The lncRNA1-miR6288b-3p-PpTCP4-PpD2 module regulates peach branch number by affecting brassinosteroid biosynthesis.

Branch number is one of the most important agronomic traits of fruit trees such as peach. Little is known about how LncRNA and/or miRNA modules regulate branching through transcription factors. Here, we used molecular and genetic tools to clarify the molecular mechanisms underlying brassinosteroid (BR) altering plant branching. We found that the number of sylleptic branch and BR content in pillar peach ('Zhaoshouhong') was lower than those of standard type ('Okubo'), and exogenous BR application could significantly promote branching. PpTCP4 expressed great differentially comparing 'Zhaoshouhong' with 'Okubo'. PpTCP4 could directly bind to DWARF2 (PpD2) and inhibited its expression. PpD2 was the only one differentially expressed key gene in the path of BR biosynthesis. At the same time, PpTCP4 was identified as a target of miR6288b-3p. LncRNA1 could act as the endogenous target mimic of miR6288b-3p and repress expression of miR6288b-3p. Three deletions and five SNP sites of lncRNA1 promoter were found in 'Zhaoshouhong', which was an important cause of different mRNA level of PpTCP4 and BR content. Moreover, overexpressed PpTCP4 significantly inhibited branching. A novel mechanism in which the lncRNA1-miR6288b-3p-PpTCP4-PpD2 module regulates peach branching number was proposed.

Development and Application of a Multi-Component Exercise Training Program for Elderly COPD Patients with Skeletal Muscle Dysfunction.

Objective: To develop a multi-component exercise training program for elderly patients with chronic obstructive pulmonary disease (COPD) and skeletal muscle dysfunction (SMD), and to explore its application value. Methods: This study involved 68 elderly patients with both COPD and SMD who were hospitalized at The First Affiliated Hospital of Hainan Medical College from September 2019 to August 2021. They were randomly divided into an intervention group and a control group. The intervention group received a 12-month multi-component exercise training program, while the control group received routine intervention. We compared skeletal muscle function, strength, lung function, and oxygen saturation between the groups before and at 3, 6, and 12 months after the intervention. The patient's daily living activities and dyspnea severity were assessed using the Modified Barthel Index (MBI) and the modified British Medical Research Council (mMRC) dyspnea scale. Flow cytometry was used to determine changes in the proportion of CD3+ T cells, CD3+CD4+ T cells, CD3+CD8+ T cells, and the CD3+CD4+/CD3+CD8+ ratio in the patients' lymphocytes before and after the intervention. Results: The six-minute walk distance of the intervention group at 6 and 12 months after the intervention was longer than that of the control group. The peak torque of elbow flexion and extension and knee flexion and extension muscles of the intervention group at 12 months after intervention was higher than that of the control group (P < .05). The forced expiratory volume in one second (FEV1), forced vital capacity (FVC), and FEV1/FVC of the intervention group at 6 and 12 months after intervention were higher than those of the control group. The oxygen saturation (SaO2) and partial pressure of oxygen (PaO2) of the intervention group at 6 and 12 months after the intervention were higher than those of the control group (P < .05). The MBI and mMRC scores of the intervention group at 6 and 12 months after intervention were higher than those of the control group (P < .05). The indexes of T lymphocyte immune function, CD3+T cells, CD3+CD4+T cells, and the ratio of CD3+CD4+/CD3+CD8+ in the intervention group were also higher than those in the control group at 3, 6, and 12 months (P < .05). Conclusion: Multi-component exercise training program intervention can effectively improve skeletal muscle function and lung function in elderly COPD patients with SMD, reduce the degree of hypoxia and dyspnea, and improve patients' daily living activities and immune function recovery.

De novo chromosome-level genome of a semi-dwarf cultivar of Prunus persica identifies the aquaporin PpTIP2 as responsible for temperature-sensitive semi-dwarf trait and PpB3-1 for flower type and size.

