Longitudinal variation of serum PCSK9 in ulcerative colitis: association with disease activity, T helper 1/2/17 cells, and clinical response of tumor necrosis factor inhibitor.

AIMS: Proprotein convertase subtilisin/kexin type 9 (PCSK9) modulates CD4+ T cell differentiation and inflammatory response, the latter ones mediate ulcerative colitis (UC) initiation. This study intended to explore the correlation of serum PCSK9 with disease activity, T helper (h)1/Th2/Th17 cells, and clinical response of tumor necrosis factor inhibitor (TNFi) in UC patients. METHODS: In 65 UC patients underwent TNFi treatment, serum PCSK9 was evaluated at baseline (W0), week (W)2, W6, and W12 by enzyme-linked immunosorbent assays; meanwhile, Th1/Th2/Th17 cells were determined at W0 by flow cytometry. Besides, serum PCSK9 was detected in 65 healthy controls (HCs). RESULTS: Serum PCSK9 was increased in UC patients compared to HCs (P<0.001), which also positively correlated with C-reactive protein (P=0.009), total Mayo score (P=0.018), Mayo-defined disease activity (P=0.020), Th1 (P=0.033), and Th17 (P=0.003) cells, but not Th2 cells (P=0.086) in UC patients. Interestingly, serum PCSK9 was steadily declined from W0 to W12 (P<0.001). W2-W0, W6-W0, and W12-W0 serum PCSK9 change (PCSK9 at W2, W6, or W12 minus PCSK9 at W0, respectively) was gradually becoming greater during TNFi treatment (P<0.001). Furthermore, forty-five (69.2%) patients achieved clinical response at W12, whose serum PCSK9 at W6 (P=0.041) and W12 (P=0.001) was lower, and W6-W0 (P=0.043), W12-W0 (P=0.019) serum PCSK9 change was more obvious compared to patients without clinical response at W12. CONCLUSIONS: Serum PCSK9 is positively related to disease activity, Th1, and Th17 cells in UC patients; further, its decline correlates with TNFi response achievement in these patients.

[Retracted] microRNA­145 inhibits osteosarcoma cell proliferation and invasion by targeting ROCK1.

Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that several of the data panels shown in Fig. 4A were overlapping with panels contained within the same figure, where the panels were intended to portray the results from a range of different cell migration assay experiments. Given the number of overlaps of data that have been identified, the Editor of Molecular Medicine Reports has decided that this paper should be retracted from the Journal on account of a lack of confidence in the presented data. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a satisfactory reply. The Editor apologizes to the readership for any inconvenience caused. [Molecular Medicine Reports 10: 155­160, 2014; DOI: 10.3892/mmr.2014.2195].

[Retracted] Knockdown of PREX2a inhibits the malignant phenotype of osteosarcoma cells.

Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that the Transwell cell migration assay data shown in Fig. 4A and B were strikingly similar to data appearing in different form in other articles by different authors. Owing to the fact that the contentious data in the above article had already been published elsewhere, or were already under consideration for publication, prior to its submission to Molecular Medicine Reports, the Editor has decided that this paper should be retracted from the Journal. The authors were asked for an explanation to account for these concerns, but the Editorial Office did not receive a satisfactory reply. The Editor apologizes to the readership for any inconvenience caused. [the original article was published on Molecular Medicine Reports 13: 1930­1936, 2016; DOI: 10.3892/mmr.2015.4728].

Surface coating prolongs the degradation and maintains the mechanical strength of surgical suture in vivo.

Biodegradable and absorbable sutures have been widely used in surgical procedures. However, for the repair of ligament and tendon injures, the biodegradable suture cannot provide sufficient mechanical support to close the wound for a long period of time which is important to completely heal the tissue. Herein, we develop a simple method that makes a surface coating to prolong the degradation of the suture in vivo. Polylactic acid (PLLA) and Polycaprolactone (PCL) were successfully coated to a commercial degradable polydioxanone (PDO) suture in this study, which was confirmed by Fourier transform infrared spectra (FTIR). Scanning electron microscopy (SEM) was used to observe the smooth surface of the coated sutures. Moreover, live/dead assay of human fibroblasts after co-culturing with the modified/unmodified sutures showed fairly good cellular activity. In vivo study demonstrates the degradation properties of sutures were significantly changed after the surface coating. The raw suture exhibited the fastest degradation in 12 weeks, showing significantly decline in mechanical strength. Interestingly, the PCL-coated suture was able to maintain more than 20% of its original tensile strength after 12 weeks' implantation. In addition, in vivo results of PCL-coated sutures also showed less inflammatory cell infiltration and less surface inflammation. These findings indicate the one step suture-coating method could be feasibly for the development of clinical equipment.

