Cytochalasans from the Endophytic Fungus Aspergillus sp. LE2: Their Structures, Antibacterial and NO Production Inhibitory Activities.

Six new cytochalasans-namely, aspergicytochalasins A-F (1-6)-together with five known analogs were isolated and characterized from the endophytic fungus Aspergillus sp. from the medicinal plant Lonicera japonica. The structures of the new compounds were established by NMR and MS methods as well as single crystal X-ray diffractions. Compounds 3 and 4 showed weak antibacterial activities to Staphylococcus aureus, with MIC values of 128 and 64 µg/mL, respectively. Compounds 1, 3, 5 and 6 showed inhibitory activities on NO production, with IC50 values less than 40 µM.

Application of a new anatomic hook-rod-pedicle screw system in young patients with lumbar spondylolysis: A pilot study.

BACKGROUND: The pedicle screw-laminar hook system has strong fixation and is conducive to bone graft fusion for lumbar spondylolysis. However, the current pedicle screw-laminar hook fixation system is not specifically designed for lumbar spondylolysis. AIM: To investigate the clinical effects of a new anatomical hook-rod-pedicle screw system in the treatment of lumbar spondylolysis in young adults. METHODS: We designed a new anatomic hook-rod-pedicle screw system for young patients with lumbar spondylolysis. The isthmus and the corresponding pedicle screw entry point were exposed through the intermuscular approach. Autogenous iliac bone graft was obtained to bridge the isthmus defect, and then the anatomic hook-rod-pedicle screw system was used to fix the isthmus in 15 young patients. RESULTS: At 24 mo follow-up, the visual analogue scale score of low back pain decreased from 6.73 ± 0.88 to 0.73 ± 0.59, and the Oswestry disability index score decreased from 58.20 ± 8.99 to 7.87 ± 4.97. Computed tomography showed bilateral isthmic bone healing in 14 cases and unilateral isthmic bone healing in 1 case. Magnetic resonance imaging showed that the lumbar disc signal of diseased segment and adjacent segments had no change compared with that before surgery. The pain visual analogue scale score of the donor site was 0.20 ± 0.41 at the last follow-up. According to the Modified Macnab score, the excellent and good rate was 100%. CONCLUSION: The application of this new anatomical hook-rod-pedicle screw system to treat young patients with lumbar spondylolysis has the advantages of less trauma, a simple operation and satisfactory clinical effects.

Synergistic Effects of APOE and CLU May Increase the Risk of Alzheimer's Disease: Acceleration of Atrophy in the Volumes and Shapes of the Hippocampus and Amygdala.

BACKGROUND: The volume loss of the hippocampus and amygdala in non-demented individuals has been reported to increase the risk of developing Alzheimer's disease (AD). Many neuroimaging genetics studies mainly focused on the individual effects of APOE and CLU on neuroimaging to understand their neural mechanisms, whereas their synergistic effects have been rarely studied. OBJECTIVE: To assess whether APOE and CLU have synergetic effects, we investigated the epistatic interaction and combined effects of the two genetic variants on morphological degeneration of hippocampus and amygdala in the non-demented elderly at baseline and 2-year follow-up. METHODS: Besides the widely-used volume indicator, the surface-based morphometry method was also adopted in this study to evaluate shape alterations. RESULTS: Our results showed a synergistic effect of homozygosity for the CLU risk allele C in rs11136000 and APOEɛ4 on the hippocampal and amygdalar volumes during a 2-year follow-up. Moreover, the combined effects of APOEɛ4 and CLU C were stronger than either of the individual effects in the atrophy progress of the amygdala. CONCLUSION: These findings indicate that brain morphological changes are caused by more than one gene variant, which may help us to better understand the complex endogenous mechanism of AD.

Assessment of load-sharing thoracolumbar injury: A modified scoring system.

