Long noncoding RNA SNHG12 promotes vascular smooth muscle cell proliferation and migration via regulating miR-199a-5p/HIF-1α.

The dysregulation of proliferation and migration of vascular smooth muscle cells (VSMCs) contributes to atherosclerosis (AS) and accumulating reports indicate the crucial role of long noncoding RNA in AS. However, the role of small nucleolar RNA host gene 12 (SNHG12) in regulating the phenotypes of VSMCs and AS remains largely unknown. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) was used to detect the expression levels of SNHG12 and miR-199a-5p in an in vivo AS model and VSMCs treated by oxidized low-density lipoprotein (ox-LDL). The proliferation ability, migration ability, and apoptosis of VSMCs were tested by cell counting kit-8, Transwell assay, and terminal deoxynucleotidyl transferase dUTP nick end labeling assay, respectively. StarBase database was used to predict the binding sites between miR-199a-5p and SNHG12. The interaction between miR-199a-5p and SNHG12 was validated by qRT-PCR, western blot, and luciferase reporter assay. Western blot was used to examine the effects of SNHG12 and miR-199a-5p on the expression of hypoxia-inducible factor 1α (HIF-1α). We found that the expression level of SNHG12 was significantly increased in the animal model and VSMCs treated by ox-LDL. Knockdown of SNHG12 suppressed the proliferation and migration abilities of VSMCs, while overexpression of SNHG12 had the opposite effects. Mechanically, we validated that miR-199a-5p was a target of SNHG12, and the target gene of miR-199a-5p, HIF-1α could be indirectly and positively regulated by SNHG12. In conclusion, SHNG12 targeting miR-199a-5p/HIF-1α contributed to the pathophysiological process of AS by regulating the phenotypes of VSMCs, and could be a potential therapy target for this disease.

Increasing the homologous recombination efficiency of eukaryotic microorganisms for enhanced genome engineering.

In recent years, eukaryotic microorganisms have been widely applied to offer many solutions for everyday life and have come to play important roles in agriculture, food, health care, and the fine-chemicals industry. However, the complex genetic background and low homologous recombination efficiency have hampered the implementation of large-scale and high-throughput gene editing in many eukaryotic microorganisms. The low efficiency of homologous recombination (HR) not only makes the modification process labor-intensive but also completely precludes the application of many otherwise very useful genome editing techniques. Thus, increasing the efficiency of HR is clearly an enabling technology for basic research and gene editing in eukaryotic microorganisms. In this review, we summarize the current strategies for enhancing the efficiency of HR in eukaryotic microorganisms (particularly yeasts and filamentous fungi), list some small molecules and candidate genes associated with homologous and non-homologous recombination, and briefly discuss the further development prospects of these strategies.

Effect of sterilization methods on cell binding activity of surface-immobilized fibronectin.

The effect of sterilization methods on biological activity of fibronectin on the surface of biomaterials was elaborated in the present study. Sterile protein- modified biomaterials were fabricated by microfilter filtration and UV irradiation, respectively. UV irradiation altered the conformation of surface- adsorbed fibronectin and further affected the attachment, morphology and biological function of endothelial cells. However, microfilter filtration did not to change the normal conformation of fibronectin, or the proliferation and biological function of endothelial cells, indicating that microfilter filtration sterilization is the most suitable method for protein-substrate.

[Effects of intrathecal injection of methylprednisolone sodium succinate in acute spinal cord injury rabbits].

