Comparison of Fusion Rate and Clinical Outcomes in Minimally Invasive and Conventional Posterior Fusion for Lumbar Degenerative Disease: A Network Meta-Analysis.

BACKGROUND: The fusion rate, clinical efficacy, and complications of minimally invasive fusion surgery and open fusion surgery in the treatment of lumbar degenerative disease are still unclear. METHODS: We conducted a literature search using PubMed, Embase, Cochrane Library, CNKI, and WANFANG databases. RESULTS: This study included 38 retrospective studies involving 3097 patients. Five intervention modalities were considered: unilateral biportal endoscopic-lumbar interbody fusion (UBE-LIF), percutaneous endoscopic-lumbar interbody fusion (PE-LIF), minimally invasive-transforaminal lumbar interbody fusion (MIS-TLIF), transforaminal lumbar interbody fusion (TLIF), and posterior lumbar interbody fusion (PLIF). Quality assessment indicated that each study met acceptable quality standards. PE-LIF demonstrated reduced low back pain (Odds Ratio = 0.50, Confidence Interval: 0.38-0.65) and lower complication rate (Odds Ratio = 0.46, Confidence Interval: 0.25-0.87) compared to PLIF. However, in indirect comparisons, PE-LIF showed the lowest fusion rates, with the ranking as follows: UBE-LIF (83.2%) > MIS-TLIF (59.6%) > TLIF (44.3%) > PLIF (39.8%) > PE-LIF (23.1%). With respect to low back pain relief, PE-LIF yielded the best results, with the order of relief as follows: PE-LIF (96.4%) > MIS-TLIF (64.8%) > UBE-LIF (62.6%) > TLIF (23.0%) > PLIF (3.2%). Global and local consistency tests showed satisfactory results, and heterogeneity tests indicated good stability. CONCLUSIONS: Compared to conventional open surgery, minimally invasive fusion surgery offered better scores for low back pain and Oswestry Disability Index, lower complication rates, reduced bleeding, and shorter hospital stays. However, minimally invasive fusion surgery did not show a significant advantage in terms of fusion rate and had a longer operative time.

TLR4 Inhibition Protects against Retinal Ganglion Cell Damage in Rats with Chronic Ocular Hypertension.

BACKGROUND: This study aims to investigate the protective effect of Toll-like receptor 4 (TLR4) inhibitor Resatorvid (TAK-242) on retinal ganglion cells (RGCs) in a chronic ocular hypertension (COH) rat model, as well as to explore the potential involved mechanisms. METHODS: COH model was built up in rats with a single intracameral administration of cross-linking hydrogel. The expression levels of TLR4, NLR family pyrin domain containing 3 (NLRP3), microglial activation and pro-inflammatory cytokines were evaluated in COH retinas and COH retinas treated with TAK-242 using immunofluorescence staining and Western blot. Additionally, retrograde labeling and neuronal nuclear protein (NeuN) staining were performed to count RGCs. RESULTS: Activated microglia and increased TLR4 expression were observed in the retinas of COH rats. This was accompanied by upregulated expressions of NLRP3, tumor necrosis factor alpha (TNF-α), cytokine interleukin-1ß (IL-1ß) and Interleukin-6 (IL-6). Intravitreal injection of TAK-242 promoted the survival of RGCs by attenuating microglial activation, interfering with the TLR4-NLRP3 pathway and regulating pro-inflammatory cytokines. CONCLUSIONS: Targeting TLR4 inhibition could be a potential therapeutic strategy to protect RGCs from COH damage.

FOXI3 pathogenic variants cause one form of craniofacial microsomia.

Craniofacial microsomia (CFM; also known as Goldenhar syndrome), is a craniofacial developmental disorder of variable expressivity and severity with a recognizable set of abnormalities. These birth defects are associated with structures derived from the first and second pharyngeal arches, can occur unilaterally and include ear dysplasia, microtia, preauricular tags and pits, facial asymmetry and other malformations. The inheritance pattern is controversial, and the molecular etiology of this syndrome is largely unknown. A total of 670 patients belonging to unrelated pedigrees with European and Chinese ancestry with CFM, are investigated. We identify 18 likely pathogenic variants in 21 probands (3.1%) in FOXI3. Biochemical experiments on transcriptional activity and subcellular localization of the likely pathogenic FOXI3 variants, and knock-in mouse studies strongly support the involvement of FOXI3 in CFM. Our findings indicate autosomal dominant inheritance with reduced penetrance, and/or autosomal recessive inheritance. The phenotypic expression of the FOXI3 variants is variable. The penetrance of the likely pathogenic variants in the seemingly dominant form is reduced, since a considerable number of such variants in affected individuals were inherited from non-affected parents. Here we provide suggestive evidence that common variation in the FOXI3 allele in trans with the pathogenic variant could modify the phenotypic severity and accounts for the incomplete penetrance.

