[Effect of dual fluorescence imaging in identifying central lymph nodes and parathyroid glands during thyroid cancer surgery].

Objective: To investigate the effect of dual fluorescence imaging in identifying central lymph nodes and parathyroid glands during thyroid cancer surgery. Methods: This study was a cross-sectional study. Patients who underwent surgery for papillary thyroid cancer (PTC) at the Department of Head and Neck Surgery, Beijing Tongren Hospital, Capital Medical University between January 2022 and September 2023 were included. All patients underwent thyroid lobectomy or total resection, and central lymph node dissection was performed at the same time. During the operation, tracing injection of mitoxantrone hydrochloride and 785 nm and 660 nm dual fluorescence imaging technique were used to measure the fluorescence intensity (FI) of parathyroid glands, central lymph nodes and background. After correcting to obtain the standardized FI, the paired t-test was used to compare the standardized FI of the parathyroid glands and central lymph nodes, and the Spearman's rank correlation analysis was used to analyze the relationship between the standardized FI and various clinical indicators. Results: The study included 30 patients (8 males and 22 females), with a mean age of (41.8±10.4) years. A total of 76 parathyroid glands and 234 central lymph nodes were identified under dual fluorescence imaging, and the standardized FI of parathyroid glands was less than that of central lymph nodes (44.7±16.8 vs 99.5±28.4, P<0.001). The visualization rate, false rate and miscut rate of parathyroid glands under 785 nm wavelength excitation light were 98.7% (76/77), 0 (0/77) and 1.3% (1/77), respectively (one case with no visualization and miscutting parathyroid gland was the encapsulated type). The visualization rate of central lymph nodes under 660 nm wavelength excitation light was 98.7% (234/237). There was no significant correlation between FI and clinical indicators such as gender, age, height, weight, body mass index, preoperative thyroid stimulating hormone, thyroglobulin antibody, thyroid microsomal antibody, serum calcium, parathyroid hormone level and surgical procedure (all P>0.05). Conclusion: Dual fluorescence imaging of central lymph nodes and parathyroid glands can improve the ability to identify parathyroid gland while assisting central lymph node dissection.

[Analysis of the change in human resources of centers for disease control and prevention in different regions of China from 2010 to 2020].

Objective: To analyze the change in human resources within China's Centers for Disease Control and Prevention (CDC) from 2010 to 2020. Methods: The self-reported information from provincial, prefectural (city), and county (district) levels of China's CDC, covering employee counts, staff composition, professional qualifications, educational backgrounds, technical titles, and tenure, were extracted from the China Disease Prevention and Control Information System. The demographic context was provided by the annual population figures from the China Statistical Yearbook (2010-2020). The profile of CDC personnel was described, and the average annual percentage rate change (AAPC), average annual percentage rate change (APC), human resource agglomeration degree (HRAD) and the difference between HRAD and population agglomeration degree (PAD) were calculated. The Joinpoint regression model was used to analyze the time trend. Results: The decade under review witnessed a net increase of 17 300 active and 18 300 enrolled personnel in the CDC, surpassing the national population growth rate with AAPCs of 0.93% and 1.03%, respectively. This upward trajectory was statistically significant (P<0.05). The ratio of disease control personnel per 10 000 population escalated from 1.14 to 1.21. An initial decline in active CDC workforce density (from 1.31 to 1.27 per 10 000 population between 2010 and 2017) was followed by an increase (from 1.28 to 1.37 between 2018 and 2020), with APCs of -0.40% and 3.73%, respectively. The proportion of professional and technical staff in 2019 was highest in the eastern region (86.01%), followed by the western (83.75%) and central regions (79.54%). The period also saw an enhancement in the average academic degree (from 1.91 to 2.43 points) and professional title scores (from 1.39 to 1.53 points) of CDC personnel. While the average tenure in the eastern and western regions showed a slight decline, the central region experienced an increase, with HRAD values indicating a higher concentration in the eastern and central regions compared to the western region. The HRAD-PAD discrepancy revealed a negative value in the eastern region, nearing zero in the central and western regions. Conclusion: Between 2010 and 2020, China's CDC experienced notable growth in human resources and underwent structural optimization, albeit with significant regional disparities in concentration.

[Analysis on the management of type 2 inflammatory asthma from the guideline of Global Strategy for Asthma Management and Prevention].

