Basis For Further Clinical Application Research Articles

Network Pharmacology-Based and Molecular Docking combined with GEO gene chips to Investigate the Potential Mechanism of Duhuo Jisheng Decoction against Rheumatoid Arthritis.

Background： Rheumatoid Arthritis (RA) is a chronic autoimmune disease with various symptoms in patients. Duhuo Jisheng Decoction (DHJSD) has been used to treat RA in China for a long history as a classic TCM formula. However, the underlying pharmacological mechanism still needs to be elucidated. Purpose： In the current study, we combined network pharmacology with molecular docking to investigate the potential mechanism of DHJSD treating RA. Methods: The active compounds and related targets of DHJSD were obtained from the TCMSP database. The RA targets were retrieved from the GEO database. The PPI network of overlapping targets was constructed, whereas the core genes were selected by CytoNCA for molecular docking. GO and KEGG enrichment analyses were used to further explore the biological process and pathways of overlapping targets. On this basis, molecular docking was carried out to verify the interrelations of the main compounds and core targets. Results： In this study, we found 81 active components corresponding to 225 targets of DHJSD. Moreover, 775 RA-related targets were obtained, of which 12 were shared between DHJSD targets and RA genes. From GO and KEGG analysis, there were 346 GO items and 18 signaling pathways. As the molecular docking showed, the binding of components was stable with the core gene. Conclusion： In conclusion, our works revealed the underlying mechanism of DHJSD for treating RA using network pharmacology and molecular docking, which provided a theoretical basis for further clinical application in the future.

Newly synthesized acridone derivatives targeting lung cancer: A toxicity and xenograft model study.

AKT is one of the overexpressed targets in nonsmall cell lung cancer (NSCLC) and plays an important role in its progression and offers an attractive target for the therapy. The PI3K/AKT/mTOR pathway is upregulated in NSCLC. Acridone is an important heterocycle compound which treats cancer through various mechanisms including AKT as a target. In the present work, the study was designed to evaluate the safety profile of three acridone derivatives (AC-2, AC-7, and AC-26) by acute and repeated dose oral toxicity. In addition to this, we also checked the pAKT overexpression and its control by these derivatives in tumor xenograft model. The results from acute and repeated dose toxicity showed these compounds to be highly safe and free from any toxicity, mortality, or significant alteration in body weight, food, and water intake in the rats. In the repeated dose toxicity, compounds showed negligible variations in a few hematological parameters at 400 mg/kg. The histopathology, biochemical, and urine parameters remained unchanged. The xenograft model study demonstrated AC-2 to be inhibiting HOP-62 induced tumor via reduction in p-AKT1 (Ser473) expression significantly. In immunofluorescence staining AC-2 treated tissue section showed 2.5 fold reduction in the expression of p-AKT1 (Ser473). Histopathology studies showed the destruction of tumor cells with increased necrosis after treatment. The study concluded that AC-2 causes cell necrosis in tumor cells via blocking the p-AKT1 expression. The findings may provide a strong basis for further clinical applications of acridone derivatives in NSCLC.

Analysis of mechanism of core points in acupuncture and moxibustion treatment for epilepsy based on data mining.

