Phylogeny and expression patterns of ERF genes that are potential reproductive inducers in hybrid larch.

BACKGROUND: Larch is an important component of northern forests and a major cultivated tree species in restoration of forest cover using improved seed material. In recent years, the continuous low seed production has severely affected the production of improved variety seedlings and natural regeneration. However, research on the reproductive growth of gymnosperms is extremely scarce. RESULTS: In this study, based on differential transcriptome analysis of two asexual reproductive phases, namely high-yield and low-yield, we further screened 5 ERF family genes that may affect the reproductive development of larch. We analyzed their genetic relationships and predicted their physicochemical properties. The expression patterns of these genes were analyzed in different tissues, developmental stages, hormone treatments, and environmental conditions in hybrid larch. CONCLUSION: The results showed that all 5 genes were induced by low temperature and ABA, and their expression patterns in different tissues suggested a suppressive role in the development of female cones in larch. Among them, LkoERF3-like1 and LkoERF071 may be involved in the flowering age pathway. This study enriches the scarce research on reproductive development in gymnosperms and provides a theoretical basis and research direction for regulating the reproductive development of larch in seed orchards.

The effect of heat stress on the epidermal growth factor (EGF)-mediated intracellular signaling, and changes cell behavior on swine testicular cell.

At present, high environmental temperature is the main factor endangering animal production, growth and development. Therefore, the harmful effects of heat stress led by hot environment on livestock have attracted much attention. In this work, the cellar property and signaling property of epidermal growth factor (EGF) below heat stress remains unclear in swine testicular cells. Here, the effect of heat stress on EGF-induced intracellular signaling and cell behavior was explored in the ST (a porcine testis cell line). A series of experiments were done to study the cellular behavior and signaling properties of EGF under heat stress. It can be discovered which high ambient temperature changed the cellular characteristics of EGF/EGFR. The eventuates displayed when cells were exposed to thermal environment, EGF/EGFR basically did not internalize, mainly gathered on the cell membrane. Our group also researched the effect of EGF's signaling properties, and the results showed that the ability of EGF to activate EGFR-mediated intracellular signaling decreased significantly under heat stress. Finally, this study illustrated that EGF's cell behavior and signaling profile are obviously altered, indicating that heat stress seriously affected the biological activity of EGF/EGFR, which establish a solid foundation for studying the effect of the EGF on testicular tissue under heat environment.

Jumping Rope Improves the Physical Fitness of Preadolescents Aged 10-12 Years: A Meta-Analysis.

To systematically review the effects of jumping rope on physical fitness among 10 to 12-year-old preadolescents, and to provide evidence-based support for its use in school physical education curriculum work. The PubMed, Web of Science, SPORTDiscus, ScienceDirect, and CNKI databases were searched for randomized controlled trials that assessed jumping rope and physical fitness among preadolescents aged 10-12 years. Meta-analyses were used to calculate the standardized mean difference (SMD) values and 95% confidence intervals (CI), and subgroup analyses were conducted on intervention duration, frequency and period. A total of 1048 subjects from 15 studies were included. Compared to regular physical education courses, jumping rope did not show a significant advantage in terms of body morphology. In terms of physical function, boys showed greater improvements in vital capacity, and girls showed greater improvements in resting HR. In terms of physical performance, boys showed greater improvements in speed, upper-body strength, lower-body strength, muscular endurance and agility, while girls showed greater improvements in coordination and balance. Boys experienced a minimal improvement in flexibility, while girls did not show a significant difference. Combining the results of the subgroup analyses, the optimal session time, frequency and intervention length of jumping rope for significantly improving the physical fitness of preadolescents were >40 min, 2 times/week and 8-12 weeks, respectively. In conclusion, jumping rope provides small to large benefits over regular physical education for both boys and girls aged 10-12 years in terms of physical function and physical performance indicators other than flexibility, while showing no significant advantage in terms of body morphology. Based on the available research, it is recommended that children aged 10-12 years participate in jump rope sessions for at least 40 minutes once or twice a week for 8-12 weeks to better develop physical fitness.

Function of MYB8 in larch under PEG simulated drought stress.

