Biological functions and clinic significance of SAF­A (Review).

As one member of the heterogeneous ribonucleoprotein (hnRNP) family, scaffold attachment factor A (SAF-A) or hnRNP U, is an abundant nuclear protein. With RNA and DNA binding activities, SAF-A has multiple functions. The present review focused on the biological structure and different roles of SAF-A and SAF-A-related diseases. It was found that SAF-A maintains the higher-order chromatin organization via RNA and DNA, and regulates transcription at the initiation and elongation stages. In addition to regulating pre-mRNA splicing, mRNA transportation and stabilization, SAF-A participates in double-strand breaks and mitosis repair. Therefore, the aberrant expression and mutation of SAF-A results in tumors and impaired neurodevelopment. Moreover, SAF-A may play a role in the anti-virus system. In conclusion, due to its essential biological functions, SAF-A may be a valuable clinical prediction factor or therapeutic target. Since the role of SAF-A in tumors and viral infections may be controversial, more animal experiments and clinical assays are needed.

EBF1, PAX5, and MYC: regulation on B cell development and association with hematologic neoplasms.

During lymphocyte development, a diverse repertoire of lymphocyte antigen receptors is produced to battle against pathogens, which is the basis of adaptive immunity. The diversity of the lymphocyte antigen receptors arises primarily from recombination-activated gene (RAG) protein-mediated V(D)J rearrangement in early lymphocytes. Furthermore, transcription factors (TFs), such as early B cell factor 1 (EBF1), paired box gene 5 (PAX5), and proto-oncogene myelocytomatosis oncogene (MYC), play critical roles in regulating recombination and maintaining normal B cell development. Therefore, the aberrant expression of these TFs may lead to hematologic neoplasms.

Development and identification of a fully human single-chain variable fragment 29 against TSLP.

Thymic stromal lymphopoietin is a key initiator for inducing Th2-type inflammation and a potential therapeutic target for allergic disease. In the present study, the naive human antibody library was enriched using human thymic stromal lymphopoietin (hTSLP) as an antigen by phage display. Single clones were randomly picked from the enriched antibody library after three rounds of selection, and these were expressed for enzyme-linked immunosorbent assay (ELISA). The positive single-chain fragment variables (scFvs) determined by ELISA were further identified by Western blot, Biacore, and flow cytometry. After three rounds of phage display, 35% of the scFv clones were positive by ELISA and could bind well with hTSLP. Further identification revealed that scFv29 had satisfactory characteristics. The scFv29 was specific to hTSLP, and had no cross-reaction with hIL-33, hIL-4, and hIL-13. The scFv29 could bind to hTSLP in competition with the TSLP receptor and could also bind to mouse TSLP. Cellular experiments revealed that mTSLP could stimulate myeloid dendritic cell (DC) to mature, and scFv29 blocking could reduce the maturation rate of DC. These findings suggest that scFv29 could be used as a neutralizing antibody to block the signaling of TSLP, and this work provides the foundation for further study of the therapeutic roles of TSLP in allergic inflammation diseases.

Changes in correlation characteristics of time consumption and mind-reading performance in pre-onset and post-onset psychosis.

There is a strong correlation between neurocognition and social cognition. However, none of these studies have examined the key role of time consumption during social cognition tasks. Participants included 84 individuals with clinical high risk of psychosis (CHR), 95 healthy controls (HC), and 66 case controls (schizophrenia patients, SZ), who were assessed through the Reading-Mind-in-Eyes Tasks (RMET) with computerized recording of the response time (RT). Neurocognitive tests were also performed for the HC and CHR groups. A comparison of RMET performance revealed significantly lower scores in the SZ group compared to the HC group, with CHR individuals scoring between these two. However, both CHR and SZ subjects spent almost twice as long of the time on RMET compared to the HC subjects. Significant positive correlation was found between RMET accuracy and RT, though only in SZ patients. Taking the RT into consideration, the RMET performances were impacted by different neurocognition domains. Our findings provide new evidence about how time consumption in mind-reading may impact the relationship between social cognition and neurocognition, and we discuss the potential importance of recording the response time during social cognition assessment in individuals with early psychosis.