BACKGROUND: Many classification systems of thoracolumbar spinal fractures have been proposed to enhance treatment protocols, but none have achieved universal adoption. AIM: To develop a new patient scoring system for cases with thoracolumbar injury classification and severity score (TLICS) = 4, namely the load-sharing thoracolumbar injury score (LSTLIS). METHODS: Based on thoracolumbar injury classification and severity score, this study proposes the use of the established load-sharing classification (LSC) to develop an improved classification system (LSTLIS). To prove the reliability and reproducibility of LSTLIS, a retrospective analysis for patients with thoracolumbar vertebral fractures has been conducted. RESULTS: A total of 102 cases were enrolled in the study. The scoring trend of LSTLIS is roughly similar as the LSC scoring, however, the average deviation based on the former method is relatively smaller than that of the latter. Thus, the robustness of the LSTLIS scoring method is better than that of LSC. LSTLIS can further classify patients with TLICS = 4, so as to assess more accurately this particular circumstance, and the majority of LSTLIS recommendations are consistent with actual clinical decisions. CONCLUSION: LSTLIS is a scoring system that combines LSC and TLICS to compensate for the lack of appropriate inclusion of anterior and middle column compression fractures with TLICS. Following preliminary clinical verification, LSTLIS has greater feasibility and reliability value, is more practical in comprehensively assessing certain clinical circumstances, and has better accuracy with clinically significant guidelines.

Treatment of Kümmell's disease with sequential infusion of bone cement: A retrospective study.

BACKGROUND: Percutaneous vertebroplasty (PVP) is an effective method for the treatment of neurologically intact Kümmell's disease, but bone cement leakage during surgery is a problem that deserves attention. AIM: To reduce bone cement leakage and evaluate the effect of the sequential infusion of bone cement during PVP for the treatment of stage I or II Kümmell's disease. METHODS: Patients with Kümmell's disease treated in our hospital from September 2015 to September 2018 were retrospectively analyzed. Patients meeting the inclusion and exclusion criteria were divided into two groups: Traditional single infusion and sequential infusion (SI). The visual analog scale (VAS) and Oswestry disability index (ODI) were evaluated and compared, and duration of operation, bone cement content and complications were recorded. RESULTS: Forty-five patients were included in this study; there were 24 in the traditional single infusion group and 21 in the SI group. The VAS and ODI were significantly different for both groups when compared pre- and postoperatively, whereas the differences between 1 wk postoperatively and at the final follow-up were not statistically. When the VAS and ODI of the two groups were compared, there were no significant differences at any time point. The leakage rate of bone cement was significantly lower in the SI group (14.3%, 3 of 21) than that in the traditional single infusion group (41.7%, 10 of 24). CONCLUSION: SI in unipedicular PVP is a safe and effective procedure for neurologically intact Kümmell's disease, and this technique could decrease the incidence of bone cement leakage.

Oral P. gingivalis impairs gut permeability and mediates immune responses associated with neurodegeneration in LRRK2 R1441G mice.

BACKGROUND: The R1441G mutation in the leucine-rich repeat kinase 2 (LRRK2) gene results in late-onset Parkinson's disease (PD). Peripheral inflammation and gut microbiota are closely associated with the pathogenesis of PD. Chronic periodontitis is a common type of peripheral inflammation, which is associated with PD. Porphyromonas gingivalis (Pg), the most common bacterium causing chronic periodontitis, can cause alteration of gut microbiota. It is not known whether Pg-induced dysbiosis plays a role in the pathophysiology of PD. METHODS: In this study, live Pg were orally administrated to animals, three times a week for 1 month. Pg-derived lipopolysaccharide (LPS) was used to stimulate mononuclear cells in vitro. The effects of oral Pg administration on the gut and brain were evaluated through behaviors, morphology, and cytokine expression. RESULTS: Dopaminergic neurons in the substantia nigra were reduced, and activated microglial cells were increased in R1441G mice given oral Pg. In addition, an increase in mRNA expression of tumor necrosis factor (TNF-α) and interleukin-1ß (IL-1ß) as well as protein level of α-synuclein together with a decrease in zonula occludens-1 (Zo-1) was detected in the colon in Pg-treated R1441G mice. Furthermore, serum interleukin-17A (IL-17A) and brain IL-17 receptor A (IL-17RA) were increased in Pg-treated R1441G mice. CONCLUSIONS: These findings suggest that oral Pg-induced inflammation may play an important role in the pathophysiology of LRRK2-associated PD.

