Objectives Examine whether osteoarthritis (OA) progression can be delayed by halofuginone in anterior cruciate ligament transection (ACLT) rodent models. Methods 3-month-old male C57BL/6J (wild type; WT) mice and Lewis rats were randomised to sham-operated, ACLT-operated, treated with vehicle, or ACLT-operated, treated with halofuginone. Articular cartilage degeneration was graded using the Osteoarthritis Research Society International (OARSI)-modified Mankin criteria. Immunostaining, flow cytometry, RT-PCR and western blot analyses were conducted to detect relative protein and RNA expression. Bone micro CT (μCT) and CT-based microangiography were quantitated to detect alterations of microarchitecture and vasculature in tibial subchondral bone. Results Halofuginone attenuated articular cartilage degeneration and subchondral bone deterioration, resulting in substantially lower OARSI scores. Specifically, we found that proteoglycan loss and calcification of articular cartilage were significantly decreased in halofuginone-treated ACLT rodents compared with vehicle-treated ACLT controls. Halofuginone reduced collagen X (Col X), matrix metalloproteinase-13 and A disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS 5) and increased lubricin, collagen II and aggrecan. In parallel, halofuginone-attenuated uncoupled subchondral bone remodelling as defined by reduced subchondral bone tissue volume, lower trabecular pattern factor (Tb.pf) and increased thickness of subchondral bone plate compared with vehicle-treated ACLT controls. We found that halofuginone exerted protective effects in part by suppressing Th17-induced osteoclastic bone resorption, inhibiting Smad2/3-dependent TGF-β signalling to restore coupled bone remodelling and attenuating excessive angiogenesis in subchondral bone. Conclusions Halofuginone attenuates OA progression by inhibition of subchondral bone TGF-β activity and aberrant angiogenesis as a potential preventive therapy for OA.
The present study aimed to investigate the molecular mechanisms underlying non‑syndromic cleft lip, with or without cleft palate (NSCL/P), and the association between this disease and cancer. The GSE42589 data set was downloaded from the Gene Expression Omnibus database, and contained seven dental pulp stem cell samples from children with NSCL/P in the exfoliation period, and six controls. Differentially expressed genes (DEGs) were screened using the RankProd method, and their potential functions were revealed by pathway enrichment analysis and construction of a pathway interaction network. Subsequently, cancer genes were obtained from six cancer databases, and the cancer‑associated protein‑protein interaction network for the DEGs was visualized using Cytoscape. In total, 452 upregulated and 1,288 downregulated DEGs were screened. The upregulated DEGs were significantly enriched in the arachidonic acid metabolism pathway, including PTGDS, CYP4F2 and PLA2G16; and transforming growth factor (TGF)‑β signaling pathway, including SMAD3 and TGFB2. The downregulated DEGs were distinctly involved in the pathways of DNA replication, including MCM2 and POLA1; cell cycle, including CDK1 and STAG1; and viral carcinogenesis, including PIK3CA and HIST1H2BF. Furthermore, the pathways of cell cycle and viral carcinogenesis, with higher degrees of interaction were found to interact with other pathways, including DNA replication, transcriptional misregulation in cancer, and the TGF‑β signaling pathway. Additionally, TP53, CDK1, SMAD3, PIK3R1 and CASP3, with higher degrees, interacted with the cancer genes. In conclusion, the DEGs for NSCL/P were implicated predominantly in the TGF‑β signaling pathway, the cell cycle and in viral carcinogenesis. The TP53, CDK1, SMAD3, PIK3R1 and CASP3 genes were found to be associated, not only with NSCL/P, but also with cancer. These results may contribute to a better understanding of the molecular mechanisms of NSCL/P.
