Molecular pathways of osmoregulation in response to salinity stress in the gills of the scalloped spiny lobster (Panulirus homarus) within survival salinity.

Scalloped spiny lobster (Panulirus homarus) aquaculture is the preferred strategy to resolve the conflict between supply and demand for lobster. Environmental conditions, such as salinity, are key to the success of lobster aquaculture. However, physiological responses of P. homarus to salinity stress have not been well studied. This study investigated the gill histology, osmoregulation and gill transcriptome of the early juvenile P. homarus (weight 19.04 ± 3.95 g) cultured at salinity 28 (control), 18, and 38 for 6 weeks. The results showed that the gill filaments of P. homarus exposed to low salinity showed severe separation of the cuticle and epithelial cells due to water absorption and swelling, as well as the dissolution and thinning of the cuticle and the rupture of the septum that separates the afferent and efferent channels. The serum osmolarity of P. homarus varied proportionately with external medium salinity and remained consistently above ambient osmolarity. The serum Na+, Cl-, K+, and Mg2+ concentrations P. homarus exhibited a pattern similar to that of serum osmolality, while the concentration of Ca2+ remained unaffected at salinity 18 but significantly increased at salinity 38. Gill Na+/K+-ATPase activity of P. homarus increased (p < 0.05) under the both salinity stress. Salinity 18 significantly increased Glutamate dehydrogenase (GDH) and Glutamicpyruvic transaminase (GPT) activity in the hepatopancreas of P. homarus (p < 0.05). According to transcriptome analysis, versus control group (salinity 28), 929 and 1095 differentially expressed genes (DEGs) were obtained in the gills of P. homarus at salinity 18 and 38, respectively, with these DEGs were mainly involved in energy metabolism, transmembrane transport and oxidative stress and substance metabolism. In addition, the expression patterns of 8 key DEGs mainly related to amino acid metabolism, transmembrane transport and oxidative stress were verified by quantitative real-time PCR (RT-qPCR). The present study suggests that salinity 18 has a greater impact on P. homarus than salinity 38, and P. homarus demonstrates effective osmoregulation and handle with salinity fluctuations (18 to 38) through physiological and functional adaptations. This study provides an improved understanding of the physiological response strategies of P. homarus facing salinity stress, which is crucial for optimizing aquaculture practices for this species.

Distinct planting patterns exert legacy effects on the networks and assembly of root-associated microbiomes in subsequent crops.

Soil legacy effects from previous crops can significantly influence plant-soil interactions in crop rotations. However, the microbial mechanism underlying this effect in subsequent root-associated compartments remains unclear. We investigated the effects of planting patterns (four-year continuous maize [MM], three-year winter wheat and one-year maize rotation [WM], and three-year potato and one-year maize rotation [PM]) on the microbial composition and structure of root-associated compartments, the effect of distinct crops on subsequent microbial co-occurrence patterns, and the assembly mechanism by which the root-associated compartments (bulk soil, rhizosphere, and roots) in subsequent crops regulate the microbiome habitat. Compared with MM, the relative abundance of Acidobacteria in WM was 29.7 % lower, whereas that of Bacteroidota in PM was 37.9 % higher in all three compartments. The co-occurrence patterns of the microbial communities exhibited varied responses to different planting patterns. Indicator taxon analysis revealed less shared and specific species in the root bacterial and fungal networks. The planting pattern elicited specific responses from modules within bacterial and fungal co-occurrence networks in all three compartments. Moreover, the planting patterns and root-associated compartments collectively drove the assembly process of root-associated microorganisms. The neutral model showed that, compared with MM, the stochasticity of bacterial assembly decreased under WM and PM but increased for fungal assembly. WM and PM increased the relative effects of the homogenized dispersal of fungal assemblies in roots. We conclude that previous crops exhibit marked legacy effects in the root-associated microbiome. Therefore, soil heritage should not be ignored when discussing microbiome recruitment strategies and co-occurrence patterns in subsequent crops.