To explore the mechanism of core points in acupuncture and moxibustion treatment for epilepsy by using data mining technique, so as to provide a reference for clinical practice and experimental research. The data comes from relevant documents collected from CNKI, Wanfang, SinoMed, VIP, PubMed, Embase, Cochrane Library, EBSCO, Web of Science databases. The selected acupoints were analyzed in descriptive statistics, high-frequency acupoints group and core acupoint prescription. Further, potential target mining, "core acupoint prescription-target-epilepsy" network construction, protein-protein interactions (PPI) network establishment and core target extraction, gene ontology (GO) and KEGG gene enrichment analysis of the core acupoint prescription were carried out to predict its anti-epileptic potential mechanism. A total of 122 acupoint prescriptions were included. The core acupoint prescriptions were Baihui (GV20), Hegu (LI4), Neiguan (PC6), Shuigou (GV26) and Taichong (LR3). 277 potential targets were identified, among which 134 were shared with epilepsy. The core targets were extracted by PPI network topology analysis, including signal transducer and activator of transcription 3, tumor necrosis factor (TNF), interleukin (IL)-6, protein kinase B1, c-Jun N-terminal kinase, brain-derived neurotrophic factor, tumor protein 53, vascular endothelial growth factor A, Caspase-3, epidermal growth factor receptor, etc. The main anti-epileptic pathways of the core acupoints were predicted by KEGG enrichment, including lipid and atherosclerosis, neurodegeneration, phosphatidylinositol-3-kinase/protein B kinase signaling pathway, mitogen-activated protein kinase signaling pathway, cyclic adenosine monophosphate signaling pathway, TNF signaling pathway, IL-17 signaling pathway, hypoxia-inducible factor-1 signaling pathway, apoptosis, etc., involving neuronal death, synaptic plasticity, oxidative stress, inflammation and other related biological process. The core acupoint prescription of acupuncture and moxibustion intervention for epilepsy can act on multiple targets and multiple pathways to exert anti-epileptic effects, which can provide a theoretical basis for further clinical application and mechanism research.

Potential roles of HSYA in attenuating sepsis-induced liver injury through multi-omics analysis

Liver injury is a strong independent predictor of mortality in patients with sepsis, in which gut dysbiosis plays a crucial role. Hydroxyl safflower yellow A (HSYA), an important component of safflower, has been used to treat liver injury in animal models. However, its role in sepsis-induced liver dysfunction and the specific molecular mechanisms remain unclear. In the current study, we first discussed the discrepancy in the gut microbiota between the cecal ligation puncture (CLP) and HSYA groups using 16 S RNA sequencing. Our data demonstrated that HSYA supplementation significantly decreased the relative abundance of Proteobacteria, Firmicutes, and Campylobacterota, and further decreased the abundance of Bacteroidota, suggesting that the protective effects of HSYA against sepsis-induced liver injury may be partially attributed to the alteration of these bacteria. In addition, the metabolomic data identified 823 differentially expressed metabolites associated with sepsis-induced liver injury. After HSYA supplementation, the levels of 56 metabolites were restored to sham-like levels. Transcriptomic analysis revealed 4990 differentially expressed genes (DEGs) between the sham and CLP groups, and after HSYA injection, 1613 genes were modulated. Comprehensive analysis demonstrated that the enrichment pathways of the 903 DEGs mainly focused on inflammatory responses, amino acid metabolism, and Lipid reactions. In conclusion, our study revealed the potential mechanism of action of HSYA in sepsis-induced liver injury through a comprehensive analysis of 16 S RNA sequencing, metabolomics, and transcriptomics, thus providing a theoretical basis for further clinical applications of HSYA.

Clostridium butyricum improves cognitive dysfunction in ICV-STZ-induced Alzheimer's disease mice via suppressing TLR4 signaling pathway through the gut-brain axis.

In recent years, the relationship between gut-brain axis and Alzheimer's disease (AD) attracted increasing attention. The aim of this study is to investigate the therapeutic effect of Clostridium butyricum (CB) on intraventricular injection of streptozotocin (ICV-STZ)-induced mice and the potential mechanisms. ICV-STZ mice were treated with CB by gavage for 21 consecutive days. The pharmacological effect of CB was assessed by behavior test, brain tissue H&E staining and tau protein phosphorylation levels of hippocampus tissues. The expression levels of TLR4, MYD88, NF-κB p65, TNF-α, iNOS, Occludin and ZO-1 in hippocampal and colonic tissues were detected by Western-blot method. 16S rRNA gene sequencing analysis was used to analyze the intestinal microbiota of mice. The results showed that CB improved the cognitive dysfunction of ICV-STZ mice, restored the structure and cell number of hippocampal and cortical neurons, decreased the protein levels of pSer404-tau protein in hippocampal tissues and TLR4, MYD88, NF-κB p65 and iNOS in hippocampal and colonic tissues, and increased the protein levels of Occludin and ZO-1 in colonic tissues. Meanwhile, CB reversed the changes of intestinal microbiota in AD mice. Therefore, the mechanisms of cognitive function and brain pathological changes in AD mice improved by CB may be related to the regulation of TLR4 signaling pathway and intestinal microbiota. This study supports the potential anti-AD effect of CB and initially revealed its pharmacological mechanism of CB, providing a theoretical basis for further clinical application of CB.