Application of molybdenum target X-ray photography in imaging analysis of caudal intervertebral disc degeneration in rats.

BACKGROUND: Conventional plain X-ray images of rats, the most common animals used as degeneration models, exhibit unclear vertebral structure and blurry intervertebral disc spaces due to their small size, slender vertebral bodies. AIM: To apply molybdenum target X-ray photography in the evaluation of caudal intervertebral disc (IVD) degeneration in rat models. METHODS: Two types of rat caudal IVD degeneration models (needle-punctured model and endplate-destructed model) were established, and their effectiveness was verified using nuclear magnetic resonance imaging. Molybdenum target inspection and routine plain X-ray were then performed on these models. Additionally, four observers were assigned to measure the intervertebral height of degenerated segments on molybdenum target plain X-ray images and routine plain X-ray images, respectively. The degeneration was evaluated and statistical analysis was subsequently conducted. RESULTS: Nine rats in the needle-punctured model and 10 rats in the endplate-destructed model were effective. Compared with routine plain X-ray images, molybdenum target plain X-ray images showed higher clarity, stronger contrast, as well as clearer and more accurate structural development. The McNemar test confirmed that the difference was statistically significant (P = 0.031). In the two models, the reliability of the intervertebral height measured by the four observers on routine plain X-ray images was poor (ICC < 0.4), while the data obtained from the molybdenum target plain X-ray images were more reliable. CONCLUSION: Molybdenum target inspection can obtain clearer images and display fine calcification in the imaging evaluation of caudal IVD degeneration in rats, thus ensuring a more accurate evaluation of degeneration.

Lumbar disc rehydration in the bridged segment using the BioFlex dynamic stabilization system: A case report and literature review.

BACKGROUND: In recent years, the mechanical concept of intervertebral disc regeneration has become more and more popular due to the increasing awareness of the importance of preservation of spine movement. Interestingly, there is increasing evidence, however, that dynamic stabilization systems may compensate non-physiological loads, limit pathological movement, normalize disc height and intradiscal pressure, and provide an adaptive environment for disc regeneration. CASE SUMMARY: The patient was a 54-year-old man, who presented with a 10-year history of mechanical back pain, which had become progressively serious and radiated into the left lower limb with numbness 3 mo prior. He had decreased muscle strength (class IV) of the left dorsal extensor and plantar flexor. Magnetic resonance imaging scans showed L3-S1 disc degeneration and L4-L5 disc herniation. Because the patient did not respond to various conservative treatments, he underwent a posterior L4-5 discectomy with fixation of the BioFlex dynamic stabilization system (Bio-Spine, Seoul, Korea). Preoperative symptoms were relieved and lumbar function was markedly improved after the operation. L4-L5 disc rehydration of instrumented segment was noted on magnetic resonance imaging at the 2-year follow-up. CONCLUSION: Rehydration of the degenerated disc in our patient indicates that the BioFlex dynamic stabilization system may promote disc regeneration. Further research is needed to provide more evidence to support lumbar disc rehydration in the bridged segment using this system.

Choice and management of negative pressure drainage in anterior cervical surgery.

BACKGROUND: Postoperative unobstructed drainage is an important measure for avoiding hematoma formation and preventing complications from anterior cervical surgery. AIM: To discuss the characteristics and key points of clinical management of two types of commonly used negative pressure drainage systems in clinical settings. METHODS: Two types of commonly used silica gel negative pressure drainage balls and a type of gastrointestinal decompression apparatus were fully emptied and then injected with different amounts of water and air. Following this, the negative pressure values of the three devices were measured. Meanwhile, we undertook a retrospective analysis of the clinical data of 1328 patients who had been treated with different negative pressure drainage apparatuses during their anterior cervical surgery in our department between January 2007 and January 2018. RESULTS: As the amount of injected air or water increased, the negative pressure of the silica gel negative pressure drainage ball decreased rapidly, dropping to zero when 150 mL of water or air was injected. In contrast, the negative pressure of gastrointestinal decompression apparatus decreased slowly, maintaining an ideal value even when 300 mL of water or air was injected. And statistical analysis demonstrated that patients who had been treated with the gastrointestinal decompression apparatus were less likely to develop severe complications than those who had been treated with the silica gel negative pressure drainage ball (P < 0.05). CONCLUSION: This study showed that the gastrointestinal decompression apparatus has the advantages of large suction capacity, long duration of continuous negative pressure, and good drainage effect, all of which are the favorable factors for the use of this apparatus for negative pressure drainage in anterior cervical surgery.

microRNA-4270-5p inhibits cancer cell proliferation and metastasis in hepatocellular carcinoma by targeting SATB2.