Synergistic Effect of Layered Double Hydroxides Nanodosage Form to Induce Apoptosis and Ferroptosis in Breast Cancer.

Background: Breast cancer is the most common cancer in women and one of the leading causes of cancer death worldwide. Ferroptosis, a promising mechanism of killing cancer cells, has become a research hotspot in cancer therapy. Simvastatin (SIM), as a potential new anti-breast cancer drug, has been shown to cause ferroptosis of cancer cells and inhibit breast cancer metastasis and recurrence. The purpose of this study is to develop a novel strategy boosting ferroptotic cascade for synergistic cancer therapy. Methods: In this paper, iron base form of layered double hydroxide supported simvastatin (LDHs-SIM) was synthesized by hydrothermal co-precipitation method. The characterization of LDHs-SIM were assessed by various analytical techniques, including ultraviolet-visible (UV-vis) spectroscopy, X-ray diffraction (XRD), Fourier transform infrared (FTIR) spectroscopy, and transmission electron microscopy (TEM). Biological activity, ferroptosis mechanism and biocompatibility were analyzed through in vivo and in vitro analysis, so as to evaluate its therapeutic effect on breast cancer. Results: The constructed LDHs-SIM nanosystem can not only release SIM through mevalonate (MVA) pathway, inhibit the expression of glutathione peroxidase 4 (GPX4), inhibit the expression of SLC7A11 and reduce the synthesis efficiency of GSH, but also promote the accumulation of Fe2+ in cells through the release of Fe3+, and increase the intracellular ROS content. In addition, LDHs-SIM nanosystem can induce apoptosis of breast cancer cells to a certain extent, and achieve the synergistic effect of apoptosis and ferroptosis. Conclusion: In the present study, we demonstrated that nanoparticles of layered double hydroxides (LDHs) loaded with simvastatin were more effective than a free drug at inhibiting breast cancer cell growth, In addition, superior anticancer therapeutic effects were achieved with little systemic toxicity, indicating that LDHs-SIM could serve as a safe and high-performance platform for ferroptosis-apoptosis combined anticancer therapy.

Hidden blood loss in three different endoscopic spinal procedures for lumbar disc herniation.

Purpose: This study compared hidden blood loss (HBL) among three different endoscopic spinal procedures and investigated its risk factors. Patients and methods: This single-centre retrospective analysis collected data from consecutive hospitalized patients with single-segment lumbar disc herniation (LDH) undergoing unilateral biportal endoscopic discectomy (UBE), percutaneous endoscopic transforaminal discectomy (PETD), or percutaneous endoscopic interlaminar discectomy (PEID) from December 2020 to October 2022. HBL was calculated using Nadler's and Gross's formulas. The authors used Pearson's or Spearman's correlation analysis to explore the relationship between patient characteristics and HBL. Multivariate linear regression analysis was used to identify independent risk factors for HBL. Results: In total, 122 consecutive patients (68 females and 54 males) were enroled in this study. The average HBL was 381.87±218.01 ml in the UBE group, 252.05±118.44 ml in the PETD group and 229.63±143.9 ml in the PEID group (P<0.05). Pearson's or Spearman's correlation analysis showed that operative time, preoperative haemoglobin, preoperative haematocrit, and preoperative Albumin (ALB) were correlated with HBL in the UBE group, while sex, age, operative time, postoperative ALB, and patients' blood volume (PBV) were related to HBL in the PETD group (P<0.05). Operative time and preoperative activated partial thromboplastin time were related to HBL in the PEID group (P<0.05). Multiple linear regression analysis showed a positive correlation between HBL and operative time in all three groups (P<0.001, P<0.001, P<0.05). Conclusion: HBL was higher in the UBE group than in the PETD and PEID groups, and operative time may be a common risk factor for the three groups.

Common carp (Cyprinus carpio) CD81 promoting CyHV-3 virus replication via regulating autophagy and RLRs-interferon signaling pathway.