The Valsa Mali effector Vm1G-1794 protects the aggregated MdEF-Tu from autophagic degradation to promote infection in apple.

Macroautophagy/autophagy is a conserved degradation pathway in eukaryotes that is required for recycling unwanted intracellular components, maintaining homeostasis, and coping with biotic and abiotic stresses. Pathogens have evolved to subvert autophagic machinery by secreting host cell-entering effector proteins. Here, we provided evidence that an apple autophagy-related gene MdATG8i, activated autophagy and contributed to resistance against Valsa canker caused by Valsa Mali (Vm) when being overexpressed in apple. MdATG8i interacted with a plastid elongation factor Tu (MdEF-Tu) which became insoluble and aggregated during Vm infection and was degraded through the autophagy pathway. Intriguingly, we identified a highly-induced effector secreted from Vm, Vm1G-1794, which competitively interacted with MdATG8i, suppressed autophagy, and depleted MdEF-Tu out of MdATG8i complexes. The formation of stable MdEF-Tu aggregates caused by Vm1G-1794 promoted the susceptibility of apple to Vm. Overall, our study demonstrated that MdATG8i contributed to Vm resistance by targeting and degrading MdEF-Tu, and Vm1G-1794 competed with MdEF-Tu to target MdATG8i and prevent MdEF-Tu degradation, thus favoring infection.Abbreviations: 35S: cauliflower mosaic virus 35S promoter; AIM: ATG8-interacting motif; ATG8-PE: ATG8 conjugated with phosphatidylethanolamine; BiFC: biomolecular fluorescence complementation; Con A: concanamycin A; Co-IP: co-immunoprecipitation; DEPs: differentially expressed proteins; DMSO: dimethyl sulfoxide; GFP: green fluorescent protein; hpt: hours post-treatment; LCI: luciferase complementation imaging; MdATG8i: autophagy-related protein 8i in Malus domestica; MDC: monodansylcadaverine; MdEF-Tu: elongation factor Tu in Malus domestica; MdNBR1: neighbor of BRCA1 in Malus domestica; N. benthamiana: Nicotiana benthamiana; OE: overexpression; PAMP: pathogen-associated molecular pattern; PTI: pattern-triggered immunity; qRT-PCR: quantitative reverse transcription PCR; RFP: red fluorescent protein; RNAi: RNA interference; ROS: reactive oxygen species; Ub: ubiquitin; V. Mali: Valsa Mali; WT: wild-type plant; YFP: yellow fluorescent protein.

Quantitative evaluation of retinal and choroidal changes in Fabry disease using optical coherence tomography angiography.

To examine the retinal and choroidal changes in patients with Fabry disease (FD) using optical coherence tomography angiography (OCTA). FD patients and age- and sex-matched healthy subjects were enrolled. A detailed ophthalmological examination was performed for all participants. The retinal thickness, ganglion cell layer with inner plexiform layer (GCIPL) thickness, choroidal thickness (CT), vessel length density (VLD), vessel perfusion density (VPD), and foveal avascular zone (FAZ) were analyzed in a detailed way with OCTA. Moreover, all FD patients underwent several laboratory tests to evaluate systemic conditions. A total of 54 subjects comprising 26 FD patients and 28 normal controls were enrolled. The retinal thickness, GCIPL thickness, and FAZ area showed no significant differences between the two groups (all P > 0.05). Only the superior CT in FD patients was significantly thinner than that in the normal subjects (P = 0.040). The macular VLD and VPD in the FD group were significantly reduced compared with the healthy controls (P = 0.026, P = 0.008). The macular VLD in FD patients had no significant correlations with different laboratory results (all P > 0.05), while the macular VPD were negatively correlated with creatinine (r = - 0.432, P = 0.028) and cystatin C (r = - 0.422, P = 0.032). FD patients may have retinal vascular dropout and choroidal vascular alterations. Analysis of vessel density using OCTA might be useful in the clinical assessment in FD patients.