[Retracted] Antitumor effect of resveratrol on chondrosarcoma cells via phosphoinositide 3­kinase/AKT and p38 mitogen­activated protein kinase pathways.

Following the publication of this paper, it was drawn to the Editors' attention by a concerned reader that the cell apoptotic data shown in Fig. 2 were strikingly similar to data appearing in different form in other articles by different authors. Owing to the fact that the contentious data in the above article had already been published elsewhere, or were already under consideration for publication, prior to its submission to Molecular Medicine Reports, the Editor has decided that this paper should be retracted from the Journal. After having been in contact with the authors, they agreed with the decision to retract the paper. The Editor apologizes to the readership for any inconvenience caused. [the original article was published in Molecular Medicine Reports 12: 3151-3155, 2015; DOI: 10.3892/mmr.2015.3683].

3D printing guide plate for accurate hemicortical bone tumor resection in metaphysis of distal femoral: a technical note.

BACKGROUND: Surgical resection and reconstruction for low-grade bone sarcoma in the metaphysis of the distal femur remain challenging. We hypothesized that 3D printing osteotomy guide plate could assist to accurately resect the tumor lesion and save the joint function. METHODS: From January 2017 to August 2019, five patients diagnosed with low-grade bone sarcoma in the metaphysis of the distal femur were treated with hemicortical resection using 3D printing guide plate. Autologous bone graft was inactivated in a high-temperature water bath and re-implanted in situ fixed with plate and screw. Patients were followed up from 17 to 33 months. The Musculoskeletal Tumor Society Score was used to evaluate the joint function. X-ray was used to evaluate the bone union. RESULTS: One patient was paracorticular osteosarcoma, and four cases had highly differentiated chondrosarcoma. All cases were involved in the metaphysis of the distal femur. Patients were followed up from 13 to 33 months, with an average of 23.6 months. There was neither post-operation infection, internal fixation loosening, nor fracture occurrence in any of the patients. The Musculoskeletal Tumor Society Score averaged at 28.1, while the International Society of Limb Salvage imaging score examination averaged 89.8%. CONCLUSIONS: Here, we demonstrate that the 3D printing osteotomy guide plate-assisted hemicortical bone resection is a beneficial strategy to effectively resect the primary low-grade malignant bone tumors in the metaphysis of the distal femur and retained satisfied joint function.

Characterizations of Gene Alterations in Melanoma Patients from Chinese Population.

Melanoma is a human skin malignant tumor with high invasion and poor prognosis. The limited understanding of genomic alterations in melanomas in China impedes the diagnosis and therapeutic strategy selection. We conducted comprehensive genomic profiling of melanomas from 39 primary and metastatic formalin-fixed paraffin-embedded (FFPE) samples from 27 patients in China based on an NGS panel of 223 genes. No significant difference in gene alterations was found between primary and metastasis melanomas. The status of germline mutation, CNV, and somatic mutation in our cohort was quite different from that reported in Western populations. We further delineated the mutation patterns of 4 molecular subgroups (BRAF, RAS, NF1, and Triple-WT) of melanoma in our cohort. BRAF mutations were more frequently identified in melanomas without chromic sun-induced damage (non-CSD), while RAS mutations were more likely observed in acral melanomas. NF1 and Triple-WT subgroups were unbiased between melanomas arising in non-CSD and acral skin. BRAF, RAS, and NF1 mutations were significantly associated with lymph node metastasis or presence of ulceration, implying that these cancer driver genes were independent prognostic factors. In summary, our results suggest that mutational profiles of malignant melanomas in China are significantly different from Western countries, and both gene mutation and amplification play an important role in the development and progression of melanomas.

Macrophage inhibits the osteogenesis of fibroblasts in ultrahigh molecular weight polyethylene (UHMWPE) wear particle-induced osteolysis.