Peach (Prunus persica) is one of the most important fruit crops globally, but its cultivation can be hindered by large tree size. 'Zhongyoutao 14' (CN14) is a temperature-sensitive semi-dwarf (TSSD) cultivar which might be useful as breeding stock. The genome of CN14 was sequenced and assembled de novo using single-molecule real-time sequencing and chromosome conformation capture assembly. A high-quality genome was assembled and annotated, with 228.82 Mb mapped to eight chromosomes. Eighty-six re-sequenced F1 individuals and 334 previously re-sequenced accessions were used to identify candidate genes controlling TSSD and flower type and size. An aquaporin tonoplast intrinsic protein (PpTIP2) was a strong candidate gene for control of TSSD. Sequence variations in the upstream regulatory region of PpTIP2 correlated with different transcriptional activity at different temperatures. PpB3-1, a candidate gene for flower type (SH) and flower size, contributed to petal development and promoted petal enlargement. The locus of another 12 agronomic traits was identified through genome-wide association study. Most of these loci exhibited consistent and precise association signals, except for flesh texture and flesh adhesion. A 6015-bp insertion in exon 3 and a 26-bp insertion upstream of PpMYB25 were associated with fruit hairless. Along with a 70.5-Kb gap at the F-M locus in CN14, another two new alleles were identified in peach accessions. Our findings will not only promote genomic research and agronomic breeding in peach but also provide a foundation for the peach pan-genome.

Parallel Optimisation and Implementation of a Real-Time Back Projection (BP) Algorithm for SAR Based on FPGA.

This study conducts an in-depth evaluation of imaging algorithms and software and hardware architectures to meet the capability requirements of real-time image acquisition systems, such as spaceborne and airborne synthetic aperture radar (SAR) systems. By analysing the principles and models of SAR imaging, this research creatively puts forward the fully parallel processing architecture for the back projection (BP) algorithm based on Field-Programmable Gate Array (FPGA). The processing time consumption has significant advantages compared with existing methods. This article describes the BP imaging algorithm, which stands out with its high processing accuracy and two-dimensional decoupling of distance and azimuth, and analyses the algorithmic flow, operation, and storage requirements. The algorithm is divided into five core operations: range pulse compression, upsampling, oblique distance calculation, data reading, and phase accumulation. The architecture and optimisation of the algorithm are presented, and the optimisation methods are described in detail from the perspective of algorithm flow, fixed-point operation, parallel processing, and distributed storage. Next, the maximum resource utilisation rate of the hardware platform in this study is found to be more than 80%, the system power consumption is 21.073 W, and the processing time efficiency is better than designs with other FPGA, DSP, GPU, and CPU. Finally, the correctness of the processing results is verified using actual data. The experimental results showed that 1.1 s were required to generate an image with a size of 900 × 900 pixels at a 200 MHz clock rate. This technology can solve the multi-mode, multi-resolution, and multi-geometry signal processing problems in an integrated manner, thus laying a foundation for the development of a new, high-performance, SAR system for real-time imaging processing.

Phylogenetic and Transcriptional Analyses of the HSP20 Gene Family in Peach Revealed That PpHSP20-32 Is Involved in Plant Height and Heat Tolerance.

The heat shock protein 20 (HSP20) proteins comprise an ancient, diverse, and crucial family of proteins that exists in all organisms. As a family, the HSP20s play an obvious role in thermotolerance, but little is known about their molecular functions in addition to heat acclimation. In this study, 42 PpHSP20 genes were detected in the peach genome and were randomly distributed onto the eight chromosomes. The primary modes of gene duplication of the PpHSP20s were dispersed gene duplication (DSD) and tandem duplication (TD). PpHSP20s in the same class shared similar motifs. Based on phylogenetic analysis of HSP20s in peach, Arabidopsis thaliana, Glycine max, and Oryza sativa, the PpHSP20s were classified into 11 subclasses, except for two unclassified PpHSP20s. cis-elements related to stress and hormone responses were detected in the promoter regions of most PpHSP20s. Gene expression analysis of 42 PpHSP20 genes revealed that the expression pattern of PpHSP20-32 was highly consistent with shoot length changes in the cultivar 'Zhongyoutao 14', which is a temperature-sensitive semi-dwarf. PpHSP20-32 was selected for further functional analysis. The plant heights of three transgenic Arabidopsis lines overexpressing PpHSP20-32 were significantly higher than WT, although there was no significant difference in the number of nodes. In addition, the seeds of three over-expressing lines of PpHSP20-32 treated with high temperature showed enhanced thermotolerance. These results provide a foundation for the functional characterization of PpHSP20 genes and their potential use in the growth and development of peach.

