Objective This study aimed to identify PAX9 variants in non-syndromic tooth agenesis families of Chi-na,as well as to analyze the genotype-phenotype of non-syndromic tooth agenesis caused by PAX9 variants,which can provide a basis for the genetic diagnosis of tooth agenesis.Methods We collected the data of 44 patients with non-syn-dromic oligodontia who underwent treatment at Stomatological Hospital of Hebei Medical University between 2018 and 2023.Whole-exome sequencing was performed on the peripheral blood of the proband and its core family members,and the variants were verified by Sanger sequencing.Pathogenicity analysis and function prediction of the variants were per-formed using bioinformatics tools.The correlation between the genotype of PAX9 variant and its corresponding pheno-type was examined by reviewing 55 publications retrieved from PubMed.The studies involved 232 tooth agenesis pa-tients with PAX9 variants.Results A novel PAX9 c.447delG(p.Pro150Argfs*62)and a reported PAX9 c.406C>T(p.Gln136*)were identified in two Chinese families.Through bioinformatics analysis and three-dimensional structural mod-eling,we postulated that the frameshift variant was pathogenic.The outcome was the premature cessation of PAX9 pro-tein,which caused severe structural and functional deficiencies.Summarizing the PAX9 genotype-phenotype relationship revealed that patients carrying the PAX9 variant commonly led to loss of the second molars.Conclusion We identified the novel PAX9 c.447delG(p.Pro150Argfs*62)in a Chinese family of non-syndromic oligodontia,expanding the known variant spectrum of PAX9.The most susceptible tooth position for PAX9 variants of tooth agenesis was the second mo-lars and the deciduous molars during the deciduous dentition.

Acyl-CoA synthetase long chain family member 5 (ACSL5), is a member of the acyl-CoA synthetases (ACSs) family that activates long chain fatty acids by catalyzing the synthesis of fatty acyl-CoAs. The dysregulation of ACSL5 has been reported in some cancers, such as glioma and colon cancers. However, little is known about the role of ACSL5 in acute myeloid leukemia (AML). We found that the expression of ACSL5 was higher in bone marrow cells from AML patients compared with that from healthy donors. ACSL5 level could serve as an independent prognostic predictor of the overall survival of AML patients. In AML cells, the ACSL5 knockdown inhibited cell growth both in vitro and in vivo. Mechanistically, the knockdown of ACSL5 suppressed the activation of the Wnt/β-catenin pathway by suppressing the palmitoylation modification of Wnt3a. Additionally, triacsin c, a pan-ACS family inhibitor, inhibited cell growth and robustly induced cell apoptosis when combined with ABT-199, the FDA approved BCL-2 inhibitor for AML therapy. Our results indicate that ACSL5 is a potential prognosis marker for AML and a promising pharmacological target for the treatment of molecularly stratified AML.
Humans ; Antineoplastic Agents/therapeutic use* ; Apoptosis ; beta Catenin/metabolism* ; Biomarkers, Tumor/metabolism* ; Cell Line, Tumor ; Coenzyme A Ligases/metabolism* ; Leukemia, Myeloid, Acute/metabolism* ; Lipoylation ; Prognosis ; Wnt Signaling Pathway

