Ameloblastoma is a benign tumor characterized by locally invasive phenotypes,leading to facial bone destruction and a high recurrence rate.However,the mechanisms governing tumor initiation and recurrence are poorly understood.Here,we uncovered cellular landscapes and mechanisms that underlie tumor recurrence in ameloblastoma at single-cell resolution.Our results revealed that ameloblastoma exhibits five tumor subpopulations varying with respect to immune response(IR),bone remodeling(BR),tooth development(TD),epithelial development(ED),and cell cycle(CC)signatures.Of note,we found that CC ameloblastoma cells were endowed with stemness and contributed to tumor recurrence,which was dominated by the EZH2-mediated program.Targeting EZH2 effectively eliminated CC ameloblastoma cells and inhibited tumor growth in ameloblastoma patient-derived organoids.These data described the tumor subpopulation and clarified the identity,function,and regulatory mechanism of CC ameloblastoma cells,providing a potential therapeutic target for ameloblastoma.

Pleomorphic adenoma (PA) is the most common benign tumour in the salivary gland and has high morphological complexity. However, the origin and intratumoral heterogeneity of PA are largely unknown. Here, we constructed a comprehensive atlas of PA at single-cell resolution and showed that PA exhibited five tumour subpopulations, three recapitulating the epithelial states of the normal parotid gland, and two PA-specific epithelial cell (PASE) populations unique to tumours. Then, six subgroups of PASE cells were identified, which varied in epithelium, bone, immune, metabolism, stemness and cell cycle signatures. Moreover, we revealed that CD36⁺ myoepithelial cells were the tumour-initiating cells (TICs) in PA, and were dominated by the PI3K-AKT pathway. Targeting the PI3K-AKT pathway significantly inhibited CD36⁺ myoepithelial cell-derived tumour spheres and the growth of PA organoids. Our results provide new insights into the diversity and origin of PA, offering an important clinical implication for targeting the PI3K-AKT signalling pathway in PA treatment.
Humans ; Adenoma, Pleomorphic/genetics* ; Phosphatidylinositol 3-Kinases ; Proto-Oncogene Proteins c-akt ; Transcriptome ; Myoepithelioma

【Objective】 To explore the effect of B lymphocytes on cardiac structure and function and myocardial immune cells during heart development. 【Methods】 Echocardiography, immunofluorescence staining and flow cytometry were used to evaluate the composition of immune cells of the heart and the cardiac structure and function in wild-type (WT) mice and B-lymphocyte-deficient (μMT) mice, respectively. 【Results】 Compared with those of μMT mice, the ratio of heart weight to mouse weight (P<0.05), left ventricular mass (P<0.05) and the cross-sectional area of myocardial cells WT mice were significantly increased, while the ventricular ejection fraction was significantly decreased (P<0.05). The results of mRNA sequencing showed that WT mice and μMT mice differentially expressed genes were mainly enriched in the signal pathway of heart development and hypertrophic cardiomyopathy. The results of flow cytometry showed that WT mice had more Ly6g⁺ neutrophils, CD4⁺ positive T cells (P<0.001) and CD8⁺T cells (P<0.05) compared with μMT mice. 【Conclusion】 B-lymphocyte depletion alters the composition of cardiomyocyte immune cells, reduces left ventricular mass, and increases myocardial contractility.

【Objective】 To explore B lymphocytes’ role and mechanisms in angiotensinⅡ/phenylephrine (AngⅡ/PE) induced cardiomyopathy so as to understand the role of inflammatory cells in myocardial injury. 【Methods】 AngⅡ/PE was administered to wild-type (WT) and B cells deficiency (μMT) mice for 14 days or 28 days. The mice were analyzed by blood pressure measurement, echocardiography imaging, flow cytometry, and histology. Cardiac fibrosis was evaluated by Masson staining. 【Results】 Compared with the control group, the left ventricular mass (P<0.01), heart mass/tibia length ratio (P<0.01) and cross-sectional area of cardiomyocytes in AngⅡ/PE group were significantly increased (P<0.01). After 2 weeks of AngⅡ/PE treatment, B lymphocytes (P<0.05), CD45⁺ leukocytes (P<0.05), CD64^-Ly6C⁺ monocytes (P<0.05), CD64⁺Ly6C^-macrophages (P<0.05) and Ly6g⁺ neutrophils (P<0.05) were recruited in myocardial tissue. Compared with WT_AngⅡ/PE group, the heart weight/tibia length ratio (P<0.05), left ventricular weight (P<0.05) and myocardial cell cross-sectional area (P<0.05) of μMT_AngⅡ/PE mice were significantly improved. CD45⁺Ly6C⁺CD64^- monocytes (P<0.05) and CD45⁺Ly6C^-CD64⁺ macrophages (P<0.05) were significantly decreased. 【Conclusion】 B lymphocytes deficiency improves AngⅡ/PE induced cardiac hypertrophy by reducing the infiltration of CD45⁺Ly6C⁺CD64^- monocytes and CD45⁺Ly6C^- CD64⁺ macrophages.