OBJECTIVE: To investigate the effects and apoptosis of intrathecal injection of Methylprednisolone Sodium Succinate (MPss) for acute spinal cord injury (SCI) in New Zealand rabbits. METHODS: Seventy-two healthy New Zealand rabbits were used for the procedure and were randomly divided into two groups: SCI group and SHAM group, which was both divided into 6 subgroups, such as the vehicle group, the MPss intrathecal injection groups (1.5 mg/kg, 3.0 mg/kg, 6.0 mg/kg group), the MPss intravenous injection group and the combined injection group. TARLOV score was tested daily to evaluate the motor function. The rabbits were sacrificed 7 days after the surgery and the thoracic spinal cord sections and the sacral sections where MPss was injected were harvested for HE and TUNEL staining. Two-Factors Repeated Measures analysis of variance for TARLOV scores tested at various times and One-Way ANOVA analysis of variance for data between groups were used. RESULT: Seven days after surgery in SCI group, there was no statistical difference between the TARLOV scores of intrathecal injection of MPss 3.0 mg/kg group, 6.0 mg/kg group and MPss intravenous injection group (P > 0.05), which were all better than the vehicle group (F = 4.762, P < 0.05). Referring to the lymphocyte infiltration at the injury site in SCI group, there was statistical difference between MPss intrathecal injection 6.0 mg/kg group (1.33 ± 0.21) and the vehicle group (2.67 ± 0.21) (F = 5.793, P < 0.05) and no statistical difference between intrathecal injection of MPss 6.0 mg/kg group and MPss intravenous injection group (P > 0.05). As for the lymphocyte infiltration at the intrathecal injection site in SHAM group, there was statistical difference between MPss intrathecal injection 6.0 mg/kg group (2.50 ± 0.55) and the vehicle group (0.50 ± 0.55) (F = 17.333, P < 0.05). TUNEL staining in SCI group showed statistical difference between MPss intrathecal injection 6.0 mg/kg group (6.3 ± 1.5) and the vehicle group (20.3 ± 2.2) (F = 71.279, P < 0.05). CONCLUSIONS: Intrathecal injection of MPss can improve the functional recovery of lower limb and decrease apoptosis of neuron cells,which can provide same effects as the traditional intravenous injection of MPss in New Zealand rabbits.

Facile synthesis of phosphorus and nitrogen co-doped carbon dots with excellent fluorescence emission towards cellular imaging.

Fluorescent carbon nanomaterials have attracted increasing attention owing to their unique photoluminescence properties, good biocompatibility and low toxicity in bioimaging as well as biosensing. Heteroatom doping is usually used to improve photoluminescence properties by tuning the functional groups and the particle size domain effect, thus leading to redshifted emission. Here, we report a straightforward strategy for the fabrication of a mixture of fluorescent phosphorus and nitrogen carbon nanodots (P,N-CDs) followed by separating two kinds of fluorescent fractions based on their different negative charges. Such a one-pot hydrothermal method using formamide, urea and hydroxyethylidene diphosphonic acid as the precursor yields fluorescent P,N-CDs. Specifically, blue-emitting CDs (bCDs) and green-emitting CDs (gCDs) were separated by using column chromatography. The quantum yields of bCDs and gCDs were 20.33% and 1.92%, respectively. And the fluorescence lifetimes of bCDs and gCDs were 6.194 ns and 2.09 ns, respectively. What is more, the resultant P,N-CDs exhibited low toxicity and excellent biocompatibility. Confocal fluorescence microscopy images were obtained successfully, suggesting that P,N-CDs have excellent cell membrane permeability and cellular imaging. This work provides a promising fluorescent carbon nanomaterial with tunable emission as a probe for versatile applications in bioimaging, sensing and drug delivery.

Leopard predation on wild Sichuan snub-nosed monkeys.

Predation is widely recognized as a powerful selective pressure on primate behavior and ecology, although knowledge of predator-prey relationships remains limited partly due to the rarity of directly observed attacks on primates. Here, we describe four confirmed or suspected instances of leopard (Panthera pardus) predation on free-ranging Sichuan (golden) snub-nosed monkeys (Rhinopithecus roxellana), a highly endangered colobine species endemic to China. We recorded predation events and the reactions of monkey group members. We suggest that the evolution of a multilevel society may be an adaptive response by Sichuan snub-nosed monkeys to the risk from leopards as well as other potential predators, one that balances the pressures of predation and intra-species competition and conflict.

The Assessment of Indoor Formaldehyde and Bioaerosol Removal by Using Negative Discharge Electrostatic Air Cleaners.

This study investigated the single-pass performance of a negative corona electrostatic precipitators (ESP) in removing suspended particulates (PM2.5 and PM10), formaldehyde (HCHO), and bioaerosols (bacteria and fungi) and measured the ozone (O3) concentration generated by ESP. The experimental results revealed that if the operational conditions for the ESP were set to high voltage (-10.5 kV) and low air flow rate (2.4 m3/min), ESP had optimal air pollutant removal efficiency. In the laboratory system, its PM2.5 and PM10 removal rates both reached 99% at optimal conditions, and its HCHO removal rate was 55%. In field tests, its PM2.5, PM10, HCHO, bacteria, and fungi removal rates reached 89%, 90%, 46%, 69%, and 85% respectively. The ESP in the laboratory system (-10.5 kV and 2.4 m3/min) generated 7.374 ppm of O3 under optimal conditions. Under the same operational conditions, O3 generated by ESP in the food waste storage room and the meeting room were 1.347 ppm and 1.749 ppm, respectively. The removal of HCHO and bioaerosols was primarily attributed to their destruction in the corona, as well as ozone oxidation, and collection on the dust collection plate.

