Background Arabidopsis protein disulfide isomerase 1 ( At PDI1) has been demonstrated to have disulfide isomerase activity and to be involved in the stress response. However, whether the anti-stress function is directly related to the activities of thiol-disulfide exchange remains to be elucidated. Results In the present study, encoding sequences of At PDI1 of wild-type (WT) and double-cysteine-mutants were transformed into an At PDI1 knockdown Arabidopsis line ( pdi ), and homozygous transgenic plants named pdi - At PDI1, pdi - At PDI1 m1 and pdi - At PDI1 m2 were obtained. Compared with the WT and pdi - At PDI1, the respective germination ratios of pdi - At PDI1 m1 and pdi - At PDI1 m2 were significantly lower under abiotic stresses and exogenous ABA treatment, whereas the highest germination rate was obtained with At PDI1 overexpression in the WT (WT- At PDI1). The root length among different lines was consistent with the germination rate; a higher germination rate was observed with a longer root length. When seedlings were treated with salt, drought, cold and high temperature stresses, pdi - At PDI1 m1 , pdi - At PDI1 m2 and pdi displayed lower survival rates than WT and At PDI1 overexpression plants. The transcriptional levels of ABA-responsive genes and genes encoding ROS-quenching enzymes were lower in pdi - At PDI1 m1 and pdi - At PDI1 m2 than in pdi - At PDI1. Conclusion Taken together, these results clearly suggest that the anti-stress function of At PDI1 is directly related to the activity of disulfide isomerase.

Osteosarcoma represents one of the most aggressive tumors of bone among adolescents and young adults. Despite improvements in treatment, osteosarcoma has a grave prognosis. The identification of prognostic factors is still in its infancy. Weighted gene correlation network analysis (WGCNA) was conducted on mRNA-sequencing and clinical information (gender, survival and metastasis) of osteosarcoma patients from the TARGET database to obtain genes in modules associated with metastasis of osteosarcoma. The Cox regression analysis was then performed on the gene expression profile from TARGET to screen genes associated with patients’ survival. Known genes related to osteosarcoma were obtained by intersecting osteosarcoma-related genes from DisGeNET and DiGSeE, followed by the construction of PPI network of osteosarcoma-related genes and survival-related genes in modules. The screened key genes were subject to multi-factor Cox proportional hazards model, and osteosarcoma patients were classified into high- and low- risk groups according to the risk score to evaluate the potential of key genes to predict the survival of osteosarcoma patients. The WGCNA showed that 4 genes in tan and 19 genes in pink modules were related to the survival of osteosarcoma patients. Osteosarcoma-related known genes (9) were obtained in intersection of DisGeNET and DiGSeE. PPI network identified 4 key genes (KRT5, HIPK2, MAP3K5 and CD5) closely associated with survival of osteosarcoma patients. HIPK2, MAP3K5 and CD5 expression was inversely correlated with survival risk, while KRT5 expression was positively correlated with survival risk. These results show KRT5, HIPK2, MAP3K5 and CD5 serve as prognostic factors of osteosarcoma patients.

Objective To study the immunogenicity of an envelope protein derived from a chronic infection with human immunodeficiency virus type I (HIV-1). Methods Balb/c mice and New Zealand white rabbits were immunized with 06044 gp140T.The specific binding antibody titers of the immunized rabbit serum were determined by enzyme-linked immunosorbent assay (ELISA), and the specific neutralizing antibody titer of rabbit serum was determined by TZM-bl pseudovirus neutralization assay.The cytokine secretion by the spleen cells of immunized mince was determined. Results Rabbit immune sera showed broadly binding breadth and endpoint titers to SF162 gp140, JRCSF gp120 and Bal gp120 were 83176.38±1.35、186208.71±2.57 respectively.After the third immunization, rabbits had high titer of specific neutralizing antibodies.The neutralizing antibody titer against SF162 pseudovirusat at two week after the 3rd, 4th, 5th, 6th immunization was 177, 183, 265, 252 respectively.Interleukin(IL)-2, IL-4, IL-6, IL-13 and interferon(IFN)-γ significantly highly expressed in immunized mice than that in the control group at day 14 after the last immunization(P values were 0.0015, 0.0009, 0.0117, 0.0001, 0.0051, respectively). Conclusion 06044 gp140T trimeric envelope protein has good immunogenicity and can induce high specific antibody levels after early immunizations. Key words: Human immunodeficiency virus type 1; gp140T; Trimer; Immunogenicity