Objective:To understand the overall satisfaction rate with vaccination services in parents of children, and the impact of additional time consumed for vaccination service on overall satisfaction rate.Methods:From December 2019 to January 2020, a total of 3 178 parents of 0-3 years old children were investigated to collect the information about their basic characteristics, additional time spent for vaccination service and overall satisfaction through questionnaires. Binary logistic regression model and restricted cubic spline model were used to evaluate the impact of additional time spend on the overall satisfaction rate.Results:The overall satisfaction rate of parents with vaccination services was 92.32%. The median time for parents to move from home to vaccination clinic was 10.00 (10.00, 20.00) minutes, the median waiting time to make an appointment was 10.00 (5.00, 15.00) minutes, the median waiting time for vaccination was 5.00 (3.00, 10.00) minutes, and the median total additional time spent was 30.00 (20.00, 45.00) minutes. The binary logistic regression analysis showed that after adjusting the relevant factors, the main factors affecting the overall satisfaction rate were the waiting time for making an appointment (the 4- minutes group vs. 8- minutes group: OR=1.863, 95% CI: 1.307-2.657), waiting time for vaccination (the <4 minutes group vs. 8- minutes group: OR=1.529, 95% CI: 1.102-2.120; the 4- minutes group vs. 8- minutes group: OR=1.534, 95% CI: 1.104-2.130), total additional time spent (the 15- minutes group vs. 30- minutes group: OR=1.470, 95% CI: 1.094-1.976). Restricted cubic spline analysis showed that the waiting time for making an appointment (non-linear: χ2=13.18, P=0.001), the waiting time for vaccination (non-linear: χ2=13.50, P=0.001), and the total additional time consumed (non-linear: χ2=9.38, P=0.009) showed a non-linear inverted "V" dose response relationship to the overall satisfaction of vaccination services. Conclusions:The waiting time for parents to make an appointment, the waiting time for vaccination and the total additional time spent for receiving vaccination services affected the overall satisfaction rate of the vaccination services. And the waiting time for making an appointment was the most important factor, and it is necessary to shorten the waiting time for appointment. It is suggested that the vaccination clinic should make use of information technology (such as WeChat public account, APP) to make accurate appointments, make appointments to the time period to control the number of people within time period.

Objective:To understand the time for observation and related factors in the clinics after vaccination among children's parents.Methods:From December 2019 to January 2020, parents of children aged 0-3 years were recruited by multiple-stage sampling from 34 vaccination clinics in 12 districts and counties in 6 provinces (Shandong, Guangdong, Henan, Sichuan, Inner Mongolia, and Liaoning). A questionnaire survey on the time of observation after vaccination was conducted. A multivariate logistic regression model was used to analyze the related factors of parental observation time after vaccination.Results:A total of 3 292 parents of 0-3 year's old children were selected, and 3 178 parents were finally included in the analysis. 87.85%(2 792/3 178) of the parents reported that the observation time after vaccination at clinics was ≥30 minutes. Multivariate logistic regression analysis showed that, after adjusting for the regions, the main factors affecting the observation time at clinics after vaccination among parents appeared as observation time informed by physicians at the clinic appeared ≥30 minutes ( OR=31.622, 95% CI: 19.847-50.384), parents were medical personnel ( OR=2.779, 95% CI: 1.505-5.133), parents being volunteers working on vaccination-related publicity and education activities ( OR=1.986, 95% CI: 1.438-2.743), parents aged 35 years old or above ( OR=1.900, 95% CI: 1.215-2.971), being parents of the first child ( OR=1.663, 95% CI: 1.282-2.156), per capita annual income of the family as 8 000- Yuan ( OR=1.646, 95% CI: 1.168-2.319), children aged 0-12 months old ( OR=1.646, 95% CI: 1.203-2.252) or 13-24 months old ( OR=1.506, 95% CI: 1.064-2.133), obedient to physicians' advice at the clinic ( OR=1.481, 95% CI: 1.067-2.055). Conclusions:The proportions of parents observed for ≥30 minutes at the clinics of vaccination were high. When the information was from the physicians at the vaccination clinic, the observation time was the most critical factor for parents to observe at clinics as required.

To observe the immunogenicity of hPDGF-B immunogens that were synthesized with the fusional expression vector pET28-Trx and to test the suppressive effect of these specific antibodies induced by both of immunogens on proliferation of human HepG2 hepatoma cells. First, we chose 2 antigenic epitopes hPDGF-BΔ103-118aa and hPDGF-BΔ152-167aa from human PDGF-B and inserted these 2 coding regions into the empty vector plasmid pET28-Trx, separately. Second, mice were immunized with purified recombinant proteins to generate polyclonal antibody. Then we intraperitoneally injected mice bearing hepatoma 22 (H22) tumor cells to prepare antibody ascites. ELISA and Western blot were used to detect the titer and the utility of the antibody, respectively. Finally, HepG2 cells were exposed to PDGF-BB protein or anti-PDGF-B ascite antibody in different dilution concentrations groups and the proliferation of HepG2 cells was quantified by CCK8 assay. As the results, we identified mice that could produce high drop of neutralizing antibodies against hPDGF-B induced by both two recombinant proteins. Two anti-PDGF-B ascite antibodies could markedly inhibit the proliferation of HepG2 cells by blocking the stimulating effect of PDGF-BB protein. Our results suggest that Trx-PDGF-B recombinant protein as immunogen provides a new method for the preparation of PDGF-B vaccine, and also a new idea for the treatment of hepatocellular carcinoma in clinical practice.