Endophytic fungi stimulate the concentration of medicinal secondary metabolites in houttuynia cordata thunb.

Endophytic fungi usually establish a symbiotic relationship with the host plant and affect its growth. In order to evaluate the impact of endophytic fungi on the Chinese herbal medicinal plant Houttuynia cordata Thunb., three endophytes isolated from the rhizomes of H. cordata, namely Ilyonectria liriodendra (IL), unidentified fungal sp. (UF), and Penicillium citrinum (PC), were co-cultured individually with H. cordata in sterile soil for 60 days. Analysis of the results showed that the endophytes stimulated the host plant in different ways: IL increased the growth of rhizomes and the accumulation of most of the phenolics and volatiles, UF promoted the accumulation of the medicinal compounds afzelin, decanal, 2-undecanone, and borneol without influencing host plant growth, and PC increased the fresh weight, total leaf area and height of the plants, as well as the growth of the rhizomes, but had only a small effect on the concentration of major secondary metabolites. Our results proved that the endophytic fungi had potential practical value in terms of the production of Chinese herbal medicines, having the ability to improve the yield and accumulation of medicinal metabolites.

Blood-Spinal Cord Barrier in Spinal Cord Injury: A Review.

The blood-spinal cord barrier (BSCB), a physical barrier between the blood and spinal cord parenchyma, prevents the toxins, blood cells, and pathogens from entering the spinal cord and maintains a tightly controlled chemical balance in the spinal environment, which is necessary for proper neural function. A BSCB disruption, however, plays an important role in primary and secondary injury processes related to spinal cord injury (SCI). After SCI, the structure of the BSCB is broken down, which leads directly to leakage of blood components. At the same time, the permeability of the BSCB is also increased. Repairing the disruption of the BSCB could alleviate the SCI pathology. We review the morphology and pathology of the BSCB and progression of therapeutic methods targeting BSCB in SCI.

The metastasis of hepatocarcinoma can be inhibited by T lymphocytes reconstitution in nude mice model.

BACKGROUND/AIMS: The metastasis of hepatic carcinoma is correlated with the body's immune status. T lymphocytes play a big part role in tumor immune. The aim of the present study is to investigate the inhibition effects of metastatsis in nude mice bearing hepatic carcinoma after T lymphocytes reconstitution. METHODOLOGY: An immune reconstitution model was established in nude mice. Then, 42 nude mice were distributed into 4 groups for T lymphocytes reconstitution. The lymph nodes of each group were obtained to investigate the tumor metastasis. And the secretion of cytokines and the apoptosis of tumor cells in each group were also detected. RESULTS: The ratio of CD3, CD4, CD8, CD4/CD8 in reconstituted groups were higher than controlled groups. The average time of tumor formation in Balb/c nu/nu mice was 7.7 +/- 0.6 days and in Balb/c mice was 11.5 +/- 1.3 days. After active T lymphocytes reconstitution, the extent lymph nodes metastasis in reconstitution groups was lower than control groups (p < 0.05), but proximal metastasis has no significant difference. The level of serum IL-10 in nude mice after immune reconstitution was significantly lower and VEGF was significantly higher than that of the control group (p < 0.05). Apoptosis of the hepatic carcinoma cells was increased significantly after immune reconstitution (p < 0.05). CONCLUSIONS: The metastasis of hepatic carcinoma can be inhibited by reconstitution of actived T lymphocytes in nude mice, which indicated that tumor metastasis can be affected by the immune status in host body.

Draft genome sequence of tetracycline-resistant Klebsiella oxytoca CCTCC M207023 producing 2,3-butanediol isolated from China.

OBJECTIVES: Recently, the Gram-negative bacterium Klebsiella oxytoca has been identified as an emerging pathogen. Here we report the draft genome of a 2,3-butanediol-producing strain, K. oxytoca CCTCC M207023, isolated from soil in Nanjing, China. The tetracycline-resistant phenotype and the high yield of 2,3-butanediol was demonstrated. METHODS: The draft genome of K. oxytoca CCTCC M207023 was determined using an Illumina NovaSeq™ 6000 next-generation DNA sequencing platform. Clean sequencing data were subsequently assembled using SOAPdenovo. RESULTS: The draft genome of K. oxytoca CCTCC M207023, comprising 5 658 144bp and with a GC content of 56.50%, was assembled into 5262 open-reading frames (ORFs). Antimicrobial resistance genes were also annotated. CONCLUSIONS: The draft genome sequence of K. oxytoca CCTCC M207023 reported here will be a reference for comparative analysis with the antimicrobial resistance mechanisms for the safety of 2,3-butanediol industrial production.