Redox-Responsive Lipidic Prodrug Nano-Delivery System Improves Antitumor Effect of Curcumin Derivative C210.

The poor bioavailability of curcumin and its derivatives limits their antitumor efficacy and clinical translation. Although curcumin derivative C210 has more potent antitumor activity than curcumin, it has a similar deficiency to curcumin. In order to improve its bioavailability and accordingly enhance its antitumor activity in vivo, we developed a redox-responsive lipidic prodrug nano-delivery system of C210. Briefly, we synthesized three conjugates of C210 and oleyl alcohol (OA) via different linkages containing single sulfur/disulfide/carbon bonds and prepared their nanoparticles using a nanoprecipitation method. The prodrugs required only a very small amount of DSPE-PEG2000 as a stabilizer to self-assemble in aqueous solution to form nanoparticles (NPs) with a high drug loading capacity (~50%). Among them, the prodrug (single sulfur bond) nanoparticles (C210-S-OA NPs) were the most sensitive to the intracellular redox level of cancer cells; therefore, they could rapidly release C210 in cancer cells and thus had the strongest cytotoxicity to cancer cells. Furthermore, C210-S-OA NPs exerted a dramatic improvement in its pharmacokinetic behavior; that is, the area under the curve (AUC), mean retention time and accumulation in tumor tissue were 10, 7 and 3 folds that of free C210, respectively. Thus, C210-S-OA NPs exhibited the strongest antitumor activity in vivo than C210 or other prodrug NPs in mouse models of breast cancer and liver cancer. The results demonstrated that the novel prodrug self-assembled redox-responsive nano-delivery platform was able to improve the bioavailability and antitumor activity of curcumin derivative C210, which provides a basis for further clinical applications of curcumin and its derivatives.

Radix Rehmanniae Praeparata (Shu Dihuang) exerts neuroprotective effects on ICV-STZ-induced Alzheimer's disease mice through modulation of INSR/IRS-1/AKT/GSK-3β signaling pathway and intestinal microbiota.

Radix Rehmanniae Praeparata (RRP, Shu Dihuang in Cinese) is widely used as primal medicine in Chinese herbal formula for the treatment of Alzheimer's disease (AD). However, the underlying mechanism of RRP for AD remains unclear. The aim of this study was to investigate the therapeutic effect of RRP on intracerebroventricular injection of streptozotocin (ICV-STZ)-induced AD model mice and its potential mechanism. ICV-STZ mice were continuously gavaged with RRP for 21days. The pharmacological effects of RRP were evaluated by behavioral tests, brain tissue H&E staining and hippocampal tau protein phosphorylation levels. The expression levels of insulin receptor (INSR), IRS-1, pSer473-AKT/AKT and pSer9-GSK-3β/GSK-3β proteins in hippocampal and cortical tissues were detected by Western-blot method. The 16S rRNA gene sequencing was used to analyze the changes of intestinal microbiota in mice. The compounds in RRP were analyzed by mass spectrometry and their binding ability to INSR proteins was detected by molecular docking. The results showed that RRP ameliorated cognitive dysfunction and neuronal pathological changes of brain tissue in ICV-STZ mice, reduced tau protein hyperphosphorylation, INSR, IRS-1, pSer473-AKT/AKT, and pSer9-GSK-3β/GSK-3β levels in hippocampal and cortical tissues. Meanwhile, RRP reversed ICV-STZ-induced dysregulation of intestinal microbiota in AD mice. Mass spectrometry analysis showed that the RRP consisted mainly of seven compounds, namely Acteoside (Verbascoside), 5-Hydroxymethyl-2-furaldehyde (5-HMF), Apigenin7-O-glucuronide, Icariin, Gallic acid, Quercetin-3β-D-glucoside, and Geniposide. Molecular docking results further indicated that the compounds in RRP have binding ability to INSR protein and potential multiple synergistic effects. RRP ameliorates cognitive dysfunction and brain histopathological changes in AD mice. The mechanism of RRP ameliorating AD may be related to the regulation of INSR/IRS-1/AKT/GSK-3β signaling pathway and intestinal microbiota. This study supports the potential anti-AD efficacy of RRP and initially reveals the pharmacological mechanism of RRP, providing a theoretical basis for further clinical application of RRP.