Effect of granulocyte colony-stimulating factor priming combined with low-dose cytarabine and homoharringtonine in higher risk myelodysplastic syndrome patients.

As sensitization of leukemia cells with granulocyte colony-stimulating factor (G-CSF) can enhance the cytotoxicity of chemotherapy in myeloid malignancies, a pilot study was conducted in order to evaluate the effect of G-CSF priming combined with low-dose chemotherapy in patients with higher risk myelodysplastic syndrome (MDS). The regimen, G-HA, consisted of cytarabine (Ara-C) 7.5mg/m(2)/12h by subcutaneous injection, days 1-14, homoharringtonine (HHT) 1.5mg/m(2)/day by intravenous continuous infusion, days 1-14, and G-CSF 150mg/m(2)/day by subcutaneous injection, days 0-14. 56 patients were enrolled, 34 patients (61%, 95% confidence interval: 51.44-70.56%) achieved complete remission (CR). Median duration of neutropenia was 7days (ranging from 2 to 16days). Grade 1-2 nonhematologic toxicities were documented, including nausea and vomiting (5%), liver function abnormality (5%), and heart function abnormality (2%). No central nervous system toxicity was found. Mortality within the first 4 weeks was 4%. The G-HA regimen is effective in remission induction for higher risk MDS patients and well tolerated due to the acceptable toxicity in maintenance therapy in the patients who cannot undergo Hematopoietic cell transplantation (HCT).

[Selection and identification of completely humanized anti-IL-4 single-chain antibodies].

OBJECTIVE: To express human interleukin 4 (IL-4), select completely humanized anti-IL-4 single-chain antibodies (scFvs) from the completely humanized scFv library and identify their specificity. METHODS: With IPTG-induced pET102/IL-4/BL21, human IL-4 was synthesized, and then purified and identified. Completely humanized scFvs against IL-4 were expressed from the completely humanized antibody phage library, and positive clones were selected with human IL-4 as antigen. The positive clones were tested by PCR, enzyme cleavage and sequencing. The specificity of the completely humanized anti-IL-4 scFvs was determined by dot blot hybridization. RESULTS: The relative molecular mass (Mr) of IL-4 fusion protein was about 27 000. With IL-4 as antigen, we obtained the positive clones expressing anti-IL-4 antibodies. PCR showed amplified target bands of about 1000 bp. Enzyme cleavage analysis revealed that fingerprint maps of the selected clones were diverse except 2 clones. Among 4 clones expressing scFvs that had different fingerprint maps and strong binding to IL-4, sequencing showed the sequence of 3 was correct. Then, dot blot hybridization demonstrated that the scFvs expressed by the 3 clones were specific. CONCLUSION: Completely humanized scFvs against IL-4 has been obtained successfully.

[Small interfering RNA targeting C-erbB-2 gene increases the radiosensitivity of lung adenocarcinoma cells in vitro].

OBJECTIVE: To investigate the effect of small interfering RNA (siRNA) targeting C-erbb-2 oncogene on the radiosensitivity of C-erbb-2-overexpressing lung adenocarcinoma cell line. METHODS: Four pairs of siRNA targeting the coding sequence of C-erbb-2 mRNA were synthesized and their interference effects were evaluated using quantitative real-time fluorescence PCR. The siRNA with the best interference effects was transfected into Calu-3 cells, which were then exposed to 2 or 5 Gy irradiation, with the cells with transfection or irradiation alone as the control groups. The cell apoptosis after the treatment was detected using annexin V-FITC Kit. RESULTS: The apoptosis rate of the Calu cells was 7.767-/+0.551 in the blank group, 14.400-/+1.114 in the interference group, 11.867-/+0.737 in 2 Gy irradition group, 23.000-/+1.664 in 2 Gy irradiation + interference group, 16.100-/+0.624 in 5 Gy irradiation group, and 27.900-/+1.709 in 5 Gy irradiation+interference group. CONCLUSION: The siRNA targeting C-erbb-2 gene can enhance the radiosensitivity of Calu-3 cells to gamma-ray and increase their apoptosis rate following gamma-ray exposure in vitro.