Efficacy of prostaglandina E1 for the treatment of patients with thrombo-occlusive vasculitis: A protocol of systematic review and meta-analysis.

BACKGROUND: This study aims to explore the efficacy and safety of prostaglandina E1 (PE1) for the treatment of patients with thrombo-occlusive vasculitis (TOV). METHODS: Electronic databases (Cochrane Library, PUBMED, EMBASE, Web of Science, Scopus, the Allied and Complementary Medicine Database, Chinese Biomedical Literature Database, and China National Knowledge Infrastructure) will be sought from onset to the March 1, 2020 without language and publication status restrictions. We will include any potential randomized controlled trials that examined the efficacy of PE1 for the treatment of patients with TOV. We will appraise study quality using Cochrane risk of bias tool, and will assess the evidence quality using Grading of Recommendations Assessment Development and Evaluation. We will use RevMan 5.3 Software for statistical analysis. RESULTS: A high-quality synthesis of present evidence of PE1 for the treatment of patients with TOV will be provided in this study. CONCLUSION: This study will provide evidence to judge whether PE1 is an effective intervention for TOV. SYSTEMATIC REVIEW REGISTRATION: INPLASY202040081.

Roles of extra-cellular signal-regulated protein kinase 5 signaling pathway in the development of spinal cord injury.

BACKGROUND: In consideration of characteristics and functions, extra-cellular signal-regulated protein kinase 5 (ERK5) signaling pathway could be a new target for spinal cord injury (SCI) treatment. Our study aimed to evaluate the roles of ERK5 signaling pathway in secondary damage of SCI. METHODS: We randomly divided 70 healthy Wistar rats into five groups: ten in the blank group, 15 in the sham surgery + BIX02188 (sham + B) group, 15 in the sham surgery + dimethyl sulfoxide (DMSO; sham + D) group, 15 in the SCI + BIX02188 (SCI + B) group, and 15 in the SCI + DMSO (SCI + D) group. BIX02188 is a specific inhibitor of the ERK5 signaling pathway. SCI was induced by the application of vascular clips (with the force of 30 g) to the dura on T10 level, while rats in the sham surgery group underwent only T9-T11 laminectomy. BIX02188 or DMSO was intra-thecally injected at 1, 6, and 12 h after surgery or SCI. Spinal cord samples were taken for testing at 24 h after surgery or SCI. RESULTS: Expression of phosphorylated-ERK5 (p-ERK5) significantly increased after SCI. Application of BIX02188 indeed inhibited ERK5 signaling pathway and reduced the degree of spinal cord tissue injury, neutrophil infiltration and proinflammatory cytokine expression, nuclear factor-κB (NF-κB) activation and apoptosis (measured by TdT-mediated 2'-deoxyuridine 5'-triphosphate nick-end labeling, expression of Fas-ligand, BCL2-associated X [Bax], and B-cell lymphoma-2 [Bcl-2]). Double immunofluorescence revealed activation of ERK5 in neurons and microglia after SCI. CONCLUSION: ERK5 signaling pathway was activated in spinal neurons and microglia, contributing to secondary injury of SCI. Moreover, inhibition of ERK5 signaling pathway could alleviate the degree of SCI, which might be related to its regulation of infiltration of inflammatory cells and release of inflammatory cytokines, expression of NF-κB and cell apoptosis.

Advances in the metabolic engineering of Yarrowia lipolytica for the production of terpenoids.

Terpenoids are a large class of natural compounds based on the C5 isoprene unit, with many biological effects such activity against cancer and allergies, while some also have an agreeable aroma. Consequently, they have received extensive attention in the food, pharmaceutical and cosmetic fields. With the identification and analysis of the underlying natural product synthesis pathways, current microbial-based metabolic engineering approaches have yielded new strategies for the production of highly valuable terpenoids. Yarrowia lipolytica is a non-conventional oleaginous yeast that is rapidly emerging as a valuable host for the production of terpenoids due to its own endogenous mevalonate pathway and high oil production capacity. This review aims to summarize the status and strategies of metabolic engineering for the heterologous synthesis of terpenoids in Y. lipolytica in recent years and proposes new methods aiming towards further improvement of terpenoid production.