The Mechanical and Antibacterial Properties of Boron Nitride/Silver Nanocomposite Enhanced Polymethyl Methacrylate Resin for Application in Oral Denture Bases.

The introduction of nanomaterials into polymethyl methacrylate (PMMA) resin has been effective for mechanically reinforcing PMMA for application in oral denture bases. However, these methods cannot simultaneously improve the mechanical and antibacterial properties, which limits widespread clinical application. Here, we self-assembled binary nanocomposites of boron nitride nanosheets (h-BNNs) and silver nanoparticles (AgNPs) as nanofillers and incorporated the nanofillers into PMMA. The aim of this study was to achieve antibacterial effects while significantly improving the mechanical properties of PMMA and provide a theoretical basis for further clinical application. We employed scanning electron microscopy (SEM), X-ray diffraction (XRD), Raman spectroscopy (Raman), Ultraviolet visible spectrum (UV) and atomic force microscopy (AFM) to investigate the microscopic morphology and composition of PMMA containing nanocomposites with different mass fraction. In addition, the content of the h-BNNs/AgNPs was 1 wt%, and the compressive strength and flexural strength of pure PMMA were improved by 53.5% and 56.7%, respectively. When the concentration of the nanocomposite in the PMMA resin was 1.4 wt%, the antibacterial rate was 92.1%. Overall, synergistically reinforcing PMMA composite resin with a multi-dimensional nanocomposite structure provided a new perspective for expanding not only the application of resins in clinical settings but also the research and development of new composite resins.

Development and biomechanical test of a new pedicle screw for thoracolumbar spinal surgery

Rupture of the medial pedicle wall often occurs during pedicle screw insertion. This allows the pedicle screw to compress or cut the nerve root and/or spinal cord. In this paper, we designed a new double-threaded pedicle screw that has a cylindrical outer diameter and a conical core diameter, with a wide thread at the front, no thread in the middle, and a narrow thread at the rear, according to an analysis of the shortcomings of the conventional pedicle screw and anatomical parameters in 300 healthy adult volunteers. After the screw was placed, the non-threaded portion of the screw was located at the vertebral pedicle. No nerve root cutting occurred if the screw was misplaced and the medial wall of the vertebral pedicle was broken. We then performed biomechanical tests using a static universal testing machine and compared the new double-threaded screws with the conventional full-threaded pedicle screws, however, the differences were not obvious. In compression bending fatigue tests, the novel double-threaded pedicle screws were subjected to 5 million fatigue cycle loads without failure. The current study demonstrated that the new pedicle screw possesses similar biomechanical features as those of the conventional fully threaded pedicle screw. This provides a basis for further clinical applications.

Multiple Roles of Paeoniflorin in Alzheimer's Disease.

Alzheimer's disease (AD) is a geriatric disease with the morbidity and mortality continuing to grow, partly due to the aging of the world population. As one of the most common types of primary neurodegenerative dementia, it is mainly due to environmental, epigenetic, immunological, and genetic factors. Paeoniflorin (PF), the main component of paeony extract, plays a more and more important role in the prevention and treatment of AD, including regulating protein, anti-inflammation, antioxidation, and antiapoptosis, protecting glial cells, regulating neurotransmitters and related enzymes and receptors, and inhibiting or activating related signal pathways. This article summarizes the latest researches on the multiple effects and the mechanisms of PF in the treatment to cure AD, providing new insights and research basis for further clinical application of traditional Chinese medicine (TCM) in the treatment of AD.

