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 establish an ultra-performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determination of tobramycin in human serum,and examine the utility of the method in a clinical pharmacokinetic study of tobramycin inhalation.Methods Serum samples were pretreated by solid phase extraction with tobramycin-D12 as internal standard.Chromatographic separation was performed on a TitankHilic(2.1 mm × 100 mm,3 μm)column.The mobile phase consisted of0.1％formic acid-acetonitrile and 0.1％formic acid aqueous solution at a flow rate of 0.4 mL/min.Electrospray ionization source and multiple reaction monitoring(MRM)scanning were used for monitoring the quantitative ion pairs with m/z 468.3→m/z 163.3(tobramycin)and m/z 480.6→m/z 166.2(tobramycin-D12).The established method was investigated in terms of selectivity,interaction,concomitant medication,standard curve and lower limit of quantitation,precision and accuracy,recovery,matrix effect,and stability of tobramycinin.Results The linear range of tobramycin was 0.050 0-10.0 mg/L(R2=0.999 5).The intra-and inter-batch precision was satisfactory(coefficient of variation[CV]≤3.6％).The accuracy ranged from-0.4％to 6.0％.The matrix effect factor(MF)in human serum samples(including hemolysis and lipemia)ranged from 92.2％to 94.9％(CV≤2.7％).The recovery of tobramycinin was 79.5％-81.9％in serum samples,while the recovery of internal standard was 78.9％.The analyte was stable in serum samples for 72 h at room temperature and for 274 days at-20℃/-70℃.The pharmacokinetic study of tobramycin inhalation in bronchiectasis patients showed that after continuous administration of tobramycin 300 mg twice a day to 3 patients,the mean Cmax of tobramycin was(0.72±0.61)mg/L on Day 1 and(0.76±0.73)mg/L on Day 28,respectively.The corresponding Tmax was(1.83±0.61)h and(1.50±0.50)h,respectively.Conclusions The UPLC-MS/MS method established in this study is sensitive,accurate and rapid.It is successfully applied to the clinical pharmacokinetic study of tobramycin inhalation.The method may be suitable for therapeutic drug monitoring of tobramycin in clinical practice.

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 explore the effects of Carolina care model in standardized training of nurses.Methods:Convenience sampling was used to select 35 nurses who entered the Department of Cardiology for standardized training (referred to as training nurses) from October 2018 to March 2019 in a Class Ⅲ Grade A hospital in Changchun as control group, and 37 nurses who entered the Department of Cardiology from May to October 2019 were selected as observation group. Control group carried out routine training, and observation group implemented a training plan based on the Carolina care model. Training effects were evaluated with the Nursing Caring Characters Assessment Tool, Clinical Communication Competence Scale and Patient Satisfaction Questionnaire.Results:After training, the scores of all dimensions of the Nursing Caring Characters Assessment Tool of nurses in observation group were higher than those in control group with statistically significant differences ( P<0.05) . After training, the scores of all dimensions of the Clinical Communication Competence Scale of nurses in observation group were higher than those in control group, and the differences were statistically significant ( P<0.05) . In observation group, patients' satisfaction with training nurses was higher than that in control group with a statistically significant difference ( P<0.05) . Conclusions:The application of Carolina care model in standardized training of nurses can not only improve nurses' humanistic care and communication competence, but also improve patient satisfaction.

Objective To investigate the changes in the percentages of CD4+T lymphocyte subsets and the homeostasis of T lymphocytes among MSM ( men who have sex with men) population with different stages of HIV-1 infection. Methods A total of 166 untreated MSM with HIV infection were enrolled and di-vided into three groups including early HIV infection (EHI, n=38) , HIV (n=94) and AIDS (n=34) groups. Sixty-two MSM negative for anti-HIV antibody were selected as healthy controls. Blood samples were collected into EDTA tubes and detected to analyze the changes in the distribution of CD4+ T cells and CD8+T lymphocyte subsets ( CD4+ CD45RA+, CD8+ CD28+, CD4+ CD25+ CD127-) and the percentages of activated (CD38, HLA-DR) and apoptotic cells (CD95) with disease progression by flow cytometry. Re-sults The expression of CD4+CD45RA+ T lymphocytes gradually decreased with the progression of AIDS. The percentage of CD4+CD45RA+ T lymphocytes in HIV group was lower than that of the control group, but higher than that of the AIDS group (P=0. 015, P=0. 000). No significant difference was found between the EHI and the control groups (P>0. 05). CD8+CD28+T cells were significantly reduced in the EHI group and remained at a low level with disease progression. No significant difference in the proportion of CD4+CD25+CD127- T cells was observed among all groups (P>0. 05). The percentage of CD4+CD38+HLA-DR+T cells increased gradually and the highest level was detected in the AIDS group, followed by those in the HIV, EHI and control groups (P<0. 01). The percentages of CD8+CD38+, CD8+HLA-DR+, CD8+ CD38+HLA-DR+and CD8+CD95+T cells in the EHI, HIV and AIDS groups were significantly higher than those in the control group (P<0. 01), but there was no significant difference among the former three groups (P>0. 05). Con-clusion HIV infection caused the changes in the numbers and functions of T lymphocyte subsets and accel-erated the activation and apoptosis of T lymphocytes, which aggravated the T lymphocyte immune dysfunction even further. A comprehensive analysis of the alterations in different T cell subsets would be conducive to re-flect the immune deficiency and the severity of disease. CD4+ and CD8+ T cells were activated in the early stage of HIV infection, which indicated that studying the immune response during that stage might help to understand their roles in disease progression.