CRISPR/Cas9-Based Genome Editing in the Filamentous Fungus Fusarium fujikuroi and Its Application in Strain Engineering for Gibberellic Acid Production.

The filamentous fungus Fusarium fujikuroi is well-known for its production of natural plant growth hormones: a series of gibberellic acids (GAs). Some GAs, including GA1, GA3, GA4, and GA7, are biologically active and have been widely applied in agriculture. However, the low efficiency of traditional genetic tools limits the further research toward making this fungus more efficient and able to produce tailor-made GAs. Here, we established an efficient CRISPR/Cas9-based genome editing tool for F. fujikuroi. First, we compared three different nuclear localization signals (NLS) and selected an efficient NLS from histone H2B (HTBNLS) to enable the import of the Cas9 protein into the fungal nucleus. Then, different sgRNA expression strategies, both in vitro and different promoter-based in vivo strategies, were explored. The promoters of the U6 small nuclear RNA and 5S rRNA, which were identified in F. fujikuroi, had the highest editing efficiency. The 5S rRNA-promoter-driven genome editing efficiency reached up to 79.2%. What's more, multigene editing was also explored and showed good results. Finally, we used the developed genome editing tool to engineer the metabolic pathways responsible for the accumulation of a series GAs in the filamentous fungus F. fujikuroi, and successfully changed its GA product profile, from GA3 to tailor-made GA4 and GA7 mixtures. Since these mixtures are more efficient for agricultural use, especially for fruit growth, the developed strains will greatly improve industrial GA production.

[Functional localization of metastasis suppressor genes for HCC on human chromosome 8].

OBJECTIVE: We previously showed that introduction of a normal, neomycin-tagged human chromosome 8 reduced the metastatic capacity of C5F rat liver cancer cell line, which had high metastatic potential without affecting tumorigenicity, suggesting the presence of one or more metastasis suppressor genes encoded on human chromosome 8. We proceeded to define further the region harboring the metastasis suppressor gene(s) and to determine the random loss of human chromosome 8 by PCR amplification of sequence tag site (STS) markers. METHODS: The national Center for Biotechnology Information (NCBI) databases were used as references of the relative genetic distances of the STS markers. C5F genomic DNA and A9/neo8 genomic DNA were used as negative and positive controls for chromosome 8 amplification, respectively. Genomic DNA was isolated and quantified from cultured hybrid clones (A9/C5F-1 and A9/C5F-2 microcell hybrid clones served as metastasis-unsuppressed groups; A9/C5F-4, A9/C5F-8 and A9/C5F-10 microcell hybrid clones served as metastasis suppressed groups). STS-PCR products were separated by electrophoresis through 2% agarose gel. RESULTS: Metastasis-suppressed microcell hybrid clones (A9/C5F-4, A9/C5F-8 and A9/C5F-10) conserved STS markers between D8S542 --> D8S1973 (8p21.1-23.1). In contrast, metastasis-unsuppressed clones (A9/C5F-1 and A9/C5F-2) lacked several markers in this region. In attempts to refine the region retained in the microcell suppressed clones, more densely spaced STS markers in the human chromosome 8p21.1-23.1 were used. We found that the metastasis-suppressed clones retained 18cM region between D8S542 and D8S1973 (8P21.1-23.1), where as the metastasis-unsuppressed clones lacked the region. CONCLUSION: Our results suggest that a metastasis suppressor gene is located within the interval between D8S542 and D8S1973 on human chromosome 8p21.1-23.1.

Biomechanical effect of bone resorption of the spinous process after single-segment interspinous dynamic stabilization device implantation: A finite element analysis.

This study aims to explore the influence of bone resorption of the spinous process after single-segment interspinous process device (IPD) implantation on the biomechanics of the lumbar spine.The 3D finite element model of the lumbar spine (L3-L5) was modified, and 2 models that simulated the presence and absence of bone resorption of the spinous process were developed using an IPD (Wallis). Its biomechanical effects, such as change in range of motion (ROM) and intervertebral disc and facet stress, were introduced at operative (L4/5) and adjacent (L3/4) levels.Compared with the INT model, the Wallis model and Wallis-BR model had similar ROMs in lateral flexion and rotation. However, the Wallis model had a lower L3-5 ROM in flexion (20.4% lower) and extension (26.4% lower), and L4-L5 ROM in flexion (74.1% lower) and extension (70.8% lower), while the overall ROM of the Wallis-BR model was greater than that of the Wallis model. The stress on the L3/L4 intervertebral disc and facets was similar for all 3 models. Compared with the INT model and Wallis-BR model, the stress on the L4/L5 intervertebral disc and facets under all movements significantly decreased in the Wallis model. The stress on the L5 process was greater than that on the L4 process in both the Wallis model and Wallis-BR model, and the load on the processes that underwent bone resorption was lower than that of the Wallis model.The function of the IPD slowly decreased with the occurrence of bone resorption of the interspinous process. This bone remodeling may be associated with high stress after IPD implantation.