Cyprinid herpesvirus type 3 (CyHV-3), also called Koi herpesvirus (KHV), which leads to mass cyprinid mortality and enormous economic losses. To establish an infection, CyHV-3 needs to counteract host antiviral responses. CD81 belongs to the evolutionary conserved tetraspanin family of proteins. Several studies have shown that different members of the tetraspanin superfamily modulates different virus infectious processes. Here we aimed at analysing the role of CD81 in CyHV-3 infection. In this study, we cloned and characterized the CD81 of Common Carp, the open reading frame of CcCD81 gene was 702 bp, which encoded 234 amino acids with four transmembrane domains (TM1 to TM4), a small extracellular loop (SEL), and a large extracellular loop (LEL). Tissue distribution analysis showed that CcCD81 was widely expressed in all the tested tissues with the highest expression in head kidney, followed by a high expression in brain. Subsequently, expression levels of CcCD81 were significantly increased in CCB cells within the first 3h after infection, meanwhile, the expression of viral gene VP136 was reduced after CcCD81 knockdown in CCB cells post CyHV-3 infection. Furthermore, CcCD81 knockdown can significantly reduce the autophagy process and increase the promoter activity of ISRE and IFN-1 in the CCB cells after viral infection, as well as other genes involved in the IFN signaling pathway, including RIG-1、MDA5、MAVS、TBK1 and IRF3. Taking the data together, we revealed that CcCD81 mediates autophagy and blocks RIG-1-mediated antiviral signaling and negatively regulates the promoter activity of type I interferon (IFN) promoting virus replication. These results reveal a new link between autophagy and four-transmembrane-domain protein superfamily and contribute to elucidate the mechanism of CyHV-3 infection.

Autophagy induced by Cyprinid herpesvirus 3 (CyHV-3) facilitated intracellular viral replication and extracellular viral yields in common carp brain cells.

Autophagy is a conservative and important process that exists in all eukaryotic cells in nature. Cyprinid herpesvirus 3 (CyHV-3), also known as KHV (Koi Herpesvirus), is a pathogen that mainly infecting common carp and koi. In the present study, we identified the CcLC3B gene, with a length of 379 bp and displaying a close evolutionary relationship with other sixteen different species, the tissue distribution and expression pattern of CcLC3 were also identified. We found that CyHV-3 infection could promote autophagy in CCB cells at the early stage but inhibit autophagy at the late stage by using confocal fluorescence microscopy, transmission electron microscopy and western blotting. And we measured the protein levels associated with the Akt/mTOR signalling pathway, intracellular replication of CyHV-3 at the mRNA and protein levels as well as viral titters. Collectively, the results taken together suggested that CyHV-3 infection could promote autophagy in CCB cells at the early stage but inhibit autophagy at the late stage via mTOR and that promoting autophagy could facilitate CyHV-3 intracellular replication and extracellular viral yields in CCB cells. These findings revealed the relationship between CyHV-3 and autophagy and provided a novel treatment strategy targeting the autophagy signalling pathway against CyHV-3 infection.

The function and community structure of arbuscular mycorrhizal fungi in ecological floating beds used for remediation of Pb contaminated wastewater.