RGD-modified multifunctional nanoparticles encapsulating salvianolic acid A for targeted treatment of choroidal neovascularization.

BACKGROUND: The development of alternative anti-angiogenesis therapy for choroidal neovascularization (CNV) remains a great challenge. Nanoparticle systems have emerged as a new form of drug delivery in ocular diseases. Here, we report the construction and characterization of arginine-glycine-aspartic acid (RGD)-conjugated polyethyleneimine (PEI) as a vehicle to load antioxidant salvianolic acid A (SAA) for targeted anti-angiogenesis therapy of CNV. In this study, PEI was consecutively modified with polyethylene glycol (PEG) conjugated RGD segments, 3-(4'-hydroxyphenyl) propionic acid-Osu (HPAO), and fluorescein isothiocyanate (FI), followed by acetylation of the remaining PEI surface amines to generate the multifunctional PEI vehicle PEI.NHAc-FI-HPAO-(PEG-RGD) (for short, RGD-PEI). The formed RGD-PEI was utilized as an effective vehicle platform to load SAA. RESULTS: We showed that RGD-PEI/SAA complexes displayed desirable water dispersibility, low cytotoxicity, and sustainable release of SAA under different pH conditions. It could be specifically taken up by retinal pigment epithelium (RPE) cells which highly expressed ɑvß5 integrin receptors in vitro and selectively accumulated in CNV lesions in vivo. Moreover, the complexes displayed specific therapeutic efficacy in a mouse model of laser induced CNV, and the slow elimination of the complexes in the vitreous cavity was verified by SPECT imaging after 131I radiolabeling. The histological examinations further confirmed the biocompatibility of RGD-PEI/SAA. CONCLUSIONS: The results suggest that the designed RGD-PEI/SAA complexes may be a potential alternative anti-angiogenesis therapy for posterior ocular neovascular diseases.

[Channel-assisted fixation and interbody fusion in treating recurrent lumbar disc herniation by muscle-splitting approach].

OBJECTIVE: To explore the clinical value and safety of unilateral pedicle screw fixation combined with contralateral translaminar facet screw fixation and interbody fusion by muscle-splitting approach treatment of recurrent lumbar disc herniation. METHODS: The clinical data of 51 patients with recurrent lumbar disc herniation treated from June 2012 to December 2017 were retrospectively analyzed. There were 32 males and 19 females, aged 34 to 64 years with an average of (51.11± 7.28) years. Lesions invoved L4,5 in 38 cases and L5S1 in 13 cases. All patients had a history of lower back pain and radiation pain of lower limbs(3 bilateral and 48 unilateral)and underwent unilateral pedicle screw combined with contralateral translaminar facet screw fixation and interbody fusion, among which 24 patients were treated through median incision approach (median incision group);other 27 patients were treated through muscle-splitting approach with channel-assisted exposure(muscle-splitting approach group). Operation time, intraoperative blood loss, postoperative drainage and incision length of the two groups were recorded. Visual analogue scale (VAS) was used to score the pain of lumbar incision at 72 h after operation, and JOA low back pain scoring system was used to evaluate the lumbar function preoperatively and at final follow-up. Imaging data were analyzed, including the changes in the height of intervertebral space of diseased segment before operation, 3 to 5 days after operation, and at final follow-up;Cobb angle changes in the coronal and sagittal planes of lumbar spine preoperatively and at final follow-up;multifidus area and multifidus fatty tissue deposition grade before and 12 months after operation; postoperative pedicle screw and laminar process screw position and intervertebral fusion condition. The complications of the two groups were compared. RESULTS: There was no statistical difference in operation time between two groups (P>0.05). Muscle-splitting approach group was better than median incision group in light of incision length, intraoperative blood loss and postoperative drainage volume (P<0.05). VAS score of lumbar incision pain at 72 h after operation was 1.61±0.54 in median incision group and 0.76±0.28 in muscle-splitting approach group(P<0.05). All patients were followed up for 12 to 84 (43.50±15.84) months. At final follow-up, the JOA scores of the two groups were significantly improved compared with those before operation(P<0.05). The rate of pedicle screw malposition was 6.25%(3/48) in medianincision group and 9.26%(5/54) in muscle-splitting approach group, there was no statistically significant difference between two groups (P>0.05). Rate of translaminar facet screw malposition in median incision group (12.50%) was significant less than the muscle-splitting approach group (18.52%)(P< 0.05). The height of the intervertebral space of the two groups was significantly restored 3 to 5 days after operation (P<0.05), and there was also a significant loss of height at final follow-up (P<0.05). At final follow-up, the balance of lumbar coronal plane and sagittal plane in two groups were improved very well (P<0.05). The comparison of the area and grade of the multifidus muscle in two groups 12 months after operation showed that obvious damage to the multifidus muscle were present in the median incision, while the multifidus muscle was less damaged by muscle-splitting approach (P<0.05). The fusion rate was 91.7%(22/24) in the median incision group and 92.6%(25/27) in muscle-splitting approach group(P>0.05). In median incision group, there were 1 case of intraoperative pedicle entry point fracture, 1 case of intraoperative dural tear and 1 case of postoperative nerve root injury;in muscle-splitting approach group, there were 1 case of intraoperative pedicle entry point fracture, 2 cases of intraoperative dural tear, 1 case of postoperative nerve root injury, 2 cases of incision epidermal necrosis and 1 case of poor incision healing. Nerve root injuries in the two groups were caused by incorrect positions of pedicle screws, the screws were immediately adjusted upon discovery. The nerve root symptoms were completely recovered 3 and 6 months after surgery. No incision infection was occurred in two groups. During the follow-up, no pedicle screw and laminar facet screw were loosened, displaced, broken, or intervertebral fusion cage moved forward and backward. The complication rate of 25.93% in muscle-splitting approach group was higher than 12.50% in the median incision group (P<0.05). CONCLUSION: Muscle-splitting approach is feasible for thetreatment of recurrent lumbar disc herniation with pedicle screw fixation combined with contralateral translaminar facet screw fixation and interbody fusion. Compared with the median incision approach, the muscle-splitting approach has the advantages of small incision, less trauma, less bleeding, rapid recovery. Also it can protect multifidus and do not increase the incidence of serious complications. Thus, it can be used as a choice for fixation and fusion of recurrent lumbar disc herniation.

