This article reported a prenatally diagnosed case of complete ring chromosome 15. A 38-year-old woman who conceived by in vitro fertilization and frozen embryo transfer underwent amniocentesis for prenatal diagnosis at 18 +5 weeks of gestation due to advanced maternal age. The result of G-banding karyotyping was mos 46,XX,r(15)[88]/45,X,-15[11]/46,XX,r(15;15)[1]. No numerical abnormalities of chromosomes or definite pathogenic copy number variations (CNVs) were detected by chromosomal microarray analysis. Amniocentesis was performed again at 31 +6 weeks of gestation. The result of genome copy number variation sequencing indicated no pathogenic CNV and fluorescence in situ hybridization on cultured amniocytes revealed nuc ish(15q)×1[15]/(15q)×3[5]/(15q)×2[80]. Based on all the prenatal diagnosis results, it was suggested that the fetus carried a complete ring chromosome 15. As the peripheral blood chromosomes of the couple were normal and no obvious abnormalities were detected by the prenatal ultrasound either in our hospital or another hospital, the pregnant woman decided to continue the pregnancy after genetic counseling and delivered a baby girl at 41 weeks of gestation. The girl showed no physical abnormalities during a seven-month follow-up.

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.

With the continuous development of maxillary sinus floor elevation technology, the osteogenesis mechanism of maxillary sinus floor elevation has always been a concern of scholars. The membrane of the maxillary sinus is an indispensable physiological structure in the process of space osteogenesis under the sinus floor after elevation of the sinus floor. In recent years, the role of the maxillary sinus floor mucosa in sinus floor space osteogenesis has been a research hotspot. Recent studies have found that the maxillary sinus floor membrane plays a role as a natural biological barrier membrane in the process of sinus floor space osteogenesis after maxillary sinus floor elevation; in addition, it has the ability to undergo osteogenesis. It has also been found that maxillary sinus membrane stem cells (MSMSCs) derived from the maxillary sinus floor membrane have characteristics of mesenchymal stem cells, which can differentiate into osteoblasts and participate in sinus floor space osteogenesis after maxillary sinus floor elevation. New studies have also found that small RNAs such as microRNAs, long noncoding RNAs and circular RNAs can regulate the osteogenic differentiation of MSMSCs, which may be important biological targets for promoting osteogenesis in the sinus floor space. In this paper, the relationship between the maxillary sinus floor mucosa and bone formation after maxillary sinus floor elevation, the barrier and osteogenic function of the maxillary sinus floor mucosa, the sources of osteoblasts involved in osteogenesis of the sinus floor space, and the molecular regulatory mechanisms of stem cells derived from maxillary sinus mucosa will be elucidated step by step.

Objective: To investigate the role of the bone morphogenetic protein 2 (BMP2)⁃Smad1/5 and p38MAPK signaling pathways in the osteogenic differentiation of MSMSCs by insulin⁃like growth factor 1 (IGF1). Methods : A re⁃ combinant adenovirus (RAD) and IGF1 expressing IGF1 gene were constructed. After osteogenic induction, qRT⁃PCR and Western blot were used to detect the phosphorylation level of Smad1/5 and the expression of the BMP⁃2 protein in the BMP⁃Smad signaling pathway; immunohistochemistry was used to observe the nuclear translocation of Smad1/5; qRT⁃PCR and Western blot were used to detect IGF with Noggin and SB203580, inhibitors of the p38MAPK signaling path⁃ way 1⁃mediated osteogenic differentiation of MSMSCs Results: The recombinant IGF1 adenovirus was constructed suc⁃ cessfully. MSMSCs were cultured in inductive medium after infection with different concentrations of Ad⁃IGF1, and then, the protein levels of BMP2 and p⁃Smad1/5 increased. IGF1 can also induce nuclear translocation of Smad1/5. In addition, Noggin significantly reduced the phosphorylation level of Smad1/5 and the formation of mineralized nodules in the MSMSCs. The mRNA levels of Runx2, OPN and ALP also decreased. In contrast, SB203580 decreased neither the phosphorylation level of p38 nor the mRNA expression of Runx2, OPN and ALP in the Ad⁃IGF1 MSMSCs Conclu⁃sion IGF1 can promote the osteogenic differentiation of MSMSCs via the BMP2⁃Smad1/5 signaling pathway. In con⁃ trast, IGF1 may not promote the osteogenic differentiation of MSMSCs via the p38MAPK signaling pathway.