Bronchial asthma is a common chronic respiratory disease, which is involved in a variety of cells and cellular components. In 2019, the guidelines for the diagnosis and treatment of asthma issued by the Global Initiative for Asthma (GINA) Committee put forward the concept of type 2 inflammatory asthma for the first time. The updated evolution of GINA guidelines has promoted the development of biological agents and disease treatment, providing effective prevention and treatment for patients with severe asthma and improving disease outcome. This paper expounds the disease mechanism and management suggestions of type 2 inflammatory asthma in GINA guidelines, and analyzes the relevant clinical studies on targeted treatment of type 2 inflammatory asthma in recent years, in order to provide reference for in-depth understanding of level 3 prevention and management of patients with type 2 inflammatory asthma.

[Using clinical research methods to answer the clinical and scientific questions].

The advantages and disadvantages of the article "influenza/pneumonia vaccination status and influencing factors in patients with chronic obstructive pulmonary disease: a real-world cross-sectional study" were interpreted by methodological perspective. The importance of topic selection closely combined with clinical problems and needs was emphasized, and reasonable and feasible research methods to escort clinical research. This paper puts forward the necessity and practicality of improving the internal quality of research and paper writing under the framework of "one paper research and answering one question", analyzes the objectives, tasks and reasonable technical route of influencing factor research, and emphasizes that clinical research methods serve to answer clinical and scientific questions.

[How to write a clinical research paper that can be published].

A clinical research paper which could be published should be based on a qualified clinical research, which includes a reasonable topic from clinical settings, concentrating resources to answer a question, having new findings and drawing concrete conclusions. The author should prepare a scientific story, which originates from clinical scenarios, with working hypothesis and research design, standardized organization and implementation, reasonable analysis and evaluation. The conclusion should be based on the research results, and go back to the problems and needs to form a closed loop. The key point of writing a clinical research paper is to provide complete information and show a complete research process according to the writing format and specification of the paper. Finally, the key points of the current paper help avoid detours and improve the probability of paper publication.

[Investigation of noise annoyance among the exposed anesthesiologists in a hospital].

Objective: To investigation noise annoyance among the exposed anesthesiologists, and to analyze the influencing factors of noise annoyance. Methods: From January to May 2017, cluster sampling was used to select 62 anesthesiologists from Peking University Third Hospital as the survey subjects. The Likert Scale (five-level) and the Visual Analog Scale (0-10 points) were used to conduct noise annoyance and noise sensitivity survey. Linear regression was used for multi-factor analysis to explore the influencing factors of noise annoyance. Results: The age of the research subjects was (32.3±6.1) years old, the working experience was 4.9 (2.0, 8.9) years, and the daily working hours were (9.8±2.8) h. The noise level in the operating room on the measurement day was (64.4±1.7) dB (A) . In the Likert Scale and the Visual Analog Scale, 88.7% (55/62) and 95.2% (59/62) anesthesiologists believed that operating room noise had adverse reactions, 6.5% (4/62) and 14.5% (9/62) anesthesiologists measured a high degree of annoyance. The results of linear regression analysis showed that noise sensitivity was an independent factor influencing the anesthesiologists' noise annoyance in the Likert Scale and the Visual Analog Scale (r=0.524, 0.700, P<0.05) . Conclusion: Noise in the operating room can cause anesthesiologists to feel annoyed and is a hidden danger to the quality of medical services, which should be paid attention to by the academic and management departments.

Inactivation efficacy and mechanisms of plasma activated water on bacteria in planktonic state.

AIMS: The study aimed to investigate the inactivation efficacy and mechanisms of plasma activated water (PAW) on selected bacteria in planktonic state. METHODS AND RESULTS: Plasma activated water was generated using an atmospheric cold plasma jet at 15, 22 and 30 kV for 5 min. Escherichia coli, Listeria innocua, Staphylococcus aureus, Aeromonas hydrophila, Pseudomonas fluorescens and Shewanella putrefaciens were selected as the representative bacterial species. Each bacterial suspension was inoculated into PAW immediately after generation, and the viable counts at different exposure times of 0·5, 1, 3, 5 and 24 h during 4°C storage were measured to determine the inactivation efficacy. Scanning electron microscopy images of the bacteria were conducted to examine the structural changes. Physicochemical properties of PAW, including pH, conductivity, oxidation reduction potential (ORP), and reactive species of H2 O2 , NO2 - and NO3 - were measured. The results demonstrated that inactivation efficacy was in positive correlation with voltage and exposure time. Gram-negative bacteria were more susceptible to PAW than Gram-positive bacteria. Morphology damage was observed for all the bacterial species. PAW was significantly acidified, conductivity and ORP were significantly increased, and reactive species were detectable after 48 h. CONCLUSIONS: This study offered a better understanding of the inactivation mechanisms of PAW, and the inactivation efficacy can be affected by voltage, exposure time and bacterial species. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrated the potential usage of PAW as an alternative disinfectant.