Novel risk factors for craniofacial microsomia and assessment of their utility in clinic diagnosis.

Craniofacial microsomia (CFM, OMIM%164 210) is one of the most common congenital facial abnormalities worldwide, but it's genetic risk factors and environmental threats are poorly investigated, as well as their interaction, making the diagnosis and prenatal screening of CFM impossible. We perform a comprehensive association study on the largest CFM cohort of 6074 samples. We identify 15 significant (P < 5 × 10-8) associated genomic loci (including eight previously reported) and decipher 107 candidates based on multi-omics data. Gene Ontology term enrichment found that these candidates are mainly enriched in neural crest cell (NCC) development and hypoxic environment. Single-cell RNA-seq data of mouse embryo demonstrate that nine of them show dramatic expression change during early cranial NCC development whose dysplasia is involved in pathogeny of CFM. Furthermore, we construct a well-performed CFM risk-predicting model based on polygenic risk score (PRS) method and estimate seven environmental risk factors that interacting with PRS. Single-nucleotide polymorphism-based PRS is significantly associated with CFM [P = 7.22 × 10-58, odds ratio = 3.15, 95% confidence interval (CI) 2.74-3.63], and the top fifth percentile has a 6.8-fold CFM risk comparing with the 10th percentile. Father's smoking increases CFM risk as evidenced by interaction parameter of -0.324 (95% CI -0.578 to -0.070, P = 0.011) with PRS. In conclusion, the newly identified risk loci will significantly improve our understandings of genetics contribution to CFM. The risk prediction model is promising for CFM prediction, and father's smoking is a key environmental risk factor for CFM through interacting with genetic factors.

Tetramethylpyrazine protects mice retinas against sodium iodate-induced oxidative injury.

Purpose: To observe the effects of tetramethylpyrazine (TMP) on mice retinas injured by sodium iodate (NaIO3). Methods: Male mice (n = 45) were randomly divided into three groups: the control group (Group C), the NaIO3-degenerated group (Group I), and the TMP-treated group (TMP group). The Group I mice were intraperitoneally injected with 35 mg/kg NaIO3. The Group C mice were injected with similar volumes of PBS. The TMP group mice were intraperitoneally injected with 80 mg/kg TMP starting 24 h after NaIO3 administration once a day for 14 days. Fundus photography, optical coherence tomography (OCT), electroretinography (ERG), hematoxylin and eosin (H&E) staining, terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assay, and western blotting were used to assess the effects of TMP on mice retinas at day 3, 7, and 14 after NaIO3 administration. Results: TMP effectively prevented the decrease in the thicknesses of the retinas and the outer nuclear layer (ONL), and effectively alleviated the functional decline in the retinas after NaIO3 administration. TMP significantly decreased the number of TUNEL-positive cells in retinas. In addition, TMP rapidly increased the expression of Nrf2 and HO-1 and decreased BAX expression in mice retinas after NaIO3 injection. Conclusions: TMP alleviates morphological and functional retinal damage in mice exposed to NaIO3 and reduces retinal apoptosis.