BACKGROUND: In the ultrahigh molecular weight polyethylene (UHMWPE) prosthetic environment, fibroblasts affected by wear particles have the capacity of osteogenesis to reduce osteolysis. We aimed to assess the effects of macrophages on the osteogenic capability of fibroblasts treated with UHMWPE wear particles. METHODS: The effect of different concentrations of UHMWPE (0, 0.01, 0.1, and 1 mg/ml, respectively) on macrophage proliferation were validated by MTT assay to determine the optimum one. The fibroblasts viability was further determined in the co-culture system of UHMWPE particles and macrophage supernatants. The experiment was designed as seven groups: (A) fibroblasts only; (B) fibroblasts + 1 mg/ml UHMWPE particles; and (C1-C5) fibroblasts + 1/16, 1/8, 1/4, 1/2, and 1/1 supernatants of macrophage cultures stimulated by 1 mg/ml UHMWPE particles vs. fibroblast complete media, respectively. Alizarin red staining was used to detect calcium accumulation. The expression levels of osteogenic proteins were detected by Western blot and ELISA, including alkaline phosphatase (ALP) and osteocalcin (OCN). RESULTS: The concentration of 0.1 mg/ml was considered as the optimum concentration for macrophage proliferation due to the survival rate and was highest among the four concentrations. Fibroblast viability was better in the group of fibroblasts + 1/16 ratio of macrophage supernatants stimulated by 1 mg/ml of UHMWPE particles than the other groups (1:8, 1:4, 1:2, 1:1). ALP and OCN expressions were significantly decreased in the group of fibroblasts + 1/4, 1/2, and 1/1 supernatants stimulated by 1 mg/ml of UHMWPE particles compared with other groups (1/8, 1/16) and the group of fibroblasts + 1 mg/ml UHMWPE (p < 0.5). CONCLUSIONS: Macrophages are potentially involved in the periprosthetic osteolysis by reducing the osteogenic capability of fibroblasts treated with wear particles generated from UHMWPE materials in total hip arthroplasty.

MicroRNA-200b acts as a tumor suppressor in osteosarcoma via targeting ZEB1.

Osteosarcoma is the most common type of cancer that develops in bone, mainly arising from the metaphysis of the long bones. MicroRNA (miR)-200b has been found to generally act as a tumor suppressor in multiple types of human cancers. However, the detailed role of miR-200b in osteosarcoma still remains to be fully understood. This study aimed to investigate the exact role of miR-200b in the progression of osteosarcoma and the underlying mechanism. Real-time reverse transcription-polymerase chain reaction data showed that miR-200b was significantly downregulated in osteosarcoma tissues compared to their matched adjacent nontumor tissues. Low miR-200b level was associated with the advanced clinical stage and positive distant metastasis. Besides, it was also downregulated in osteosarcoma cell lines (U2OS, Saos2, HOS, and MG63) compared to normal osteoblast cell line NHOst. In vitro study showed that restoration of miR-200b led to a significant decrease in proliferation, migration, and invasion of osteosarcoma cells. Moreover, ZEB1 was identified as a target gene of miR-200b, and its expression levels were negatively mediated by miR-200b in osteosarcoma cells. In addition, ZEB1 was significantly upregulated in osteosarcoma cells compared to the normal osteoblast cell line NHOst, and inhibition of ZEB1 expression also suppressed the proliferation, migration, and invasion in osteosarcoma cells. Finally, we showed that ZEB1 was frequently upregulated in osteosarcoma tissues compared to their matched adjacent normal tissues, and its expression was reversely correlated to the miR-200b levels in osteosarcoma tissues. Based on these findings, our study suggests that miR-200b inhibits the proliferation, migration, and invasion of osteosarcoma cells, probably via the inhibition of ZEB1 expression. Therefore, miR-200b/ZEB1 may become a potential target for the treatment of osteosarcoma.

Knockdown of PREX2a inhibits the malignant phenotype of osteosarcoma cells.

Phosphatidylinositol­3,4,5­trisphosphate­dependent Rac exchange factor 2a (PREX2a), which is a regulator of the small guanosine triphosphatase Rac, has recently been reported to have an oncogenic role via the suppression of phosphatase and tensin homolog deleted on chromosome ten (PTEN) activity, and the subsequent activation of phosphoinositide 3­kinase (PI3K) signaling. However, the detailed role of PREX2a in osteosarcoma (OS) remains unclear. Reverse transcription quantitative polymerase chain reaction and western blotting was used to detect mRNA and protein levels. MTT assay was performed to examine cell proliferation and a Transwell assay and wound healing assay were conducted to examine cell invasion and migration. The present study demonstrated that PREX2a was markedly upregulated in OS cell lines, as compared with in normal osteoblast hFOB1.19 cells. In addition, knockdown of PREX2a expression significantly inhibited OS cell proliferation, whereas overexpression of PREX2a markedly promoted OS cell proliferation. Inhibition of PREX2a also markedly suppressed the invasion and migration of OS cells, at least partly by suppressing the protein expression of matrix metalloproteinase (MMP)2 and MMP9. Conversely, upregulation of PREX2a enhanced OS cell invasion and migration. In addition, PI3K signaling activity was significantly reduced following knockdown of PREX2a, and this was accompanied by an upregulation of PTEN activity. The results of the present study suggested that PREX2a may act as an oncogene in OS via the inhibition of PTEN activity and activation of PI3K signaling.

