Genome-wide identification, classification, and analysis of heat shock transcription factor family in Chinese cabbage (Brassica rapa pekinensis).

Chinese cabbage (Brassica rapa ssp. pekinensis) is one of the most important vegetable crops grown worldwide, and various methods exist for selection, propagation, and cultivation. The entire Chinese cabbage genome has been sequenced, and the heat shock transcription factor family (Hsfs) has been found to play a central role in plant growth and development and in the response to biotic and abiotic stress conditions, particularly in acquired thermotolerance. We analyzed heat tolerance mechanisms in Chinese cabbage. In this study, 30 Hsfs were identified from the Chinese cabbage genome database. The classification, phylogenetic reconstruction, chromosome distribution, conserved motifs, expression analysis, and interaction networks of the Hsfs were predicted and analyzed. Thirty BrHsfs were classified into 3 major classes (class A, B, and C) according to their structural characteristics and phylogenetic comparisons, and class A was further subdivided into 8 subclasses. Distribution mapping results showed that Hsf genes were located on 10 Chinese cabbage chromosomes. The expression profile indicated that Hsfs play differential roles in 5 organs in Chinese cabbage, and likely participate in the development of underground parts and regulation of reproductive growth. An orthologous gene interaction network was constructed, and included MBF1C, ROF1, TBP2, CDC2, and HSP70 5 genes, which are closely related to heat stress. Our results contribute to the understanding of the complexity of Hsfs in Chinese cabbage and provide a basis for further functional gene research.

Genome-wide identification, classification, and expression analysis of sHSP genes in Chinese cabbage (Brassica rapa ssp pekinensis).

Small heat shock proteins (sHSPs) are essential for the plant's normal development and stress responses, especially the heat stress response. The information regarding sHSP genes in Chinese cabbage (Brassica rapa ssp pekinensis) is sparse, hence we performed a genome-wide analysis to identify sHSP genes in this species. We identified 26 non-redundant sHSP genes distributed on all chromosomes, except chromosome A7, with one additional sHSP gene identified from an expressed sequence tag library. Chinese cabbage was found to contain more sHSP genes than Arabidopsis. The 27 sHSP genes were classified into 11 subfamilies. We identified 22 groups of sHSP syntenic orthologous genes between Chinese cabbage and Arabidopsis. In addition, eight groups of paralogous genes were uncovered in Chinese cabbage. Protein structures of the 27 Chinese cabbage sHSPs were modeled using Phyre2, which revealed that all of them contain several conserved ß strands across different subfamilies. In general, gene structure was conserved within each subfamily between Chinese cabbage and Arabidopsis, except for peroxisome sHSP. Analysis of promoter motifs showed that most sHSP genes contain heat shock elements or variants. We also found that biased gene loss has occurred during the evolution of the sHSP subfamily in Chinese cabbage. Expression analysis indicated that the greatest transcript abundance of most Chinese cabbage sHSP genes was found in siliques and early cotyledon embryos. Thus, genome-wide identification and characterization of sHSP genes is a first and important step in the investigation of sHSPs in Chinese cabbage.

Biochemical composition of the ovarian fluid and its effects on the fertilization capacity of turbot Scophthalmus maximus during the spawning season.

This study investigated the biochemical composition of ovarian fluid and its effect on the fertilization capacity of turbot Scophthalmus maximus during the spawning season. The fertilization rate and pH of ovarian fluid varied throughout the spawning season, with the highest values recorded at the mid-season. Positive correlations were found between the fertilization rate and the ovarian fluid pH. The composition of major inorganic ions (Na(+) , K(+) , Ca(2+) and Cl(-) ) showed no significant changes during the spawning season. Alkaline phosphatase (AKP) activity was significantly higher during mid-season than other seasons. The lowest levels of protein, acid phosphatase (ACP) and aspartate aminotransferase (AAT) were in the ovarian fluid released at the mid-season. Moreover, significant relationships were observed between the fertilization rate and the levels of protein, ACP, AKP and AAT. These observations suggest that the biochemical profile of ovarian fluid affects the insemination microenvironment as well as the fertilization capacity of S. maximus eggs. Determination of such profiles may prove to be a useful strategy to improve S. maximus breeding techniques.

Isolation and Characterization of a Moderately Virulent Classical Swine Fever Virus Emerging in China.