LncRNA SAMMSON overexpression distinguished glioblastoma patients from patients with diffuse neurosarcoidosis.

The MRI characteristics of diffuse neurosarcoidosis are similar to those of glioblastoma. Therefore, identification of novel biomarkers to distinguish these two diseases is needed. We found that lncRNA Survival Associated Mitochondrial Melanoma-Specific Oncogenic Non-Coding RNA (SAMMSON) was upregulated in plasma of glioblastoma patients but not in diffuse neurosarcoidosis patients comparing to healthy controls. Upregulated SAMMSON distinguished glioblastoma patients from diffuse neurosarcoidosis patients and healthy controls. MiR-622 in glioblastoma patients was inversely correlated with SAMMSON. SAMMSON overexpression caused the downregulated expression of miR-622 in glioblastoma cells, while miR-622 overexpression did not affect SAMMSON expression. SAMMSON overexpression mediated the increased proliferation rate of glioblastoma cells. MiR-622 overexpression played an opposite role and reduced the effects of SAMMSON overexpression. Therefore, plasma SAMMSON has diagnostic value for glioblastoma and SAMMSON overexpression may promote glioblastoma cell proliferation by downregulating miR-622.

Baicalein and baicalin inhibit colon cancer using two distinct fashions of apoptosis and senescence.

Baicalein and baicalin are active components of the Scutellaria baicalensis Georgi and both have broad anti-tumor activity. However, how and whether baicalein and baicalin inhibit colon cancer is unclear. Here we demonstrate that baicalein and baicalin can significantly inhibit human colon cancer cell growth and proliferation. Furthermore, both can induce cell cycle arrest, and suppress cancer cell colony formation and migration. The suppressive effects are mechanistically due to the induction of colon cancer cell apoptosis and senescence mediated by baicalein and baicalin, respectively. Furthermore, we revealed that baicalin-induced senescence in tumor cells is due to its inhibition of telomerase reverse transcriptase expression in tumor cells, and that MAPK ERK and p38 signaling pathways are causatively involved in the regulation of colon cancer cell apoptosis and senescence mediated by baicalein and baicalin. In addition, our in vivo studies using human colon cancer cells in humanized mouse xenograft models, further demonstrated that baicalein and baicalin can induce tumor cell apoptosis and senescence, resulting in inhibition of tumorigenesis and growth of colon cancer in vivo. These data clearly suggest that baicalein and baicalin have potent anti-cancer effects against human colon cancer and could be potential novel and effective target drugs for cancer therapy.

Structural and functional analyses of genes encoding VQ proteins in apple.

Recent studies with Arabidopsis and soybean have shown that a class of valine-glutamine (VQ) motif-containing proteins interacts with some WRKY transcription factors. However, little is known about the evolution, structures, and functions of those proteins in apple. Here, we examined their features and identified 49 apple VQ genes. Our evolutional analysis revealed that the proteins could be clustered into nine groups together with their homologues in 33 species. Historically, the main characteristics of proteins in Groups I, V, VI, VII, IX, and X were thought to have been generated before the monocot-dicot split, whereas those in Groups II, III + IV, and VIII were generated after that split. In the structural analysis, apple MdVQ proteins appeared to bind only with Group I and IIc MdWRKY proteins. Meanwhile, MdVQ1, MdVQ10, MdVQ15, and MdVQ36 interacted with multiple MdVQ proteins to form heterodimers but MdVQ15 formed a homodimer. The functional analysis indicated that overexpression of some apple MdVQs in Arabidopsis and tobacco plants effected their vegetative and reproductive growth. These results provide important information about the characteristics of apple MdVQ genes and can serve as a solid foundation for further studies about the role of WRKY-VQ interactions in regulating apple developmental and defense mechanisms.

Baicalin induces cellular senescence in human colon cancer cells via upregulation of DEPP and the activation of Ras/Raf/MEK/ERK signaling.