The expression of MdBBX10 was significantly induced by different stresses and ABA treatments. Overexpression of MdBBX10 in Arabidopsis significantly enhanced abiotic stresses tolerance by ABA signalling. The roles of B-box domain(s) containing proteins (BBXs) in regulation of flowering and light morphogenesis of plants were intensively studied. However, the roles of plant BBXs in abiotic stresses are poorly understood. A B-box protein encoding gene from apple (MdBBX10) was found to be up-regulated from gene expression profile under salt stress. qRT-PCR analysis indicated that the expression of MdBBX10 was significantly induced by different stresses and exogenous abscisic acid (ABA) in apple roots and leaves. The β-glucuronidase activity driven by the promoter of MdBBX10 was also strongly induced by NaCl, H2O2, polyethylene glycol and exogenous ABA, which was consistent to the existence of rich cis-acting elements related to the abiotic stresses in the promoter sequence. Over-expression of MdBBX10 in Arabidopsis significantly enhanced tolerance to abiotic stresses, with higher germination ratio and longer length of roots than the wild type plants. Transgenic plants of over-expressing MdBBX10 lines were more sensitive to exogenous ABA than the wild type plants. Under abiotic stress treatments, the transcript levels of ABA- and stress-related genes were higher in MdBBX10-overexpressing plants than wild type plants. Over-expression of MdBBX10 could enhance plant's ability to scavenge reactive oxygen species (ROS) under stresses, which is correlated with the expression of ROS-scavenging genes. These results provided the evidences that MdBBX10 plays an important role in enhanced plant tolerance to abiotic stresses, which were involved in ABA-mediated response and ROS response.

目的:探讨巨细胞病毒(CMV)再活化对于完全缓解期行异基因造血干细胞移植术(allo-HSCT)的急性髓系白血病(AML)患者复发和生存的影响.方法:对106例于完全缓解期行allo-HSCT的成人AML患者的资料进行回顾性分析,包括移植术后CMV再活化的发生情况和影响因素,以及CMV再活化对于复发和生存的影响.结果:67.0％(71/106)的患者在移植后12个月内出现了CMV再活化,中位时间为移植后46(1～117)d.HLA不全相合和预处理应用兔抗人胸腺细胞免疫球蛋白(ATG)是移植后CMV再活化的影响因素.106例患者移植后的中位随访时间为36(1～171)个月,19例患者移植后出现复发,中位复发时间为4.5(2～38)个月,3年累积复发率为17.8％.单因素分析显示,Ⅱ～Ⅳ度急性移植物抗宿主病和慢性移植物抗宿主病的发生可降低患者的复发率,而CMV再活化对于移植后的复发率无明显影响.106例患者的3年总生存率(OS)为77.2％,无病生存率(DFS)为76.0％.单因素分析显示,年龄＞40岁和移植后CMV再活化影响患者的OS,但不影响DFS.Cox多因素回归分析显示,移植后CMV再活化是影响患者3年OS的独立危险因素.结论:HLA不全相合和预处理过程中应用ATG是导致移植后CMV再活化的危险因素,而CMV再活化对于完全缓解期AML移植后的复发并没有影响,但仍是影响此类患者生存的独立危险因素.