Hepatocellular carcinoma (HCC) remains a lethal cancer type for both males and females. MicroRNAs (miRNAs) contribute to the initiation, development and metastasis of cancer. Although several miRNAs have been identified as drivers or suppressors of HCC, the molecular mechanisms of many miRNAs have not been investigated. Currently, we discovered that miR-4270-5p was a significantly downregulated miRNA in HCC. We revealed that miR-4270-5p overexpression inhibited cell proliferation and invasion of HCC cells. The data manifested that miR-4270-5p directly targeted SATB2, a key regulator of epithelial mesenchymal transition (EMT), in HCC cells and reversed the EMT process. The rescue experiments suggested that SATB2 overexpression reversed the biological function of miR-4270-5p in HCC cells. Clinical data indicated that SATB2 expression was negatively correlated with miR-4270-5p levels in HCC patients. Our findings provided potential targets for prognosis and treatment of patients with HCC.

Characterization of the complete chloroplast genome of Melampyrum roseum.

In this study, the complete chloroplast (cp) genome of Melampyrum roseum was determined through Illumina sequencing method. The complete cp genome of M. roseum was 143,896 bp in length and contained a pair of IR regions (25,210 bp) separated by a small single-copy region (10,292 bp) and a large single-copy region (83,184 bp). The cp genome of M. roseum encoded 117 genes including 78 protein-coding genes, 31 tRNA genes, and 8 rRNA genes. The overall GC content of M. roseum cp genome is 38.1%. By phylogenetic analysis using maximum-likelihood (ML) method, M. roseum was placed in Rhinantheae clade as expected.

Characterization of the complete chloroplast genome of Phtheirospermum japonicum (Rhinantheae).

In this study, we report the complete chloroplast (cp) genome of Phtheirospermum japonicum was determined through Illumina sequencing method. The complete chloroplast genome of Ph. japonicum was 153,397 bp in length and contained a pair of IR regions (25,601 bp) separated by a small single copy region (17,728 bp) and a large single copy region (84,467 bp). The cp genome of Ph. japonicum encoded 125 genes including 86 protein-coding genes, 31 tRNA genes and eight ribosomal RNA genes. The overall GC content of Ph. japonicum cp genome is 38.3%. By phylogenetic analysis using ML method, Ph. japonicum was placed in family Orobanchaceae and showed the closest relationship with Castilleja paramensis.

The Effect of LncRNA H19/miR-194-5p Axis on the Epithelial-Mesenchymal Transition of Colorectal Adenocarcinoma.

BACKGROUND/AIMS: Since the combined actions of lncRNAs and miRNAs have been considered to be involved in the occurrence and development of various neoplasms, the main purpose of this study was to discover whether and how lncRNA H19 and miR-194 influenced the epithelial-mesenchymal transition (EMT) process of colorectal adenocarcinoma (CRA). METHODS: Totally 214 pairs of CRA and adjacent normal tissues were collected, and 5 human CRA cell lines (i.e. HCT116, HT-29, RKO SW280 and Lovo) were purchased. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was adopted to quantify the H19 and miR-194-5p expressions in cells and tissues. The expressions of FoxM1, E-cadherin, vimentin, N-cadherin were determined using western blot. On the side, si-H19, si-NC, miR-194-5p mimic, miR-194-5p inhibitor and negative control (NC) were transfected into CRA cell lines. Meanwhile, the invasive, migratory and proliferative conditions of the cells were assessed through transwell, wound healing and colony-forming experiments, with final verification of the relationship between H19 and miR-194-5p employing dual-luciferase reporter gene assay. RESULTS: Highly-expressed H19, lowly-expressed miR-194-5p, low-grade differentiation and lymph node metastasis appeared as the independent predictors of unfavorable prognosis in CRA patients' (all P< 0.05). It indicated that FoxM1 expression displayed positive correlations with H19 expression, yet negative associations with miR-194-5p expression within CRA tissues (P< 0.05). In addition, transfection of H19-siRNA and miR-145-5p mimic triggered a conspicuous increase in E-cadherin expression, as well as an evidently down-regulation in vimentin and N-cadherin expressions within HT29 and RKO cells (P< 0.05). On the other hand, the invasive and migratory capacities of CRA cells were significantly hindered (P< 0.05). Moreover, the luciferase reporter gene assay confirmed that H19 modified miR-194-5p expression through directly targeting at it (P< 0.05). Ultimately, FoxM1 could reverse the role of miR-194-5p in inhibiting invasion, migration and EMT of CRA cells (P< 0.05). CONCLUSION: LncRNA H19/miR-194/FoxM1 axis could serve as a profound target for the diagnosis and treatment of CRA.