Background The use of heating and ventilation air conditioning (HVAC) in public places is conducive to improving indoor air quality and increasing the users comfort level. However, HVAC may also become potential carriers of indoor airborne microbial contamination. Objective To understand the characteristics of microbial pollution and distribution of related pathogenic microorganisms in HVAC of star-rated hotels, and to provide a basis for effective control of such pollution. Methods According to the requirements of the Hygienic specification of central air conditioning ventilation system in public buildings (WS 394-2012), two sets of HVAC in two star-rated hotels (A and B, inaugurated in 2002 and 1998, respectively) in the central area of Shanghai were randomly selected on September 9, 2020 for the hygienic evaluation of microorganisms in the air supply and respirable particulate matter (PM₁₀) in the air supply, dust accumulation and microorganisms on the inner surface of the ducts, as well as Legionella pneumophila in cooling water and condensate water. At the same time, 3 samples from the inner surface of ducts, 1 sample from the surface of the filter, 1 sample from the condensate, and 1 sample from the cooling water were collected from each set of HVAC, a total of 12 sample from 2 sets of HVAC, for Illumina HiSeq metagenomic sequencing，and the samples are divided into 3 groups according to their types: duct group, filter group, and water sample group. The α-diversity indices (Shannon index, Simpson index, Chao1 index, ACE index, and goods_coverage index) were calculated to reflect the microbial community composition; and the β-diversity of the three groups were studied by principal component analysis to determine the similarity of the microbial communities. Results The maximum total number of bacteria and fungi in the air supply of the HVAC were 1158 CFU·m⁻³ and 344 CFU·m⁻³ for Hotel A respectively; and 2000 CFU·m⁻³ and 532 CFU·m⁻³ for Botel B respectively. β-hemolytic streptococci were negative in all samples; the respirable particulate matter, microorganisms and dust accumulation on the inner surface of air ducts, Legionella pneumophila IN cooling water and condensate samples all met the standards. The results of Illumina HiSeq sequencing showed that a total of 17322 microorganisms were reported in the 12 samples, with bacterial microbiota accounting for 97.31% of the classified genes and the remaining 2.69% were from fungi, viruses, and parasites. At the species level, Staphylococcus epidermidis, Pseudomonas, Alternaria, and Malassezia were the dominant microbial taxa measured in this survey. The results of α-diversity analysis showed that the values of Shannon index, Simpson index, and Chao1 index for the three groups of samples were duct > filter > water sample. The goods coverage indices of all sample groups s were close to 1. The principal component analysis showed that the contributions of two principal components were 19.27% and 14.25%, respectively, in which the samples of the filter and duct groups were better clustered into one category. Conclusion The overall hygiene conditions of the two hotels are good, except for the serious microbial contamination in the air supply of HVAC. Metagenomic sequencing reveals complex microbial communities of HVAC, including bacteria, fungi, viruses, and parasites. The species composition vary by sample groups, particularly the species compositions of the samples from filters and ducts are close and dominated by pathogenic microorganisms of human origin, suggesting that the potential biosafety hazards of HVAC should not be ignored.

ObjectiveTo understand the correlation between air quality and pollutants in three underground subway stations in Shanghai and their influencing factors， in order to provide a reference for improving the ambient air quality of rail transit and enhancing control measures. MethodsAir quality was monitored in summer， autumn， and winter of 2021 at the halls， platforms， carriages and fresh air wells of three underground stations of Shanghai metro line 9， with indicators including temperature， relative humidity， wind speed， CO₂， PM₁， PM_2.5， PM₁₀， total airborne bacteria， total airborne fungi， and passenger flow. Correlation analysis and multiple regression analysis were used to explore the association between pollutant indicators and environmental indicators. ResultsThe relative humidity of air at three stations did not meet the national health standards （GB 37488‒2019）， with an exceedance rate of 66.7%； the PM₁₀ detection index at individual stations in autumn and winter did not meet the national health standards （GB 37488‒2019）， with an exceedance rate of 20.6%， while the rest of the indexes basically met the requirements of the standards. Spatially， the CO₂ and wind speed in the carriage were higher than in other areas of the station； the concentration of particulate matter in the fresh air well and the total number of airborne fungi were higher than in other areas. Seasonally， the concentration of particulate matter was higher in winter than in summer and autumn， while the microbial indexes were higher in summer than in autumn and winter. The correlation and regression analysis showed that the passenger flow was significantly and positively correlated with the CO₂ concentration； the temperature and humidity were significantly and negatively correlated with the particulate matter concentration and positively correlated with the microbial index. The correlation and multiple regression analysis showed that the subway station health indicators were significantly affected by the ambient temperature， humidity， seasonal and monitoring regional factors， while the influence of sampling time had no significant effect on the differences in the health indicators. ConclusionThe air quality of the three stations in this survey generally meets the requirements of health standards. The changes of air pollutants in subway stations show obvious spatial and temporal characteristics. The passenger flow and temperature and humidity， as well as sample sites are important factors affecting the concentration of particulate matter and the level of air microorganisms， which need to be considered when formulating the corresponding protection strategies.