Homeobox proteins (HOXs) comprise a large family in eukaryotes and share a highly conserved DNA-binding motif, the homeodomain (HD). HOXs play an important role in the regulation of plant growth, development and stress response. However, systematic analysis and expression profiling of these genes have not been reported in Malus domestica. In this study, a total of 207 HOXs of M. domestica (MdHOXs) were identified and classified into 11 distinct subfamilies, and an unclassified group according to their functional domains. The MdHOXs were localized in all 17 chromosomes with various densities and a majority of them tended to formgene clusters. Analysis of the Ka/Ks ratios suggested that the duplicated genes of MdHOXs mainly underwent purifying selection with restrictive functional divergence after the duplication events. The expression of MdHOXs has organ specific characteristics and were divided into seven different groups. Stress-related cis-acting elements were prevalent in the upstream sequence of MdHOXs by systematic analysis. To explore the response to abiotic stress, eight MdHOXs were randomly selected to investigate their expression using quantitative real-time polymerase chain reaction. Transcription levels of MdHOXs were upregulated in the leaves and roots under cold, osmotic, high salinity or exogenous ABA treatments, which suggested that they may take part in the plant response to abiotic stress. These results provided basic information of HOXs in apple and will further contribute to the functional research of MdHOXs, especially the response to abiotic stress.

The B-box proteins (BBXs) are zinc finger proteins containing one or two B-box domain(s) and involved in regulation of development processes as transcription factors in plants. Here, seven BBX genes in Malus domestica genome (MdBBXs) were identified and found to be up-regulated under abiotic stresses, with 2–12 folds in roots. All recombinant MdBBXs expressed in Escherichia coli (E. coli) enhanced the cell’s tolerance to salt and osmotic stresses, respectively. Deficiency of B-box domain of MdBBX10 led to the loss of anti-stress functions. Five conservative cysteines in B-box domain played crucial roles in stress resistance, which are involved in two of metal iron binding sites of zinc finger motifs in BBXs. All the above results suggested MdBBXs confer stress tolerance to E. coli cell against abiotic stresses.

目的 探讨无血清内皮细胞生长培养基套装(EGM-2 BulletKit)对骨髓间充质干细胞(MSC)向血管内皮细胞分化的诱导作用.方法 通过贴壁培养获取MSC,用流式细胞仪检测MSC表面标志物表达.将获得的MSC随机分为对照组与诱导组.对照组用无血清EGM-2基础培养基进行培养,诱导组选用无血清EGM-2基础培养基+BulletKit进行诱导.于诱导第2、6、10、14天用流式细胞仪、细胞免疫荧光法检测内皮细胞表面分子CD31表达.采用DiI标记的乙酰化低密度脂蛋白吞噬实验检测MSC诱导分化后的吞噬功能,用基底膜基质胶成管实验检测细胞体外成管功能.结果 贴壁培养获得的MSC呈长梭形,流式细胞仪检测结果显示MSC标志物表达阳性的比例均大于95％,而内皮细胞标志物无表达.诱导组诱导第2天CD31开始表达,随诱导时间增加,CD31表达量逐渐升高,诱导第14天CD31表达量最高,CD31表达阳性的内皮细胞比例为55.8％;对照组无CD31表达.诱导第14天,诱导组能够摄取脂质,而对照组无摄取现象.诱导组细胞能够形成典型的管状结构;对照组细胞未形成典型的线状、管状结构.结论 无血清EGM-2Bullet Kit可诱导MSC分化为血管内皮细胞,且具有内皮细胞的特性、功能.

Protein disulfide isomerase (PDI) catalyzes the conversion of thiol-disulfide and plays an important role in various physiological events in animals. A PDI (OaPDI) from a tropical plant was detailed studied and it was found to be involved in response of biotic stress (Gruber et al., 2007). However, the activities of PDI related to physiological functions in plants are poorly understood. In the present study, a homolog of human PDI in Arabidopsis (AtPDI1), encoded by the gene (At3g54960), was characterized. The recombinant AtPDI1 protein had disulfide isomerase activity in vitro and two pairs of conservative cysteines in catalytic domains play a crucial role in the PDI activities. Expression of AtPDI1 in Escherichia coli significantly enhanced stress tolerance of cells and the mutations of critical cysteines almost lose this function. In plants, AtPDI1 was strongly induced by abiotic stresses and exogenous abscisic acid. An Arabidopsis AtPDI1 knockdown mutant (pdi1) and overexpression lines of transgenic plants obtained by this investigation were used to further examine the function of AtPDI1. The mutant line was more sensitive to stresses than the wild-type, while overexpressing AtPDI1 increased tolerance of seedlings to abiotic stresses, with a higher germination ratio and longer length of roots than the wild-type. Our results suggested AtPDI1 played roles in anti-stresses in Arabidopsis, which relate to the activities of PDI.