Alteration of the RANKL/RANK/OPG System in Periprosthetic Osteolysis with Septic Loosening.

The pathogenesis of periprosthetic osteolysis with septic loosening remains incompletely understood. The purpose of this study was to investigate whether expression of the RANKL/RANK/OPG system is altered in septic interface membranes (SIMs). Seventeen cases with a SIM, 26 cases with an aseptic interface membrane (AIM), and 12 cases with a normal synovium (NS) were assessed. Scanning and transmission electron microscopy (SEM and TEM, respectively) were used to observe the microscopic morphology of three tissue conditions. Differences in RANKL, RANK, and OPG expression at the mRNA level were assessed by real-time quantitative PCR, and differences at the protein level were assessed by immunohistochemical staining and Western blotting. SEM showed wear debris widely distributed on the AIM surface, and TEM showed Bacillus activity in the SIM. RANKL expression and the RANKL/OPG ratio were significantly increased in SIMs. Imbalance in the RANKL/RANK/OPG system is related to periprosthetic osteolysis with septic loosening but is not the only possible pathogenic mechanism.

Antitumor effect of resveratrol on chondrosarcoma cells via phosphoinositide 3-kinase/AKT and p38 mitogen-activated protein kinase pathways.

Chondrosarcoma is one of the most common types of primary bone cancer that develops in cartilage cells. Resveratrol (Res), a natural polyphenol compound isolated from various fruits, has a suppressive effect on various human malignancies. It has been shown that Res inhibits matrix metalloproteinase (MMP)-induced differentiation in chondrosarcoma cells. However, the effects of Res on cell proliferation, apoptosis and invasion of chondrosarcoma cells, as well as the underlying mechanisms, remain largely unknown. To the best of our knowledge, the present study showed for the first time that Res inhibited proliferation and induced apoptosis in chondrosarcoma cells in a dose-dependent manner. Furthermore, it was shown that Res also suppressed chondrosarcoma cell invasion in a dose-dependent manner, probably via the inhibition of MMP2 and MMP9 protein expression. Molecular mechanism investigations revealed that Res could inhibit the activity of phosphoinositide 3-kinase/AKT and p38 mitogen-activated protein kinase signaling pathways, which has been demonstrated to be important in the regulation of proliferation, apoptosis and invasion in various cancer cell types. In conclusion, this study suggests that Res may serve as a promising agent for the treatment of chondrosarcoma.

microRNA-145 inhibits osteosarcoma cell proliferation and invasion by targeting ROCK1.

Osteosarcoma (OS), a malignant mesenchymal sarcoma, is the most frequent primary bone tumor, with a peak incidence in young children and adolescents. The downregulation of microRNA­145 (miRNA/miR­145) has previously been identified to be associated with the aggressiveness and metastasis of OS. However, the detailed regulatory mechanism by which miR­145 inhibits OS remains largely unknown. The present study demonstrated that miR­145 was significantly downregulated in OS tissues and KHOS and U2OS cell lines. Rho­associated protein kinase 1 (ROCK1), a key regulator of actin cytoskeleton reorganization, was identified as a novel target of miR­145. Ectopic expression of miR­145 notably suppressed the protein expression of ROCK1 without affecting its mRNA level. Furthermore, the expression of ROCK1 was significantly increased in the OS tissues and in the KHOS and U2OS cells. It was further demonstrated that the overexpression of miR­145 downregulated KHOS and U2OS cell proliferation and invasion, which was reversed by restoration of ROCK1. To the best of our knowledge, the present study demonstrates for the first time that, as a tumor suppressor, miRNA­145 inhibits OS cell proliferation and invasion, at least in part by directly targeting ROCK1. These results indicate that miR­145 may be a potential candidate for the diagnosis and treatment of OS.