Arbuscular mycorrhizal fungi (AMF) have been demonstrated to be ubiquitous in aquatic ecosystems. However, their distributions and ecological functions are rarely studied. To date, a few studies have combined sewage treatment facilities with AMF to improve removal efficiency, but appropriate and highly tolerant AMF strains have not been explored, and the purification mechanisms remain unclear. In this study, three ecological floating-bed (EFB) installations inoculated with different AMF inocula (mine AMF inoculum, commercial AMF inoculum and non-AMF inoculated) were constructed to investigate their removal efficiency for Pb-contaminated wastewater. The AMF community structure shifts in the roots of Canna indica inhabiting EFBs during the three phases (pot culture phase, hydroponic phase and hydroponic phase with Pb stress) were tracked utilizing quantitative real-time polymerase chain reaction and Illumina sequencing techniques. Furthermore, transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDS) were used to detect the Pb location in mycorrhizal structures. The results showed that AMF could promote host plant growth and enhance the Pb removal efficiency of the EFBs. The higher the AMF abundance, the better the effect of the AMF on Pb purification by EFBs. Both flooding and Pb stress decreased the AMF diversity but did not significantly inhibit the abundance. The three inoculation treatments showed different community compositions with different dominant AMF taxa in different phases, and an uncultured Paraglomus species (Paraglomus sp. LC516188.1) was found to be the most dominant (99.65 %) AMF in the hydroponic phase with Pb stress. The TEM and EDS analysis results showed that the Paraglomus sp. could accumulate Pb in plant roots through their fungal structures (intercellular mycelium, intracellular mycelium, etc.), which alleviated the toxic effect of Pb on plant cells and limited Pb translocation. The new findings provide a theoretical basis for the application of AMF in plant-based bioremediation of wastewater and polluted waterbodies.

D-beta-hydroxybutyrate protects against microglial activation in lipopolysaccharide-treated mice and BV-2 cells.

Microglial activation is a key event in neuroinflammation, which, in turn, is a central process in neurological disorders. In this study, we investigated the protective effects of D-beta-hydroxybutyrate (BHB) against microglial activation in lipopolysaccharide (LPS)-treated mice and BV-2 cells. The effects of BHB in mice were assessed using behavioral testing, morphological analysis and immunofluorescence labeling for the microglial marker ionizing calcium-binding adaptor molecule 1 (IBA-1) and the inflammatory cytokine interleukin-6 (IL-6) in the hippocampus. Moreover, we examined the levels of the inflammatory IL-6 and tumor necrosis factor-α (TNF-α), as well as those of the neuroprotective brain-derived neurotrophic factor (BDNF) and transforming growth factor-ß (TGF-ß) in the brain. In addition, we examined the effects of BHB on IL-6, TNF-α, BDNF, TGF-ß, reactive oxygen species (ROS) level and cell viability in LPS-stimulated BV-2 cells. BHB treatments attenuated behavioral abnormalities, reduced the number of IBA-1-positive cells and the intensity of IL-6 fluorescence in the hippocampus, with amelioration of microglia morphological changes in the LPS-treated mice. Furthermore, BHB inhibited IL-6 and TNF-α generation, but promoted BDNF and TGF-ß production in the brain of LPS-treated mice. In vitro, BHB inhibited IL-6 and TNF-α generation, increased BDNF and TGF-ß production, reduced ROS level, ameliorated morphological changes and elevated cell viability of LPS-stimulated BV-2 cells. Together, our findings suggest that BHB exerts protective effects against microglial activation in vitro and in vivo, thereby reducing neuroinflammation.

Genome-Wide Analysis of Differentially Expressed mRNAs and lncRNAs in Koi Carp Infected with Koi Herpesvirus.

