[Research progress of C-X-C motif chemokine ligand 12 and its receptor signaling axis in the regulation of pulmonary fibrosis].

Pulmonary fibrosis is an irreversible interstitial lung disease characterized by lung parenchyma remodeling and collagen deposition. In recent years, the incidence and mortality of pulmonary fibrosis caused by unknown causes have risen. However, its pathogenesis is still unclear. C-X-C motif chemokine ligand 12 (CXCL12)/C-X-C chemokine receptor 4 (CXCR4)/CXCR7 signal axis plays a critical regulatory role in pulmonary fibrosis disease. In addition, the signal axis has been shown to regulate recruitment and migration of circulating fibrocytes, mesenchymal stem cells to the damage lung tissue, the migration of endothelial cells, the proliferation and differentiation of fibroblasts and endothelial cells, which further affects the occurrence and progression of pulmonary fibrosis. In this review, we summarized the pathogenesis and treatment research progress of CXCL12 and its receptor CXCR4/CXCR7 in the occurrence and progression of pulmonary fibrosis.

Molecular cloning, functional verification, and evolution of TmPm3, the powdery mildew resistance gene of Triticum monococcum L.

Powdery mildew (Pm) is one of the most harmful diseases in wheat. Three Pm-resistance genes, Pm3, Pm21, and Pm8, have been cloned but most Pm3/Pm8 alleles have lost their resistance to Pm in hexaploid wheat. In this study, a new Pm3 homolog gene (TmPm3) was isolated from Triticum monococcum L. using a homology-based cloning strategy, being the first report of a functional Pm3 homolog gene from a diploid wheat species. The transient expression of TmPm3 in leaf epidermal cells showed that over-expressed TmPm3 could significantly inhibit the penetration of Blumeria graminis f. sp tritici conidia spores and the formation of haustoria. Sequence analysis of Pm3 alleles shed new light on the evolution of Pm3 genes, providing a better understanding of the molecular basis of disease resistance. This study also suggested that homology-based cloning of resistance genes is a feasible method for the isolation of functional resistance genes from wheat germplasm.

Molecular cloning, characterization and expression analysis of WAG-1 in the pistillody line of common wheat.

Wheat WAG-1 is a C-class MADS-box gene, which is orthologous to AGAMOUS in Arabidopsis. In this study, we report the cloning, characterization, and expression patterns of WAG-1 in the pistillody mutant HTS-1 and its sib-line CSTP. The cDNA of WAG-1 was found to be 765 bp in length, which was equal to the length of its open reading frame, encoding 254 amino acids. The location of WAG-1 revealed that it has three homologous genes from the short arm of chromosome 1A, 1B, and 1D. Their genomic sequences were determined to be 5864, 6454, and 6447 bp long, respectively, and possessed seven exons and six introns. Young spikes from HTS-1 contained higher levels of WAG-1 transcript than did those from CSTP, and the transcript levels in the young spikes (7-10 mm in length) of HTS-1 increased 3.3-fold relative to those of the CSTP line. The transcript level in the pistil and pistil-like stamens of HTS-1 was over 2-fold higher than that in the stamens of CSTP, and expression in the pistil-like stamens of HTS-1 was slightly higher than that in its pistils. These data provide a basis for future research into the function of WAG-1, and offer further insight into the molecular mechanism of the pistillody mutation in common wheat.

Chloroplast ultrastructure regeneration with protection of photosystem II is responsible for the functional 'stay-green' trait in wheat.

CN17 is a functional stay-green wheat variety that exhibits delayed leaf senescence and enhanced photosynthetic competence. To better understand these valuable traits, levels of chlorophyll a and b, soluble proteins, unsaturated fatty acids, and other components of CN17 were assayed. In addition, chloroplast ultrastructure, chloroplast number, and differences in gene expression between CN17 and a control variety, MY11, were examined. By 21 d post-anthesis (DPA), CN17 leaves exhibited a significantly higher maximal photochemical efficiency for photosystem II (PSII) (F(v) /F(m) ) and a significantly higher efficiency of excitation capture by open PSII reaction centres (F(v) '/F(m) '). In addition, chlorophyll degradation in CN17 was delayed by approximately 14 d, and was not blocked as observed in cosmetic stay-green phenotypes. The soluble protein content (Ps) of CN17 was higher than MY11 at all timepoints assayed, and the ratio of unsaturated to saturated fatty acids was significantly higher. CN17 also exhibited isolated granal lamellae associated with vesicles and diminished peroxidation, and between 35 and 42 DPA, a sharp decrease in chloroplast number was detected. Taken together, these results strongly support the hypothesis that chloroplast ultrastructure regeneration is responsible for the functional stay-green trait of CN17, and gene expression data provide insight into the mechanistic details.