Long-term infection of SARS-CoV-2 changed the body's immune status.

The outbreak of SARS-CoV-2-associated pneumonia, a disease called COVID-19, has caused a pandemic worldwide. To investigate the immune responses after infection of SARS-CoV-2 in non-critical patients may help to better understand the disease progression. We collected 334 confirmed COVID-19 cases including 212 still in hospital with nucleic acid test positive on halfway for SARS-CoV-2 and 122 discharged from hospital, compared specific antibodies, immune cells, and cytokine changes between the hospitalized and discharged patients. The hospitalized patients had a longer illness time compared with discharged patients. Analysis of viral loads explained long-term or persistent infection of SARS-CoV-2, which existed with the median time of 18.5 days of the positive nucleic acid test. Serum analysis showed that the specific anti-N IgG antibody was positive in all detected patients after infection of two weeks. Neutrophils, Monocytes, NK cells, and CD4+ T cells significantly increased, while total lymphocytes and CD8+ T cells decreased from non-critical hospitalized patients after longer-term infection. Further analysis of the cytokines showed that IL-6, TNF-α, IFN-γ, IL-2, IL-4, and IL-10 from the hospitalized patients were significantly higher, indicating a potential of the increased CD4+ T cell differentiation.

Head-to-head comparative pharmacokinetic and biodistribution (PK/BD) study of two dexamethasone prodrug nanomedicines on lupus-prone NZB/WF1 mice.

HPMA copolymer-based dexamethasone prodrug (P-Dex) and PEG-based dexamethasone prodrug (PEG-Dex, ZSJ-0228) were previously found to passively target the inflamed kidney and provide potent and sustained resolution of nephritis in NZB/WF1 lupus-prone mice. While both prodrug nanomedicines effectively ameliorate lupus nephritis, they have demonstrated distinctively different safety profiles. To explore the underlining mechanisms of these differences, we conducted a head-to-head comparative PK/BD study of P-Dex and PEG-Dex on NZB/WF1 mice. Overall, the systemic organ/tissue exposures to P-Dex and Dex released from P-Dex were found to be significantly higher than those of PEG-Dex. The high prodrug concentrations were sustained in kidney for only 24 h, which cannot explain their lasting therapeutic efficacy (>1 month). P-Dex showed sustained presence in liver, spleen and adrenal gland, while the presence of PEG-Dex in these organs was transient. This difference in PK/BD profiles may explain PEG-Dex' superior safety than P-Dex.

Breastfeeding practices and associated factors at the individual, family, health facility and environmental levels in China.

We examined the association between breastfeeding practices and associated factors using cross-sectional data from face-to-face interviews with 9,745 mother-child dyads in China. The study collected information on breastfeeding practices and potential associated factors at the individual, family, health facility and environmental levels in China. We used survey commands in Stata to consider sampling weight and survey design effects. Although breastfeeding was the norm (97.4% ever breastfed), the prevalence of early initiation of breastfeeding (EIBF) in 0-11 months old was 8.2%, exclusive breastfeeding (EBF) in 0-5 months old was 27.8% and breastfeeding on the previous day in 6-11 months old was 77.5%. The prevalence of EIBF was lower for caesarean delivery and among mothers belonging to ethnic minority groups. The prevalence of EBF was higher among mothers who practiced EIBF, received information that encouraged breastfeeding and knew that a baby should be breastfed on demand and exclusively. By contrast, the prevalence of EBF was lower in mothers who received infant formula advice or felt uneasy breastfeeding in public places. The prevalence of breastfeeding on the previous day was higher among mothers whose partners supported breastfeeding and who knew about timing of colostrum production, EBF for 6 months, and to nurse more to stimulate milk production. The prevalence of breastfeeding on the previous day was lower in mothers who received infant formula advice or felt uneasy breastfeeding in public places. In conclusion, we found that the prevalence of EIBF and EBF practices in China was low and associated with factors at individual, family, health facility and environmental levels.