In this study we investigated the effects of fucoidan on the proliferation of fibroblasts and the reconstruction of a skin equivalent (SE). Fucoidan significantly stimulated the proliferation of CCD-25Sk human fibroblasts and Western blot analysis demonstrated that fucoidan markedly increased the expression of cyclin D1 and decreased the expression of p27. Fucoidan was used to reconstruct SE. Immunohistochemical staining showed that the addition of fucoidan to dermal equivalents increased expression of proliferating cell nuclear antigen (PCNA) and p63. In addition, expression of alpha6-integrin was significantly increased by fucoidan, whereas expression of beta1-integrin, type 1 collagen, elastin, fibronectin did not markedly change. These results suggest that fucoidan has positive effects on epidermal reconstruction and will therefore be beneficial in the reconstruction of SE.
Blotting, Western ; Collagen Type I ; Cyclin D1 ; Elastin ; Fibroblasts ; Fibronectins ; Humans ; Proliferating Cell Nuclear Antigen ; Skin*

According to the independent component analysis (ICA) theory, in order to seperate the fetal electrocardiograms (FECG) from the observed data, we must use the larger number of observed signals than that of independent components. This requires that the number of the sentors on the electrocardiograph must be larger than a particular value, which is in practice hard to be satisfied. So we proposed another algorithm using fewer channels of abdominal electrocardiograms, which combines ICA and adaptive noise cancellation to extract the FECG from two leads of abdominal electrocardiograms. The experiment results showed that we could obtain clear FECG by the method we proposed.
Algorithms ; Electrocardiography ; methods ; Female ; Fetal Heart ; physiology ; Humans ; Pregnancy ; Principal Component Analysis ; Signal Processing, Computer-Assisted

Objective To compare the concordance among RECIST, EASL and modified RECIST criteria for the evaluation of tumor response after transarterial chemoembolization of primary liver cancer.Methods Fifty patients with primary liver cancer underwent 2 TACE cycles separated by 30-40 days.Triphasic helical CT or MRI scans were performed at baseline, at 4 weeks after TACE procedure, and 2 independent radiologists evaluated tumor response according to above-mentioned three different criteria. Chisquare test was used to compare the response rate, and kappa coefficients were used to evaluate the coherence. Results When tumor responses were evaluated using the RECIST-EASL and modified RECIST criteria, the numbers of the patients achieved complete response, partial response, stable disease,progressive disease were 0, 10, 30, 10; 6,21,14,9; 6,21,13,10 respectively. The objective response rates for three different criteria were 20%, 54%, 54% respectively ( P ＜ 0. 01 ). Kappa coefficients between RECIST and EASL, between RECIST and modified RECIST, between EASL and modified RECIST were 0. 382, 0. 170, and 0. 857 (P = 0. 000). Conclusions RECIST criteria underestimates the extent of tumor response after TACE in primary liver cancer. Both EASL and modified RECIST criteria appear to agree with each other in determining treatment response. Furthermore, the modified RECIST is more convenient in clinical practice compared with EASL criteria.

Objective To screen serum biomarkers in patients with hepatocelhlar carcinoma by using surface-enhanced laser desorption and ionization time-of-flight mass spectrometry(SELDI-TOF-MS)technique and to evaluate its clinical implication in the patients whose alpha-fetoprotein were negative in the sera. Methods Proteomic spectra were generated by mass spectroscopy in 112 cases, including 57 cases AFP-negative hepatocellular carcinoma,and 55 cases of healthy control. The consequence was analyzed and the characteristic preteomic peaks was selected by using Biomarker Wizard. Results Seven low expressed potential biomarker were indentified with the mass-to-charge ratio of 4.2×103, 4.1×103, 6.7×103, 5.7×103, 6.5×103, 6.9×103, 5.8×103. The sensitivity for diagnosing hepatic cancer were 88.23 % and specificity was 92.31%. These peaks were not correlated with age, sex, tumor mass size, pathology grading and cirrhosis in hepatocellular carcinoma. Conclusion SELDI-TOF-MS offers a unique platform for the proteomic detection of hepatocellular carcinoma.It also offers an auxiliary diagnosis method to the patients whose alpha-fetoprotein are negative in the serum.

Objective Adhesion or too highly located folds upper eyelid and even hlepharoptosis are common complications after double eyelid operation. The method was designed to correct these de-formities. Methods The incision of upper eyelid was designed under the scar. Skin was resected between the former and the new double line, if there was excessive skin. Completely relieve the adhesion until nor-mal tissue was exposed. Expose lateral extension of retro-orbicularis oculi fat pad. An upper pedicle fat flap was formed after cutting tissues off along bilateral borders. Appropriately and transversely cutting lateral portion of the pedicle made it rotate inside downward with enough blood supply. The fat flap was fixed with released orbital fat and the lower edge of fat flat fixed with upper border of supratarsal to restorate the volume. For severe condition, fat granule injection could be used after the skin was sutured. Results 42 cases (80 eyes) were treated in the past two years. The deformities of 32 cases were corrected by the application of retro-orbicularis oculi fat pad. Fat granule injection was applied for the other 10 pa-tients meanwhile. 30 cases (60 eyes) were followed up for 6 to 24 months. The results were satisfactory. Conclusion The methods are performed easily and the results are satisfied. It is worth of recommenda-tion.