[Prognosis analysis of 112 cases with Japanese encephalitis in adults].

Objective: To analyze the improvement of clinical symptoms and recovery of neurological function in adult Japanese encephalitis, and study the prognostic factors. Methods: Follow-up was conducted for 112 hospitalized patients with Japanese encephalitis (JE) in adults at the Department of Neurology of three hospitals in Gansu province from July to October 2016, from July to October 2017, 6 months and 1 year after onset, respectively. The neurological functional recovery was evaluated by modified Ranking Scale (mRS).The influencing factors were analyzed by logistic regression model. Results: Among the 112 adult patients with JE after 1year follow-up, 57% (64/112) were completely recovered (mRS score=0), and 14%(16/112) had mild neurological dysfunction (mRS score=1 or 2 points), 20% (22/112) had moderate to severe neurological dysfunction (mRS score 3 to 5), and 9% (10/112) died. In 102 survivors, decreased consciousness were fully recovered (100%), 75% of the mental and behavior disorders, 64% of cognitive/memory impairment, 71% of language function disorder, 61% of paralysis, 73% of extrapyramidal symptoms were fully recovered, and 92% of the seizures were controlled. Comparison of clinical data of initial on-set between good prognosis group (mRS score≤2, 80 cases) and poor prognosis group (mRS score>2, 32 cases) showed that initial clinical manifestation with seizures, consciousness (GCS score), cerebrospinal fluid pressure, and lesion of MRI involved in midbrain had statistically significant differences (all P<0.05) . Multivariate analysis demonstrated that cerebrospinal fluid (CSF) pressure>250 mmH(2)O and lesion of midbrain in MRI were independent risk factors of poor prognosis in adult patients with JE. Conclusion: JE is an acute and infectious viral encephalitis of the central nervous system with high disability and mortality. Most patients were completely recovered, and some had neurological sequelae. CSF pressure>250 mmH(2)O and lesion of midbrain in MRI are independent risk factors for poor prognosis.

[Investigation and analysis of reproductive health status of female street cleaners in a district].

Objective: To explore the reproductive health status of the female street cleaners in Chaoyang district of Beijing and its influencing factors. Methods: In July 2018, a total of 647 questionnaires were sent out to female road cleaning and sanitation workers in the whole district, 613 of which were valid, with an effective rate of 94.7%. Frequency or percentage (%) is used for statistical description of counting data, and mean standard deviation is used for measurement data. The influencing factors were analyzed by 2 test. Results: The average age of 613 female road sweepers in Chaoyang District of Beijing was 42.01 (SD=6.69) years old, including 535 married female workers (87.28%) , 356 middle school and below educated (58.08%) , 292 non Beijing registered (47.63%) , accounting for (/613) , (/613) female workers working hours >8 hours per day 110 (17.94%) , weekly rest <2 days 341 (55.63%) . 144 (23.49%) women workers did not have regular gynecological examination, 119 (19.41%) had reproductive tract infections, 177 (28.87%) had abnormal menstruation in recent 6 months. Drinking, sexual behavior outside marriage, night shift and job satisfaction were all the influencing factors (P<0.05) . The increasing age, working years, drinking, household registration in other places and the decreasing satisfaction of women workers＇ rights and interests protection were all the influencing factors (P<0.05) . Conclusion: The reproductive health status of female road sweepers in Chaoyang District of Beijing is not optimistic.

A biological study of supernumerary teeth derived dental pulp stem cells based on RNA-seq analysis.