Objective: To investigate the effect of fractionated radiotherapy on the immune system of mice with subcutaneously transplanted hepatocellular carcinoma. Methods: Logarithmic growth of mouse hepatocellular carcinoma cells Hepa 1-6 were inoculated subcutaneously on the right side of C57BL/6 J mice (1×10⁷ cells /mice). The tumor-bearing mice were randomly divided into control group (Ctrl) and irradiation group (IR), 20 mice in each group. Additionally, 10 healthy mice were set as normal control group. Local fractionated X-ray irradiation of 8 Gy×3 fraction was given to the subcutaneous tumors, and the dose rate was 0.883 Gy/min. At 7 and 14 d after irradiation, the tumor organ index, spleen organ index, spleen pathological changes, and splenic T lymphocyte subsets, B lymphocyte subsets, and NK cells were detected. Results: Compared with Ctrl, at 7 and 14 d after irradiation, the tumor organ index decreased (t=4.649, 26.34, P<0.05), and the percentage of NK cells increased significantly (t=3.952, 3.633, P<0.05). The percentages of CD3+ , CD4+ , CD3+ CD4+ lymphocytes and the ratio of CD4+ /CD8+ lymphocyte decreased at 7 d after irradiation (t=3.193, 3.656, 3.219, 2.641, P<0.05), and the percentage of CD3+ lymphocyte decreased at 14 d after irradiation (t=3.031, P<0.05). But after irradiation, there were no significant changes in spleen organ index, B lymphocyte, CD3+ CD4+ lymphocyte and CD8+ lymphocyte. Conclusions Local hepatoma radiotherapy causes imbalance of lymphocytes in distal spleen of mice and hence reduces immunity, which provides a novel mechanism of radiological immunity damage.

Objective: To detect the expression level of miR-27a during the osteogenic differentiation of beagle maxillary sinus membrane stem cells (MSMSCs) and explore the role of miR-27a in the osteogenic differentiation of MSMSCs. Methods: Beagle MSMSCs were cultured in vitro. The expression level of miR-27a was detected via RT-PCR after an osteogenic inductive culture was prepared. The mRNA expression levels of Runx2 and OPN were examined via RT-PCR, and the protein expression levels of Runx2 and OPN were examined via Western blot after the cells were transfected with pre-miR-27a or anti-miR-27a. Finally, osteoprogenitor cells transfected with pre-miR-27a were composited with Bio-Oss particles and subcutaneously implanted into nude mice to form ectopic bone formation models, and then the inhibition of bone formation from miR-27a was observed in vivo. Results: The expression level of miR-27a in the beagle MSMSCs decreased after osteogenic inductive culturing. The relative miR-27a levels were significantly decreased at day 1 (t=3.795, P=0.023), day 3 (t=4.493, P=0.011), day 7 (t=11.591, P < 0.001), day 14 (t=12.542, P < 0.001), and day 21 (t=5.621, P=0.008) compared with day 0. In addition, the expression levels of Runx2 mRNA (t=4.923, P=0.007) and protein (t=4.425, P=0.008) were reduced after the cells were transfected with pre-miR-27a. The expression levels OPN mRNA (t=5.253, P=0.006) and protein (t=5.132, P=0.006) were also reduced. In contrast, the mRNA expression levels of Runx2 (t=3.925, P=0.013) and OPN (t=3.712, P=0.019) were increased after the cells were transfected with anti-miR-27a, and bone formation was observed after the subcutaneous implantation of beagle MSMSCs composited with Bio-Oss in nude mice. Nevertheless, ectopic bone formation was inhibited by pre-miR-27a-transfected beagle MSMSCs composited with Bio-Oss (t=7.219, P=0.0020). Conclusion MiR-27a negatively regulates the osteogenic differentiation of MSMSCs.