Clinical and Radiologic Features of 3 Reconstructive Procedures for the Surgical Management of Patients with Bilevel Cervical Degenerative Disc Disease at a Minimum Follow-Up Period of 5 Years: A Comparative Study.

OBJECTIVE: To assess the mid-long-term follow-up of the safety and efficacy of anterior cervical discectomy and fusion (ACDF), cervical artificial disc replacement (CADR) and hybrid surgery (HS) for bilevel cervical degenerative disc disease (cDDD). METHODS: 77 patients who underwent ACDF, HS, and CADR were retrospectively reviewed. Clinical effects were evaluated based on Neck Disability Index (NDI), Visual Analog Scale (VAS), and Japanese Orthopedic Association (JOA) scores and the Odom criteria. Radiographic outcomes were evaluated, including cervical range of motion (ROM), ROM in the operative and adjacent segments, incidence of degeneration in the adjacent segments (ASD), and heterotopic ossification (HO). RESULTS: NDI, VAS, and JOA scores significantly improved in all patients after surgery without significant differences between groups. The excellent-to-good ratio in the Odom scale was 28/30 for the HS group, 30/33 for the ACDF group, and 13/14 for the CADR group. No significant differences in clinical outcomes or complication were found between groups (P > 0.05). Furthermore, the HS and CADR groups had less decreased ROM in the cervical and operative segments and less compensatory ROM in adjacent segments (P < 0.05). By contrast, the ACDF group had decreased ROM in the cervical and operative segments and significantly increased ROM in adjacent segments (P < 0.05). Moreover, the incidence of ASD was higher in the ACDF group, but the difference was not statistically significant (P > 0.05). HO was found in 10 patients (33.3%) in the HS group and 5 patients (35.7%) in the CADR group. CONCLUSION: HS was superior to ACDF with regard to equivalent clinical outcomes in the mid-long-term follow-up. Furthermore, HS was superior in the maintenance of ROM and had less impact on its adjacent segments. The efficacy of HS is similar to that of CADR.

[Relation between changes of dendritic cell function and down-regulation of beta-centractin in hepatocellular carcinoma].

OBJECTIVE: To investigate the relation between the changes of dendritic cell (DC) function and down-regulation of beta-centractin in hepatocellular carcinoma. METHODS: DC derived from peripheral blood were cultured and then pulsed by lysates from hepatocarcinoma cells (HCC) with high, low, or none metastatic potential of the lines HCCLM6, MHCC97L, and Hep3B, and from normal human liver cell of the line Chang liver. DC not pulsed were used as control group. Three days later scanning electron microscopy and inverted microscopy were used to observe the morphology of the DC. Flow cytometry was used to observe the phenotype. The protein expression of beta-centractin was detected by Western blotting and immunocytochemistry. RESULTS: Scanning electron microscopy and inverted microscopy showed no change in the morphology of the DC pulsed by different antigens. The expression levels of HLA-DR, CD80, CD83, and CD86 of the 4 pulsed groups were all significantly higher than that of the un-pulsed group (all P < 0.05). The expression of CD86 of the DC + LM6 group was significantly lower than those of the DC + Chang, DC + Hep3B, and DC + 97L groups (all P < 0.05). The mixed lymphocyte reaction (MLR) levels of the 4 pulsed groups were all significantly higher than that of the control group (all P < 0.05). The MLR level of the DC + LM6 group was significantly lower than those of the DC + Chang, DC + Hep3B, and DC + 97L groups (all P < 0.05). Western blotting showed that beta-centractin was not expressed in the control DC and was expressed in the 3 pulsed DC groups, and the beta-centractin expression levels of the DC + Chang, DC + Hep3B, and DC + 97L groups were all higher than that of the DC + LM6 group. The results of immunocytochemistry were similar to that of Western blotting. CONCLUSION: The down-regulation of beta-centractin in the DC pulsed with high metastatic potential HVV cell lysates is associated DC dysfunction and may be one of the mechanisms of HCC immune escape.