LncRNA DANCR affected cell growth, EMT and angiogenesis by sponging miR-345-5p through modulating Twist1 in cholangiocarcinoma.

OBJECTIVE: LncRNA DANCR has been reported to play an important role in various cancers. Therefore, this study aimed at exploring the function and regulatory mechanism of DANCR in Cholangiocarcinoma (CCA). PATIENTS AND METHODS: qRT-PCR was used to measure the expression of DANCR, miR-345-5p in tissues and cells. Western blot was applied to measure the protein expression of Twist, N-cadherin, Vimentin, E-cadherin, VEGF-A, VEGF-C, PCNA and C-caspase 3. The relationship between DANCR and miR-345-5p was determined by luciferase reporter assay. MTT assay and flow cytometry were used to assess cell proliferation and apoptosis, respectively. Transwell assay was performed to detect cell invasion and migration. RESULTS: We found that the expression of DANCR was significantly induced in CCA tissues and cells. Inhibition of DANCR remarkably suppressed CCA cell proliferation, migration, invasion, EMT and angiogenesis as well as induced cell apoptosis in vitro and in vivo. Luciferase reporter assay determined that DANCR directly targeted miR-345-5p and Twist1 was a target mRNA of miR-345-5p. Otherwise, miR-345-5p down-expression partially reversed the effect induced by the suppression of DANCR in CCA. Moreover, the suppressive effects of high miR-345-5p expression on CCA cells were reversed by improving Twist1 expression. CONCLUSIONS: In this study, we verified that LncRNA DANCR affected cell proliferation, migration, invasion, angiogenesis, epithelial-mesenchymal transition (EMT) and induced apoptosis through modulating miR-345-5p/Twist1 axis in Cholangiocarcinoma.

Pharmacology of radioiodinated hexadecenoic acid--a myocardial imaging agent.

123I- and 131I-labeled hexadecenoic acid (IHDA, radiochemical purity over 92%, dissolved in 6% bovine serum albumin solution) was investigated in vivo. ICR mice were administered IHDA via the tail vein. Maximum myocardial uptake (27.3 +/- 5.1%) was reached about 0.5 min after the injection. The ratio of uptake in the heart to that in the lungs was 2.3, to that in liver 1.5 and to that in other organs 2.4 to 6.4. The dog myocardium was visualized distinctly within 3-5 min with a gamma camera after i.v. 131I-IHDA, and not interfered with by activities in the lungs, liver and other organs. The low blood levels at 20 min had little effect on the quality of the heart images.

A study of 11C-benzoate as a radiopharmaceutical for renal imaging.

11C-benzoic acid prepared in a radiochemical purity over 90% was studied radiopharmacologically in mice and rabbits. The uptake of 11C-benzoate in ICR mice increased quickly. The ratio of kidney uptake rate to that in other organs reached values between 9 and 55 with a maximum at 10 min after i.v. injection. Gamma camera imaging of rabbits showed that uptake in the kidneys began at 2 min after injection and that activity began to appear in the bladder 4 min later. Rabbits with left renal artery ligature showed no uptake in the left kidney but the right kidney was imaged to the same extent as that of a rabbit without artery ligature. The kidney imaging of 11C-benzoic acid may be a useful method for renal diagnosis.

[Relationship between differential expression patterns of multigene families and heterosis in a wheat diallel crosses].

In order to understand molecular basis of heterosis, the patterns of differential gene expression of multigene families between wheat hybrids and their parents in seedling leaves were analyzed by using mRNA differential display. Relationships between differential gene expression patterns, heterosis and F1 hybrid performance were determined. Four patterns of differential gene expression were observed, which include: (1) bands observed in both parents but not in the F1; (2) bands occurring in only one parent but not in the F1 or the other parent; (3) bands detected in only the F1 but neither of the parents; (4) bands present in one parent and F1 but absent in the other parent. The analysis showed that patterns of differential gene expression were not correlated with the F1 hybrid performance for all the eight agronomic traits. However, differentially expressed fragments that occurred only in the F1 but neither of the parents were found to be positively correlated with heterosis. On the contrary, fragments observed in both parents but not in the F1 were negatively correlated with heterosis. It is concluded that the differential expression of regulatory genes plays an important role in heterosis.

[Molecular tagging of a major powdery mildew resistance gene MlG in wheat derived from wild Emmer by using microsatellite marker].