A novel endogenous badnavirus exists in Alhagi sparsifolia.

We report the recovery of a 7068-nt viral sequence from the "viral fossils" embedded in the genome of Alhagi sparsifolia, a typical desert plant. Although the full viral genome remains to be completed, the putative genome structure, the deduced amino acids and phylogenetic analysis unambiguously demonstrate that this viral sequence represents a novel species of the genus Badnavirus. The putative virus is tentatively termed Alhagi bacilliform virus (ABV). Southern blotting and inverse polymerase chain reaction (PCR) data indicate that the ABV-related sequence is integrated into the A. sparsifolia genome, and probably does not give rise to functional episomal virus. Molecular evidence that the ABV sequence exists widely in A. sparsifolia is also presented. To our knowledge, this is the first endogenous badnavirus identified from plants in the Gobi desert, and may provide new clues on the evolution, geographical distribution as well as the host range of the badnaviruses.

Procaine is a specific DNA methylation inhibitor with anti-tumor effect for human gastric cancer.

DNA hypermethylation and the silencing of tumor suppressor genes caused by DNA hypermethylation is considered as a molecular hallmark of many kinds of cancers. Procaine, a local anesthetic, has been shown as a potential DNA methylation inhibitor in some types of cancers. However, the influence of procaine on DNA methylation regulation as well as the biological function in gastric cancer is still unknown. We report here that procaine represses the DNA-methylation level and promotes the proliferation arrest and apoptosis of gastric cancer cells. Global DNA methylation measurement demonstrates that procaine significantly reduces the global DNA methylation level. Analyses of the DNMTs expression and activity show procaine represses the activity, but not the expression, of DNMT1/DNMT3A. Further evidence on specific genes shows that procaine reduces the DNA methylation level in the promoter regions of CDKN2A and RARß genes through abrogating the binding of DNMT1/DNMT3A toward these regions. This repression would not be reversed by the overexpression of DNMT1/DNMT3A. Moreover, RT-qPCR and luciferase report assays demonstrate that procaine leads to the upregulation of CDKN2A and RARß due to the activation of the promoter of these genes. In the end, we test the function of procaine toward gastric cancer cells and find that procaine has the growth inhibitory and apoptosis inducement effect toward gastric cancer cells. Collectively, our data not only uncovers the regulation mechanisms of procaine to DNA methylation but also suggests an anti-tumor potential of procaine specific to the gastric carcinoma and provides a new therapeutic strategy for gastric carcinoma.

Two thymol derivatives from the flower buds of Lonicera japonica and their antibacterial activity.

A new monoterpenoid, 7-acetyl-8,9-dihydroxy thymol (1), together with a known one 7,8-dihydroxy-9-buyryl thymol (2), were isolated from the dried flower buds of Lonicera japonica Thunb. Their structures were elucidated on the basis of extensive 1D and 2D NMR spectroscopic data analyses. The potential antibacterial effects of these compounds on Staphylococcus aureus, Escherichia coli, Micrococcus luteus, and Bacillus cereus were evaluated. Interestingly, both compounds 1 and 2 exhibited significant antibacterial activities with IC50 values range from 27.64 ± 2.26 to 128.58 ± 13.26 µg/mL.