This study aimed to investigate the effect of small-conductance calcium-activated potassium channels (SK channels) on the dopaminergic (DA) neuron pathways in the ventral tegmental area (VTA) during the pathogenesis of post-stroke depression (PSD) and explore the improvement of PSD by inhibiting the SK channels.Four groups of Sprague-Dawley rats were randomly divided: Control, PSD, SK channel inhibitor (apamin) and SK channel activator (CyPPA) groups. In both control and CyPPA groups, sham surgery was performed. In the other two groups, middle cerebral arteries were occluded. The behavioral indicators related to depression in different groups were compared. Immunofluorescence was used to measure the activity of DA neurons in the VTA, while qRT-PCR was used to assess the expression of SK channel genes.The results showed that apamin treatment improved behavioral indicators related to depression compared to the PSD group. Furthermore, the qRT-PCR analysis revealed differential expression of the KCNN1 and KCNN3 subgenes of the SK channels in each group. Immunofluorescence analysis revealed an increase in the expression of DA neurons in the VTA of the PSD group, which was subsequently reduced upon apamin intervention.This study suggests that SK channel activation following stroke contributes to depression-related behaviors in PSD rats through increased expression of DA neurons in the VTA. And depression-related behavior is improved in PSD rats by inhibiting the SK channels. The results of this study provide a new understanding of PSD pathogenesis and the possibility of developing new strategies to prevent PSD by targeting SK channels.
Thermoacoustic imaging (TAI) has been shown to be promising for breast cancer detection. Contrast-enhanced TAI (CETAI) applying exogenous contrast agents can enhance the contrast in the dielectric properties between the tumor and its surrounding tissues. Combining compressive sensing (CS) and CET AI can make the measurements much more time-efficient. This work proposes a new approach to enable the advancing of the CS based CETAI from 2-D to 3-D. Simulation result using a realistic breast phantom shows that the measurement of CS based CET AI can be expedited by a factor more than 10 compared with the conventional CET AI.
Objective To investigate the killing effects of DEIC/Dkk-3 on renal cancer cell line 786-0 and its potential mechanism.Methods We transfected the pEZ-M29-REIC/Dkk-3 plasmid into 786-0 cells to overexpress REIC/Dkk-3.The inhibitory effects of REIC/Dkk-3 on cell proliferation was assessed using cell countiong kit-8 (CCK-8) at 24,48,72 and 96 h after transfection.Apoptosis of 786-0 cells was detected by AnnexinV/PI double staining,and cell cycle was detected by PI staining using flow cytometry.Changes of protein expression related to apoptosis and cell cycle arrest were measured by Westem blotting.Results The results of immunoblotting (Western blotting) and real-time polymerase chain reaction (Real-time PCR) showed that REIC/Dkk-3 gene expression was increased five folds after transfection as compared with control group.The results of CCK-8 method indicated that with the prolongation of transfection time,inhibitory effect of 786-0 cell proliferation was increased.At 24,48,72 and 96 h,cell survival rate in gene overexpression group and control vector group was 92.0％,90.0％,84.0％ and 80.5％,and 89.50％,73.75％,63.00％ and 58.50％,respectively (P ＜0.05).The apoptosis rate in gene overexpression group,control vector group and blank control group was 28.5％,7.2％ and 0.6％ respectively.Overexpression of REIC/Dkk-3 could up-regulate the expression of Cleaved-Caspase-3,and down-regulate the expression of B cell lymphoma/leukemia-2 (bcl-2).In addition,overexpression of REIC/Dkk-3 caused G1 phase arrest of 786-0 cells.The percentage of G1 phase in pEZ-M29-REIC/Dkk-3 group,pEZ-M29-Flag group and control group was (69.90 ± 3.40) ％,(59.57 ± 1.20) ％ and (52.57 ± 1.20) ％ respectively.Results of Western blotting showed that overexpression of REIC/Dkk-3 could increase the expression of p21 and decrease the expression of Cyclin D1.Conclusion REIC/Dkk-3 can inhibit the proliferation of renal cancer cell 786-0 probably by inducing cell apoptosis and G1 phase arrest.
Key words:
REIC/Dkk-3; Renal cell cancer; Proliferation
Objective
To explore the diagnosis and treatment of BKV nephropathy after renal transplantation.
Methods
A total of 62 patients with progressive creatinine elevation were routinely examined by blood and urine BKV-DNA. And 21 patients with positive results underwent graft biopsies for confirming a diagnosis.