AIM: To identify the basic characteristics and gene expression profiles of supernumerary teeth derived stem cells (SNTSCs) and compare them with those of normal dental pulp stem cells (DPSCs). METHODOLOGY: Flow cytometry was conducted to identify the protein expression of stem cell markers. Cell proliferation, migration and differentiation abilities of both SNTSCs and DPSCs were determined by CCK8, transwell and differentiation assays, respectively. Gene expression profiles were studied by RNA sequencing analyses. After knocking down the expression of certain differential expression genes (DEGs), the function of DEGs was investigated by CCK8 and transwell assays. Statistical differences were determined using a two-tailed t-test and P values below 0.05 were considered significant. RESULTS: Supernumerary teeth derived stem cells were capable of differentiating into adipocyte, chondrocyte and osteoblast lineage cells, and compared to ordinary DPSCs, SNTSCs had a significantly higher cell proliferation rate (P < 0.01) and significantly lower migration rate (P < 0.01). RNA-seq results revealed the differential expression genes (DEGs) between SNTSCs and DPSCs. A principal component analysis (PCA) and cluster analysis revealed that the gene expression patterns of SNTSCs and DPSCs were different from each other. A total of 12 861 genes were differentially expressed at a significant P value (P ≤ 0.01), and 5292 of these increased in SNTSCs and 7569 decreased. Further study on the selected DEGs revealed that FUT11, FAM155A and BRD2 inhibited the cell proliferation rate of SNTSCs, and FUT11 and GLUD1 inhibited the cell migration rate, whilst FAM155A promoted the migration rate. CONCLUSIONS: The biological characteristics and gene expression profile of SNTSCs was revealed. The stem cell properties of SNTSCs were similar to normal DPSCs but they had a high cell proliferation rate and may have greater potential for cell differentiation.

[Effects of mineral trioxide aggregate and ethanolic extracts of Shandong propolis on the biological properties of human dental pulp fibroblasts].

OBJECTIVE: To evaluate the effect of mineral trioxide aggregate (MTA) and propolis from Shangdong province on the cell viability, mineralization and migration and anti-inflammatory ability of dental pulp fibroblasts. METHODS: The human dental pulp fibroblasts were cultured and subjected to 10 mg/L of propolis and 1:8 dilution of MTA extraction. The cell viability was evaluated with cell counting kit-8 (CCK-8) after 1, 5, 7 and 9 days. The cells in the upper inserts and the test culture media on the bottoms of 24-well plates interacted for 15 hours. Then the numbers of cells migrated through the permeable membranes were compared. The cells seeded in the 24-well plates were incubated in osteogenic medium with different materials for 21 days and stained with alizarin red S, then photographed. To evaluate the deposition of calcified matrix, the wells were destained with 100 mmol/L cetylpyridinium chloride. Finally, the cells were exposed to 1 mg/L lipopolysaccharide (LPS) to induce an inflammatory response, in the presence of propolis, MTA extraction. The cells were collected after 3 h, and the expressions of tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) were determined using real-time polymerase chain reaction (real-time PCR). Statistical analysis was performed by one-way ANOVA and nonparametric tests (P<0.05). RESULTS: The cell viability of propolis group was significantly lower than those of MTA and control groups on days 5, 7 and 9, while MTA significantly increased the numbers of the viable cells on days 7 and 9. The migration cells of propolis group (26.67±2.52) were fewer than control group (61.33±4.93), and the cells of MTA group (80.00±2.65) were statistically more than those of the other two groups. The propolis group significantly induced more calcified matrix deposition than MTA group after 21 days of culture. Propolis significantly suppressed the expressions of IL-1ß and IL-6 after LPS exposure compared with MTA and control groups. CONCLUSION: The propolis from Shandong compared with MTA showed a certain degree of cytotoxicity, and had no significant effect on cell migration. On the other hand, propolis exhibited significant anti-inflammatory and mineralization promotion effect, suggesting that the active ingredients of propolis could be introduced as a supplement of pulp capping materials, or used as an irrigant or intracanal medicament due to its excellent anti-inflammatory effect. Propolis may have potential in vital pulp treatment of young permanent tooth suffering pulp inflammation.

[Comparative study of differentiation potential of mesenchymal stem cells derived from orofacial system into vascular endothelial cells].

