[18F] AlF-NOTA-FAPI-04 PET/CT can predict treatment response and survival in patients receiving chemotherapy for inoperable pancreatic ductal adenocarcinoma.

PURPOSE: We investigated whether uptake of [18F] AlF-NOTA-FAPI-04 on positron emission tomography/computed tomography (PET/CT) could predict treatment response and survival in patients with pancreatic ductal adenocarcinoma (PDAC). METHODS: We prospectively evaluated 47 patients with histopathologically confirmed primary PDAC who provided pretreatment [18F] AlF-NOTA-FAPI-04 scans to detect fibroblast activation protein (FAP) on the tumor surface by uptake of [18F] AlF-NOTA-FAPI-04. PDAC specimens were immunohistochemically stained with cancer-associated fibroblast (CAF) markers. We obtained a second PET scan after one cycle of chemotherapy to study changes in FAPI uptake variables from before to during treatment. Correlations between baseline PET variables and CAF-related immunohistochemical markers were assessed with Spearman's rank test. Cox regression and Kaplan-Meier methods were used to assess relationships between disease progression and potential predictors. Receiver operating characteristic (ROC) curve analysis was used to define the optimal cut-off points for distinguishing patients according to good response vs. poor response per RECIST v.1.1. RESULTS: The FAPI PET variables maximum and mean standardized uptake values (SUVmax, SUVmean), metabolic tumor volume (MTV), and total lesion FAP expression (TLF) were positively correlated with CAF markers (FAP, α-smooth muscle actin, vimentin, S100A4, and platelet-derived growth factor receptor α/ß, all P < 0.05). MTV was associated with survival in patients with inoperable PDAC (all P < 0.05). Cox multivariate regression showed that MTV was associated with overall survival (MTV hazard ratio [HR] = 1.016, P = 0.016). Greater changes from before to during chemotherapy in SUVmax, MTV, and TLF were associated with good treatment response (all P < 0.05). ΔMTV, ΔTLF, and ΔSUVmax had larger areas under the curve than ΔCA19-9 for predicting treatment response. Kaplan-Meier analysis showed that the extent of change in MTV and TLF from before to after treatment predicted progression-free survival, with cut-off values (based on medians) of - 4.95 for ΔMTV (HR = 8.09, P = 0.013) and - 77.83 for ΔTLF (HR = 4.62, P = 0.012). CONCLUSIONS: A higher baseline MTV on [18F] AlF-NOTA-FAPI-04 scans was associated with poorer survival in patients with inoperable PDAC. ΔMTV was more sensitive for predicting response than ΔCA19-9. These results are clinically meaningful for identifying patients with PDAC who are at high risk of disease progression.

A pilot study on the use of a novel digital real-time evaluation system in undergraduate preclinical training of tooth preparation in fixed prosthodontics.

INTRODUCTION: In the light of the digital teaching, it is necessary that the effectiveness of a new digital real-time evaluation system in the preclinical training of tooth preparation be evaluated. MATERIALS AND METHODS: Forty undergraduate dental students of the fourth year were randomly divided into the control group and the experimental group to complete the training task of tooth preparation for porcelain fused to metal (PFM) crown restoring the upper right central incisor. The control students received conventional training with instructor's guidance. The experimental students received training with the digital system without instructor's guidance. Every student exercised preparation in two resin incisors in 3 h training by respective training methods. A third incisor was prepared on a dental model in the simulated head phantom by each student as the test on the next day. All students' tooth preparations were scored by the same two experienced experts. The experimental students were asked to answer a questionnaire regarding their attitudes and opinions on the digital evaluation system in preclinical training. RESULTS: There was no significant difference between the scores of the experimental group and the control group (p > .05). The students of two groups obtained the similar scores in the test (p > .05). Most of the students were supportive of the application of digital training system in the preclinical tooth preparation training course. CONCLUSIONS: The digital real-time evaluation system could provide effective training effects for the dental undergraduate students in the preclinical training of tooth preparation in fixed prosthodontics.

Effect of Aging Time and Thickness on Low-Temperature Degradation of Dental Zirconia.

