Human with bi-allelic WNT10A mutations and epithelial Wnt10a knockout mice present enlarged pulp chamber and apical displacement of the root furcation of multi-rooted teeth,known as taurodontism;thus,indicating the critical role of Wnt10a in tooth root morphogenesis.However,the endogenous mechanism by which epithelial Wnt10a regulates Hertwig's epithelial root sheath(HERS)cellular behaviors and contributes to root furcation patterning remains unclear.In this study,we found that HERS in the presumptive root furcating region failed to elongate at an appropriate horizontal level in K14-Cre;Wnt10afl/flmice from post-natal day 0.5(PN0.5)to PN4.5.EdU assays and immunofluorescent staining of cyclin D1 revealed significantly decreased proliferation activity of inner enamel epithelial(IEE)cells of HERS in K14-Cre;Wnt10afl/flmice at PN2.5 and PN3.5.Immunofluorescent staining of E-Cadherin and acetyl-α-Tubulin demonstrated that the IEE cells of HERS tended to divide perpendicularly to the horizontal plane,which impaired the horizontal extension of HERS in the presumptive root furcating region of K14-Cre;Wnt10afl/flmice.RNA-seq and immunofluorescence showed that the expressions of Jag1 and Notch2 were downregulated in IEE cells of HERS in K14-Cre;Wnt10afl/fl mice.Furthermore,after activation of Notch signaling in K14-Cre;Wnt10afl/flmolars by Notch2 adenovirus and kidney capsule grafts,the root furcation defect was partially rescued.Taken together,our study demonstrates that an epithelial Wnt10a-Notch signaling axis is crucial for modulating HERS cell proper proliferation and horizontal-oriented division during tooth root furcation morphogenesis.

Objective To evaluate the inhibitory activity of fluoxetine alone and in combination with fluconazole on Candida albicans.Methods This study used standard strains of Candida albicans,clinical isolates of Candida albicans,and drug-resistant strains of Candida albicans for experiments to prepare the planktonic and biofilm states of Candida albicans,respectively.The effect of fluoxetine on Candida albicans was detected by XTT reduction method.Pharmacodynamic properties of fluoxetine against Candida albicans biofilm were determined by improved time-kill test.The morphological changes of fluoxetine on Candida albicans biofilm were observed by scanning electron microscopy,and the synergistic effect of fluoxetine combined with fluconazole on Candida albicans biofilms was detec-ted by the checkerboard dilution method.Results Fluoxetine had inhibitory effects on three types of Candida albicans biofilms.The minimum drug concentration that inhibited 50%activity(SMIC50)of biofilms of standard strains of Candida albicans and drug-resistant strains was 32 μg/mL;the SMIC50 of clinical strains of Candida albicans biofilm was 64 μg/mL.Time-kill test displayed that the bio-film activity of three types of Candida albicans significantly decreased compared to the control group at the fluoxetine concentration of 1×SMIC50 and 2×SMIC50.Scanning electron microscopy observation showed that compared with the control group,the number of fungal hyphae in the biofilm of Candida albicans treated with fluoxetine decreased and their morphology wrinkled.The checkerboard microdilu-tion method showed that the combination of fluoxetine and fluconazole exhibited a synergistic effect on the standard strain of Candida al-bicans bioflim,while showing no or antagonistic effects on the other two strains.Conclusion Fluoxetine alone has inhibitory effects on biofilms of different types of Candida albicans and has a synergistic inhibitory effect on biofilms of standard strains of Candida albicans combined with fluconazole.

Objective To evaluate the effects of chloroquine alone and in combination with traditional antifungal agents on the Candi-da albicans biofilms and its drug resistance.Methods This study used standard strains of Candida albicans,and drug-resistant strains of Candida albicans.The inhibitory effects of chloroquine alone and in combination with antifungal drugs on biofilms of Candida albi-cans were detected by XTT reduction method and chessboard dilution method respectively.The morphological characteristics of biofilms were observed under scanning electron microscopy.Results Chloroquine at the concentration of 50 μmol/L or above showed a direct inhibitory effect and increased with concentration.Chloroquine combined with amphotericin B had a synergistic inhibitory effect.Results of the time-killing curve showed that the growth trends of biofilms treated with chloroquine alone and combined with amphotericin B var-ied in different time periods during the experimental culture.Morphological observation also revealed that chloroquine alone and in com-bination with amphotericin B could reduce the ability of Candida albicans to form hyphae and biofilms.Conclusion Chloroquine has an inhibitory effect on Candida albicans biofilms and can reduce its drug resistance.Furthermore,chloroquine shows a synergistic anti-fungal effect when combined with amphotericin B.