OBJECTIVE: To compare the proliferation and capacity of differentiation to vascular endothelial cells and angiogenesis induction among stem cells from human exfoliated deciduous teeth (SHED), dental pulp stem cells (DPSC) and human bone marrow mesenchymal stem cells (BMSC) from orofacial bone. METHODS: SHED and DPSC were isolated from pulp tissue of the patients. BMSC were isolated from orthognathic or alveolar surgical sites. The surface markers of the cells were detected by flowcytometry. Cell counting kit-8 (CCK-8) assays were conducted to detect the proliferation ability of the cells. The cells were induced into endothelial cells with conditional medium and then the induced cells were cultured in Matrigel medium. The expression of angiogenesis-related genes such as platelet endothelial cell adhesion molecule-1 (PECAM-1/CD31), vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 1 (VEGFR1), vascular endothelial growth factor receptor 2 (VEGFR2) and von Willebrand Factor (vWF) were quantified by real-time PCR. The cells were cultured in chick embryo chorioallantoic membrane (CAM) and the vessels were counted after 5 days. RESULTS: The cell surface markers CD73, CD90, CD105 and CD146 of all the stem cells were positive, CD34 and CD45 were negative. The CD146 positive rate of SHED and DPSC was higher than that of BMSC. SHED had a higher proliferation rate than DPSC and BMSC. After angiogenic induction for 14 d, 3 kinds of cells emanated pseudopodia formed grid structure long vasculature in Matrigel media. The total length of tube formation of induced BMSC (7 759.7 µm) and SHED (7 734.3 µm) was higher than DPSC (5 541.0 µm). The meshes number of induced SHED (70.7) was higher than DPSC (60) and BMSC (53.7) in Matrigel medium. The expression of CD31, VEGFR2 and vWF genes of SHED were higher than those of BMSC and DPSC. VEGFR1 gene expression of BMSC was higher than that of the other groups, and SHED was higher than DPSC. The expression of VEGF showed no difference among the cells. No deference was showed between the effect of the stem cells and negative control on new formed vessels in CAM. The total length of vessels of SHED (30.4 mm) was higher than that of the negative control (20.9 mm) and BMSC (28.0 mm). CONCLUSION: SHED, DPSC and BMSC can differentiate into vascular endothelial cells. SHED showed a stronger angiogenesis differentiation and proliferation potential compared with DPSC and BMSC.

[Correlation between the expression of STOX1 in placenta of patients with early onset preeclampsia].

Objective: To investigate the relationship of STOX1expression and pathogenesis of early onset preeclampsia. Methods: 65 cases of preeclampsia women who delivered in Shanghai Pudong Hospital from October 2015 to June 2018, were recruited, which included 31 cases with early onset preeclampsia (early onset group, gestational week<34 weeks) and 34 patients with late onset preeclampsia (late onset group, gestational week ≥34 weeks). 34 cases women who received caesarean section because of pelvic structural deformities, breech presentation, macrosomia and social factors were included as the control group(gestational week ≥34 weeks) were selected as control group.The expression and localization of STOX1 mRNA and protein in placenta of three groups of maternal were evaluated by immunohistochemistry SP, RT-qPCR and Western blotting. Results: (1) The expression of STOX1 in placenta mainly distributed in the cytoplasm of placental syncytiotrophoblasts, cytotrophoblasts, vascular endothelial and mesenchymal cells, a few in the cell nucleus.The staining intensity of STOX1 in early onset group was significantly stronger than that in late onset group, the staining intensity of the late onset group was similar to that of the control group. The positive expression rates of STOX1protein in early onset group, late onset group and control group were 96.8%(30/31), 70.6%(24/34), 67.6%(23/34) respectively, which was higher in early onset group than that in late onset group(P=0.005). There was no statistical difference of STOX1 level between the late onset group and the control group(P=0.793). (2)Relative expression of STOX1 mRNA in early onset group, late onset group and control group were 0.054 3±0.003 5,0.037 5±0.000 7,0.035 2±0.000 4 respectively, which was significantly higher in early onset group than that in late onset group(P<0.05), while there was no statistical difference between the late onset group and the control group(P>0.05).(3)Relative expression level of STOX1 protein in early onset group, late onset group and control group were 0.78±0.04,0.59±0.020 and 0.54±0.018 respectively, which is higher in early onset group than that in late onset group(P<0.05). There was no statistical difference of STOX1 level between the late onset group and the control group(P>0.05). Conclusions: The pathogenesis of early onset and late onset preeclampsia may be different. Up-regulated expression of STOX1 in placenta may be associated with the pathogenesis of early onset preeclampsia.

[Expression and significance of STOX1 in villi and placenta at different stages of normal gestation].