PURPOSE: To evaluate the influence of specimen thickness and low-temperature degradation (LTD) on yttrium-stabilized tetragonal zirconia polycrystals (Y-TZP). MATERIALS AND METHODS: Thin discs of Y-TZP from four manufacturers were sintered according to each manufacturer's recommendations, cut into 23 mm (length) × 4 mm (width) × 0.8 mm/1.5 mm (thickness) specimens, artificially aged under standard autoclave sterilization conditions (34°C at 0.2 MPa for 10 and 20 hours), and finally ground and polished. Tetragonal to monoclinic transformation was confirmed by X-ray diffraction (XRD) analysis. Flexural strength was measured by 3-point bending tests and Vickers hardness measurements. Fracture surfaces were examined by scanning electron microscopy (SEM). RESULTS: SEM investigation revealed that with increasing aging time, the surface defects and grain size increased, particularly in the 20-hour group. Compared with the 1.5 mm group, the 0.8 mm group showed more significant defects, irrespective of aging time. The flexural strengths of Y-TZP materials decreased with a decrease in the thickness. Moreover, LTD of Y-TZP can cause significant tetragonal to monoclinic transformation, which also results in a statistically significant decrease in the flexural strength. CONCLUSION: A thinner Y-TZP specimen was likely to present surface defects and microcracks after aging. In addition, the flexural strengths decreased with a decrease in the thickness, a notable fact for further studies.

Clinical Evaluation and Patient Satisfaction of Single Zirconia-Based and High-Noble Alloy Porcelain-Fused-to-Metal Crowns in the Esthetic Area: A Retrospective Cohort Study.

PURPOSE: The aim of this study was to assess clinical survival, complications, and patient satisfaction with single zirconia-based crowns (ZC) or high-noble alloy PFM crowns (HC) in esthetic areas. MATERIALS AND METHODS: The study was a retrospective cohort study with up to 3.8 years follow-up (mean 2.5 years). Patients who met the inclusion criteria were reviewed. Survival rates, complications, satisfaction rates (color and form), and overall satisfaction (visual analogue scale, VAS) were evaluated by follow-up examination. Chi-square test (survival, complication, and satisfaction rate) and unpaired t-test (overall satisfaction: VAS) were used to test the difference between the ZC and HC groups. Survival rates and complications were assessed both at subject-level (patient as statistical unit) and at tooth-level (tooth as statistical unit). RESULTS: One hundred and thirteen patients met the inclusion criteria; 95 patients (ZC: 45, HC: 50) with 132 crowns (ZC: 61, HC: 71) were enrolled in the study. The overall survival rate was 93.3% for ZC crowns and 96.0% for HC crowns at subject-level and 93.4% for ZC crowns and 97.2% for HC crowns at tooth-level (p = 0.56 and 0.30, respectively). Veneer chipping was the most frequently seen complication. One HC crown lost retention because of poor cementation. One patient in the ZC group showed progression of periodontal disease. No significant difference of complications was found between the two groups at subject-level and tooth-level (p = 0.37 and 0.34, respectively). The overall satisfaction (VAS) was 8.18 for ZC crowns and 8.46 for HC crowns (p = 0.34). CONCLUSIONS: The results of the study showed that a high survival rate could be achieved in both ZC and HC groups. Chipping of the veneering porcelain was the most frequently seen event that led to short-term (less than 3 years) failure and complications. The patients' rating of ZC crowns was not superior to HC crowns. Well-designed studies with high evidence level and large sample size are still needed to further explore the clinical success of ZC and HC crowns.

Enhanced portable detection for Sars-CoV-2 utilizing DNA tetrahedron-tethered aptamers and a pressure meter.