Classical swine fever (CSF) is a devastating infectious disease of pigs caused by classical swine fever virus (CSFV). In China, CSF has been under control owing to extensive vaccination with the lapinized attenuated vaccine (C-strain) since 1950s, despite sporadic or endemic in many regions. However, recently, CSF outbreaks occurred in a large number of swine herds in China. Here, we isolated 15 CSFV strains from diverse C-strain-vaccinated pig farms in China and characterized the genetic variations and antigenicity of the new isolates. The new strains showed unique variations in the E2 protein and were clustered to the subgenotype 2.1d of CSFV recently emerging in China in the phylogenetic tree. Cross-neutralization test showed that the neutralizing titres of porcine anti-C-strain sera against the new isolates were substantially lower than those against both the highly virulent Shimen strain and the subgenotype 2.1b strains that were isolated in China in 2006 and 2009, respectively. In addition, experimental animal infection showed that the HLJZZ2014 strain-infected pigs displayed lower mortality and less severe clinical signs and pathological changes compared with the Shimen strain-infected pigs. The HLJZZ2014 strain was defined to be moderately virulent based on a previously established assessment system for CSFV virulence evaluation, and the virus shedding and the viral load in various tissues of the CSFV HLJZZ2014 strain-infected pigs were significantly lower than those of the Shimen strain-infected pigs. Taken together, the subgenotype 2.1d isolate of CSFV is a moderately virulent strain with molecular variations and antigenic alterations.

Effect of physical training on the age-related changes of acetylcholinesterase-positive fibers in the hippocampal formation and parietal cortex in the C57BL/6J mouse.

We investigated the effect of a moderate amount of prolonged physical training initiated at 3 months of age on the age-related changes of the hippocampal and cortical cholinergic fibers. A total of 80 male C57BL/6J mice were divided into five groups which were trained (including adult and old trained, AT and OT), sedentary (adult and old sedentary, AS and OS) and young (Y). From 3 months old, the mice of the trained groups were treated with a voluntary running wheel for 1 h each day, 5 days per week. AT had been trained up to 13-month-old whereas OT up to 24 months old. At the same time, the mice of the sedentary groups were put in immobilized wheels. We set the criterion for effective training in the trained mice such that the heart-to-body weight ratio should be at least 2 S.D. above the mean in the age-matched groups. Using AChE histochemistry and stereology, the AChE-positive fibers were analyzed quantitatively in the molecular layers in CA1, CA3 and the dentate gyrus of the hippocampal formation, and in III, V layers in the motor and somatosensory cortex. Comparison of Y, AS and OS (3, 13 and 24 months of age) showed minimum AChE-positive fiber density in the hippocampal formation and the cortex in OS (P < 0.01). After 10 and 21 months of running, the AChE-positive fibers in all regions examined in the trained groups were significantly increased compared to their age-matched controls (P < 0.05 or 0.01). In the hippocampal formation, the increase was about 17% in AT and 23% in OT, whereas, in the cortex, it was 13% in AT and 22% in OT. These results indicated that a moderate amount of prolonged physical training could modify the age-related loss of cholinergic fibers in the hippocampal formation and cortex, furthermore the modified loss of cholinergic fibers might be associated with the regeneration of hippocampal and cortical cholinergic fibers stimulated by chronic running.

Physical training modifies the age-related decrease of GAP-43 and synaptophysin in the hippocampal formation in C57BL/6J mouse.

We investigated the effect of a moderate amount of prolonged physical training initiated at 3 months of age on the expression of GAP-43 and synaptophysin in the hippocampal formation. C57BL/6J mice were divided into three groups which were trained (24 months old), sedentary (24 months old) and young (3 months old). From 3 months of age on, mice of trained group were treated with voluntary running wheel for 1 h each day (5 days per week) until 24 months of age (21 months running), whereas mice of sedentary group were put in immobilized wheels for the same time. Using immunohistochemistry and image analysis system, GAP-43 and synaptophysin were analysed quantitatively in the CA1, CA3 areas and the dentate gyrus of the hippocampal formation. As compared with young mice, the densities of GAP-43 and synaptophysin immunostaining showed a significant decrease in the hippocampal formation in sedentary group (P<0.01). After 21 months of running, the densities of GAP-43 and synaptophysin immunostaining significantly increased in the examined areas of the hippocampal formation in trained mice compared to their age-matched sedentary controls (P<0.05, 0.01). These results indicate that a moderate amount of prolonged physical training could modify the age-related decrease of the expression of GAP-43 and synaptophysin in the hippocampal formation, and that the increased expression of GAP-43 and synaptophysin might be associated with the anatomical sprouting and synaptogenesis.