OBJECTIVE：To prepare chelerythrine nanoparticles（CHE-NPs），optimize their formulation ，and evaluate its drug release behavior in vitro and its inhibitory effect on melanoma. METHODS ：Using methoxy polyethylene glycol-poly （lactic-co- glycolic acid ）（mPEG-PLGA）as carrier ，CHE-NPs were prepared by the nano-precipitation method. HPLC method and dialysis bag method were used to determine entrapment efficiency and drug loading. The formulation of CHE-NPs was optimized by Box-Behnken response surface design using overall desirability （OD）of them as dependent variables ，CHE dosage ，mPEG-PLGA concentration and poloxamer 188（F68）concentration as independent variables. The particle size and Zeta potential of CHE-NPs prepared by the optimal formulation were detected ；the characteristics of drug release in vitro were investigated ；the effects of CHE and CHE-NPs on survival rate of mice B 16 melanoma cells were compared ，and median inhibition concentrations （IC50）of them were calculated. RESULTS ：The optimal formulation included CHE of 2 mg，mPEG-PLGA of 13 mg/mL，F68 of 1.8％. Average entrapment efficiency rate of CHE-NPs prepared by the optimal formulation was （80.18±1.11）％，average drug loading was （11.36±0.28）％，average OD value was 0.96±0.04 [the relative deviation from predicted value （0.90）of OD was 6.67％]； particle size was （113.1±1.40）nm，and Zeta potential was （－21.6±0.29）mV；polydispersity index was 0.07±0.01（n＝3）； accumulative release rates of CHE control and CHE-NPs were 90.87％ and 68.68％ within 8 h，and drug release behavior in vitro of the latter was in accordance with Weibull kinetic model. Inhibitory effect of CHE-NPs on B 16 melanoma cells was significantly stronger than that of CHE ；the 24 h IC 50 of CHE-NPs and CHEwere 69.35 and 107.36 μg/mL，respectively. CONCLUSIONS ：The prepared CHE-NPs show good sustained-effect and high capacity of drug loading ，and strengthen the inhibitory effect of CHE on melanoma.

Objective:To explore the clinical characteristics and establish a corresponding prognostic scoring model in patients with early-stage clinical features of hepatitis B-induced acute-on-chronic liver failure (HBV-ACLF).Methods:Clinical characteristics of 725 cases with hepatitis B-related acute-on-chronic hepatic dysfunction (HBV-ACHD) were retrospectively analyzed using Chinese group on the study of severe hepatitis B (COSSH). The independent risk factors associated with 90-day prognosis to establish a prognostic scoring model was analyzed by multivariate Cox regression, and was validated by 500 internal and 390 external HBV-ACHD patients.Results:Among 725 cases with HBV-ACHD, 76.8% were male, 96.8% had cirrhosis base,66.5% had complications of ascites, 4.1% had coagulation failure in respect to organ failure, and 9.2% had 90-day mortality rate. Multivariate Cox regression analysis showed that TBil, WBC and ALP were the best predictors of 90-day mortality rate in HBV-ACHD patients. The established scoring model was COSS-HACHADs = 0.75 × ln(WBC) + 0.57 × ln(TBil)-0.94 × ln(ALP) +10. The area under the receiver operating characteristic curve (AUROC) of subjects was significantly higher than MELD, MELD-Na, CTP and CLIF-C ADs( P < 0.05). An analysis of 500 and 390 cases of internal random selection group and external group had similar verified results. Conclusion:HBV-ACHD patients are a group of people with decompensated cirrhosis combined with small number of organ failure, and the 90-day mortality rate is 9.2%. COSSH-ACHDs have a higher predictive effect on HBV-ACHD patients' 90-day prognosis, and thus provide evidence-based medicine for early clinical diagnosis and treatment.

Objective To analyze the abnormalities of local chemokines in patients with vitiligo and to explore the effect of tacrolimus on the secretion of chemokines in keratinocytes.Methods Blister fluids of 50 patients with vitiligo were collected,including lesion areas and normal areas.Luminex was used to analyze the concentration of local chemokines in patients with vitiligo to determine whether the chemokines were closely related to vitiligo.The effect of tacrolimus on chemokine secretion of was analyzed by Western blot in HaCaT cells.Results By Luminex analysis of blister fluid,it was found that CXCL9 and CXCL10 were significantly higher in the leukoplakia of vitiligo,and there was a significant difference,compared with the blister fluid in the normal site (P＜0.01).IFN-γ significantly stimulated the keratinocyte cell line HaCat to express CXCL9 and CXCL10.After pretreatment of HaCaT cells with 20 mg tacrolimus,the expression of CXCL9 and CXCL10 was significantly decreased,compared with the blank control (P＜0.01).Conclusions The leukoplakia chemokines CXCL9 and CXCL10 are highly expressed in vitiligo patients.The tacrolimus can significantly reduce the expression of CXCL9 and CXCL10 in keratinocytes under stress,and it therefore plays a therapeutic role in vitiligo.

The fourth affiliated hospital of Zhejiang university is responsible for training general practitioners in view of the basics of local general practitioners and the characteristics of chronic and frequently-occurring diseases in Yiwu city. This program is carried out relying on the high-quality faculty of medical alliances within Zhejiang university, and funded by the municipal government of Yiwu. The "4+6"training program is designed to elevate the competence of primary general practitioners. Such training fit the overall medical level of general practitioners, enhances people′s trust of general practitioners and the rate of diagnosis at primary level.