Tethering oligonucleotide aptamers to a DNA tetrahedron structure can enhance the recognition of SARS-CoV-2 spike protein to effectively overcome challenges with its detection in current diagnostic assays. Building on this framework, we have developed a unique portable detection method for COVID-19 that provides exceptional sensitivity and selectivity via pressure meter readout. This innovative assay streamlines the detection process, providing a rapid, sensitive, cost-effective, and user-friendly diagnostic tool. This point-of-care test exhibits high sensitivity and specificity, achieving an impressive detection limit of 0.1 pg mL-1 for the spike protein. The effectiveness of this method was validated using pseudoviruses and oropharyngeal swab samples, and its utility for environmental monitoring is demonstrated by testing sewage samples. With a wide linear range and strong potential for clinical or home application, our assay represents a major innovation in point-of-care diagnostics and provides a vital contribution to the current toolkit for controlling the impacts of COVID-19.

PARP inhibitor plus radiotherapy reshape the immune suppressive microenvironment and potentiate the efficacy of immune checkpoint inhibitors in tumors with IDH1 mutation.

Isocitrate dehydrogenase 1 mutant (IDH1mut) tumors respond poorly to immunotherapy, but are more sensitive to chemoradiotherapy and poly (ADP-ribose) polymerase inhibition (PARPi). Accordingly, some efforts have aimed to capitalize on the IDH1 mutation rather than reverse it. Moreover, radiotherapy (RT) and PARPi can stimulate antitumor immunity, raising the possibility of reversing the immunosuppression caused by IDH1 mutation while killing the tumor. To assess this possibility, we treated IDH1mut tumors and cells with RT + PARPi. RT + PARPi showed enhanced efficacy over either modality alone both in vitro and in vivo. RT + PARPi induced more DNA damage and activated the cGAS-STING pathway more. IFNß, CXCL10, and CCL5 were also more highly expressed at both the mRNA and protein levels. In two different tumor models, RT + PARPi increased infiltration and cytolytic function of CD8+ T cells, with one model also showing increased CD8+T cell proliferation. RT+PARPi also increased PD-L1 expression and enhanced checkpoint inhibition. Knocking out cGAS reversed the increased CD8+ T cell infiltration and the antitumor effect of RT+PARPi. We conclude that RT + PARPi reshapes the IDH1mut tumor immunosuppressive microenvironment, thereby augmenting checkpoint inhibition.

Simultaneous detection of multiple influenza virus subtypes based on microbead-encoded microfluidic chip.

Influenza virus, existing many subtypes, causes a huge risk of people health and life. Different subtypes bring a huge challenge for detection and treatment, thus simultaneous detection of multiple influenza virus subtypes plays a key role in fight against this disease. In this work, three kinds of influenza virus subtypes are one-step detection based on microbead-encoded microfluidic chip. HIN1, H3N2 and H7N3 were simultaneously captured only by microbeads of different magnetism and sizes, and they were further treated by magnetic separation and enriched through the magnetism and size-dependent microfluidic structure. Different subtypes of influenza virus could be linearly encoded in different detection zones of microfluidic chip according to microbeads of magnetism and size differences. With the high-brightness quantum dots (QDs) as label, the enriched fluorescence detection signals were further read online from linearly encoded strips, obtaining high sensitivity with detection limit of HIN1, H3N2, H7N3 about 2.2 ng/mL, 3.4 ng/mL and 2.9 ng/mL. Moreover, a visual operation interface, microcontroller unit and two-way syringe pump were consisted of a miniaturized detection device, improving the detection process automation. And this assay showed strong specificity. This method improves a new way of multiple pathogens detection using microbead-encoded technologies in the microfluidic chip.

Analysis of the Relationship between National Image Building and International Communication Ability from the Perspective of Ecology under the Media Environment.

With the gradual expansion of China's political, economic, and military strength, the rapid rise of China has become a globally recognized fact. At present, although China's international image as the world's second largest economy, military power, and political power has been increasingly accepted by the global community, China's image construction, as an important part of national soft power, is still facing many problems. This chapter focuses on defining the basic concepts of international publicity, China's soft power, and China's image construction in the context of new media integration, analyzes the international development trend of international publicity and the important functions and limitations of foreign publicity in establishing China's image construction, and analyzes the ecological environment of media integration development. Considering that there are great unfairness in the environment and the conflict of communication values is not conducive to the establishment of China's urban image, the article focuses on how to use global publicity values to establish China's national image and puts forward corresponding countermeasures.