Larch, a prominent afforestation, and timber species in northeastern China, faces growth limitations due to drought. To further investigate the mechanism of larch's drought resistance, we conducted full-length sequencing on embryonic callus subjected to PEG-simulated drought stress. The sequencing results revealed that the differentially expressed genes (DEGs) primarily played roles in cellular activities and cell components, with molecular functions such as binding, catalytic activity, and transport activity. Furthermore, the DEGs showed significant enrichment in pathways related to protein processing, starch and sucrose metabolism, benzose-glucuronic acid interconversion, phenylpropyl biology, flavonoid biosynthesis, as well as nitrogen metabolism and alanine, aspartic acid, and glutamic acid metabolism. Consequently, the transcription factor T_transcript_77027, which is involved in multiple pathways, was selected as a candidate gene for subsequent drought stress resistance tests. Under PEG-simulated drought stress, the LoMYB8 gene was induced and showed significantly upregulated expression compared to the control. Physiological indices demonstrated an improved drought resistance in the transgenic plants. After 48 h of PEG stress, the transcriptome sequencing results of the transiently transformed LoMYB8 plants and control plants exhibited that genes were significantly enriched in biological process, cellular component and molecular function. Function analyses indicated for the enrichment of multiple KEGG pathways, including energy synthesis, metabolic pathways, antioxidant pathways, and other relevant processes. The pathways annotated by the differential metabolites mainly encompassed signal transduction, carbohydrate metabolism, amino acid metabolism, and flavonoid metabolism.

Analysis of oxidase activity and transcriptomic changes related to cutting propagation of hybrid larch.

Hybrid larch is the main timber and afforestation tree species in Northeast China. To solve the problem of rooting difficulties in larch cutting propagation, enzyme activity determination and transcriptome sequencing were carried out on the rooting tissues at five timepoints after cutting. peroxidase (POD), indole acetic acid oxidase (IAAO) and polyphenol oxidase (PPO) play important roles in the larch rooting process after cutting. A total of 101.20 Gb of clean data was obtained by transcriptome sequencing, and 43,246 unigenes were obtained after further screening and assembly. According to GO analysis and KEGG enrichment analysis, we think that plant hormones play an important role in the rooting process of larch stem cuttings. in the plant hormone signal transduction pathway, a larch gene c141104.graph_c0 that is homologous to the Arabidopsis AUX1 was found to be significantly up-regulated. We suggest that AUX1 may promote IAA transport in larch, thus affecting adventitious root development. According to the results of POD, PPO IAAO indexes and GO analysis, we think s1 and s2 periods may be important periods in the rooting process of larch stem cuttings, so we built a gene regulatory network, a total of 14genes, including LBD, NAC, AP2/ERF, bHLH and etc., may be important in different stages of cutting propagation. As the rooting rate after cutting inhibits the development of larch clone propagation, identifying the genes that regulate rooting could help us to preliminarily understand the molecular mechanism of adventitious root formation and select a better treatment method for cutting propagation.

The effect of heat stress on the cellular behavior, intracellular signaling profile of porcine growth hormone (pGH) in swine testicular cells.

At present, heat stress caused by the thermal environment is the main factor that endangers the reproductive function of animals. Growth hormone (GH) is a polypeptide hormone, the biological function of reproductive organs has been reported, and it has many important physiological functions in the body. However, so far, the behavior and signal transduction of GH in testicular cells under heat stress are still unclear. To this end, in the current work, we use a swine testicular cell line (ST) as an in vitro model to explore the cell behavior and intracellular signaling profile of porcine growth hormone (pGH) under heat stress; the results showed that when cells were under heat stress, pGH and GHR were basically not internalized, and a large number of them accumulated on the cell membrane. In addition, we also studied the effect of pGH on the JAK2-STATs signaling pathway and IGF-1 expression under heat stress, we found that the ability of pGH to activate the JAK-STATs signaling pathway and IGF-1 under heat stress was greatly reduced (p < 0.05). In conclusion, our research shows that when cells undergo heat stress, the internalization of pGH and GHR were inhibited, and the activation of the JAK2-STATs signaling pathway and IGF-1 expression were reduced; this lays a solid foundation for further research on the effect of pGH on swine testicular tissue under thermal environment.

Genetic transformation of LoHDZ2 and analysis of its function to enhance stress resistance in Larix olgensis.