Baicalin is a natural flavonoid glycoside which has potent anti-tumor and antioxidant activity in cancer cells. In the present study, we found that baicalin treatment significantly induced senescence in colon cancer cells. Furthermore, baicalin upregulated the expression of decidual protein induced by progesterone (DEPP) in HCT116 colon cancer cells, which accompanied with the activation of Ras/Raf/MEK/ERK and p16INK4A/Rb signaling pathways. Meanwhile, these phenomena also appeared under the anti-oxidation effect exerted by baicalin. In addition, ectopic expression of DEPP in HCT116 cells significantly induced the activity of senescence-associated ß-galactosidase (SA-ß-Gal) in tumor cells regulated by Ras/Raf/MEK/ERK signaling pathway. Knockdown of DEPP by RNA interference efficiently counteracted the baicalin-mediated growth inhibition, senescence and cell cycle arrest in cancer cells. Importantly, in a xenograft mouse model of human colon cancer, we further confirmed that baicalin treatment dramatically inhibited tumor growth, which was due to the induction of tumor cellular senescence via the upregulation of DEPP and the activation of Ras/Raf/MEK/ERK signaling in vivo. In addition to baicalin treatment, we found that the hypoxia-response protein DEPP functions as a positive regulator involving the regulations of Ras/Raf/MEK/ERK signaling pathway and inhibition of human colon cancer by other anti-oxidative drugs, such as curcumin and sulforaphane, resulting in tumor cellular senescence. These results collectively suggest that baicalin upregulates the expression of DEPP and activates its downstream Ras/Raf/MEK/ERK and p16INK4A/Rb pathways by acting as an antioxidant, leading to senescence in colon cancer cells.

Risk Factors Analyses for Seizure Recurrence in Different Periods After Refractory Epilepsy Surgery: A Prospective Single-Center Study.

OBJECTIVE: To evaluate the potential risk factors associated with seizure recurrence in different periods after epilepsy surgery. METHODS: A total of 303 patients with refractory epilepsy after epilepsy surgery were included. The Kaplan-Meier method with log-rank test and univariate and multivariate Cox proportional hazards model were performed to calculate the comparison of survival curves between groups and identify the risk factors associated with seizure recurrence in different periods after surgery. RESULTS: The significant predictors of seizure recurrence were determined, including duration of epilepsy (P = 0.018), seizure types (P = 0.009), magnetic resonance imaging findings (P = 0.007), intracranial electroencephalographic recordings (P = 0.002), sides of epileptogenic zone (P = 0.025), and types of surgery (P = 0.002). Moreover, the significant predictors of seizure recurrence within 12 months after surgery were also included, such as gender (P = 0.007), duration of epilepsy (P = 0.013), intracranial electroencephalographic recordings (P = 0.003), and types of surgery (P < 0.001). Our results indicated that the variables of magnetic resonance imaging findings (P = 0.015), sides of epileptogenic zone (P = 0.004), and seizure relapse within 12 months after surgery (P < 0.001) were significantly associated with seizure recurrence in 12-36 months after surgery. Seizure relapse within 12 months after surgery (P < 0.001) was also associated with seizure recurrence >36 months after surgery. CONCLUSIONS: We reconfirmed the well-known risk factors associated with seizure recurrence and also identified the controversial variables. In addition, we found that the risk factors associated with seizure recurrence were different in different periods after epilepsy surgery.

Salvianolic Acid A Inhibits OX-LDL Effects on Exacerbating Choroidal Neovascularization via Downregulating CYLD.

BACKGROUNDS: Age-related macular degeneration is closely related to lipid oxidation, while relationship between OX-LDL and choroidal neovascularization is unclear. Recently, cylindromatosis is proved to regulate angiogenesis. However, its role in CNV progression remained unclear. Salvianolic acid A is widely used in vascular diseases. We investigated the relationship between OX-LDL and CNV and explore antineovascularization mechanism of Sal A. METHODS: C57BL6/J mice were randomized into four groups and injected with PBS or OX-LDL, together with Sal A for one week. CNV was induced by laser; CNV severity was analyzed by fundus fluorescein angiography, H&E staining, and choroid flat mount after 1 week. In in vitro experiments, ARPE-19 and HUVECs were cultured with OX-LDL (with or without Sal A) for 48 hours. Angiogenic proteins, cell junction integrity, and tube formation were measured. CYLD siRNA and specific inhibitors were used to explore mechanisms of CYLD in promoting OX-LDL-induced CNV progression. RESULTS: OX-LDL promoted laser-induced CNV volume by increasing VEGF, PDGF, and CYLD levels. Sal A antagonized OX-LDL effects and restrained CNV progression by decreasing VEGF/PDGF/CYLD, increasing antiangiostatin levels, and promoting P62-CYLD-TRAF6 interaction. CONCLUSIONS: We demonstrated oxidation damage exacerbates CNV progression, and Sal A could be a clinical therapeutic reagent to exudative AMD.