[Corrigendum] Overexpression of HES6 has prognostic value and promotes metastasis via the Wnt/ß-catenin signaling pathway in colorectal cancer.

Subsequently to the publication of the above article, an interested reader drew to the authors' attention that Fig. 2 on p. 1266 and Fig. 5 on p. 1269 contained some apparent errors in terms of the assembly of the various data panels. Specifically, Fig. 2D appeared to contain a pair of overlapping images, and Figs. 5D and 8A also appeared to include overlapping images. However, the authors were able to consult their original data, and assess where the errors had been made during the compilation of these figures. The corrected versions of Figs. 2 (showing the correct data for the '5T' panel in Fig. 2D) and 5 (showing alternative data) are shown on the subsequent pages. The authors regret the errors that were made during the preparation of the published figures, and confirm that these errors did not grossly affect the conclusions reported in the study. The authors are grateful to the Editor of Oncology Reports for allowing them the opportunity to publish a Corrigendum, and all the authors agree to this Corrigendum. Furthermore, they apologize to the readership for any inconvenience caused. [the original article was published in Oncology Reports 40: 1261­1274, 2018; DOI: 10.3892/or.2018.6539].

ALG3 contributes to stemness and radioresistance through regulating glycosylation of TGF-ß receptor II in breast cancer.

BACKGROUND: Radiotherapy is a conventional and effective local treatment for breast cancer. However, residual or recurrent tumors appears frequently because of radioresistance. Novel predictive marker and the potential therapeutic targets of breast cancer radioresistance needs to be investigated. METHODS: In this study, we screened all 10 asparagine-linked glycosylation (ALG) members in breast cancer patients' samples by RT-PCR. Cell viability after irradiation (IR) was determined by CCK-8 assay and flow cytometry. The radiosensitivity of cell lines with different ALG3 expression was determined with the colony formation assay by fitting the multi-target single hit model to the surviving fractions. Cancer stem-like traits were assessed by RT-PCR, Western blot, and flow cytometry. The mechanisms of ALG3 influencing radiosensitivity was detected by Western blot and immunoprecipitation. And the effect of ALG3 on tumor growth after IR was verified in an orthotopic xenograft tumor models. The association of ALG3 with prognosis of breast cancer patients was confirmed by immunohistochemistry. RESULTS: ALG3 was the most significantly overexpressing gene among ALG family in radioresistant breast cancer tissue. Overexpression of ALG3 predicted poor clinicopathological characteristics and overall survival (OS), and early local recurrence-free survival (LRFS) in breast cancer patients. Upregulating ALG3 enhanced radioresistance and cancer stemness in vitro and in vivo. Conversely, silencing ALG3 increased the radiosensitivity and repressed cancer stemness in vitro, and more importantly inhibition of ALG3 effectively increased the radiosensitivity of breast cancer cells in vivo. Mechanistically, our results further revealed ALG3 promoted radioresistance and cancer stemness by inducing glycosylation of TGF-ß receptor II (TGFBR2). Importantly, both attenuation of glycosylation using tunicamycin and inhibition of TGFBR2 using LY2109761 differentially abrogated the stimulatory effect of ALG3 overexpression on cancer stemness and radioresistance. Finally, our findings showed that radiation played an important role in preventing early recurrence in breast cancer patients with low ALG3 levels, but it had limited efficacy in ALG3-overexpressing breast cancer patients. CONCLUSION: Our results suggest that ALG3 may serve as a potential radiosensitive marker, and an effective target to decrease radioresistance by regulating glycosylation of TGFBR2 in breast cancer. For patients with low ALG3 levels, radiation remains an effective mainstay therapy to prevent early recurrence in breast cancer.

Study of Sr-Ca-Si-based scaffolds for bone regeneration in osteoporotic models.