To study the function of LoHDZ2 in larch, we first constructed a VB191103-LoHDZ2::GUS overexpression vector. Through Agrobacterium-mediated infection, the expression vector was transferred into a larch embryogenic cell line. A stable resistant cell line was subsequently screened, and mature embryos were induced to grow until they developed into seedlings. Antagonistic cell lines were identified at both the DNA and RNA levels. The transgenic cell lines were then subjected to GUS staining, and transgenic cell lines were ultimately identified and obtained. These transgenic cell lines were sequenced to identify differentially expressed genes, and a cluster analysis was performed. The resistant cell lines were cultured under stress conditions involving 20% PEG6000 and 200 mM NaCl proliferation media (1/10-BM). After the stress treatment, the contents of peroxidase (POD), malondialdehyde (MDA) and superoxide dismutase (SOD) in both wild-type and transgenic cell lines were measured. The results are summarized below: (1) When the specific fragment of the target gene in the genome of the resistant cell line was amplified. At the RNA level, the expression of the fragment in four resistant lines increased. In addition, GUS staining showed a blue reaction, indicating that LoHDZ2 was successfully integrated into the larch embryonic cell lines. (2) To verify the accuracy and reliability of the transcriptome data, 10 differentially expressed genes (5 upregulated and 5 down regulated genes) were subjected to qRT-PCR verification. The results showed that the expression trend of the 10 differentially expressed genes was the same as that revealed by RNA-Seq, indicating that the transcriptome data were reliable. (3) The transcriptome sequencing showed that 176 genes were upregulated and that 140 genes were down regulated. Through GO enrichment analysis and KEGG metabolic pathway analysis, the screened differentially expressed genes were related to biological processes such as larch metabolism and response to stimuli, indicating that these genes may be closely involved in the regulation of the larch response to external stimuli, including heat stress, drought stress, metal ion stress and bacterial infection, and may participate in the growth process. (4) After 20% PEG6000 treatment, the POD enzyme activity of the transgenic cell line was greater than that of the wild-type; this activity could effectively remove the amount of peroxide produced. The MDA content of the transgenic cell lines was lower than that of the wild-type cell lines, and the accumulation degree of harmful substances was low, indicating that the degree of oxidative damage of the transgenic cell lines was lower than that of the wild-type cell lines. The SOD content of the transgenic cell lines was lower than that of the wild-type cell lines, indicating that the drought resistance of the transgenic cell lines was enhanced. After 200 mM NaCl treatment, although the increase in SOD content was not obvious, the same trend was detected, indicating that the resistance of the transgenic cell lines was indeed stronger than that of the wild-type cell lines. According to the results of previous experiments, after this gene was overexpressed in tobacco, the transformed plants showed obvious dwarfing, which may indicate that the stress resistance of the plant was enhanced. In conclusion, a transgenic larch cell line was successfully obtained, and transgenic larch seedlings were successfully induced. LoHDZ2 may participate in the response of plants to the external environment, and may participate in the growth and development of Larix olgensis by affecting plant metabolic pathways.

The Cellular Behavior, Intracellular Signaling Profile and Nuclear-Targeted Potential Functions of Porcine Growth Hormone (pGH) in Swine Testicular Cells.

Porcine growth hormone (pGH) has many important biological functions and roles, and the biological activity of pGH is closely related with its cell behavior and characteristics. However, so far, the behavior of pGH in swine testicular cell remains unclear. For this, in the current work, the swine testicular cell line (ST) was used as an in vitro model, and CLSM (Confocal laser scanning microscope), IFA (Indirect immunofluorescence assay), FCM (Flow cytometry) and WB (Western-blotting) were used to explore the pGH's cell behivior and function, and the results showed that pGH and GHR could internalize into ST cell and transported to the nucleus. Furthermore, we studied the internalization kinetics of pGH and GHR on ST cell, and found that pGH and GHR internalizes into ST cell in a time-dependent manner. More importantly, we also investigated the potential molecular functions of pGH-GHR after it entered into the cell nuclei. The results indicated that nuclear-localized GHR could participate in cell proliferation by regulating the signal intensity of STAT5. In summary, our current research shows that the nuclear-localized pGH-GHR participates in the cell proliferation of ST cell, which lays a solid foundation for further research on the regulatory effect of pGH on testicular tissue.

The effect of growth hormone-induced cellular behavior and signaling properties on induced cellular senescence in human mesenchymal stem cells.