Objective: To investigate the expression and significance of STOX1 in different stages of gestation villi and placenta. Methods: Totally 137 cases of normal villi and placenta of pregnant women were collected from the Department of Obstetrics of Shanghai Pudong Hospital from October 1(st) 2015 to February 28(th) 2018, including 64 cases of early pregnancy (early pregnancy group) which consists of 32 cases of 5-7(+6) weeks gestation (early pregnancy group A) and 32 cases of 8-11(+3) weeks gestation (early pregnancy group B), 28 cases of 14-26 weeks gestation(middle pregnancy group) and 45 cases of 37-41 weeks gestation (late pregnancy group). The expression and localization of STOX1 mRNA and protein in placenta were evaluated by RT-qPCR, Western blotting and immunohistochemistry. Results: (1)STOX1 was positively expressed in the cytotrophoblasts and syncytiotrophoblasts as well as interstitial and vascular endothelial cells of all groups. (2)STOX1 mRNA expression in each group was significantly different (P<0.05), the lowest was in the early pregnancy group A(0.007 8±0.000 4), which increased along with the progression of gestational age(P<0.05),and reached the highest level in the third trimester(0.064 4±0.001 3). (3)The protein level of STOX1 in different stages of normal pregnancy was 0.53±0.20 in early pregnancy group A;0.62±0.37 in early pregnancy group B;0.70±0.03 in middle pregnancy group and 0.81±0.04 in late pregnancy group respectively; which was positively related with the progression of gestational age (P<0.05). Conclusion: The expressions of STOX1 is gradually increasing along with the normal pregnancy progression, suggesting that it might be involved in proliferation, differentiation and infiltration and (or) apoptosis of trophoblast cells and the development of the placenta.

[Effect of monoacylglycerol lipase with proliferation of MHCC97H human liver cancer cells in vivo].

Objective: To investigate the effects of different expression of monoacylglycerol lipase (MAGL) in tumor-associated macrophages (TAMs) with the proliferation of MHCC97H human liver cancer cells in vivo and its mechanism. Methods: Human peripheral blood-derived monocyte was induced to differentiate into M2-type TAMs and was identified by flow cytometry. The co-culture model of TAMs and MHCC97H human liver cancer cells was established, and the expression of MAGL in TAMs cells was detected by qRT-PCR. The expression of MAGL in TAMs cells was detected by plasmid transfection. ELISA and qRT-PCR was used to detect the mRNA expression levels and secretion levels of inflammatory factors in TAMs cells. The subcutaneous tumor model of MHCC97H mice was constructed to observe the effect of different expression of MAGL in TAMs cells with the proliferation of MHCC97H human liver cancer cells in vivo. F-test was used for the measurement of homogeneity of variance between two independent samples. A t-test was used for homogeneity of variance, and the corrected t-test was used for non-homogeneity of variance. Results: Human peripheral blood-derived monocytes were successfully induced to differentiate into M2-type TAMs. An in vitro co-culture model was established. qRT-PCR showed that MHCC97H human liver cancer cells significantly down-regulated the expressional level of MAGL in TAMs cells. The constructed subcutaneous tumor model of mice demonstrated that up-regulation up-regulation of MAGL expression in M2-type TAMs inhibited the proliferation of MHCC97H human liver cancer cells in vivo. Furthermore, the mechanistic study illustrated that the high expression of MAGL promoted the transcription and secretion of inflammatory factors such as interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha in M2-type TAMs cells. Conclusion: The overexpression of MAGL inhibits the proliferation of MHCC97H hepatocellular carcinoma cells in vivo, and its mechanism may be associated to the release of inflammatory factors that from TAMs cells.

Effect of propolis on preserving human periodontal ligament cells and regulating pro-inflammatory cytokines.

BACKGROUND/AIM: Propolis has been suggested as a storage medium for avulsed teeth. The aim of this study was to compare the effectiveness of Brazilian propolis with Hank's balanced salt solution and milk in maintaining the viability of human periodontal ligament cells, their osteogenic differentiation potential, and pro-inflammatory cytokine expression. MATERIAL AND METHODS: Cell Counting Kit 8 assays were performed to test human periodontal ligament cell viability in different storage media. The preservative effect on osteogenic differentiation was evaluated using alkaline phosphatase staining and activity assays, Alizarin Red S staining, and western blotting. Quantification of pro-inflammatory cytokines was performed using real-time PCR and enzyme-linked immunosorbent assays. RESULTS: Brazilian propolis at 10 µg/ml was not cytotoxic toward human periodontal ligament cells. The milk group showed the highest cell viability. Brazilian propolis and Hank's balanced salt solution groups showed similar cell viabilities. Alkaline phosphatase staining and activity were similar in all groups. Calcium deposition and mineralization nodule formation were similar in the Brazilian propolis and Hank's balanced salt solution groups, but were higher in the milk group. Osteogenic marker gene and protein levels were similar in all groups. The genes and protein expression levels of IL1ß, IL6, and IL8 decreased significantly after treatment with Brazilian propolis. TNFα mRNA expression showed no significant difference among the experimental groups. Pro-inflammatory cytokine levels in the milk group were higher than in the Brazilian propolis and Hank's balanced salt solution groups. CONCLUSIONS: Brazilian propolis, Hank's balanced salt solution, and milk maintained the viability of human periodontal ligament cells and preserved their osteogenic differentiation ability similarly. However, Brazilian propolis showed a better anti-inflammatory effect. This article is protected by copyright. All rights reserved.