Bone tissue engineering has emerged as a promising alternative therapy for patients who suffer bone fractures or defects caused by trauma, congenital diseases or tumours. However, the reconstruction of bone defects combined with osteoporosis remains a great challenge for clinicians and researchers. Based on our previous study, Ca-Si-based bioceramics (MSCs) showed enhanced bone formation capabilities under normal conditions, and strontium was demonstrated to be therapeutic in promoting bone quality in osteoporosis patients. Therefore, in the present study, we attempted to enlarge the application range of MSCs with Sr incorporation in an osteoporotic bone regeneration model to evaluate whether Sr could assist in regeneration outcomes. In vitro readout suggested that Sr-incorporated MSC scaffolds could enhance the expression level of osteogenic and angiogenic markers of osteoporotic bone mesenchymal stem cells (OVX BMSCs). Animal experiments showed a larger new bone area; in particular, there was a tendency for blood vessel formation to be enhanced in the Sr-MSC scaffold group, showing its positive osteogenic capacity in bone regeneration. This study systematically illustrated the effective delivery of a low-cost therapeutic Sr agent in an osteoporotic model and provided new insight into the treatment of bone defects in osteoporosis patients.

Efficacy of Acupuncture and Moxibustion as a Subsequent Treatment after Second-Line Chemotherapy in Advanced Gastric Cancer.

OBJECTIVE: To explore whether acupuncture and moxibustion can prevent disease progression of advanced gastric cancer patients completing second-line chemotherapy and, if so, the related mechanism. METHOD: Progression-free survival (PFS) and overall survival (OS) were main outcome measures. The real-time quantitative PCR was used to detect the expression of genes including T-bet, IFN-γ, GATA3, and IL-4 in peripheral blood mononuclear cells (PBMCs). IL-4, IL-6, Ca199, CRP, and IFN-γ in plasma levels were checked. RESULTS: 170 patients were randomly assigned in a 3 : 2 ratio to receive either acupuncture and moxibustion or sham acupuncture until progression. 135 patients were included in the primary analysis. Both PFS and OS in treatment group were proven to be better than control group. Acupuncture and moxibustion promoted typical Th1 cells drifting, as confirmed by increased T-bet/IFN-γ and decreased GATA3/IL-4 in mRNA levels from PBMCs, as well as upregulating IFN-γ and downregulating IL-4 in plasma levels. IL-6, Ca199, and CRP in plasma levels were also reduced by acupuncture and moxibustion. CONCLUSIONS: Acupuncture and moxibustion can prolong PFS and OS of advanced gastric cancer patients completing second-line chemotherapy by reversing Th1/Th2 shift and attenuating inflammatory responses.

Challenges and Experiences of Dental Medicine in Combating the COVID-19 Pandemic.

The world has been suffering incredible loss due to a pandemic caused by a novel coronavirus called severe acute respiratory syndrome coronavirus 2' (SARS-CoV-2; 2019-nCoV). The disease was later named the coronavirus disease-19 (COVID-19). The transmission routes of COVID-19 include respiratory transmission, aerosol transmission, and contact transmission. Many dental diagnosis and treatment procedures generate droplets and aerosols, and thus both dental staff and patients are at a high risk of becoming infected and transmitting COVID-19 to others. Shanghai Ninth People's Hospital is a comprehensive hospital with 18 craniofacial-/dental-related departments. During the outbreak of COVID-19 and up to the present date, there have been no confirmed cases of COVID-19 in this hospital thanks to strict protocols for infection prevention and control. In this communication, we would like to share with the prosthodontic community our experience in the prevention and control of COVID-19 in our dental departments and hope it will contribute to the worldwide efforts to overcome the global COVID-19 pandemic.

Overexpression of HES6 has prognostic value and promotes metastasis via the Wnt/ß-catenin signaling pathway in colorectal cancer.