Identification of a p.Trp403* nonsense variant in PHEX causing X-linked hypophosphatemia by inhibiting p38 MAPK signaling.

X-linked hypophosphatemia (XLH) is the most common hereditary rickets, caused by mutations in PHEX encoding the phosphate regulating endopeptidase homolog X-linked. Here, we report a nonsense variant in exon 11 of PHEX (c.1209G>A p.Trp403*) cosegregating with XLH in a Chinese family with a LOD score of 2.70. Real-time reverse transcription polymerase chain reaction analysis demonstrated that p.Trp403* variant did not cause nonsense-mediated mRNA decay (NMD), but significantly increased the expression level of FGF23 mRNA in the patients. Interestingly, p.Trp403* significantly reduced phosphorylation of p38 mitogen-activated protein kinase (MAPK) but not ERK1/2. Moreover, overexpression of FGF23 significantly decreased phosphorylation of p38 MAPK, whereas knockdown of FGF23 by siRNA significantly increased phosphorylation of p38 MAPK. These data suggest that p.Trp403* may not function via an NMD mechanism, and instead causes XLH via a novel signaling mechanism involving PHEX, FGF23, and p38 MAPK. This finding provides important insights into genetic and molecular mechanisms for the pathogenesis of XLH.

The Development of a Macromolecular Analgesic for Arthritic Pain.

The addictive potential of clinically used opioids as a result of their direct action on the dopaminergic reward system in the brain has limited their application. In an attempt to reduce negative side effects as well as to improve the overall effectiveness of these analgesics, we have designed, synthesized, and evaluated an N-(2-hydroxypropyl)methacrylamide (HPMA)-based macromolecular prodrug of hydromorphone (HMP), a commonly used opioid. To this end, P-HMP was synthesized via RAFT polymerization and a subsequent polymer analogous reaction. Its interaction with inflammatory cells in arthritic joints was evaluated in vitro using a RAW 264.7 cell culture, and subsequent confocal microscopy analysis confirmed that P-HMP could be internalized by the cells via endocytosis. In vivo imaging studies indicated that the prodrug can passively target the arthritic joint after systemic administration in a rodent model of monoarticular adjuvant-induced arthritis (MAA). The inflammatory pain-alleviating properties of the prodrug were assessed in MAA rats using the incapacitance test and were observed to be similar to dose-equivalent HMP. Analgesia through mechanisms at the spinal cord level was further measured using the tail flick test, and it was determined that the prodrug significantly reduced spinal cord analgesia versus free HMP, further validating the peripheral restriction of the macromolecular prodrug. Immunohistochemical analysis of cellular uptake of the P-HMP within the MAA knee joint proved the internalization of the prodrug by phagocytic synoviocytes, colocalized with HMP's target receptor as well as with pain-modulating ion channels. Therefore, it can be concluded that the novel inflammation-targeting polymeric prodrug of HMP (P-HMP) has the potential to be developed as an effective and safe analgesic agent for musculoskeletal pain.

Local Application of Pyrophosphorylated Simvastatin Prevents Experimental Periodontitis.

PURPOSE: Simvastatin (SIM), a HMG-CoA reductase inhibitor widely prescribed for hypercholesterolemia, has been reported to ameliorate inflammation and promote osteogenesis. Its clinical applications on these potential secondary indications, however, have been hampered by its lack of osteotropicity and poor water solubility. To address this challenge, we propose to design and evaluate the therapeutic efficacy of a novel simvastatin prodrug with better water solubility and bone affinity. METHOD: The prodrug (SIM-PPi) was synthesized by directly conjugating a SIM trimer to a pyrophosphate (PPi). It was characterized and evaluated in vitro for its water solubility, osteotropicity, toxicity, anti-inflammatory and osteoinductive properties. It was then tested for anti-inflammatory and osteoinductive properties in vivo by three weekly injections into gingiva of a ligature-induced experimental periodontitis rat model. RESULTS: In vitro studies showed that SIM-PPi has greatly improved water-solubility of SIM and shows strong binding to hydroxyapatite (HA). In macrophage culture, SIM-PPi inhibited LPS-induced pro-inflammatory cytokines (IL-1ß, IL-6). In osteoblast culture, it was found to significantly increase alkaline phosphatase (ALP) activity with accelerated mineral deposition, confirming the osteogenic potential of SIM-PPi. When tested in vivo on an experimental periodontal bone-loss model, SIM-PPi exhibited a superior prophylactic effect compared to dose equivalent SIM in reducing inflammatory cells and in preserving alveolar bone structure, as shown in the histological and micro-CT data. CONCLUSION: SIM-PPi may have the potential to be further developed for better clinical management of bone loss associated with periodontitis.