Long noncoding RNAs (lncRNAs) constitute an emerging group of ncRNAs that modulate gene expression at the transcriptional or translational level. Koi herpesvirus (KHV), also known as Cyprinus herpesvirus type 3 (CyHV-3) and characterized by high pathogenicity and high mortality, has caused substantial economic losses in the common carp and koi carp fisheries industry. In this work, we sequenced the lncRNA and mRNA of host koi carp infected with KHV. A total of 20,178 DEmRNAs were obtained, of which 5021 mRNAs were upregulated and 15,157 mRNAs were downregulated. Both KEGG pathways and GO terms were enriched in many important immune pathways. The KEGG analysis showed that DEGs were significantly enriched in many important immune pathways, such as apoptosis, NOD-like receptor signaling pathway, Jak-STAT signaling pathway, TNF signaling pathway, IL-17 signaling pathway, NF-kappa B signaling pathway, and so on. Furthermore, a total of 32,697 novel lncRNA transcripts were obtained from koi carp immune tissues; 9459 of these genes were differentially expressed. Through antisense, cis-acting, and trans-acting analyses, the target genes of differentially expressed lncRNAs (DElncRNAs) were predicted. Gene ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that the DElncRNA expression pattern was consistent with the differential mRNA expression pattern. The lncRNA-mRNA network analysis, which included many immune pathways, showed that after KHV infection, the expression of most lncRNAs and their target mRNAs were downregulated, suggesting that these lncRNAs engage in a positive regulatory relationship with their target mRNAs. Considering that many studies have shown that herpesviruses can escape the immune system by negatively regulating these immune pathways, we speculated that these lncRNAs play a significant role in KHV's escape from host immunity. Furthermore, 10 immune-related genes and 20 lncRNAs were subsequently verified through RT-qPCR, to confirm the accuracy of the high-throughput sequencing results. In this study, we aimed to explore lncRNA functions in the immune response of lower vertebrates and provide a theoretical basis for the study of noncoding RNAs in teleosts. Therefore, exploring lncRNA expression in KHV-infected koi carp helped us better understand the biological role played by lncRNA-dependent pathways in aquaculture animal viral infection.

Comparison of four tools to identify painful new osteoporotic vertebral fractures in the postmenopausal population in Beijing.

Objectives: To validate and compare four tools, the Fracture Risk Assessment Tool (FRAX) without bone mineral density (BMD), Beijing Friendship Hospital Osteoporosis Screening Tool (BFH-OST), Osteoporosis Self-Assessment Tool for Asians (OSTA), and BMD, to identify painful new osteoporotic vertebral fractures (PNOVFs). Methods: A total of 2874 postmenopausal women treated from June 2013 to June 2022 were enrolled and divided into two groups: patients with PNOVFs who underwent percutaneous vertebroplasty (PNOVFs group, n = 644) and community-enrolled females (control group, n = 2230). Magnetic resonance and X-ray imaging were used to confirm the presence of PNOVFs. Dual-energy X-ray absorptiometry was performed to calculate the BMD T-scores. Osteoporosis was diagnosed according to WHO Health Organization criteria. Data on the clinical and demographic risk factors were self-reported using a questionnaire. The ability to identify PNOVFs using FRAX, BFH-OST, OSTA, and BMD scores was evaluated using receiver operating characteristic (ROC) curves. For this evaluation, we calculated the areas under the ROC curves (AUCs), sensitivity, specificity, and optimal cut-off points. Results: There were significant differences in FRAX (without BMD), BFH-OST, OSTA, and BMD T-scores (total hip, femoral neck, and lumbar spine) between the PNOVFs and control groups. Compared with BFH-OST, OSTA, and BMD, the FRAX score had the best identifying value for PNOVFs; the AUC of the FRAX score (optimal cutoff =3.6%) was 0.825, while the sensitivity and specificity were 82.92% and 67.09%, respectively. Conclusion: FRAX may be the preferable tool for identifying PNOVFs in postmenopausal women, while BFH-OST and OSTA can be applied as more simple screening tools for PNOVFs.

Atypical Proximal Cervical Spondylotic Amyotrophy: Case Report Demonstrating Clinical/Imaging Discrepancy.

PURPOSE: The aim of this study was to present a novel case of unilateral proximal cervical spondylotic amyotrophy (CSA) with contralateral spinal cord compression, which is subject to misdiagnosis and missed diagnosis. BACKGROUND: CSA is the rare form of cervical spondylosis, which is characterized by severe muscle atrophy in the upper extremities. It can be classified in the proximal subtype and the distal subtype. The etiology, pathophysiology and treatment of CSA are still controversial. METHODS: A rare case of atypical proximal CSA, who presented with left shoulder and arm weakness, but cervical magnetic resonance imaging (MRI) showed large right paracentral disc herniation in the C4-5 level. Twelve weeks after undergoing anterior cervical discectomy and fusion technique in C4-5 level, the patient's symptoms obviously recovered. CONCLUSION: The opposite sides between disc herniation and clinical symptoms of upper extremity may be attributed to C5 ventral rootlet becoming stretched caused by spinal cord rotation or shift to the opposite side.