Powdery mildew caused by Erysiphe graminis f. sp. tritici is one of the most important wheat diseases in many regions of the world. Breeding for resistant cultivars has been proved to be an effective and environmentally safe method to control diseases in wheat production. It is necessary to search for more resistance genes for the diversification of resistance genes in wheat breeding. An Isreali wild emmer wheat (Triticum dicoccoides) accession "G-305-M" was found resistant to the prevailing E. graminis f. sp. tritici isolate Race No. 15 in Beijing region. The powdery mildew resistance has been transferred from G-305-M into common wheat by crossing and backcrossing (G-305-M/781//Jing 411* 3). Genetic analysis showed that the resistance was controlled by a single dominant gene at the seedling stage. A segregating BC2F3 family of the cross "G-305-M/781//Jing 411* 3" with 167 plants was chosen for SSR analysis. Totally 96 wheat microsatellite primer pairs were screened, only one primer pair WMS570 could generate polymorphic DNA fragments between the resistant and susceptible plants. After evaluating this polymorphic marker in the segregating population, the microsatellite locus Xgwm570 mapped on chromosome 6AL was found to be linked to the resistance gene, with the estimated genetic distance of 14.9 +/- 3.0 cM. Based on the origin and chromosomal location of the gene, it is suggested that the resistance gene derived from G-305-M should be a novel Pm gene and is temporarily designated MlG.

[Observation on phrenic discharges induced by bicuculline or picrotoxin in spinal rabbits].

The respiratory activity of spinal cord-transected animals was reinvestigated on 71 rabbits. Under light urethane anesthesia and paralyses by gallamine, the spinal cord was transected at either C1 or C2 level, and phrenic discharges were monitored in 43 rabbits. After cordotomy, tonic phrenic discharges were observed in 25 of the 43 spinal rabbits. No spontaneous rhythmic phrenic activities appeared in any of the animal. Respiratory-like, long lasting phrenic bursts (lasting longer than 0.25s) or convulsive-like, short lasting phrenic bursts (lasting shorter than 0.1s on the average) were induced by administration of bicuculline (BCL, 20 micrograms/10 microliters-40 micrograms/20 microliters, intra-subarachnoid space, i.s.s.) in 24 spinal rabbits or picrotoxin (PIC, 20 micrograms/20 microliters, i.s.s. or 3-5 mg/kg i.v.) in 13 spinal rabbits. According to the pattern of the recruitment and de-recruitment of the phrenic discharges, the long lasting phrenic bursts may be divided into three types: Type I, average frequency 23.5 +/- 2.3 cycles/min (BCL), incidence 58.5% (BCL) or 67.7% (PIC); Type II, average frequency 33.8 +/- 4.7 cycles/min (BCL), incidence 39.3% (BCL) or 32.3% (PIC); Type III, average frequency 21.3 +/- 2.8 cycles/min (BCL), incidence 2.2% (BCL) or 3.3% (PIC). The duration of the evoked phrenic discharges was 60.0 +/- 18.9 min (BCL) or 42.0 +/- 0.8 min (PIC). The type I and type II showed respiratory-like discharges, accounting for more than 97.8% of the incidence of the long lasting phrenic bursts. It is suggested that the endogenous GABA system in spinal cord might exert a tonic inhibitory action on the spinal "respiration" activity.

[Effects of acupuncture on blood pressure, SOD,LPO and five kinds of trace elements to stenosis of renal artery caused hypertension in rats].

Changes of blood pressure, superoxide dismutase (SOD), lipid peroxidation (LPO) and concentration of five kinds of trace elements including Cu, Zn, Fe, Ca, Mg were observed before or after acupuncture treatment in the stenosis of renal artery caused hypertension in rats [correction of mice]. It was demonstrated that acupuncture in the points of Zusanli, Neiguan, Sanyinjiao and Yongquan in mice could reduce the blood pressure significantly and influence the concentrations of SOD, LPO and five kinds of trace elements in the stenosis of renal artery caused hypertension in mice. The possible mechanisms of acupuncture in reducing the blood pressure and influencing the changes of SOD, LPO and five kinds of trace elements were also discussed.

Efficacy of intratumoral chemotherapy using arsenic trioxide (As2O3) sustained release tablets for the treatment of neurogliocytoma in nude mice.

