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.

Protein aggregation is a critical issue in the production of biopharmaceuticals. During protein production, transport and storage, various factors can lead to protein aggregation. With the in-depth study, different ways of protein aggregation and various influencing factors were identified. This includes physical and chemical factors, translation modifications and protein structure. Since protein aggregation exerts major impact on the activity and homogeneity of proteins, it is of great importance to study the ways of protein aggregation and how to control it to obtain high-quality proteins. The review focuses on three ways of protein aggregation, namely 3D domain swapping, salt bridge formation, and oxidative stress, as well as methods to control protein aggregation during protein production, transport and storage. This may facilitate reducing the loss caused by the formation of protein aggregation and improving the purity and homogeneity of protein in research and commercial production.
Protein Aggregates ; Proteins/chemistry* ; Oxidative Stress

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 investigate the immunomodulatory activity of polysaccharides from the roots of Brassica rapa. Methods: The crude polysaccharide from roots of B. rapa (BRP) was extracted and purified to further investigate the active fraction of BRT for inducing macrophage phagocytosis. Results: Effects on RAW264.7 cells demonstrated that BRP behaved better phagocytic capacity and had potent immunomodulatory activity, including increasing production of nitric oxide (NO), tumor necrosis factor α (TNFα) and upregulating mRNA levels of inducible NO synthase (iNOS) and TNFα. Furthermore, modulation of macrophage by BRP was indicated to be mediated via the activation of Akt and nuclear factor-kappa B (NF-κB). Conclusion: The beneficial effects of BRP could be used as an immunotherapeutic adjuvant in treatment of inflammatory diseases.

Objective:To systematically compare the prognosis in non-compressible torso hemorrhage（NCTH）treated by resuscitative endovascular balloon occlusion of the aorta（REBOA）and resuscitation thoracotomy（RT）.Methods:Data were searched form MEDLINE, EMBASE, PubMed, WanFang, CNKI and VIP databases to collect studies on the prognosis of patients with NCTH undergone REBOA and RT from inception to December 2020. Two reviewers independently screened studies according to the inclusion and exclusion criteria, extracted data and evaluated the quality of the included studies. The Meta-analysis was performed using Revman 5.3. The patients were divided into REBOA group and RT group according to the different surgical treatment methods on admission, and the prognosis of each group was evaluated. The difference of mortality rate, reoperation rate of laparotomy after operation, reoperation rate of embolization after operation and mortality rate in different operating room area were compared between the two groups. Publication bias was assessed using the Egger test.Results:A total of 2 prospective studies and 4 retrospective studies involving 2, 588 subjects were included. There were 1, 591 patients in REBOA group and 997 patients in RT group. Significant differences were observed in the mortality rate（ I2=68%, OR=0.33, 95% CI 0.26-0.42, P<0.01）, reoperation rate of laparotomy after operation（ I2=76%, OR=1.41, 95% CI 1.11-1.77, P<0.01）and reoperation rate of embolization after operation（ I2=84%, OR=0.76, 95% CI 0.59-0.99, P<0.05）between REBOA group and RT group. Subgroup analysis showed that the mortality rate in the ICU were not statistically different between the two groups（ I2=83%, OR=0.69, 95% CI 0.45-1.05, P>0.05）, but the mortality rate in the emergency room was lower in REBOA group than that in RT group（ I2=94%, OR=0.52, 95% CI 0.38-0.70, P<0.01）. Egger test showed that publication bias had little effect on the results. Conclusions:For patients with NCTH, REBOA can reduce the mortality rate and reoperation rate of embolization after operation, but increase the reoperation rate of laparotomy after operation when compared with RT. In addition, the emergency room may be a more suitable operationg room area for REBOA.

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.

Objective To construct a prokaryotic expression vector in BL21 to secretorily expressα-Cyclodextrin Glycosyltransferase(α-CGTase). Methods α-CGT gene was amplified from Bacillus macerens genome by PCR.pET26b and α-CGT gene were connected after digested with Nco I, Xho I respectivly, and then transformed into Escherichia coli BL21 strain.α-CGTase was expressed in fermentation culture medium and AA-2G was prepared by using α-CGTase, VC and starch.Results α-CGTase was expressed secretorily and the enzyme activity was up to 120 U/mL.AA-2G was prepared by the biotransformation of VC and starch using α-CGTase which proved to be correct by HPLC.Conclusion AA-2G was prepared by using self-madeα-CGTase, after optimized the preparation conditions the yield of AA-2G was 17.46 g/L, and the conversion rate reached 58.2%(mg/mg).

Objective To construct a eukaryotic expression vector in Pichia pastoris containing human fibrinogen gene, in order to achieve high level secretory expression in extracellular.Methods Expression plasmid,pGAPZαA-FGB-FGG-FGA-AOX1,was constructed by inserting the synthesized sequence encoding human fibrinogen(FGA, FGB,FGG) and then introduced into Pichia pastoris SMD1168H by electroporation.Transformants were availably screened by Zeocin resistance,the expression of recombinant protein was identified by SDS-PAGE and Western blot analysis, the protein yield was tested by ELISA assay.After ultrafiltration and purification, the biological activity of protein was detected.Results The crude yield of human fibrinogen in Pichia pastoris supernatant reached 15 mg/L in flask and the biological aggregation activity was determined.Conclusion The human fibrinogen gene was obtained and successfully expressed in Pichia pastoris and the active products were secreted into the medium.

Aim To study the effect of rhein on renal damage induced by aristolochic acid. Methods Ze-brafish model of aristolochic acid nephropathy, genera-ted by treating zebrafish larvae with aristolochic acid for 24 h, was treated with rhein simultaneously . Mor-pholigical changes were observed and the creatinine level in larvae tissue was measured. And mRNA ex-pression levels of inflammatory factor cox2 a and fibrosis factor TGF-β1 in larvae tissue were detected using qPCR. Results Some larvae show periocular edema and circulation system defection e. g. weak heart beat, narrow cardiac vesicle, decreased blood flow and even blockage , with a dose-response relationship after expo-sure to aristolochic acid for 24 h. The creatinine level in larvae tissue of the treated group was significantly higher than that of the control larvae. And the expres-sion levels of cox2 a and TGF-β1 in larvae tissue of the treated group were also significantly increased. Per-centage of abnormal larvae and creatinine level in lar-vae tissue were decreased when treated with rhein sim-ultaneously. And the expression levels of cox2a was down-regulated by rhein compared with the aristolochic acid treated group. But rhein had no effect on TGF-β1 expression. Conclusion To some extent rhein can protect renal from damage induced by aristolochic acid.