HES6 is a member of the hairy-enhancer of the split homolog family, which has been implicated in oncogenesis and cancer progression in a variety of human cancers, including prostate and breast cancer. However, its clinical significance and biological role in colorectal cancer (CRC) remain unclear. In the present study, the expression of HES6 was significantly upregulated in CRC cell lines and CRC tissues at both the mRNA and protein levels. The present study also reported high expression of HES6 in 138/213 (64.8%) paraffin-embedded archived CRC specimens. HES6 expression was significantly correlated with T classification (P<0.001), N classification (P=0.020), and distant metastasis (P<0.001). Patients with higher HES6 expression levels exhibited a reduced overall survival (P<0.001). In addition, a multivariate analysis revealed that the expression of HES6 may be a novel prognostic marker for the survival of patients with CRC. Furthermore, the present study demonstrated that ectopic expression of HES6 enhanced the migration and invasive abilities of CRC cells. These abilities were significantly inhibited upon knockdown of endogenous HES6 expression by specific short hairpin RNAs. Additionally, the present study reported that the effects of HES6 on metastasis may be associated with the activation of the Wnt/ß-catenin signaling pathway. Collectively, the findings of the present study revealed that overexpression of HES6 played a key role in the progression of CRC, leading to a poor prognosis and clinical outcome.

Jade family PHD finger 3 (JADE3) increases cancer stem cell-like properties and tumorigenicity in colon cancer.

Jade family PHD finger 3 (JADE3) plays a role in inducing histone acetylation during transcription, and is involved in the progression of several human cancers; however, its role in colon cancer remains unclear. Herein, we found that JADE3 was markedly upregulated in colon cancer tissues and significantly correlated with cancer progression, and predicted shorter patient survival. Further, JADE3 was expressed much higher in colon cancer cell lines that are enriched with a stem-like signature. Overexpression of JADE3 increased, while silencing JADE3 reduced cancer stem cell-like traits in colon cancer cells in vitro and in vivo. Importantly, silencing of JADE3 strongly impaired the tumor initiating capacity of colon cancer cells in vivo. Furthermore, JADE3 interacted with the promoters of colon stem cell marker LGR5 and activated its transcription, by increasing the occupancy of p300 acetyltransferase and histone acetylation on the promoters. Finally, we found that JADE3 expression was substantially induced by Wnt/ß-catenin signaling. These findings suggest an oncogenic role of JADE3 by regulating cancer stem cell-like traits in the colon cancer, and therefore JADE3 might be a potential therapeutic target for the treatment of colon cancer.

miR-145 suppresses colorectal cancer cell migration and invasion by targeting an ETS-related gene.

MicroRNA-45 (miR-145) has been demonstrated to be downregulated in various cancer types including colorectal cancer (CRC). However, the function of miR­145 in CRC has not been clearly elucidated. In this study, we examined miR-145 expression by quantitative real­time PCR (qRT­PCR) in CRC cell lines as well as tumors and corresponding normal mucosa, and the results were correlated to the clinicopathological parameters. In addition, using computational algorithms we investigated putative miR­145 targets. The role of miR­145 was further examined in studies in vitro. In our study miR­145 was significantly decreased in CRC tissues and cell lines compared with non­cancerous colorectal mucosa, especially lymph node or distance metastasis cases. Based on computational algorithms, we assumed that ERG was directly modulated by miR­145 in colorectal cancer cells. For the first time, we demonstrated that ERG was highly expressed in CRC tissues compared with normal ones by qRT­PCR. The inverse correlation between the expression of miR­145 and ERG was observed in CRC tissues. Dual­Luciferase assays demonstrated the direct interaction between miR­145 and 3'­UTR of ERG mRNA. Ectopic expression of miR­145 suppressed the proliferation and invasion ability of colorectal cancer cells, while ERG knockdown partially restored the tumor suppressive effect of miR­145. These results suggested that miR­145 might act as a tumor suppressor during the process of CRC malignant transformation by interacting with ERG.

Prognostic value of decreased expression of RBM4 in human gastric cancer.