As a family of transcription factors, DNA binding with one figure (Dof) proteins play important roles in various biological processes in plants. Here, a total of 60 putative apple (Malus domestica) Dof genes (MdDof) were identified and mapped to different chromosomes. Chromosomal distribution and synteny analysis indicated that the expansion of the MdDof genes came primarily from segmental and duplication events, and from whole genome duplication, which lead to more Dof members in apples than in other plants. All 60 MdDof genes were classified into thirteen groups, according to multiple sequence alignment and the phylogenetic tree constructed of Dof genes from apple, peach (Prunus persica), Arabidopsis and rice. Within each group, the members shared a similar exon/intron and motif compositions, although the sizes of the MdDof genes and encoding proteins were quite different. Several Dof genes from the apple and peach were identified to be homologues based on their close synteny relationship, which suggested that these genes bear similar functions. Half of the MdDof genes were randomly selected to determine their responses to different stresses. The majority of MdDof genes were quite sensitive to PEG, NaCl, cold and exogenous ABA treatment. Our results suggested that MdDof family members may play important roles in plant tolerance to abiotic stress.

In comparison with general FISH for preparing probes in terms of time and cost, synthesized oligonucleotide (oligo hereafter) probes for FISH have many advantages such as ease of design, synthesis, and labeling. Low cost and high sensitivity and resolution of oligo probes greatly simplify the FISH procedure as a simple, fast, and efficient method of chromosome identification. In this study, we developed new oligo and oligo multiplex probes to accurately and efficiently distinguish wheat (Triticum aestivum, 2n = 6x, AABBDD) and Thinopyrum bessarabicum (2n = 2x = 14, JJ) chromosomes. The oligo probes contained more nucleotides or more repeat units that produced stronger signals for more efficient chromosome painting. Four Th. bessarabicum-specific oligo probes were developed based on genomic DNA sequences of Th. bessarabicum chromosome arm 4JL, and one of them (oligo DP4J27982) was pooled with the oligo multiplex #1 to simultaneously detect wheat and Th. bessarabicum chromosomes for quick and accurate identification of Chinese Spring (CS) - Th. bessarabicum alien chromosome introgression lines. Oligo multiplex #4 revealed chromosome variations among CS and eight wheat cultivars by a single round of FISH analysis. This research demonstrated the high efficiency of using oligos and oligo multiplexes in chromosome identification and manipulation.

Objective To analyze the efficacy and safety of conditioning regimen with busulfan and increased-dose of fludarabine for allogenetic hematopoietic stem cell transplantation (allo-HSCT) in intermediate and high-risk myeloid leukemia.Methods A total of 111 cases of intermediate and high-risk myeloid leukemia receiving allo-HSCT from Jan.2007 to Dec.2015 were retrospectively analyzed.Conditioning regimen consisted of busulfan (4 mg/kg.day orally or 3.2 rng/kg.day intravenously for 3 days) with total dose of 200 mg/m2 fludarabine (by intravenous infusion,divided into 5-8 days).For patients with unrelated and mismatched related donors,total dose of 7.5-10 mg/ kg rabbit antithymocyte globulin (by intravenous infusion,divided into 4 days) was added to the conditioning regimen.GVHD prophylaxis consisted of cyclosporine A,short-term MTX and mycophenolatemofetial (MMF).Results Neutrophil with ANC ＞0.5 × 109/L was achieved in 100％ of recipients with median time of 13 days (range:9-31 days) and platelet ＞20 × 109/L in 98.2％ of recipients after a median of 16 days (range:8-75 days).The cumulative incidence of grade Ⅱ to Ⅳ aGVHD was 39.6％ and that of grade Ⅲ to Ⅳ aGVHD was 10.3％,and overall cGVHD was 21.0％ with extensive cGVHD being apparent in 6.9％.The other complication included hemorrhagic cystitis (17.1％) and no hepatic veno-occlusive disease (VOD) was diagnosed.For all patients,3-year overall survival (OS) and 3-year disease-free survival (DFS) was 78.8％ and 77.6％ respectively.For patients with complete remission,the 3-year OS was 88.6％,which was compared to that of 61.2％ with advanced diseases (P =0.001).The overall cumulative rate of relapse was 18.3％ with 7.3％ for stable disease and 36.2％ for advanced diseases (P＜ 0.001).Conclusion Busulfan combined with increased-dose of fludarabine as conditioning regimen for allo-HSCT in AML patients can achieve stably hematopoietic reconstruction,has low toxicities and survival advantage.Such regimen can be used for transplantation in intermediate and high-risk myeloid leukemia and particularly recommended for patients with stable disease.