A rare lumbar pyogenic spondylodiscitis caused by staphylococcus caprae with initial misdiagnosis: case report and literature review.

BACKGROUND: Staphylococcus caprae (Sc) is an uncommon causative organism for human. Lumbar pyogenic spondylodiscitis (LPS) of Sc is extremely rare and only a few cases have been reported. As far as we know, there is no specific literature on the diagnosis and treatment for LPS of Sc with L5 nerve root irritation. CASE PRESENTATION: A 65-year-old male patient complained of chronic low back pain for 10 years, acute worsening with radiating pain to left lower extremity over a month. Physical examination revealed tenderness point on his low back, 3/5 dorsiflexor strength in his left 1st toe and decreased sensation of pin prick over the left lateral shank and medial dorsal foot. The individual was initially misdiagnosed with lumbar disc herniation (LDH) without further examination in outpatient, which was then found to be LPS of Sc with L5 nerve root irritation after admission to our hospital. Magnetic resonance images (MRI) of lumbar spine exhibited inflammation signal at L4-L5 level of the vertebral body and disc with hypointense on T1-weighted images (T1-WI) and hyperintense on T2-weighted images (T2-WI). The causative organism was confirmed by the culture of irrigation fluid obtained from L5 vertebrae by needle puncture. After systemic conservative treatment including using sensitive antimicrobial agents and immobilization, the rare infection was finally cured. The patient also showed a satisfactory recovery during the 36-month follow-up period. CONCLUSIONS: Confirming the diagnosis and identifying the causative organism as soon as possible is the key point for the treatment of LPS. LPS of Sc causing nerve root irritation is rare but curable with early diagnosis and proper therapy. The culture of irrigation fluid obtained from vertebrae by needle puncture may be an effective and sensitive attempt for potential infection of spine to identify the causative organism at early stage of the disease.

Comparison of OSTA, FRAX and BMI for Predicting Postmenopausal Osteoporosis in a Han Population in Beijing: A Cross Sectional Study.

PURPOSE: To validate the efficacies of three screening tools including the Osteoporosis Self-Assessment Tool for Asians (OSTA), Fracture Risk Assessment Tool (FRAX) without bone mineral density (BMD), and body mass index (BMI) for predicting postmenopausal osteoporosis (OP) and to define the ideal thresholds for avoidance of dual-energy X-ray absorptiometry (DXA) scanning in a Han Chinese population in Beijing. PATIENTS AND METHODS: A total of 2055 community-dwelling Han Beijing postmenopausal females aged ≥45 years were enrolled in this study. All participants completed a questionnaire, and BMD was measured by DXA. OP was defined by a T-score at least -2.5 SD less than that of average young adults in different diagnostic criteria [lumbar spine, femoral neck, total hip, worst hip, WHO]. The abilities of the OSTA, FRAX, and BMI to predict OP were analyzed by receiver operating characteristic (ROC) curves. Sensitivity, specificity, and area under the ROC curves (AUC) were calculated. Ideal thresholds for identifying OP were proposed. RESULTS: The prevalence of OP ranged from 8.1% to 28.4% according to different diagnostic criteria. The AUC range for the OSTA (0.758-0.849) was similar to the FRAX (0.728-0.855), which revealed that both tools predicted OP reliably. The AUC range for BMI was 0.643-0.682, suggesting limited predictive value. According to WHO criteria, the AUC values for the FRAX for hip fracture risk (FRAX-HF) and for the OSTA were 0.796 and 0.798, with corresponding sensitivities of 74.79% and 69.64% and specificities of 70.45% and 75.07%, respectively. At defined thresholds, the FRAX-HF and OSTA allowed avoidance of DXA in 42.4-37.6% of participants, at a cost of missing only 7.2-8.6% of individuals with OP. CONCLUSION: The OSTA and FRAX-HF may be reliable and effective tools for identifying postmenopausal OP in the Han Beijing population without BMD.