MicroRNA-485 inhibits malignant biological behaviour of glioblastoma cells by directly targeting PAK4.

Glioblastoma (GBM), which is characterised by rapid growth, cellular heterogeneity, angiogenesis, extensive invasion, hypoxia and necrosis, is the most common histological subtype of glioma in adults. MicroRNA (miRNA) dysregulation is a common feature of human cancers, including GBM. Previous studies have suggested that miRNAs are a novel class of regulatory molecules in various human cancers. Therefore, miRNAs may be investigated as a novel candidate and screening tool in the clinical diagnosis, therapy and prognosis of GBM. Recent accumulating evidence has demonstrated that miRNA­485 (miR­485) is involved in the development and progression of several types of human cancers. However, the expression level, exact role and underlying mechanisms of miR­485 in GBM remain unclear. In this study, miR­485 was downregulated in both GBM tissue specimens and cell lines. miR­485 overexpression inhibited GBM cell proliferation, colony formation, migration and invasion; increased apoptosis in vitro; and reduced tumour growth in vivo. In addition, p21­activated kinase 4 (PAK4) was demonstrated to be a direct and functional target of miR­485 in GBM. Furthermore, PAK4 was upregulated in GBM tissues and negatively correlated with miR­485 expression. Moreover, PAK4 knockdown exhibited a similar effect to miR­485 overexpression in GBM cells. Enforced expression of PAK4 rescued miR­485 tumour­suppressor functions in GBM cells. miR­485 inhibited the activation of the AKT and ERK signalling pathways in GBM. These results indicate that miR­485 acts as a tumour suppressor in GBM by, at least partially, directly targeting PAK4 and regulating the AKT and ERK signalling pathways. Thus, miR­485 may be a potential target for the treatment of patients with GBM.

Salvianolic acid A protects retinal pigment epithelium from OX-LDL-induced inflammation in an age-related macular degeneration model.

BACKGROUND: Salvianolic acid A (Sal A), an active monomer of Salvia miltiorrhiza, is a phenolic carboxylic acid derivative. The present study was performed to investigate the underlying mechanism of the anti-inflammation effect of Sal A, especially focusing on mTOR-KEAP1-Nrf2 and P2X7R-PKR-NLRP3 signaling pathways. METHODS: SD mice were divided into four groups: PBS, oxidized-low density lipoprotein (ox-LDL, 3 mg/kg), and ox-LDL (3 mg/kg) + Sal A (5 mg/kg) and + Sal A (10 mg/ml) groups. In in vitro experiments, ARPE-19 cells were cultured with serum free medium (SFM) or ox-LDL (100 mg/L), with or without Sal A (5 µM/50 µM) for 24 hours. RESULTS: Sal A attenuated ox-LDL-induced lipidosis and apoptosis in the retinal pigment epithelium (RPE) layer. Ox-LDL elevated ROS level and induced RPE inflammation, which were inhibited by Sal A pretreatment. Sal A activated PI3K/AKT/mTOR signaling pathway, which further promoted the disassociation of Keap1-Nrf2 complex and the phosphorylation of Nrf2. PI3K and mTOR chemical inhibitors abolished Sal A-induced Nrf2 activation while it had no influence on nlrp3 expression. Sal A also inhibited RPE inflammation by inactivating the P2x7r-Pkr-Nlrp3 signaling pathway. CONCLUSIONS: The above results indicate that Sal A protects RPE from lipid oxidative damage and chronic inflammation through up-regulating Nrf2 and inactivating the P2x7r-Pkr-Nlrp3 signaling pathway.

Potential regulation of glioma through the induction of apoptosis signaling via Egl-9 family hypoxia-inducible factor 3.