[Relationship between the familial nonrandom chromosome loss (NCL) and leukomogenesis].

By using R-banding karyotypic analysis technique, the bone marrow (BM) cells were performed in 223 hematopoietic malignacies and 105 diopathic throbocytopenic purpura (ITP), which served as control. The following results were obtained: (1) Nonrandom chromosome loss (NCL), such as, -11, -14, -21, etc, which were found in the affected members of leukemia families, were found in about 30% sporadic ANLL, MDS and about 50% ALL, espedislly in 100% (5/5) CLL, but not found in ITP (P < 0.001). These results indicated that the familial nonrandom chromosome loss were associated with leukomogenesis. (2) Because most of BM cells are hypodiploed and have the same kinds of NCL in each cases of CLL, which can develop into ALL, ANLL and also cancers, ALL BM hypo- and hyper-diploid and/or polyploid cells might be origin of hypodiploid cells. (3) 28% (6/21) of pediatric patients with AL, MDS, or FA (Fanconi Anemia) have one parent, who have up to 30% BM hypodiploid cells and similar kinds of NCL and also have the rearrangement of C-erbB and abnormal proliferation of BM. The NCL were found in the three consecutive generations of a family with 5 ALL among 10 members of third generation. It indicated that the familial NCL might be inherited and be coded by a unknown gene alteration, which might be related to leukomo and carcinogenesis, because there are genes or their candidates for leukemia in the chromosomes 11, 14 and 21. (4) Based on the works of my colleages and I, the model of leukomo and carcinogenesis was proposed and the relationship between chromosome monosomy, deletion, translocations and leukomogenesis were showed elswhere. The significance of monosomy 11, 14 and 21 etc. were discussed briefly.

[Relationship between the occurrences of AL, MDS and AA and abnormal BM proliferation of patient's parents].

After studying of familial leukemias, Myelodysplastic Syndrome (MDS) and aplastic anemia (AA), we observed and analysed bone marrow (BM) cells hematologically and molecular-cytogenetically in 36 persons who are first degree relatives (FDRs) of patients with acute leukemias (AL), MDS and AA. The peripheral blood (PB) lymphocyte chromosome fragility sensitive to folic acid and unstability was also analysed in 18 FDRs. The abnormal BM megakaryocystic/erythroid cellularity and the rearrangement of c-erbB were found in 66%-86.1% of parents and siblings of patients. The associations of dysplastic megakaryopoiesis, including the presence of lymphoid small megakaryocytes, with the chromosomal monosomy or/and the rearrangement/amplification of C-erbB, were found in a few parents and siblings. These results were consistent with those of MDS, Fanconi Anemia (FA) and AL. The normal karyotype and SCD positive of BM cells and PB lymphocytes, and PB lymphocyte chromosomal fragility and unstability were found in most of patients' parents, while familial chromosomal monosomy of BM cells and PB lymphocyte chromosomal fragility were found in parents and siblings of familial leukemia patients. Based on the studies of a large family with 7 cases of acute erythroleukamia and relative myeloleukemias in three consecutive generations and a family with 3 CAA and 1 AML, the rearrangement of c-erbB might be inherited. The rearrangement/amplification of c-erbB and its PCR detected results could be the indicators of gene diagnosis of preleukemia and might be useful in genetic conselling of leukemias. The common origin of AL, MDS and AA was discussed.

[Gene diagnosis and successful reversion in a patient with preleukemia].

According to the FAB classification, a patient (case 1) could not be diagnosed as MDS-RA, although she had clinical features of MDS, as compared with another patient (case 2) who was diagnosed as RAS and had abnormal karyotype (20q- and 5q-) of bone marrow (BM) cells. BM cells of the two patients were SCD (sister chromatid differentiation) negative. Rearrangement of c-erbB and c-erbA was found in the genome of the BM cells in both patients, when southern blot hybridization was performed with probe v-erbB+A. Therefore, case 1 could be diagnosed as preleukemia. During a period about 3 years of treatment with the drug stanozolol in case 1 there was good effect and successful reversion was obtained. She had then normal hematologic and cytogenetic patterns of BM and PB and the rearrangement of c-erbB of BM cells also disappeared. She has worked for two years since then. The mechanism of effective treatment and successful reversion was discussed briefly. Probe v-erbB was shown to be useful in investigation of gene diagnosis of preleukemia or MDS (shown elsewhere).