New combination of IntraSPINE device and posterior lumbar interbody fusion for rare skipped-level lumbar disc herniation: a case report and literature review.

Posterior lumbar interbody fusion is an open surgical technique that has been widely used for the treatment of degenerative lumbar disease. However, traditional lumbar spinal fusion, especially long-segment fusion surgery, is associated with several complications. The IntraSPINE (Cousin Biotech, Wervicq-Sud, France) is a new device for non-fusion lumbar spine surgery that is used as an alternative for the treatment of degenerative lumbar disease. Although the designer of the IntraSPINE proposed indications for its use, including combination of the device with lumbar spinal fusion for the treatment of degenerative lumbar disease, use of the IntraSPINE has not been reported in the clinical literature. In the present case, we boldly combined the IntraSPINE device and posterior lumbar interbody fusion for the treatment of skipped-level lumbar disc herniation to explore the indications of the IntraSPINE and report its clinical outcomes.

Molecular characterization and expression of SiFad1 in the sea urchin (Strongylocentrotus intermedius).

Fatty acid desaturases (Fads) are a key enzyme in the process of biosynthesis of highly unsaturated fatty acids (HUFAs). In this study, we cloned the full-length sequence of the SiFad1 gene (SiFad1) and analyzed its expression profiles during different developmental stages and in different tissues of Strongylocentrotus intermedius. The full-length cDNA of SiFad1 is composed of 1086 bp, with a putative open reading frame of 885 bp encoding a polypeptide of 294 amino acid (AA) residues. The predicted molecular mass of SiFad1 is 34.67 kDa and its theoretical pI is 8.41. The presence of conserved motifs including three histidine boxes (HXXXH, HXXHH, XXXHH), a FA_desaturases domain and three transmembrane domains suggests that SiFad1 belongs to the microsomal fatty acid desaturases family. Its tissue distribution showed that the highest expression of SiFad1 is in the intestine and the weakest expression is in Aristotle's lantern of S. intermedius. Time-course expression measurements in different developmental stages showed the highest expression of SiFad1 occurs in the gastrula and the weakest expression in the juvenile sea urchin. Knock-down of SiFad1 by specific siRNA revealed that the significantly depressed expression of Elovl5 had decreased in the coelomocytes, intestines and gonads at 24 h post transfection, indicating that the downstream target gene of SiFad1 is Elovl5 and SiFad1 and Elovl5 have positive regulatory effects. When we examined the changes in fatty acids in the gonads before and after interference, the results showed that after 24 h of interference, the content of C20:4n-6 produced by SiFad1 had decreased. Taken together, these results will enable us to understand the role of SiFad1 in fatty acid anabolism, which will help us to understand the fatty acid synthesis pathways and regulatory mechanisms of Strongylocentrotus intermedius and provide a theoretical experimental basis for improving the ability of sea urchins to synthesize fatty acids and cultivating sea urchins of higher quality and nutritional value.

A Novel Approach for Percutaneous Vertebroplasty Based on Preoperative Computed Tomography-Based Three-Dimensional Model Design.