Three-dimensional finite element analysis of a novel interzygapophyseal fusion device for lower cervical spine

Purpose: A three-dimensional finite element model of the lower cervical spine was established to evaluate the biomechanical stability and stress distribution of the new lower cervical interzygapophyseal fusion device (IZFD) developed by ourselves under different construct. The aim of this study was to provide theoretical basis for further clinical application. Methods: A normal fresh cadaveric specimen (male, 35 years old) was used to establish an intact three-dimensional finite element model of C3–C6. On this basis, the comparative finite element models of the lateral mass screw rod (LMSR) system and LMSR+IZFD were established. Only C4–C5 is fixed in the lateral mass. The range of motion (ROM) and stress distribution in the flexion, extension, lateral bending and rotation of the C4–C5 segment under the three constructs were analyzed. Results: The ROM and stress distribution of the three-dimensional finite element model under load construct were within a reasonable range, which proved the validity and reliability of the model. The ROM and stress distribution of C4–C5 segment was significantly decreased in both LMSR and LMSR+IZFD constructs than those in the intact construct. The ROM and stresss distribution were even smaller in LMSR+IZFD construct than in LMSR construct. Conclusions: The IZFD combined with LMSR system can provide satisfactory stability for the lower cervical spine, and the IZFD can further improve the fixation effect of the LMSR system.

Research Progress on the Antitumor Mechanism of Compound Kushen Injection

Compound Kushen Injection (CKI), as a clinical traditional Chinese medicine preparation, has prominent antitumor effect but with several side effects. A large number of studies have shown that CKI plays an antitumor role by regulating tumor cell proliferation, inducing tumor cell differentiation and apoptosis, inhibiting tumor cell invasion and metastasis, reducing tumor angiogenesis, regulating the immunity, and so on. Clinically, CKI is widely used to treat various tumors, where it is often combined with surgery, chemotherapy, radiotherapy, targeted therapy, and other antitumor treatments. This article reviews the antitumor mechanism of CKI and the progress of its clinical application in order to provide a theoretical basis for further clinical application.

Literature research and traditional Chinese medicine properties of Aspalathus linearis

Aspalathus linearis is a needle-shaped shrub that grows in the Cedarberg mountains in southern South Africa, with an extremely high medicinal value. In 2014, China has approved A. linearis as a new food material. Through retrieval in CNKI, Wanfang, VIP, Web of Science, PubMed and Scopus databases, the literatures were excluded, classified and summarized.On the basis of Chinese medicine theory, the traditional Chinese medicine properties were deducted. Finally, 264 relevant li-teratures were included and classified into 6 categories: review, planting, chemical composition, clinical study, pharmacological effects and safety. The traditional Chinese medicinal properties were deducted as sweet flavor and neutral property. It enters kidney, spleen, heart and liver channels. The major functions are to tonify the kidney and benefit the essence, nourish Qi and spleen, nourish Yin and prompt the production of body fluid, tranquilize mind, and relieve pain. It can be used for soreness of the waist and fatigue, sexual disinterest, limbs heaviness, thirst due to insufficiency of fluid and internal heat, irritability and insomnia, forget fulness, stomachache, joint pain, dysmenorrhea, headache. Preparation for external use can treat eczema itching. Water decoction(2-15 g) can also be used as tea directly. This paper defined the traditional Chinese medicine properties of A. linearis, so as to provide the theoretical basis for further clinical application.