Serum miR-146a, miR-155, and miR-210 as potential markers of Graves' disease.

BACKGROUND: Previous studies have demonstrated that dysfunctional regulatory T cells (Tregs) may be associated with Graves' disease (GD). In this study, we evaluated four serum Treg-associated miRNAs (miR-210, miR-182, miR-155, and miR-146a) expressions and assessed the potential of serum miRNAs as biomarkers of GD. METHODS: Foxp3 and serum miRNAs expressions both in GD patients and healthy controls were measured by RT-PCR. RESULTS: Serum miR-210 in GD patients was significantly higher than that of healthy controls (2.64-fold, P<.001); in contrast, miR-155 and miR-146a were lower (P<.001 and P=.008). No significant difference was found in miR-182. ROC curve analysis indicated that miR-210, miR-155, and miR-146a with the area under ROC (AUC) of 0.803 (70.0% sensitivity and 83.1% specificity), 0.796 (76.3% sensitivity and 76.9% specificity), and 0.736 (68.8% sensitivity and 73.8% specificity), respectively, could differentiate GD patients from healthy controls. Combination of three miRNAs yielded an AUC of 0.976 (91.3% sensitivity and 93.8% specificity) with 92.41% diagnostic efficiency. In addition, serum miR-210 and miR-155 in GD were associated with the extent of goiter. Three miRNAs levels were different by gender. Besides, serum miR-210 was positively correlated with free thyroxine (FT4) and thyrotrophin receptor antibody (TRAb) level. CONCLUSION: The serum levels of miR-210, miR-155, and miR-146a may be potential new markers for the diagnosis of GD and play important roles in GD pathogenesis.

Bone-targeting liposome formulation of Salvianic acid A accelerates the healing of delayed fracture Union in Mice.

Delayed fracture union is a significant clinical challenge in orthopedic practice. There are few non-surgical therapeutic options for this pathology. To address this challenge, we have developed a bone-targeting liposome (BTL) formulation of salvianic acid A (SAA), a potent bone anabolic agent, for improved treatment of delayed fracture union. Using pyrophosphorylated cholesterol as the targeting ligand, the liposome formulation (SAA-BTL) has demonstrated strong affinity to hydroxyapatite in vitro, and to bones in vivo. Locally administered SAA-BTL was found to significantly improve fracture callus formation and micro-architecture with accelerated mineralization rate in callus when compared to the dose equivalent SAA, non-targeting SAA liposome (SAA-NTL) or no treatment on a prednisone-induced delayed fracture union mouse model. Biomechanical analyses further validated the potent therapeutic efficacy of SAA-BTL. These results support SAA-BTL formulation, as a promising therapeutic candidate, to be further developed into an effective and safe clinical treatment for delayed bone fracture union.

Bioinspired Synthesis of (+)-Cinchonidine Using Cascade Reactions.

The development of efficient syntheses of complex natural products has long been a major challenge in synthetic chemistry. Designing cascade reactions and employing bioinspired transformations are an important and reliable means of achieving this goal. Presented here is a combination of these two strategies, which allow efficient asymmetric synthesis of the cinchona alkaloid (+)-cinchonidine. The key steps of this synthesis are a controllable, visible-light-induced photoredox radical cascade reaction to efficiently access the tetracyclic monoterpenoid indole alkaloid core, as well as a practical biomimetic cascade rearrangement for the indole to quinoline transformation. The use of stereoselective chemical transformations in this work makes it an efficient synthesis of (+)-cinchonidine.