目的 研究抗抑郁治疗对老年高血压合并抑郁障碍患者血压的影响及其改善生活质量的临床疗效.方法 老年高血压合并抑郁障碍患者304例,随机分为治疗组156例和对照组148例,分别给予盐酸曲唑酮和盐酸曲唑酮模拟药,2组治疗4周,观察治疗前和治疗后第1、2、3、4周患者血压、17项汉密尔顿抑郁量表(HAMD17)、汉密尔顿焦虑量表(HAMA)、健康状况问卷(SF36)和治疗药物副作用量表等评分的变化,判断2组的临床疗效.结果 治疗组第3、4周末HAMD17评分较对照组显著降低,治疗组抑郁治疗的有效率优于对照组(79.9％ vs 28.3％,P＜0.05);治疗组的目标血压达标率显著高于对照组(76.4％ vs 44.9％,P＜0.01).治疗组HAMA评分第3、4周末较对照组显著下降(P＜0.01);2组SF-36总分及各项评分治疗第4周末较治疗前均有显著提高(P＜0.05,P＜0.01),治疗组生理功能、躯体疼痛、生命活力、社会功能和健康变化等的改善优于对照组(P＜0.05).结论 抗抑郁治疗有助于改善老年高血压合并抑郁障碍患者的血压和生存质量.

This study was purposed to analyze the clinical characteristics and prognostic factors in patients with primary gastrointestinal non-Hodgkin's lymphoma (PGI-NHL). The pathological data of 101 PGI-NHL patients admitted in our hospital in the past 15 years were analyzed retrospectively. The results showed that 101 patients with PGI-NHL accounted for 14.49% of NHL in the same period, there were 64 males, 37 females, the range of ages was from 18 to 87 years old, median age was 61 years old; in disease distribution, the stomach PGI-NHL accounted for 58.42%, intestine PGI-NHL accounted for 39.60%, multiple GI involvements (MGI) accounted for 1.98%; in pathological type, diffuse large B cell lymphoma (DLBCL) accounted for 66.34%, mucosa-associated lymphoid tissue (MALT) lymphoma accounted for 17.82%, mantle cell lymphoma (MCL) accounted for 3.96%, enteropathy-associated T cell lymphoma (EATL) accounted for 7.92%, extra-nodal nasal type NK/T cell lymphoma accounted for 1.98%, follicular lymphoma (FL) accounted for 0.99%, small lymphocyte lymphoma (SLL) accounted for 0.99%. Eighty-nine out of 101 patients were followed up (49 cases live, 40 cases dead), data of the 12 patients were lost; the median survival time was 29 months (1 - 173). The three-year OS and five-year OS were 58.4% and 52.6% respectively. Univariate analysis revealed that the factors affecting OS included sex (P = 0.004), lesion site (P = 0.002), tumor size (P = 0.011), clinical Lugano staging for gastrointestinal non-Hodgkin's lymphoma (P = 0.003), IPI score (P = 0.000), pathological cell phenotype (P = 0.001), and pathological type (P = 0.006), their differences were statistically significant (P < 0.05). Multivariate Cox regression analysis indicated that clinical Lugano staging for gastrointestinal non-Hodgkin's lymphoma, IPI score, pathological type were independent prognostic risk factors affecting OS. It is concluded that clinical Lugano staging for gastrointestinal non-Hodgkin's lymphoma, IPI score and pathological type are independent risk factors affecting OS.