Combination chemotherapy with low-dose cytarabine, homoharringtonine, and granulocyte colony-stimulating factor priming in patients with relapsed or refractory acute myeloid leukemia.

As sensitization of leukemic cells with granulocyte colony-stimulating factor (G-csf) can enhance the cytotoxicity of chemotherapy in acute myeloid leukemia (AML), a pilot study was conducted in order to evaluate the effect of G-csf priming combined with low-dose chemotherapy in patients with relapsed and refractory AML. The regimen, G-HA, consisted of cytarabine 7.5 mg/m2/12 hr by subcutaneous injection, days 1-14, homoharringtonine 1.5 mg/m2/day by intravenous continuous infusion, days 1-14, and G-csf 150 microg/m2/day by subcutaneous injection, days 0-14. Thirty-six AML patients were enrolled, 23 refractory and 13 relapsed. Eighteen patients (50%, 95% confidence interval: 33-67%) achieved complete remission (CR) with a median CR duration of 7.2 months, and two elderly patients continued a regimen of maintenance therapy and remained in remission for 26.3 and 14.1 months, respectively, as of last follow-up. Eight patients (22%) experienced neutropenia (median duration: 6 days; range: 2-22 days). Thirteen of the 36 (36%) developed severe infections. Grade 1-2 nonhematologic toxicities were documented, including nausea and vomiting (20%), liver function abnormality (6%), and heart function abnormality (6%). No central nervous system and kidney toxicity was observed. The G-HA regimen is effective in remission induction for refractory and relapsed AML patients and well tolerated in maintenance therapy in some subgroups of elderly patients. Further studies are necessary to elucidate optimum dose and schedule for this regimen to enhance the treatment efficacy of relapsed or refractory AML patients.

Successful treatment of primary refractory anemia with a combination regimen of all-trans retinoic acid, calcitriol, and androgen.

We investigated the efficacy and safety of a combination regimen in 63 patients with primary refractory anemia (RA). The daily treatment protocol comprised all-trans retinoic acid (ATRA) (30 mg/m(2)), calcitriol (0.1 microg/m(2)), and androgen (stanozolol 3mg/m(2), or danazol 300 mg/m(2)) in three separate doses for eight consecutive weeks. Hematologic improvement was observed in 43 (68.3%) patients. The treatment administered was generally well tolerated, with no severe regimen-related toxicity. The overall survival rates at 3 and 5 years were 68.72% and 53.18%, respectively. These results indicate that this combination regimen is an effective and well-tolerated treatment for patients with RA.

A phase-I clinical trial of active immunotherapy for acute leukemia using inactivated autologous leukemia cells mixed with IL-2, GM-CSF, and IL-6.

UNLABELLED: We evaluated the efficacy and toxicity of vaccination in 29 patients with relapsed or refractory acute leukemia using inactivated autologous leukemia cells combined with interleukin-2 (IL-2), granulocyte-macrophage colony-stimulating factor (GM-CSF), and IL-6. MHC-I, MHC-II, and B7-1 expression status on the surface of leukemia cells and the cytokine profile of IFN-gamma and IL-10 in serum before and after vaccination was detected. RESULTS: Five achieved a complete remission (CR) and six a partial remission (PR) in this vaccination procedure. Adverse effects were erythema, swelling erosion, and even ulcers at vaccination sites and low grade fever during the first three days of vaccination. No other significant side effects were observed. The expression of MHC-I and MHC-II on leukemia cells was 100% and 90% positive, respectively. B7-1 was exclusively expressed on some cases of M4 and M5. The efficacy of the vaccine was statistically associated with the expression status of B7-1 on leukemia cells (P < 0.01). The serum level of IL-10 reduced significantly in the five patients who achieved complete remission (CR) after vaccination as compared with when they were originally diagnosed (P < 0.01). CONCLUSION: We presented here a promising immunotherapy in the treatment of acute leukemia, especially for F.A.B. M5.