Objective: To investigate the osteogenic properties of maxillary sinus membrane stem cells (MSMSCs). Methods : Beagle maxillary sinus mucosa was collected, immunomagnetic bead method was applied for isolation of CD146+ cells, and MSMSCs were harvested and cultured from the canine maxillary sinus floor mucosa. The levels of the cell surface antigens CD44, CD146, and CD34 were determined at passage one by flow cytometry. Cells at passage one were cultured in basal medium and osteogenic inductive medium. Real-time PCR, immunohistochemical staining, alkaline phosphatase activity, alizarin red staining and Von Kossa staining were used to investigate the osteogenic properties in vitro. Results: The canine MSMSCs were cultured successfully. The results of flow cytometry were positive for CD146 and CD44 expression but negative for CD34 expression. The relative mRNA expression of runt-related transcription factor 2 (RUNX2) (t = 14.44,P < 0.001), osteopontin (OPN) (t = 7.85,P = 0.001) and alkaline phosphatase alkaline phosphatase (t = 14.27,P < 0.001) was apparently higher in the osteoinductive medium group than in the basal medium group, the differences in relative mRNA expression between the groups were significant. The protein levels of RUNX2 and OPN increased in the osteoinductive medium group. The alkaline phosphatase activity of the MSMSCs increased when the cells were cultured in osteoinductive medium; the activity increased to a level that was significantly higher than that in basal medium, particularly at days 3 (t = 8.79, P < 0.001), 7 (t = 9.75,P < 0.001), 14 (t = 12.14,P < 0.001), 21 (t = 19.62,P < 0.001) and 28 (t = 17.53,P < 0.001). Obvious mineralized nodules were observed by alizarin red staining or Von Kossa staining. Conclusion Maxillary sinus membrane stem cells exhibit osteogenic ability.

To evaluate the clinical results of reconstruction orbital defect with craniomaxillofacial implant. Three patients with orbital defect were treated with ten implants. The magnetic abutments were attached six months after one stage operation and the prostheses were fabricated. Within 11 to 47 months of follow up, all implants were stable with successful osseointegration. The prosthesis fit the orbital defects well. Reconstruction of orbital defect with craniomaxillofacial implant can be considered as a viable alternative treatment.

BACKGROUND:Bone marrow mesenchymal stem cel s have been shown to be differentiated into periodontal ligament fibroblasts when co-cultured with periodontal ligament cel s. Existing studies have shown that estrogen has the ability to influence bone marrow regeneration. OBJECTIVE:To investigate the effects of estrogen on osteogenesis and fibroblast-related factors alkaline phosphatase, type I and III col agen in bone marrow mesenchymal stem cel s. METHODS:Bone marrow mesenchymal stem cel s isolated from Beagle dogs were treated with estrogen. Osteogenesis and fibroblast-related mRNA and protein expression of bone marrow mesenchymal stem cel s was determined by RT-PCR and western blot assay, respectively. RESULTS AND CONCLUSION:mRNA and protein expression of type I and III col agen in bone marrow mesenchymal stem cel s was upregulated fol owing estrogen treatment;especial y, in contrast with type III col agen, the changes of type I col agen were more obvious. Estrogen did not influence mRNA and protein expression of alkaline phosphatase. These findings suggest that estrogen promotes the differentiation of bone marrow mesenchymal stem cel s into fibroblasts, whereas does not impact the genes involved in parodontium mineralization.

BACKGROUND:Stromal cel derived factor-1 is a smal molecular protein with a wide range of biological activity that can cause immune cel chemotaxis, and it also has a chemotactic effect on bone marrow stem cels and periodontal ligament cels.
OBJECTIVE:To investigate the effect of stromal cel derived factor-1 with different concentrations on the proliferation of bone marrow stem cels and to probe the best concentration.
METHODS:Bone marrow stem cels from beagle dogs were culturedin vitro and stimulated by different concentrations of stromal cel derived factor-1 (100, 200, 300 μg/L). MTT was used to detect the influence of stromal cel derived factor-1 on the proliferation of bone marrow stem cels so as to screen the best concentration of stromal cel derived factor-1. Then, stromal cel derived factor-1 at the best concentrations was used to intervene the bone marrow stem cels, and MTT was used again to detect the proliferation of bone marrow stem cels.
RESULTS AND CONCLUSION:Stromal cel derived factor-1 at concentrations of 100, 200, 300 μg/L could promote the proliferation of bone marrow stem cels, and the effect was more notable at 200 and 300 μg/Lbut withno significant difference. Therefore, 200 μg/L was considered to be the best concentration of stromal cel derived factor-1 for intervention of bone marrow stem cels. Compared with the blank control group, 200 μg/L stromal cel derived factor-1 could significantly promote the proliferation of bone marrow stem cels. Taken together, stromal cel derived factor-1 can promote the proliferation of bone marrow stem cels, and its best concentration is 200 μg/L.