弥漫性大B细胞淋巴瘤(DLBCL)是非霍奇金淋巴瘤(NHL)中发病最多的亚型,占非霍奇金淋巴瘤的30%～40%[1].为进一步确定弥漫性大B细胞淋巴瘤预后因素及不同化疗方案对患者预后的影响,我们回顾性分析了143例DLBCL患者的临床资料,现报告如下.

目的 探讨异基因造血干细胞移植后淋巴组织增生性疾病的危险因素、临床表现、早期诊断和治疗方法.方法 回顾性分析3例亲属供者人类白细胞抗原(HLA)配型不完全相合异基因造血干细胞移植后发生淋巴组织增生性疾病患者的临床表现、实验室检查、治疗及预后.结果 2例患者分别在移植后65d和51 d出现高热,抗生素治疗无效,并很快出现全身淋巴结肿大,血清EB病毒(EBV)-DNA分别为3.50×108、1.33×107拷贝/ml;抗病毒治疗无效分别于移植后80 d和57 d死亡.另1例移植后52 d出现全身淋巴结肿大,不伴发热,血清EBV-DNA为7.24×106拷贝/ml,由于诊断早,及时治疗,抗病毒治疗15周后肿大淋巴结消退,血清EBV-DNA转阴.结论 移植后淋巴组织增生性疾病是异基因造血干细胞移植后的严重并发症之一,病死率高,临床医师应提高对其的认识,如能早期诊断、早期治疗,可明显改善预后.

OBJECTIVE To observe whether rhIL-11 could accelerate the engraftment of platelets after unrelated cord blood transplantation (CBT). METHODS Nine patients (3 children and 6 adults) were enrolled in this study. The degree of HLA disparity was 0-2 loci. Cord blood was given two units for adults and one unit for children. Conditioning regimens were CY/TBI in 1 and BU/CY in 8 cases, both with antithymocyte globulin. Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine and short-term methotrexate. On day +1, rhIL-11 was used at 50 microg x kg(-1) x d(-1) and G-CSF at 5 microg x kg(-1) x d(-1) to accelerate hematopoiesis recovery. RESULTS The median age of the patients was 22.3 years and the median weight 52.3 kg. Among the 9 patients, 8 (88.9%) experienced engraftment. The median time to neutrophil > 0.5 x 10(9)/L was 21.3 (14-37) days and to platelet > 20 x 10(9)/L was 25 (18-36) days. 42.9% of the patients developed grade I aGVHD and 33.3% developed localized chronic GVHD. Six patients were alive and disease-free at a median follow-up of 7 months. Infection was the primary cause of death. The expected 1-year survival was 77.8%, 2-year survival was 52.2%. Five of 8 patients (62.5%) who received IL-11 presented leakage syndrome. On prophylaxis with drugs containing Arnebia root extract, all patients could tolerate the treatment. CONCLUSION rhIL-11 maybe helpful for accelerating the platelet recovery and reducing aGVHD severity in unrelated CBT. The major side effect is leakage syndrome. It is well tolerated on prophylaxis with drugs containing Arnebia root.

Objective To investigate the clinical efficacy and toxicity of arsenic trioxide on treating non-APL hematologic malignancy.Methods Analyzed the treatment effects and side effects in 5 cases of NHL,1 case of MM and 3 cases of MDS.Results 2 cases achieved CR and 1 PR in 5 NHL,respectively,1 NR in MM,1 case achieved cytogenetic remission in 3 MDS.The side effects of arsenic trioxide were arrest of bone marrow,fever,skin rash,hydrothorax,impairment of renal function.Conclusion Arsenic trioxide tended to be effective on treating non-APL hematologic malignancy.

嗜酸细胞性胃肠炎是以胃肠道组织中嗜酸细胞异常浸润为特征的胃肠道疾病.Kaizser于1937年首先报道.迄今国内文献报道100余例.本病病因不明,发病率低,现报道1例如下.