Glioma is an aggressive form of brain cancer that occurs following the abnormal proliferation of glial cells. Although glioma cannot spread to other organs, the morbidity and mortality rates of the disease are high, even following surgery, radiotherapy and chemotherapy. The function of Egl-9 family hypoxia-inducible factor 3 (Egln3) in cancer is controversial, and it is debated as to whether Egln3 positively or negatively regulates tumors. In the present study, a mouse model of low-grade glioma was successfully established. Through the use of immunohistochemical and western blot analyses, it was demonstrated that Egln3 expression in glioma tissue performed an important role in regulation by amplifying the signals for apoptosis, as determined by an increase in DNA fragments. Furthermore, Egln3 expression was inhibited by the administration of dimethyloxalylglycine, and the downregulated expression of Egln3 had marked effects on the regulation of glioma through apoptosis. The present study therefore provides evidence of an association between Egln3 expression and apoptosis in low-grade glioma.

Anticonvulsant effect of piperine ameliorates memory impairment, inflammation and oxidative stress in a rat model of pilocarpine-induced epilepsy.

The primary active component of black pepper is piperine, which is purified and used to treat epilepsy, achieving higher efficiency when purified. The present study was conducted to evaluate whether the anticonvulsant effect of piperine ameliorates pilocarpine-induced epilepsy, and to investigate the mechanism underlying these effects. Epilepsy was induced in Sprague Dawley rats using pilocarpine. Pilocarpine-induced epilepsy in the rats was treated with 40 mg/kg piperine for 45 consecutive days. Status epilepticus and a Morris water maze test were used to analyze the anticonvulsant effects of piperine in the epileptic rats. Inflammation and oxidative stress were then measured using commercially-available kits following piperine treatment. Lastly, the activity of caspase-3 and the protein expression levels of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X protein (Bax) were evaluated using commercially-available kits and western blot analysis, respectively. The results demonstrated that treatment with piperine was able to reduce the status epilepticus and prevented memory impairment following pilocarpine-induced epilepsy in rats. The anticonvulsant effects of piperine decreased inflammation and oxidative stress following pilocarpine-induced epilepsy in rats. The upregulated activity of caspase-3 and expression levels of Bax/Bcl-2 were suppressed following treatment with piperine in the rats with pilocarpine-induced epilepsy. These results suggest that the anticonvulsant effects of piperine ameliorate memory impairment, inflammation and oxidative stress in a rat model of pilocarpine-induced epilepsy.

Quantitative Study of Posterior Fossa Crowdedness in Hemifacial Spasm.

OBJECTIVE: To quantitatively study the degree of posterior fossa crowdedness (PFC) in patients with hemifacial spasm (HFS) and to further investigate whether overcrowding in posterior fossa affects the efficacy and safety of microvascular decompression. METHODS: We conducted a prospective case-control study of patients diagnosed with HFS and sex- and age-matched healthy control patients. All subjects underwent high-resolution, 3-dimensional magnetic resonance imaging, and posterior fossa crowdedness index (PFCI) and cerebrospinal fluid volume (CSFV) were measured. Patients with HFS underwent primary microvascular decompression and long-term follow-up. Associations of PFCI and other factors with operative outcomes and complications were analyzed. RESULTS: The study enrolled 153 subjects: 102 patients and 51 control subjects. Compared with control subjects, patients had a more PFC (PFCI: 83.2% vs. 80.2%; P = 0.005) and smaller posterior fossa CSFV (16,248.0 mm(3) vs. 20,054.0 mm(3); P = 0.001). The mean effective space of posterior fossa cerebrospinal fluid in patients with HFS was 11.8% smaller than in control subjects (P = 0.004). Compared with men, women showed larger PFCI (83.6% vs. 82.2%; P = 0.012) and smaller PF CSFV (16,027.5 mm(3) vs. 17,299.5 mm(3); P = 0.009). Sixty-one patients (59.8%) were spasm-free immediately postoperatively, and 95 (93.1%) were spasm-free at follow-up. Complication rates were 9.8% in the short term, and 2.9% at follow-up. Lower PFCI (odds ratio [OR] 0.63; P = 0.019) and severe indention (OR 1.41; P = 0.027) were significantly associated with better short-term outcomes. Greater PFCI (OR 2.05, P = 0.008) was associated with greater early complication incidence. CONCLUSION: Patients with HFS had more PFC. PFC potentially leads to cranial nerve and vascular structure crowding, which may increase HFS risk. PFC was significantly associated with poor short-term outcomes and greater incidence of early complications but not long-term outcomes and complications.