[Retrospective study of dental treatment under general anesthesia of 62 disabled children and adolescents].

OBJECTIVE: To retrospectively figure out the oral health status, treatment and follow-ups after dental treatment under general anesthesia (DGA) of disabled children or adolescents. METHODS: Clinical data of disabled children or adolescents and normal children as control received DGA in the Department of Pediatric Dentistry, Peking University School and Hospital of Stomatology from August 2008 to September 2015 were recorded, including: gender, birth date, treatment date, disability type, oral health status before treatment, treatment content and follow-ups (in 1 year). Differences among ages and disabilities were analyzed statistically. RESULTS: Sixty-two disabled patients and fifty-seven controls were recruited, mean aged (9.38±5.22) years and (3.00±1.41) years. Most patients had 10 to 15 problem teeth with which the mean number of the disabled children and adolescents was (11.79±4.98) while that of the normal controls was (12.40±4.11). Caries, pulpitis, periapical periodontitis, dental trauma and developmental tooth anomalies of the disabled patients accounted for 67.56%, 13.54%, 15.15%, 1.07%, and 2.68%, respectively and the DMFT/dmft index was 11.55±5.56 while in the control group those were at 65.35%,19.09%,14.14%,0,1.41% and 12.23±4.42. The DMFT/dmft index of the disabled patients in the group 6-12 years (8.35±4.69) was significantly less than that of the other three groups (P<0.01) while no differences were found in disabilities (P=0.239). Resin restoration, pit and fissure sealant, preventive resin restoration, pulpotomy, pulpectomy/RCT, extraction and crown of the disabled patients were performed as 52.71%, 7.24%, 8.56%, 0.72%, 17.13%, 10.01% and 3.62% respectively whereas those made up as 56.31%, 1.27%, 0.13%, 2.29%, 19.87%, 7.90% and 12.23% in the control group. Thirty-five (56.45%) disabled patients and forty-three (75.44%) controls recalled. Problem teeth within one year after operation in diabled patients and controls were both nearly twice as much as the number within half a year. Restoration loss/fractured mainly occurred in anterior primary teeth while secondary/ recurrent caries and pulpitis/perapical periodontitis mostly occurred in primary molars. CONCLUSION: Oral health status in our disabled children and adolescents is poor. Though dental treatment under GA is an effective way to improve the oral health of disabled children and adolescents, periodic follow-ups and family oral health care are equal important for oral health maintenance.

[Comparison of the properties of CD146 positive and CD146 negative subpopulations of stem cells from human exfoliated deciduous teeth].

OBJECTIVE: Stem cells from human exfoliated teeth (SHED) were sorted by magnetically activated cell sorting (MACS) technique to obtain the CD146 positive and negative cell subpopulation. Then the biological characteristics of these subpopulations were compared to explore their specific application potential in tissue engineering. METHODS: In this study, freshly extracted deciduous teeth without any caries or dental pulp disease were obtained. SHED was isolated using enzyme digestion method and then sorted by MACS, CD146 positive cells and CD146 negative cells were obtained after cell sorting. The biological characteristics of the unsorted mixed cells, CD146 positive subpopulation and CD146 negative subpopulation were compared. The proliferation ability was detected through cell counting kit-8 (CCK-8) and colony-forming unit (CFU). After osteogenic induction, alizarin red staining was performed and the gene expression of osteogenic related markers was detected by quantitative real-time polymerase chain reaction(qPCR). After adipogenic induction, oil-red O staining was performed and the gene expression of adipogenic related markers was detected. After neurogenic differentiation induction, the expression of neural markers was detected by immunofluorescence and the gene expression of neural markers was detected by qPCR. RESULTS: SHED of the fifth passage was sorted by MACS. And the CD146 positive cell subpopulation and CD146 negative cell subpopulation were obtained. CCK8 assay showed that the proliferative tendency of the three cell groups was consistent, but the proliferation potential of CD146 positive and negative cell subpopulations was significantly lower than that of the unsorted cells. The colony forming rates of the unsorted mixed cell group, CD146 positive and negative populations were 28.6%±3%,17.1%±2.3% and 27.5%±2.5%, respectively. After 21 days of osteogenic induction, alizarin red staining and qPCR showed that the CD146 positive cell population had more mineralized nodule formation and expressed higher level of osteogenic related genes compared with the other two groups. After 21 days of adipogenic induction, oil red O staining and qPCR results showed that the CD146 negative subpopulation produced more lipid droplets and the expression of lipid related genes increased more significantly. After 14 days of neural induction, cell immunofluorescence and qPCR results showed that the unsorted mixed cell group and CD146 positive subpopulation expressed glial cell marker, and the expressions of neural precursor cells and neuronal marker increased significantly in negative subpopulation. CONCLUSION: The unsorted mixed cells showed better proliferative potential than CD146 positive and negative subpopulations. The CD146 positive subpopulation was most potent in osteogenic differentiation; it was more suitable for bone tissue engineering. The CD146 negative cells had stronger adipogenic differentiation potential than the other two cell groups; different subpopulations differed in neural differentiation.