Comparative study between 3D guide plate assisted and free-hand insertion of anterior cervical transpedicular screw

To compare accuracy of anterior cervical pedicle screws between assist of rapid prototyping 3D guide plate and free-hand insertion, and evaluate the safety of two methods. Eight adult cervical cadaver specimens after formaldehyde immersion, including 4 males and 4 females, aged 32 to 65(40.3±5.6) years old. After X-ray examination to exclude bone damage and deformity, 4 of them (3D guide plate group) randomly selected were for CT scan to obtain DICOM format data, and the data was imported into Mimics software for model, designed the ideal entry point and nail path for anterior cervicaltranspedicular screw (ATPS). After obtaining the personalized guide plate of the nail channel, it was exported as STL data, and the individual guide plate was printed by rapid prototyping and 3D printing technology. In turn, with the assistance of 3D guide plates, one-to-one personalized ATPS screws were placed on the four lower cervical cadaver specimens. Another 4 (free-hand group) lower cervical cadaver specimens were implanted with ATPS screws using free-hand technique. All specimens were performed CT thin-layer scanning and three-dimensional reconstruction after operation. The Tomasino method was used to evaluate the safety of the screws on the CT cross-sectional and sagittal images, to determine whether there was a cortical puncture of the lower and inner edges of the pedicle. According to the CT rating results, gradeⅠandⅡwere safe, and grade Ⅲ- Ⅴ were dangerous.And the accuracy of screws was recorded and analyzed between two groups. Two screws were inserted in each segment from C3 to C7 in 8 adult cadavers. A total of 80 screws were inserted, 40 in the 3D guide plate group, and 40 in the free-hand group. The Tomasino screw rating method was used to evaluate the safety of screw, 21 screws were gradeⅠ, 14 screws were gradeⅡ, 3 screws were grade Ⅲ, 1 screw was grade Ⅳ, 2 screws were grade Ⅴ in 3D guide plate group, while 14 screws were gradeⅠ, 8 screws were gradeⅡ, 8 screws were grade Ⅲ, 6 screws were grade Ⅳ, 2 screws were grade Ⅴ in free-hand group. The safety rate of 3D guide plate group was 87.5%, and 55.0% of the free hand group (χ2=8.7, P=0.003). The 3D printing rapid prototyping guide plate assisted insertion of the anterior cervical pedicle screw can significantly improve the accuracy and safety, and provide a theoretical basis for further clinical application.

Targeting delivery and minimizing epidermal diffusion of tranexamic acid by hyaluronic acid-coated liposome nanogels for topical hyperpigmentation treatment

Hyperpigmentation is a common complaint and distressing problem in dermatology, and tranexamic acid (TA) is an effective treatment agent but limited by the delivery to melanocytes in the epidermis. Herein, a novel TA naogels (named HA/TA-LP), combining the advantages of liposomes and hyaluronic acid (HA), are prepared and assessed for topical hyperpigmentation treatment with targeting delivery and minimizing epidermal diffusion. Morphological characteristics indicate numerous TA-loaded liposomes packed in HA gels. In vitro cell studies using human A375 melanoma cells show that HA/TA-LP can promote the uptake of TA by targeting delivery with resulting inhibition of tyrosinase activity and melanin production. Guinea pigs are used to construct hyperpigmentation models and investigate the topical delivery and treatment efficacy of HA/TA-LP. In vivo topical delivery studies indicate HA/TA-LP realize the effective delivery into melanocytes with an ideal balance of effective permeability and minimizing epidermal diffusion. Subsequently, hyperpigmentation treatment assessments reveal that HA/TA-LP inhibit tyrosinase activity and melanin production under the radiation of UVB. Our study identifies favorable properties of HA/TA-LP for treating hyperpigmentation, and provides an experimental basis for further clinical application.