IL-35 induces N2 phenotype of neutrophils to promote tumor growth.

IL-35 is an immunosuppressive cytokine and exerts regulatory effects on T cells, B cells, macrophages and dendritic cells. Neutrophils are important innate immune cells that play key roles in tumor development. The effect of IL-35 on neutrophils remains unknown. Here, we report that IL-35 can induce N2 neutrophil polarization (protumor phenotype) by increasing G-CSF and IL-6 production, and promote neutrophil infiltration into tumor microenvironment. The sustained expression of IL-35 could promote chronic inflammation to augment the proangiogenic and immunosuppressive function of neutrophils. IL-35 stimulated macrophages to secrete proinflammatory cytokines IL-1ß and IL-6. IL-1ß stimulated γδ T cells to produce IL-17, which in turn increased the production of G-CSF. By increasing the expression of G-CSF and IL-6, IL-35 could up-regulate the expression of MMP-9 and Bv8, and down-regulate TRAIL expression in neutrophils, thus augmenting the proangiogenic function of neutrophils. Moreover, G-CSF/IL-6 induced the enhanced activation of STAT3 and ERK pathways in neutrophils, thus increasing the expression of iNOS to suppress T cell activation. Our findings suggest that IL-35 can promote tumor progression by functioning as an up-stream cytokine to promote cancer-associated inflammation and control neutrophil polarization. Targeting IL-35 might be an important approach for designing new strategy of tumor therapy.

miR-938 promotes colorectal cancer cell proliferation via targeting tumor suppressor PHLPP2.

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths worldwide. Although the development of therapy approaches, the outcome of CRC patients still is poor, understanding the biological mechanism of CRC progression is critical to improve the treatment strategies. miRNAs regulate CRC progression, we found miR-938 was upregulated in CRC tissues and cells, MTT assay, colony formation assay and soft agar growth assay suggested miR-938 overexpression promoted CRC cell proliferation, miR-938 knockdown inhibited CRC cell proliferation. Tumor suppressor PH domain Leucine-rich-repeats Protein Phosphatase 2 (PHLPP2) was a target of miR-938, miR-938 inhibited PHLPP2, luciferase activity assay suggested miR-938 directly bound to the 3'UTR of PHLPP2, meanwhile, we found miR-938 promoted c-Myc and Cyclin D1 expression, confirming miR-938 promoted CRC cell proliferation. Double knockdown of miR-938 and PHLPP2 promoted CRC cell proliferation, suggesting miR-938 promoted CRC cell proliferation by inhibiting PHLPP2.

Circulating tumor cells promote the metastatic colonization of disseminated carcinoma cells by inducing systemic inflammation.

Circulating tumor cells (CTCs) have been studied well in the prognosis for malignant diseases as liquid biopsy, but their contribution to tumor metastasis is not clearly defined. Here we report that CTCs could promote the metastatic colonization of disseminated carcinoma cells by inducing systemic inflammation and neutrophil recruitment to pre-metastatic organs. Depletion of neutrophils in vivo could effectively abrogate the promoting effect of CTCs on tumor cell metastasis. In the presence of CTCs, the pro-tumor function of neutrophils was augmented, whereas the antitumor function of neutrophils was suppressed. Mechanically, CTC-derived ligands for TLR2 and TLR4 (TLR2/4) induced the systemic inflammation, thus increasing the production of proinflammatory cytokines such as G-CSF and IL-6 that could induce the conversion of neutrophil function from tumor-suppressing to tumor-promoting. Moreover, CTCs induced the production of endogenous TLR2/4 ligands such as S100A8, S100A9, and SAA3, which may amplify the stimulating effect that induces the expression of proinflammatory cytokines. The promoting effect of CTCs on tumor cell metastasis could be abrogated by suppressing inflammatory response with IL-37, an anti-inflammatory cytokine, or blocking CTC-derived ligands for TLR2/4. Identification of the metastatic axis of CTCs/systemic inflammation/neutrophils may provide potential targets for preventing tumor cell metastasis.