[Cytogenetic studies on 334 myelodysplastic syndrome (MDS), aplastic anemia (AA) and other hematological diseases].

In order to distinguish various types of MDS, such as RA/AA, RA/ITP or RA/HA, from AA, ITP or HA, bone marrow (BM) cells were studied by using cytogenetic techniques including R-banding karyotypic analysis and sister chromatid differentiation (SCD) assay in 334 cases of hematological diseases (160 MDS, 54 RA/AA, RA/ITP or RA/HA; 60 AA, 3 other known anemias, 38 PNH and 19 ITP). The results showed: (1) karyotypes and SCD values were both normal in more than 90% of AA, PNH, ITP and other known anemias, but they were both abnormal in about 35.6% of MDS and only 13.0% of RA/AA, RA/ITP or RA/HA. These results indicated that cytogenetic techniques were useful in hematological clinic and that RA/AA, or RA/ITP or RA/HA might be pre-RA or atypic RA. This was supported by the results of following up on some RA/AA, RA/ITP or RA/HA cases, (2) clonal abnormal karyotypes were found in 64.4% of MDS. The recurrent chromosomal alterations were +8, 20q-, -5/5q-, -7/7q-, similar to those reported in literatures. (3) 16 MDS cases were followed up and 15 MDS with SCD negative, but one with SCD positive developed leukemia in our hospital. It is suggested that change from SCD positive to negative was indicative of malignant transformation of BM cells. This was supported by the results of cytogenetic analysis in RA/AA, RA/ITP, RA, RAEB, RAEBT and leukemias. (4) Because more structural chromosome alterations occur in SCD negative than SCD positive MDS, the numerous chromosome alterations (monosomy) might occur in earliest development of MDS into leukemias.

[Gene diagnosis of familial erythroleukemia at the early stage of leukomogenesis].

Southern blot hybridization was performed with probe v-erbB + A in 4 subjects. Two of them were patients with similar erythroid alterations of bone marrow: one of them was diagnosed as erythroleukemia (EL). The other two were members of a same family (husband and wife). The husband had normal erythroid cellularity of bone marrow but the wife and their 7 living sons and daughters had erythroid hypercellularity. In the second generation of this family 4 hematological patients (1 with AML and 3 with CAA) were found in the past decade. Rearrangement of cerbB and c-erbA genes was found in 3 of the 4 subjects (2 with EL and the wife mentioned above). They had 4 new hybridization bands besides 3 germlines, but the husband had only 3 germlines. The result suggests that using Southern blot hybridization technique with probe v-erbB + A, the rearrangement of c-erbB/c-erbA genes might be a diagnostic indicator of erythroleukemia at early stage of leukomogenesis in familial and sporadic patients. The etiological significance of the rearrangement of c-erbB/c-erbA in leukomogenesis was discussed.

An analysis of 5 years of medical informatics publications.

This study describes an analysis of 1520 papers that were published in six medical informatics journals from 1992 through 1996, and indexed in NLM's MEDLINE. Of retrieved publications the countries of origin were determined, and insight in the subject of the publication was obtained by analysing the used MeSH terms. The main conclusion is that despite the fact that publications in medical informatics stem from a wide international community, scientific recognition can still not be demonstrated by high impact factors of journals.

Morphological plasticity of axotomized retinal ganglion cells following intravitreal transplantation of a peripheral nerve segment.

During normal development of retinal ganglion cells when the axons are growing, transient dendritic spines have been observed. Similar dendritic spine-like processes are also exhibited by retinal ganglion cells undergoing axonal regeneration into a peripheral nerve grafted to the damaged optic axons. Here we show, using the intracellular injection of Lucifer Yellow, that when a segment of peripheral nerve is transplanted to the vitreous body, a procedure which induces ectopic sprouting of axon-like processes from the cell bodies and dendrites of some retinal ganglion cells, similar spine-like processes appear on the dendrites of cells with ectopic sprouts. Quantitative analysis indicated that there were significant changes with posttransplantation survival time in the distributions of spine-like processes and axon-like processes on these sprouting retinal ganglion cells following the intravitreal transplantation of a piece of peripheral nerve. The remodelling of the spine-like processes and axon-like processes correlated with one another suggesting that plastic changes can occur in certain dendritic subcompartments independent of the growth activity of the other dendritic subcompartments.