OBJECTIVE: This study aimed to investigate the inhibiting effect of arsenic trioxide (As2O3) on neurogliocytoma in nude mice and the mechanism responsible for this effect. MATERIALS AND METHODS: Neurogliocytoma implantation models were constructed in nude mice, which were assigned to three groups: the control group, the sustained release tablet-polylactic acid-glycolic acid polymer (50:50) group (PLGA group) and the As2O3-polylactic acid-glycolic acid polymer (50:50) (As2O3-PLGA group). One tablet of As2O3-PLGA was implanted in the tumor of the As2O3-PLGA group. Intratumoral implantation was also performed in the other groups using a different type of tablet. The sustained releasing As2O3 had an inhibiting effect on the tumors. The TUNEL assay was used to determine the apoptosis rates in the implanted tumors. Immunohistochemical staining and Western blotting was carried out to determine the expression levels of caspase-3 and Bcl-2. RESULTS: No inhibitory effect was observed on the tumor in the PLGA group, and there was no significant difference between this group and the control group. Subcutaneous tumor growth in nude mice was significantly inhibited in the As2O3-PLGA group relative to that in the control group, and the difference was statistically significant (p < 0.01). The tumor inhibition rate was 60.8%. The percentage of apoptotic tumor cells in the As2O3-PLGA group was 30.8%, which was significantly higher than that in the control group (3.92%) and that in the PLGA group (4.08%). The expression of Bcl-2 in the implanted tumor tissue was significantly reduced, but the expression of caspase-3 increased significantly. CONCLUSIONS: As2O3 has a potent inhibiting effect on the growth of neurogliocytoma in vivo and can induce the apoptosis of tumor cells. The molecular mechanism of this effect may be related to the downregulation of Bcl-2 expression and the upregulation of caspase-3 expression.

Novel DNA variations to characterize low molecular weight glutenin Glu-D3 genes and develop STS markers in common wheat.

Low-molecular-weight glutenin subunits (LMW-GS) play an important role in bread and noodle processing quality by influencing the viscoelasticity and extensibility of dough. The objectives of this study were to characterize Glu-D3 subunit coding genes and to develop molecular markers for identifying Glu-D3 gene haplotypes. Gene specific primer sets were designed to amplify eight wheat cultivars containing Glu-D3a, b, c, d and e alleles, defined traditionally by protein electrophoretic mobility. Three novel Glu-D3 DNA sequences, designated as GluD3-4, GluD3-5 and GluD3-6, were amplified from the eight wheat cultivars. GluD3-4 showed three allelic variants or haplotypes at the DNA level in the eight cultivars, which were designated as GluD3-41, GluD3-42 and GluD3-43. Compared with GluD3-42, a single nucleotide polymorphism (SNP) was detected for GluD3-43 in the coding region, resulting in a pseudo-gene with a nonsense mutation at the 119th position of deduced peptide, and a 3-bp insertion was found in the coding region of GluD3-41, leading to a glutamine insertion at the 249th position of its deduced protein. The coding regions for GluD3-5 and GluD3-6 showed no allelic variation in the eight cultivars tested, indicating that they were relatively conservative in common wheat. Based on the 12 allelic variants of three Glu-D3 genes identified in this study and three detected previously, seven STS markers were established to amplify the corresponding gene sequences in wheat cultivars containing five Glu-D3 alleles (a, b, c, d and e). The seven primer sets M2F12/M2R12, M2F2/M2R2, M2F3/M2R3, M3F1/M3R1, M3F2/M3R2, M4F1/M4R1 and M4F3/M4R3 were specific to the allelic variants GluD3-21/22, GluD3-22, GluD3-23, GluD3-31, GluD3-32, GluD3-41 and GluD3-43, respectively, which were validated by amplifying 20 Chinese wheat cultivars containing alleles a, b, c and f based on protein electrophoretic mobility. These markers will be useful to identify the Glu-D3 gene haplotypes in wheat breeding programs.

Cloning and characterization of leaf cDNAs that are differentially expressed between wheat hybrids and their parents.

Previous studies have shown that heterosis is associated with differential gene expression between hybrids and their parents. In this study, we performed a screen for genes that are differentially expressed between wheat hybrids and their parents in jointing-stage leaves and flag leaves using the differential display technique. Twenty-four differentially expressed cDNA were cloned and sequenced, and their expression patterns were confirmed by reverse-Northern blotting. Sequence analysis and database searches revealed that among the genes that showed differential expression between hybrid and parents were transcription factor genes and genes involved in metabolism, signal transduction, disease resistance, and retrotransposons. These results indicate that hybridization between two parental lines can cause changes in the expression of a variety of genes, and it is concluded that the altered pattern of gene expression in the hybrid may be responsible for the observed heterosis.