Medium-term outcomes of cemented prostheses and cementless modular prostheses in revision total hip arthroplasty.

There is an increasing trend towards cementless modular femoral prostheses for revision hip replacement surgery, especially in patients with severe proximal femoral bone defects. However, for minor femoral bone defects, the benefit of cementless modular is not clear. We designed a retrospective cross-sectional study to compare outcomes of the two femoral implant designs. There were no significant differences in terms of visual analog pain scores, Harris hip scores, femoral bone restoration, stem subsidence, leg length correction, or overall complication rate. Three femoral reoperations (11%) occurred in the cemented group, and two (9%) in the cementless modular group. One femoral stem re-revised (4%) in the cemented group due to recurrent deep infection. Five-year survival for femoral reoperation was 88.2% for patients with the cemented implant and 91.3% for cementless group. Both groups had good clinical and radiological outcomes for femoral revision in patients with minor femoral bone defects during medium-term follow-up.

Three to seven year follow-up of a tapered modular femoral prosthesis in revision total hip arthroplasty.

PURPOSE: Modular femoral prostheses can provide independent distal fixation and intraoperative flexibility and are being used increasingly, especially in patients with proximal femoral bone defects. This retrospective clinical study evaluated whether modular prostheses were effective and reliable implants for femoral revision. METHOD: This case series consisted of 58 patients who underwent hip revision with a tapered modular femoral prosthesis at our institution between 2004 and 2008. Mean patient age at surgery was 64 years (range 18-86 years). Femoral bone defects before revision surgery were evaluated using the Paprosky classification. All patients were followed for a minimum of 3 years (mean 4.3 years, range 3-7 years) with clinical and radiographic evaluation. Re-revisions and complications were also recorded. RESULTS: Two stems required re-revision, one each for recurrent deep infection and periprosthetic fracture. At last follow-up, the Harris Hip Score and Visual Analog Pain Scores had improved significantly, the median radiographic stem migration was 1.6 mm, leg length discrepancy was corrected in 64 % of the patients and osseointegration occurred in 90 %. Complications included intraoperative fracture in 10 (17 %) patients and hip dislocation in 2 (3 %). CONCLUSION: Modular femoral components can improve hip function, provide distal fixation, equalize leg length, and result in fewer complications when used to revise failed femoral components.

[Mid-term effectiveness of two-stage hip prosthesis revision in treatment of infection after hip arthroplasty].

OBJECTIVE: To evaluate the mid-term effectiveness of two-stage hip prosthesis revision in the treatment of infection after hip arthroplasty. METHODS: Between April 2002 and November 2006, 12 cases of infection after hip arthroplasty were treated. There were 5 males and 7 females, aged from 47 to 72 years (mean, 59.8 years). The femoral head arthroplasty was performed in 2 cases and total hip arthroplasty in 10 cases. Infection occurred 1 to 67 months after arthroplasty. According to the Segawa classification criteria, infections included type 2 in 1 case, type 3 in 2 cases, and type 4 in 9 cases. The preoperative Harris score was 36.7 +/- 6.1. Nine cases had elevated C reactive protein and 10 cases had elevated erythrocyte sedimentation rate. The results of bacterial culture were positive in 8 cases and negative in 4 cases. After the removal of the infected prosthesis and thorough debridement, antibiotic-loaded cement spacers or infected therapeutic temporary prosthesis were used as placeholders, and then the anti-infection treatments were given after operations; two-stage hip prosthesis revisions were performed 3 to 10 months after debridement. RESULTS: In 1 patient who failed to control infection after debridement, infection was controlled after the second debridement and the antibiotic-loaded cement spacer as placeholder. Other patients achieved healing of incision by first intention, and no complication such as deep venous thrombosis and nerve injury occurred. All patients were followed up 3 to 8 years after revision (mean, 5.4 years). During the follow-up, no infection recurrence and joint dislocation occurred. Dull pain was present in 2 cases during activity and mild claudication in 3 cases at last follow-up. The Harris score was 81.6 +/- 4.5, showing significant difference (t = 52.696, P = 0.000) when compared with preoperative score. The X-ray films showed that no prosthesis loosening and obvious subsidence were observed, and bone graft healed. CONCLUSION: The two-stage hip prosthesis revision has good infection control rate and mid-term effectiveness in treatment of infection after hip arthroplasty.