RNA-binding motif 4 (RBM4) is a multifunctional protein that participates in regulating alternative splicing and mRNA translation. Its reduced expression has been associated with poor overall survival in lung cancer, breast cancer and ovarian cancer. We assessed RBM4 protein expression levels with immunohistochemistry in tissue microarrays containing malignant gastric cancer tissues and benign tissues from 813 patients. We also examined the expression levels of RBM4 mRNA in twenty-five paired gastric cancer samples and adjacent noncancerous tissues. Both RBM4 protein and mRNA expression levels were significantly lower in gastric cancer tissues compared with the adjacent noncancerous tissues. There was a significant association between reduced RBM4 protein expression and differentiation (P < 0.001), lymph node metastasis (P = 0.026), TNM state (P = 0.014) and distant metastasis (P = 0.036). Patients with reduced RBM4 expression (P < 0.001, CI = 0.315-0.710) and TNM stage III and IV (P < 0.001, CI = 4.757-11.166) had a poor overall survival. These findings suggest that RBM4 is a new biomarker in gastric cancer, as the reduced expression of this protein is correlated with poor differentiation, lymph node status and distant metastasis. Further, lower RBM4 expression is an independent prognostic marker for gastric cancer.

rhPDGF-BB via ERK pathway osteogenesis and adipogenesis balancing in ADSCs for critical-sized calvarial defect repair.

Adipose-derived stem cells (ADSCs) with the capacity of differentiating into osteo-like cells have been widely investigated for bone tissue engineering as a novel seed cell source. Recombinant human platelet-derived growth factor (rhPDGF-BB) is a clinically proven growth factor with the potential of promoting cell proliferation and osteogenic differentiation during the bone regeneration process. In this study, we investigated the effects of rhPDGF-BB on the proliferation and osteogenic and adipogenic differentiation of rat ADSCs and explored whether the extracellular signal-related kinase (ERK) signaling pathway might be involved. We found that rhPDGF-BB significantly enhanced ADSCs proliferation and osteogenic differentiation, as detected by MTT, real-time polymerase chain reaction (PCR), ALP activity assays, and calcium deposition in vitro, in concert with ERK pathway activation. In contrast, the adipogenesis of ADSCs, as detected by real-time PCR and Oil Red O staining, was suppressed in the presence of rhPDGF-BB. Furthermore, with the supplement of the ERK inhibitor PD98059, cell proliferation and osteogenesis were reduced; as expected, adipogenesis was enhanced. Subsequently, for the first time, we evaluated the effect of ADSCs associated with rhPDGF-BB on bone regeneration in a critical-sized rat calvarial defect model with silk scaffold as a carrier. Micro-computed tomography and histological analyses exhibited dramatically more new bone formation and trabecular number in the Silk/PDGF/ADSC group. These data indicated that rhPDGF-BB promoted cell proliferation and osteogenic differentiation and suppressed adipogenic differentiation in vitro via ERK pathway and that ADSCs associated with rhPDGF-BB could be a promising tissue-engineered construct for craniofacial bone regeneration in vivo.

Curculigoside promotes osteogenic differentiation of bone marrow stromal cells from ovariectomized rats.

OBJECTIVES: Curculigoside, a natural compound isolated from the medicinal plant Curculigo orchioides has been reported to prevent bone loss in ovariectomized rats. However, the underlying molecular mechanisms are largely unknown. This study investigated the effects of curculigoside on proliferation and osteogenic differentiation of bone marrow stromal cells (BMSCs). METHODS: The toxicity, proliferation and osteogenic differentiation of BMSCs cultured with various concentrations (0 as control, 10, 100 and 500 µm) of curculigoside were measured by viability assay, MTT analysis, alkaline phosphatase (ALP) activity assay, alizarin red staining and mineralization assay, real-time PCR analysis on osteogenic genes including ALP, type I collagen (Col I), osteocalcin (OCN) and osteoprotegerin (OPG), runt-related transcription factor 2 (Runx2), as well as OPG enzyme-linked immunosorbent assay. KEY FINDINGS: No significant cytotoxicity was observed for BMSCs after supplementation with curculigoside. The proliferation of BMSCs was enhanced after administration of curculigoside, especially 100 µm curculigoside. Moreover, the osteogenic gene expression was significantly enhanced with 100 µm curculigoside treatment. Importantly, curculigoside significantly increased OPG secretion. CONCLUSIONS: The data indicate that curculigoside could promote BMSC proliferation and induce osteogenic differentiation of BMSCs. The most profound response was observed with 100 µm curculigoside. These findings may be valuable for understanding the mechanism of the effect of curculigoside on bone, especially in relation to osteoporosis.