A six-herb Chinese medicine composition ointment as a promising candidate for treatment of hypertrophic scars

ObjectiveTo study the anti-hypertrophic scar effect of the six-herb Chinese medicine composition (SCMC) ointment on the rabbit ear hypertrophic scar models. MethodsThe optimal formulation of SCMC ointment matrix was screened by the orthogonal designs and a series of evaluation tests. The SCMC ointment was prepared through emulsifying method. The rabbit ear hypertrophic scar models were established and used to investigate the anti-hypertrophic scar effect of SCMC ointment. ResultsOur results demonstrated that all the quality control indications of the SCMC ointment met the requirements. Anti-hypertrophic scar activity results showed that all the rabbit ear scar tissues appeared different degrees of shrink and fading, and took an unobvious but palpable shift from hard to soft texture with the low, middle and high concentration SCMC ointments treatments in vivo. Additionally, on 21st day the scar area and thickness in different concentrations of SCMC ointment groups were significantly reduced than control group, in a concentration-dependent manner. The immunohistochemical results also indicated that the SCMC ointment had good anti-hypertrophic scar properties and could inhibit hypertrophic scar formation. ConclusionThe SCMC ointment could improve the blood circulation condition of hypertrophic scar tissues. Our research has demonstrated the Chinese medicine composition ointment with good anti-hypertrophic scar properties that could be used to treat hypertrophic scars. Meanwhile, it provides a theoretical basis for further clinical application.

MiR-143 inhibits renal cell carcinoma cells metastatic potential by suppressing ABL2.

Although micro RNA (miRNA) expression profiles are widely investigated in renal cell carcinoma (RCC), their potential roles for affecting RCC initiation and progression remain largely unknown. Here, we examined the aberrant expression profiles of miRNAs inhuman metastatic RCC tissues based on Gene Expression Omnibus (GSE37989). We further validated them iRNAs expression data in the largest clinical dataset: The Cancer Genome Atlas (TCGA). And cell adhesion and migration abilities and epithelial me senchymal transition (EMT) related proteins were assessed in both normal and tumor RCC cell lines. We suggest that hsa-miR-143 is a potential tumor suppressor in RCC as its down regulation positively correlated with adverse prognosis. Biologically, cell adhesion, migration, and EMT were dramatically inhibited by miR-143. Mechanistically, we found that miR-143 targets ABL proto-oncogene 2 (ABL2), which was also found to be an indicator for poor survival in TCGA database. Our results have important implications in understanding functions of miRNAs in metastatic RCC and will provide a basis for further clinical application.

Efficacy and mechanism of Wuzi Yanzong pill in the treatment of EAE

Wuzi Yanzong Pill (WYP, a classical Chinese herbal formula) has a potential therapeutic effect on experimental autoimmune encephalomyelitis (EAE), but the mechanism is not clear. To determine the therapeutic effect of WYP in EAE, and explore its mechanism. C57BL/6 mice were randomly divided into control, EAE, and high/medium/low dose WYP groups. Mice received WYP twice daily from 7 days before immunization to 14 and 28 days post immunization (p.i.) (high/medium/low dose =16/8/4 g/kg). At days 14 and 28 p.i., spinal cord and spleen were obtained. WYP treatment delayed EAE onset and relieved the severity of EAE accompanied by the inhibition of inflammatory cell infiltration and the improvement of demyelination in spinal cord. WYP treatment may more effectively induce M2 macrophage polarization that was related with decreased IFN-γ, IL-17 and TNF-α. WYP treatment also inhibited the expression/activation of ROCK and TLR-4/NF-κB signaling pathway and reduced the expression of MCP-1 and CCR-2 in brain, spinal cord and spleen compared to EAE control. Provides an experimental basis for further clinical application of WYP for the treatment of MS. (NNSF of China 81102552 and 81703978, Shanxi Scholarship Council project 2017-129 and 2017-19, Shanxi International Science and Technology cooperation project 201703D421016, Scientific and technological innovation team of integrated Chinese and western medicine for the prevention and treatment on nervous system diseases of Shanxi university of Chinese medicine 2018TD-012, Shanxi Key Laboratory project SXIDL-2018-04, Shanxi Key project 201803D31209. Ma and Xiao are corresponding authors).