OBJECTIVE: To describe a new technique for the efficient use of preoperative planning based on preoperative computed tomography-based three-dimensional (3D) model design for percutaneous vertebroplasty (PVP) in a patient with osteoporotic vertebral compression fracture. METHODS: A 76-year-old woman with acute osteoporotic vertebral compression fracture (L1 level) accepted a novel precise PVP. A 3D model of thoracolumbar vertebrae (T12-L2) based on preoperative computed tomography scanning data and a simulative PVP (via a bilateral transpedicular approach) were built in MIMICS (Materialise Interactive Medical Image Control System) software. With the help of 3 radiopaque markers located at the skin of the back and preoperative digital design by MIMICS, bilateral skin entry points, needles direction including abduction angle and head inclination angle, and needle insertion depth were established. RESULTS: During surgery, only 1 shot of fluoroscopy was required to confirm the skin entry points. The operation took only about 23 minutes and total patient exposure dose was 4.5 mSv. The intraoperative radiologic results showed that the cement distribution in the L1 vertebra was good without any puncture-related complications. The patient's visual analog scale score improved from 9 points preoperatively to 2 points postoperatively. The patient's preoperative Oswestry Disability Index score was 80 points, which improved to 57.8 points postoperatively. CONCLUSIONS: The novel precise PVP based on preoperative 3D model design allows 1) visualization of the morphology of the fractured vertebral body, 2) increased precision of puncture with decreasing incidence of puncture-related complications and reduced radiation exposure, and 3) less operation time, decreasing the learning curve of beginners with limited experience.

Revisit the Correlation between the Elastic Mechanics and Fusion of Lipid Membranes.

Membrane fusion is a vital process in key cellular events. The fusion capability of a membrane depends on its elastic properties and varies with its lipid composition. It is believed that as the composition varies, the consequent change in C0 (monolayer spontaneous curvature) is the major factor dictating fusion, owing to the associated variation in GEs (elastic energies) of the fusion intermediates (e.g. stalk). By exploring the correlations among fusion, C0 and Kcp (monolayer bending modulus), we revisit this long-held belief and re-examine the fusogenic contributions of some relevant factors. We observe that not only C0 but also Kcp variations affect fusion, with depression in Kcp leading to suppression in fusion. Variations in GEs and inter-membrane interactions cannot account for the Kcp-fusion correlation; fusion is suppressed even as the GEs decrease with Kcp, indicating the presence of factor(s) with fusogenic importance overtaking that of GE. Furthermore, analyses find that the C0 influence on fusion is effected via modulating GE of the pre-fusion planar membrane, rather than stalk. The results support a recent proposition calling for a paradigm shift from the conventional view of fusion and may reshape our understanding to the roles of fusogenic proteins in regulating cellular fusion machineries.

A low ratio of n-6/n-3 polyunsaturated fatty acids suppresses matrix metalloproteinase 13 expression and reduces adjuvant-induced arthritis in rats.

Increased expression of matrix metalloproteinase 13 (MMP13) in chondrocytes contributes to the development of osteoarthritis. The hypothesis of this study was that diet with a low ratio of n-6/n-3 polyunsaturated fatty acids (PUFAs) is associated with reduced MMP13 expression in inflammatory chondrocytes in vitro and in vivo. Human chondrocytes were cultured with different ratios of linoleic acid (LA, n-6 PUFA) to α-linolenic acid (n-3 PUFA) from 1:1 to 10:1. Proliferation of chondrocytes, MMP13 protein and mRNA levels were detected, respectively. Sprague-Dawley rats (n=30) were fed diets containing different ratios of n-6/n-3 PUFA. Freund's complete adjuvant was injected to make the model of arthritis. Paw swelling rate was measured and all rats were euthanized after 6 weeks of treatment. Serum MMP13 and IL-1 were measured by enzyme-linked immunosorbent assay. Joint histological sections were stained with safranin-O Fast Green to evaluate cartilage damage. Low ratio of LA/α-linolenic acid decreased the mRNA and protein levels of MMP13 but did not affect chondrocytes proliferation. Ratios of PUFA such as 1:1 and 2:1 significantly reduced paw swelling rate, and serum MMP13 and IL-1 levels in a rat model. Histological staining showed that ratios of 1:1 and 2:1 PUFA significantly alleviated cartilage damage in adjuvant-induced arthritis. A ratio of n-6/n-3 PUFA of 1:1 showed the strongest inhibitory effect on MMP13. Our results indicate that a low ratio of n-6/n-3 PUFA at 1:1 significantly suppressed MMP13 expression both in vitro and in vivo and reduced adjuvant-induced arthritis in rats could be a means to control and reduce the symptoms of osteoarthritis.