUTCOME MEASURES: Engraftment, GVHD incidence and severity, relapse and leukemia-free survival and the immune function of patients in months 1, 3, 6, 12 and 18 postoperatively. RESULTS: Totally 42 patients achieved complete and sustained donor-type engraftment. Nineteen patients developed acute GVHD, the 2-year cumulative incidences of acute GVHD were 50.8%, gradeⅠ acute GVHD occurred in 16 cases and grade Ⅱ in 3 cases. Thirty-one patients were followed up more than 6 months, 23 of them developed chronic GVHD with limited in 20 and extensive in 3, the 2-year cumulative incidences of chronic GVHD were 57.1%. No patients died of GVHD. There were no significant differences in the reduction and recovery of T cells and B cells between HLA haploidentical PBSCT without in vitro T cell depletion and HLA-matched PBSCT. CONCLUSION: Haploidentical PBSCT is feasible and safe for malignant hematological diseases to use myeloablative conditioning regiment combination of intensive immunosuppressants without in vitro T cell depletion. A large amount of clinical cases need to be investigated in the near future.

Objective To investigate the correlation between immature granulocyte and CD34+ cells, mononuclear cells (MNC) in donor's peripheral blood by recombinant human granulocyte colony-stimulating factor (rhG-CSF). Methods The stem cell were mobilized by rhG-CSF 7.25-10 μg·kg-1·d-1 from 122 allo-PBSCT donors. Before and after mobilization, to test CD34+ cells of peripheral blood stem cell graft and number of MNC, immature granulocyte, CD34+ cell per patient' weight were calculated. Results White blood cell count and immature granulocyte gradually increased, and reached the peak on the frith day. There was a good relationship between increased immature granulocytes and increased CD34+ cells. The patients all achieved completed donor engraftment and achieved hematopoietic recovery. The chromosome, blood type and HLA type were transformed to be donor's type. Ph1 changed to be negative in CML patients. Conclusion rhG-CSF (7.25～10 μg·kg-1·d-1) had a good effect to mobilize PBSC. There was a good relationship between in-creased immature granulocytes and increased CD34+ cells after mobilization by rhG-CSF. The number of immature granulocytes can reflect indirectly the count of stem/progenitor, so the MNC and immature granulo-cytes can become a threshold of dosage standard.