Association between FOXP3 gene polymorphisms and risk of differentiated thyroid cancer in Chinese Han population.

BACKGROUND: Foxp3 plays important roles in autoimmune and inflammatory diseases as well as human malignancies. This study aimed to investigate the association between Foxp3 gene polymorphisms and the susceptibility to differentiated thyroid cancers (DTC). METHODS: Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) in 350 DTC patients and 306 healthy controls. FOXP3 relative expression was measured by real-time quantitative PCR (qRT-PCR). RESULTS: AA/AC genotype of Foxp3-rs3761548 was associated with a higher risk of DTC. The frequency of Foxp3-rs2280883 CC/CT genotype was lower in DTC patients. Besides, the AA/AC genotype of rs3761548 was more frequent in female DTC than male DTC. The association between two single nucleotide polymorphisms (SNPs) and clinical characteristics of DTC was further analyzed. We found that rs3761548 AA/AC genotype was more frequent in severe DTC patients (tumor diameter >1 cm) compared with the relative tender DTC patients (tumor diameter <1 cm). On the contrast, the frequency of rs2280883 CC/CT genotype was lower in severe DTC patients. In addition, the Foxp3 relative expression in DTC with AA/AC genotype of rs3761548 was higher than that of DTC with CC genotype. CONCLUSION: Our findings suggested that Foxp3 polymorphisms were associated with the risk of DTC in Chinese Han population.

Dextranase from Arthrobacter oxydans KQ11-1 inhibits biofilm formation by polysaccharide hydrolysis.

Dental plaque is a biofilm of water-soluble and water-insoluble polysaccharides, produced primarily by Streptococcus mutans. Dextranase can inhibit biofilm formation. Here, a dextranase gene from the marine microorganism Arthrobacter oxydans KQ11-1 is described, and cloned and expressed using E. coli DH5α competent cells. The recombinant enzyme was then purified and its properties were characterized. The optimal temperature and pH were determined to be 60°C and 6.5, respectively. High-performance liquid chromatography data show that the final hydrolysis products were glucose, maltose, maltotriose, and maltotetraose. Thus, dextranase can inhibit the adhesive ability of S. mutans. The minimum biofilm inhibition and reduction concentrations (MBIC50 and MBRC50) of dextranase were 2 U ml-1 and 5 U ml-1, respectively. Scanning electron microscopy and confocal laser scanning microscope (CLSM) observations confirmed that dextranase inhibited biofilm formation and removed previously formed biofilms.

Total syntheses of (-)-isoschizogamine and (-)-2-hydroxyisoschizogamine.

Total syntheses of (-)-isoschizogamine and (-)-2-hydroxyisoschizogamine are described. The synthesis employs two asymmetric Michael additions to establish chiral centers at C7 and the quaternary carbon C20. Regioselective reduction of the methylthioiminium cation rather than the enamine generates an isoschizogamine-type pentacyclic skeleton. Acidic hydrolysis of the isoschizogamine-type intermediate in the absence of oxygen provides natural (-)-isoschizogamine. Conducting the reaction in the presence of oxygen leads to a multistep oxidative hydrolysis cascade that affords unnatural (-)-2-hydroxyisoschizogamine.

Enhanced production of L-sorbose from D-sorbitol by improving the mRNA abundance of sorbitol dehydrogenase in Gluconobacter oxydans WSH-003.