Senescence is associated with a decline in physiological function, which is accompanied by onset of diseases. Growth hormone (GH) is a class of growth-promoting cytokines with reduced secretion in aging populations. However, the effect of senescence on GH bioactivity is not fully understood in human mesenchymal stem cells (hMSCs). In this work, GH-induced cellular behavior and intracellular signaling transduction were explored in senescent hMSCs. Therefore, hMSCs were used to establish a senescence model by H2O2 treatment for this study. First, we investigated the effects of cellular senescence on the cell behavior of GH. The experimental results suggested that GH could not be internalized into the nucleus, and a significant reduction in GH internalization into the cytoplasm was observed in senescent hMSCs compared to the control group. Second, the effect of cellular senescence on GH-mediated intracellular signaling pathways was investigated by Western blotting. For this, the signaling molecule activation of Janus kinase 2 (JAK2)/signal transducer and activator transcription (STAT) stimulated by GH was detected. Our data indicated that the signaling intensity of p-JAK2, p-STAT5, p-STAT3 and p-STAT1 was considerably weakened. Taken together, these findings provide important insights into the impaired effects of cellular senescence on the biological activity of GH.

Genetic transformation and growth index determination of the Larix olgensis LoHDZ2 transcription factor gene in tobacco.

Homeodomain-leucine zippers (HD-Zip) are plant-specific transcription factors that participate in different plant development processes and differentially regulate metabolic processes. LoHDZ2 is an HD-ZipII subfamily transcription factor gene that we identified from a transcriptomic analysis of Larix olgensis. To understand its function, we built a LoHDZ2 expression vector and then inserted it into tobacco by genetic transformation. Transgenic plants were identified at the DNA and RNA levels. Phenotypic index analysis of transgenic tobacco showed dwarfed growth with larger leaves and earlier flowering than the wild type. LoHDZ2 was expressed differently after hormone treatment with IAA, MeJA and 2,4-D. The results suggested that LoHDZ2 may respond to hormones and be involved in regulating growth and metabolism. These results helped us better understand the function of LoHDZ2 and provided a candidate gene for Larix olgensis molecular breeding.

How to Avoid Postoperative Remaining Ossification Mass in Anterior Controllable Antedisplacement and Fusion Surgery.

OBJECTIVES: To retrospectively review the cases with ossification of the posterior longitudinal ligament (OPLL) treated with anterior controllable antedisplacement and fusion (ACAF). Patients with postoperative remaining ossification mass (PROM) are analyzed to figure out the causes and preventions of this problem. METHODS: A total of 115 patients were included. PROM was identified as remaining OPLL existed in the spinal canal other than included in the vertebral-OPLL complex on postoperative computed tomography. The Japanese Orthopaedic Association scoring system was used to evaluate the neurologic status. Surgery-related complications such as cerebrospinal fluid (CSF) leakage and spinal cord or nerve injury were all recorded. The patients with the PROM group and those without the PROM group were compared. RESULTS: There were 14 patients with wide-base OPLL (12.2%) and 10 patients (8.7%) with PROM among the 115 patients with OPLL. The 10 patients with PROM were all with wide-base OPLL. The average improvement rate of Japanese Orthopaedic Association score in patients without PROM was significantly larger than that in patients with PROM (69.5 ± 22.6% vs. 36.7 ± 22.0, P < 0.01). Incidence rate of postoperative CSF leakage and neural deterioration were significantly higher in patients with PROM than that in patients without PROM (CSF leakage, 40.0% vs. 5.9%; neural deterioration, 50.0% vs. 3.0%). No other complications were observed. CONCLUSIONS: The occurrence of PROM might cause complications and poor neural function recovery in patients treated with ACAF. Surgical techniques should be noted to avoid PROM in ACAF surgery.

[The clinical significance of serum tuberculosis specific antigen antibody in the diagnosis of tuberculosis].

OBJECTIVE: To evaluate the significance of serum tuberculosis specific antigen (TB-SA) antibody detection in the diagnosis of tuberculosis. METHODS: TB-SA antibody in the serum samples from 829 cases of tuberculosis, 278 patients with non-tuberculosis lung diseases and 125 healthy volunteers were detected using enzyme-linked immunosorbent assay (ELISA). Tuberculosis was confirmed by clinical, bacteriology, X-ray examination and pathology studies. RESULTS: The sensitivity of TB-SA antibody in diagnosis of bacteriologically positive and negative pulmonary tuberculosis and extrapulmonary tuberculosis was 75.1% (272/362), 68.9% (226/328) and 71.2% (99/139), respectively; the combined sensitivity was 72.0% (597/829), and the specificity 82.1% (331/403). The relationship between the OD(405) of TB-SA antibody and PPD positivity showed no linear relationship, suggesting that BCG vaccination did not affect the value of serum TB-SA antibody. CONCLUSION: The results suggest that measurement of serum TB-SA is a relatively sensitive and specific method for the diagnosis and differential diagnosis of tuberculosis.