[Research progress of the relationship between von Willebrand factor and hepatic diseases].

Hepatic diseases refer to acute and chronic liver disease linked to all kinds of hepatic pathological changes. Viral hepatitis, bacterial infections, endotoxins and other factors initiate the activation and injury of vascular endothelial cells in patients with liver diseases. Von Willebrand factor (vWF) represents a specific marker of endothelial dysfunction and plays a vital role in the occurrence and development of hepatic diseases. This paper reviews the research progress of the relationship between vWF and hepatic diseases.

[Recombinant human transforming growth factor ß1 promotes dental pulp stem cells proliferation and mineralization].

OBJECTIVE: To explore suitable concentration of recombinant human transforming growth factor ß1 (rhTGF-ß1) usage and study the effect of rhTGF-ß1 on differentiation of dental pulp stem cells (DPSCs). METHODS: DPSCs were isolated from the undiseased third molars of people aged 18-25 years and cultured according to instructions in vitro. Different concentrations (1 , 6 , 10 µg/L) of rhTGF-ß1 were added to the culture medium to examine DPSCs proliferation by CCK-8 (cell counting kit-8) assay.The suitable concentration was then selected. For differentiation, the DPSCs were incubated for 7 or 14 days with rhTGF-ß1 supplemented with osteo/odontoblastic induction medium containing 10 nmol/L dexamethasone, 10 mmol/L b-glycerophosphate, 50 g/L ascorbate phosphate, 10 nmol/L 1,25-dihydroxyvitamin D3 and 10% fetal bovine serum. The cells were then washed 3 times with phosphate-buffered saline and sonicated with 1%Triton X-100 for 30 minutes on ice. Cellular alkaline phosphatase (ALP) activity was assayed with p-nitrophenyl phosphate as the substrate. The enzyme activity was expressed as p-nitrophenyl produced per milligram of protein [bicinchoninic acid (BCA) protein assay kit]. To examine mineral nodule formation, the cultured cells were fixed in 4% paraformaldehyde and washed in water, and the mineralization of the extracellular matrix was assayed by 1% alizarin red S staining and elution of staining was examined as optical density (D) under microplate reader. The mean difference was considered significant at 0.05 and 95% confidence interval. RESULTS: The DPSCs had typical fibroblast morphology and could form mineral nodules after being cultured with osteo/odontoblstic induction medium for 14 days. 6 µg/L rhTGF-ß1 significantly promoted the DPSCs proliferation on the 3rd and 5th days. After the incubation of osteo/odontoblastic induction medium, the DPSCs with the 6 µg/L rhTGF-ß1 increased ALP activities compared with the control; D values in the 6 µg/L rhTGF-ß1 group was 0.31±0.03, while the control group was 0.02±0.01 (P<0.05). The total protein content in the 6 µg/L rhTGF-ß1 group was (2 775.46±83.54) mg/L, and the control group was (1 432.20±110.83) mg/L (P<0.05). To eliminate the cells proliferation influence, relative ALP activities, which was defined as the total ALP divided by the total protein content, the 6µg/L rhTGF-ß1 group was 6 times higher than the control group. Alizarin red S staining showed increased mineral nodule formation in the rhTGF-ß1 group. The elution of staining under microplate reader also showed more optical density in the 6 µg/L rhTGF-ß1-treated cells (0.83±0.02) than that in the control groups (0.55±0.05, P<0.05). CONCLUSION: 6 µg/L rhTGF-ß1 could significantly promote DPSCs proliferation and odontoblastic differentiation in vitro.