Results
Among 21 cases of BKV infection, 20 cases received leflunomide in replacing mycophenolate mofetil (MMF) and a lower dose of tacrolimus. One case with urine (-) & blood (+ ) received sirolimus in replacing tacrolimus and a lower dose of MMF. Among 11 cases with urine (+ ) and blood (-), urinary BKV-DNA turned negative & creatinine decreased markedly (n=4), urinary BKV-DNA load decreased & creatinine stablized (n=4), death from pulmonary infection with hepatic & renal failure (n=1), urine BKV-DNA load decreased & creatine increased (n=1), BKV–DNA load was not re-examined in 1 case of acute rejection and hydronephrosis with elevated creatine; Among 9 cases with urine (+ ) & blood (+ ), blood BKV-DNA turned negative with urinary BKV-DNA load & creatine decreased (n=6), blood BKV-DNA load decreased & creatine stablized (n=2) and no re-examination with a stable level of creatine (n=1); One case with urine (-) & blood (+ ) was not timely treated and ultimately leading to graft loss after an onset of acute rejection.
Conclusions
BKV nephropathy may be effectively treated by decreasing immunosuppressive intensity. However, clinicians should stay on a high alert for acute rejection due to an excessive reduction of immunosuppressive agents.
Key words:
Kidney transplantation; BK Virus; Acute rejection
Cytokine-induced killer (CIK) cells are immune effector cells characterized by co-expression of CD3 and CD56 molecules. We examined the quantities of CIK cells and the changes of these cell expressing NK cell receptors in HIV-1-positive children infected via mother-to-child transmission. The percentage of CIK cells was quantified and the changes in the surface cell receptor profiles in 18 HIV-1-infected children were examined. We found that CIK cell percentages were dramatically increased in HIV-1-infected children. Furthermore, the expressions of CD16, NKp30, NKp44, NKp46, NKp80 and CD244 on CIK cells were decreased, while the expressions of KIR3DL1 and NKG2D on CIK cells were increased in HIV-1-infected children. However, the expressions of KIR2D and NTB-A on CIK cells did not change in the HIV-1-infected children. CIK cells possessed the characteristics of promoting the maturation of dendritic cells and killing functions in HIV-1-infected children. Moreover, serum concentrations of IL-4 and IFN-γ were significantly increased in HIV-1-infected children compared with the HIV-negative controls. These changes likely occurred as a protective mechanism against transmission of maternal HIV-1 virus and thereby helped to limit viral spread, eliminate infected cells and help HIV-1-infected patients to slow the progression to AIDS.
The present study aimed to investigate the genetic effects of hydrocortisone (HC) treatment on keloids and screen medicines to be used in a combination therapy of keloids with HC. The dataset GSE7890 was downloaded from Gene Expression Omnibus. It contained data regarding 4 fibroblast samples from normal scar tissue and 5 samples from keloid tissue with HC treatment, as well as 5 samples from normal scar and 5 samples from keloids without HC treatment. Following the identification of differentially expressed genes (DEGs), the functions of these DEGs were analyzed by Gene Ontology (GO) and pathway enrichment analyses. Furthermore, adverse effects of HC were identified using WebGestalt. Additionally, candidate small molecule drugs associated with keloids were selected from a connectivity map database. A total of 166 and 41 DEGs, with and without HC treatment respectively, were only present in dermal fibroblasts from keloids (termed genesets A and B, respectively). A set of 26 DEGs was present following both treatments (geneset C). A number of DEGs in geneset B (COL18A1 and JAG1) were associated with endothelial cell differentiation. However, in genesets A and C, certain genes (CCNB1 and CCNB2) were involved in the cell cycle and p53 signaling pathways, and a number of genes (IL1R1 and COL1A1) were associated with bone loss. Additionally, numerous small molecule drugs (including acemetacin) were associated with keloids. Thus, it has been determined that HC may treat keloids by targeting genes associated to endothelial cell differentiation (COL18A1 and JAG1). However, HC has a number of adverse effects, including bone loss. Acemetacin may be applied in a combination therapy, along with HC, to treat keloids.
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