Objective:To investigate the induction and differentiation potential of ADSCs by tissue culture method,and to preliminary study on the origin of ADSCs.Methods:Using adipose tissue culture method to culture human ADSCs.The third generation of ADSCs for the adipogenic and osteogenesis differentiation,and staining by oil red O and alizarin red S.HE staining was performed after the seventh day culture of adipose tissue.Results:The primary human ADSCs were successfully cultured with adipose tissue culture method.ADSCs cultured to the eighth generation,still maintained a good proliferation ability and cell morphology.ADSCs can be successfully induced into adipose cells and bone cells.ADSCs were mainly distributed around the mesenchymal vascular and connective tissue,by HE staining of adipose tissue after seven days of culture.Conclusion:The cells that were cultured with adipose tissue have the potential to adipogenic and osteogenesis differentiation.The ADSCs were mainly distributed around the mesenchymal vascular and connective tissue.

Aim To investigate the modeling of breast cancer in zebrafish embryos and its related protein expression. Methods 48hpf wild type AB/ TG(Transgenic) zebrafishs were micro-in-jected with breast cancer cell line: MCF-7,T-47D, MDA-MB-231 respectively, the relationship between the number of tumor and model application was investigated, and the number of sub-intestinal veins(SIVs) was detected under confocal microscope, as well as the metastasis of tumor cells in embryos; then the ze-brafish xenografts of MB-231 were co-cultured with tofacitinib/ptk787 for 48 h, optical density(OD) of the cell survival and subintestinal veins(SIVs) were evaluated under confocal micro-scope, and Western blot(WB) analysis was used to test micro-circumstances related protein. Results When the number of in-oculated cells was more than 200 per embryo, xenograft model rate woule be more than 0. 90;MB-231 xenografts showed metas-tasis feature in zebrafish, which could be inhibited by tofacitinib (P < 0. 01), while the number of xenograft MB-231 cells was reduced significantly(P < 0. 01); in another zebrafish xenografts SIVs assay, the tumor could promote the proliferation of SIVs, and 4 mg·L - 1 PTK787 showed inhibiton effect( P < 0. 01). Western blot showed 4d T-47D xenograft zebrafish got more HER2 expression than AB embryos; VEGFa expression in ze-brafish MB-231 model group was higher, and model zebrafish P53 expressi was higher after treated by tofacitinib. Conclusion A zebrafish xenograft model of human brest cancer can be es-tablished, which demonstrates applicability for screening com-pounds in drug discovery studies.

BACKGROUND:Salvia miltiorrhiza bone-setting capsule is a traditional Chinese medicine for the treatment of fractures due to activating blood circulation to dissipate blood stasis, reducing sweling and pain. OBJECTIVE: To observe the effects of Salvia miltiorrhiza bone-setting capsule on the fracture healing in a rat model of closed femoral fractures. METHODS: Rats were randomly divided into salvia miltiorrhiza bone-setting capsule group, physiological saline group and normal group. In the salvia miltiorrhiza bone-setting capsule group and physiological saline group, rat models of closed femoral fractures were prepared, and then given physiological saline and salvia miltiorrhiza bone-setting capsule 2 pils by intragastric administration. In the normal group, rats were housed normaly. At 7, 14 and 28 days after fractures, hematoxylin-eosin staining conditions, serum osteocalcin, the expression of colagen type I, and the expression of protein and mRNA calus transforming growth factor-beta 1 were observed in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining demonstrated that at 7 days after fractures, no significant difference was found in pathological changes of femoral fracture in salvia miltiorrhiza bone-setting capsule group and physiological saline group. At 14 and 28 days after fractures, pathological repair was more obvious in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group. (2) At 3 and 7 days after fractures, serum osteocalcin and the expression of type I colagen were significantly increased in the salvia miltiorrhiza bone-setting capsule group and physiological saline group (P < 0.05), and the expression trend was consistent in both groups. The expression was always higher in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group, and significant differences were found at 14 and 28 days after fractures (P < 0.01). (3) Transforming growth factor beta 1 expression reached a peak at 3 days after fractures, gradualy reduced, increased at 14 days (the second peak), and diminished at 28 days in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. The expression trend of transforming growth factor beta 1 was consistent in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. At 7, 14 and 28 days, the transforming growth factor beta 1 expression was higher in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group. (4) Results showed that salvia miltiorrhiza bone-setting capsule could promote fracture healing, and its mechanism was probably associated with serum osteocalcin, the expression of colagen type I and transforming growth factor-β1.