Objective:To measure the crown conical degree of the remaining maxillary incisors in patients with congenital tooth agenesis, and to analyze the influence of different gene mutations on the crown conical degree of patients.Methods:Whole exome sequencing was performed on 85 patients with congenital tooth agenesis (50 males, 35 females, median age 19 years old) who visited the Department of Prosthodontics, Peking University School and Hospital of Stomatology from January 2019 to January 2023. The pathogenic gene was identified. The width of the crowns of the maxillary central and lateral incisors at the incisal 1/3 and gingival 1/3 were measured on the panoramic radiograph, and the ratio was defined as the crown conical degree. The smaller the ratio is, the more likely is the crown to be peg shaped teeth. The control group was matched by age and gender with 85 other patients with intact maxillary permanent incisors who were treated in the Department of Prosthodontics, Peking University School and Hospital of Stomatology from January 2019 to January 2023. The conical degree of the tooth agenesis group was compared with the control group by t-test, and the differences in the crown conical degree in different gene mutation groups were compared using one-way analysis of variance, and the multiple comparisons among gene groups were carried out using the LSD method.Results:Among the 85 tooth agenesis patients, the numbers of patients in each gene mutation group were 20 in ectodysplasin A (EDA) group, 8 in ectodysplasin A receptor (EDAR) group, 15 in wingless-type MMTV integration site family, member 10A (WNT10A) group, 16 in paired box 9 (PAX9) group, 10 in Msh homeobox 1 (MSX1) group, 10 in low-density lipoprotein receptor related protein 6 (LRP6) group, and 6 in bone morphogenetic protein4 (BMP4) group. The number of missing teeth were 1-27, median number 15 among the tooth agenesis patients. There was no significant difference in the conical degree between the left and right homonymous teeth in the congenital tooth agenesis group and the control group ( P>0.05). The crown conical degree of maxillary central incisor and lateral incisor in the congenital missing teeth group (0.95±0.24, 0.90±0.22) was significantly smaller than that in the control group (1.12±0.09, 1.13±0.09) ( t=-8.50, P<0.001; t=-11.47, P<0.001). In WNT10A mutants, the conical degree of lateral incisors (0.89±0.18) was less than that of central incisors (1.07±0.15)( t=3.68, P<0.001). The conical degree of central incisors and lateral incisors (0.70±0.23, 0.57±0.15) of EDA mutants was significantly lower than that in patients with other gene mutations ( P>0.05). Conclusions:Compared with the normal control group, the remaining maxillary central and lateral incisors of the seven gene mutation groups of patients with congenital tooth agenesis all had different degrees of conical crown. Among them, the crown conical degree of maxillary central and lateral incisors of the EDA mutation was the most severe, and the WNT10A mutation affected the maxillary lateral incisors more specifically.

The goal of this study was to identify MSX1 gene variants in multiple Chinese families with nonsyndromic oligodontia and analyse the functional influence of these variants. Whole-exome sequencing (WES) and Sanger sequencing were performed to identify the causal gene variants in five families with nonsyndromic oligodontia, and a series of bioinformatics databases were used for variant confirmation and functional prediction. Phenotypic characterization of the members of these families was described, and an in vitro analysis was performed for functional evaluation. Five novel MSX1 heterozygous variants were identified: three missense variants [c.662A>C (p.Q221P), c.670C>T (p.R224C), and c.809C>T (p.S270L)], one nonsense variant [c.364G>T (p.G122*)], and one frameshift variant [c.277delG (p.A93Rfs*67)]. Preliminary in vitro studies demonstrated that the subcellular localization of MSX1 was abnormal with the p.Q221P, p.R224C, p.G122*, and p.A93Rfs*67 variants compared to the wild type. Three variants (p.Q221P, p.G122*, and p.A93Rfs*67) were classified as pathogenic or likely pathogenic, while p.S270L and p.R224C were of uncertain significance in the current data. Moreover, we summarized and analysed the MSX1-related tooth agenesis positions and found that the type and variant locus were not related to the severity of tooth loss. Our results expand the variant spectrum of nonsyndromic oligodontia and provide valuable information for genetic counselling.
Anodontia/genetics* ; Humans ; MSX1 Transcription Factor/genetics* ; Pedigree ; Whole Exome Sequencing

Objective To clarify the effects of HBV co-infection on liver function of patients with different types of hepatic echinococcosis.Methods We recruited 409 patients diagnosed with hepatic echinococcosis at three hospitals in western regions in China from 2014 to 2015.Venous blood was withdrawn to detect to liver function indications.ELISA was performed to detect HBsAg.We analyzed liver function in patients stratified by different types of hepatic echinococcosis with or without HBV infection.Results The hepatic echinococcosis patients infected with HBV had more severe impairment in liver functions such as reduced albumin and increased transaminase.The patients with hepatic alveolar echinococcosis were more vulnerable to HBV infection compared with those with hepatic cystic echinococcosis (38.4％ vs.86.4％, P<0.05).In addition, liver injury was more severe in patients diagnosed with alveolar hepatic echinococcosis and HBV infection compared with those diagnosed with cystic hepatic echinococcosis and HBV infection (all P<0.05).Conclusion Hepatic alveolar echinococcosis patients co-infected with HBV have worse liver injury compared with those hepatic cystic with HBV. Therefore, they deserve special attention in clinical treatment.