BACKGROUND: Production of L-sorbose from D-sorbitol by Gluconobacter oxydans is the first step to produce L-ascorbic acid on industrial scale. The sldhAB gene, which encodes the sorbitol dehydrogenase (SLDH), was overexpressed in an industrial strain G. oxydans WSH-003 with a strong promoter, P tufB . To enhance the mRNA abundance, a series of artificial poly(A/T) tails were added to the 3'-terminal of sldhAB gene. Besides, their role in sldhAB overexpression and their subsequent effects on L-sorbose production were investigated. RESULTS: The mRNA abundance of the sldhAB gene could be enhanced in G. oxydans by suitable poly(A/T) tails. By self-overexpressing the sldhAB gene in G. oxydans WSH-003 with an optimal poly(A/T) tail under the constitutive promoter P tufB , the titer and the productivity of L-sorbose were enhanced by 36.3% and 25.0%, respectively, in a 1-L fermenter. Immobilization of G. oxydans-sldhAB6 cells further improved the L-sorbose titer by 33.7% after 20 days of semi-continuous fed-batch fermentation. CONCLUSIONS: The artificial poly(A/T) tails could significantly enhance the mRNA abundance of the sldhAB. Immobilized G. oxydans-sldhAB6 cells could further enlarge the positive effect caused by enhanced mRNA abundance of the sldhAB.

Investigation into the effect of hepatitis B virus on apoliprotein A1 expression and its mechanism.

BACKGROUND: Hepatitis B virus (HBV) infection poses a serious threat to human health, with China being one of the highly affected countries. However, the pathogenesis of chronic hepatitis B (CHB) is still unclear. Apolipoprotein A1 (ApoA1) which represents the major protein component of high-density lipoprotein is normally secreted by hepatocytes. When hepatocytes are infected with HBV may lead to the disruption of ApoA1 secretion. In this study, we investigated the effect of HBV on ApoA1 expression and preliminarily explored its molecular mechanism of regulation for revealing the pathogenesis of CHB. METHODS: The expression of mRNA and protein of ApoA1 in Human HepG2 hepatoblastoma cells and subline HepG2.2.15 cells were performed by reverse transcription-polymerase chain reaction (RT-PCR) and Western-blot. The serum ApoA1, by the immune turbidimetric test, and high-density lipoprotein cholesterol (HDL-C) in CHB patients and healthy controls, based on the enzymatic method, were measured with autobiochemical analyzer. The statistical difference was analyzed by SPSS 13.0. HBV infectious clone, pHBV1.3, and ApoA1 gene promoter were co-transfected into HepG2, and the luciferase activity was determined. The changes of ApoA1 mRNA and protein expression were detected by RT-PCR and Western-blot method, after HepG2 cells were transfected with pHBV1.3. RESULTS: The expression of ApoA1 mRNA and protein in HepG2.2.15 were lower than those in HepG2, and when compared with healthy controls, serum levels of ApoA1 and HDL-C in CHB patients were lower (P < 0.05). pHBV1.3 in HepG2 cells restrained the activity of ApoA1 promoter, mRNA and protein expression. CONCLUSIONS: HBV could inhibit the expression of ApoA1 in vitro and in vivo.

A simple and efficient gene functional analysis method for studying the growth and development of peach seedlings.

Stable genetic transformation of peach [Prunus persica (L.) Batsch] still faces many technical challenges, and existing transient expression methods are limited by tissue type or developmental stage, making it difficult to conduct functional analysis of genes regulating shoot growth. To overcome this dilemma, we developed a three-step method for efficient analysis of gene functions during peach seedling growth and development. This method resulted in transformation frequencies ranging from 48 to 87%, depending on the gene. From transformation of germinating seeds to phenotyping of young saplings took just 1.5 months and can be carried out any time of year. To test the applicability of this method, the function of three tree architecture-related genes, namely PpPDS, PpMAX4, and PpWEEP, and two lateral root-related genes, PpIAA14-1 and -2, were confirmed. Since functional redundancy can challenge gene functional analyses, tests were undertaken with the growth-repressor DELLA, which has three homologous genes, PpDGYLA (DG), PpDELLA1 (D1), and -2 (D2), in peach that are functionally redundant. Silencing using a triple-target vector (TRV2-DG-D1-D2) resulted in transgenic plants taller than those carrying just TRV2-DG or TRV2. Simultaneously silencing the three DELLA genes also attenuated the stature of two dwarf genotypes, 'FHSXT' and 'HSX', which normally accumulate DELLA proteins. Our study provides a method for the functional analysis of genes in peach and can be used for the study of root, stem, and leaf development. We believe this method can be replicated in other woody plants.