Objective To evaluate therapeutic effects of bone marrow mesenchymal stem cells(MSC)derived exosomes on alcohol-induced liver injury.Methods Eighteen male C57BL/6 mice aged 6 to 8 week were randomly divided into control group,model group and exosomes group,with 6 mice in each group.The mice in the model group and the exosomes group were fed with Lieber-DeCarli ad libitum diet(Dyets Inc.)for 4 weeks,followed by gavage a bolus of ethanol at day 26,27 and 28.The mice in the control group matched the alcohol-derived calories with dextran-maltose.Meanwhile,the mice in exosomes group were injected with MSC-exosomes via the tail vein at day 14 and 26.After the experiment,serum levels of alanine aminotransferase(ALT)and aspartate aminotransaminase(AST)were detected,and the pathological changes of liver tissues were observed.The expressions of nuclear factor erythroid 2-related factor 2(Nrf-2),heme oxygenase-1(HO-1),CD63,CD81,TSG101 and Cytochrome C were analyzed by Western blot,and mRNA levels of Nrf-2,HO-1,interleukin(IL)-10 and IL-17 were analyzed by real-time polymerase chain reaction(RT-PCR).The commercial kits were used to detect serum IL-10,IL-17 levels and liver tissue malondialdehyde(MDA),glutathione(GSH),superoxide dismutase(SOD)oxidative stress indicators.The numbers of regulatory T cell(Treg)and help T(Th)17 cells in the liver were analyzed by flow cytometry.One-way analysis of variance was used for comparison between groups.Results MSC-exosomes expressed positive markers CD63,CD81 and TSG101,but did not express the negative markers Cytochrome C.The serum ALT and AST levels in model group were(87.3±25.1)U/L and(223.2±43.5)U/L,respectively,while those in exosomes group were(47.7±12.0)U/L and(128.2±33.6)U/L,respectively.The differences between the two groups were both statistically significant(F=12.818 and 12.226,respectively,both P<0.05).Compared with control group,the SOD activity and GSH level in the model group significantly decreased with statistically significant differences(F=4.245 and 24.074,respectively,both P <0.05).Lieber-DeCarli ethanol feeding significantly increased intrahepatic MDA level in the model mice,which was reversed by MSC-exosomes supplementation,and the difference was statistically significant(F=36.675,P <0.05).Compared with control group,the intrahepatic protein expressions of Nrf-2 and HO-1 in model group were significantly decreased,while the expressions in exosomes group were obviously increased.The differences were statistically significant(F=33.623 and 14.960,respectively,both P <0.05).The expression trends of Nrf-2 and HO-1 mRNA were the same as those of protein expressions(F=20.784 and 276.336,respectively,both P <0.05).The proportions of liver Treg/Th17 in the control group,model group and exosomes group were 4.3±0.9,0.4±0.2,and 3.4±0.5,respectively.The differences among groups were statistically significant(F=64.227,P <0.05).Compared with control group,the serum protein and intrahepatic gene expression of IL-17 in the model group were significantly increased,which were reversed by MSC-exosomes treatment.The differences were statistically significant(F=15.581 and 40.095,respectively,both P<0.05).Serum IL-10 protein levels and intrahepatic IL-10 gene expression were significantly decreased after Lieber-DeCarli ethanol feeding,which were lower than the exosomes group.The differences were statistically significant(F=98.268 and 153.743,respectively,both P <0.05).Conclusions MSC-exosomes transplantation may relieve alcohol-induced liver injury.The mechanism could involve reduction of oxidative stress in the liver via regulating Nrf-2/HO-1 and normalizing the balance of Treg and Th17 cells.

Multi-disciplinary team(MDT) is a new mode of medical diagnosis and treatment service. Multidisciplinary discussion could provide patients with a scientific, standardized and effective individualized diagnosis and treatment plan to avoid over-treatment. This article is based on the application experience of the MDT information management platform designed by the First Affiliated Hospital of Zhejiang University during the past five years. It discussed how to build a MDT management system which could fit the medical environment of large general hospitals in China. The MDT management system simplifies the MDT process, improves the efficiency of MDT work, and enhances the overall medical quality of hospitals. Meanwhile, it also contributes to strengthen the disciplinary collaboration in such aspects as disease diagnosis and treatment, personnel training, and scientific research innovation, ultimately forming a new multi-disciplinary collaboration system in hospitals.