Identification of a 5‑microRNA signature and hub miRNA‑mRNA interactions associated with pancreatic cancer.

miRNA‑gene axes have been reported to serve an important role in the carcinogenesis of pancreatic cancer (PC). The aim of the present study was to systematically identity the microRNA signature and hub molecules, as well as hub miRNA‑gene axes, and to explore the potential biomarkers and mechanisms associated with the carcinogenesis of PC. Eleven microRNA profile datasets were obtained from the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) and ArrayExpress databases, and a meta‑analysis was performed to identify the differentially expressed miRNAs (DEMs) between tumor tissue and normal tissue. Subsequently, a diagnostic regression model was constructed to identify PC based on The Cancer Genome Atlas (TCGA) miRNA sequence data by using the least absolute shrinkage and selection operator (LASSO) method. In addition, GSE41368 was downloaded, and a weighted gene co‑expression network analysis (WGCNA) was performed to obtain the gene module associated with carcinogenesis by using the TCGAbiolinks and WGCNA packages, respectively. Finally, miRNA‑gene networks were constructed and visualized using Cytoscape software, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses based on the Database for Annotation, Visualization, and Integrated Discovery (DAVID). A total of 14 DEMs were identified, and a 5‑microRNA‑based score generated by the LASSO regression model provided a high accuracy for identifying PC [area under the curve (AUC)=0.918]. In addition, 44 miRNA‑mRNA interactions were constructed, and 4hub genes were screened on the basis of the above bioinformatic tools and databases. Furthermore, 14 biological process (BP) functions and 6 KEGG pathways were identified according to gene set enrichment analysis (GSEA). In summary, the present study applied integrated bioinformatics approaches to generate a holistic view of PC, thereby providing a basis for further clinical application of the 5‑miRNA signature and the identified hub molecules, as well as the miRNA‑gene axes, which could serve as diagnostic markers and potential treatment targets.

Sustained release of recombinant human bone morphogenetic protein-2 combined with stromal vascular fraction cells in promoting posterolateral spinal fusion in rat model

To observe the effect of stromal vascular fraction cells (SVFs) from rat fat tissue combined with sustained release of recombinant human bone morphogenetic protein-2 (rhBMP-2) in promoting the lumbar fusion in rat model. SVFs were harvested from subcutaneous fat of bilateral inguinal region of 4-month-old rat through the collagenase I digestion. The sustained release carrier was prepared via covalent bond of the rhBMP-2 and β-tricalcium phosphate (β-TCP) by the biominetic apatite coating process. The sustained release effect was measured by BCA method. Thirty-two rats were selected to establish the posterolateral lumbar fusion model and were divided into 4 groups, 8 rats each group. The decalcified bone matrix (DBX) scaffold+PBS, DBX scaffold+rhBMP-2/β-TCP sustained release carrier, DBX scaffold+SVFs, and DBX scaffold+rhBMP-2/β-TCP sustained release carrier+SVFs were implanted in groups A, B, C, and D respectively. X-ray films, manual spine palpation, and high-resolution micro-CT were used to evaluate spinal fusion at 8 weeks after operation; bone mineral density (BMD) and bone volume fraction were analyzed; the new bone formation was evaluated by HE staining and Masson's trichrome staining, osteocalcin (OCN) was detected by immunohistochemical staining. The cumulative release amount of rhBMP-2 was about 40% at 2 weeks, indicating sustained release effect of rhBMP-2; while the control group was almost released within 2 weeks. At 8 weeks, the combination of manual spine palpation, X-ray, and micro-CT evaluation showed that group D had the strongest bone formation (100%, 8/8), followed by group B (75%, 6/8), group C (37.5%, 3/8), and group A (12.5%, 1/8). Micro-CT analysis showed BMD and bone volume fraction were significantly higher in group D than groups A, B, and C ( P<0.05), and in group B than groups A and C ( P<0.05). HE staining, Masson's trichrome staining, and immunohistochemistry staining for OCN staining exhibited a large number of cartilage cells with bone matrix deposition, and an active osteogenic process similar to the mineralization of long bones in group D. The bone formation of group B was weaker than that of group D, and there was no effective new bone formation in groups A and C. The combination of sustained release of rhBMP-2 and freshly SVFs can significantly promote spinal fusion in rat model, providing a theoretical basis for further clinical applications.