Toxicity of the Mycotoxin Deoxynivalenol on Early Cleavage of Mouse Embryos by Fluorescence Intensity Analysis.

Deoxynivalenol is a mycotoxin, produced by Fusarium from contaminated corn, wheat, and other grains, that induces multiple effects in humans and animals, including cytotoxic, genotoxic, immunotoxic, and carcinogenic effects. Recent studies show that deoxynivalenol also affects the reproductive system of mammals, including oocyte quality. However, the effects of deoxynivalenol on early embryonic development have not been reported. In this study, fluorescence intensity analysis was used to show that deoxynivalenol disrupted the first cleavage of the zygote. The high deoxynivalenol dose disturbed the movement of the pronucleus after fertilization, while the low deoxynivalenol dose caused aberrant spindle morphology during the metaphase of the first cleavage. Further analysis showed that the reactive oxygen species level increased in the deoxynivalenol-exposed two-cell embryos, indicating oxidative stress. Moreover, deoxynivalenol caused DNA damage in the embryos, as positive γH2A.X signals were detected in the nucleus. These events led to the early apoptosis of mouse embryos, which was confirmed by autophagy. Taken together, our study provides evidence for the toxicity of deoxynivalenol during early embryonic development in the mouse model.

Aflatoxin B1 impairs porcine oocyte quality via disturbing intracellular membrane system and ATP production.

Aflatoxin is the most common type of mycotoxins in contaminated corn, peanuts and rice, which affects the livestock and ultimately endangers human health. Aflatoxin is reported to have carcinogenicity, mutation, growth retardation, immunosuppression and reproductive toxicity. In present study we reported the causes for the declined porcine oocyte quality under aflatoxin exposure. We set up an in vitro exposure model and showed that aflatoxin B1 disturbed cumulus cell expansion and oocyte polar body extrusion. We found that aflatoxin B1 exposure disrupted ER distribution and elevated the expression of GRP78, indicating the occurrence of ER stress, and the increased calcium storage also confirmed this. Besides, the structure of cis-Golgi apparatus, another intracellular membrane system was also affected, showing with decreased GM130 expression. The oocytes under aflatoxin B1 exposure showed aberrant lysosome accumulation and higher LAMP2 expression, a marker for lysosome membrane protection, and this might be due to the aberrant mitochondria function with low ATP production and the increase of apoptosis, since we found that BAX expression increased, and ribosomal protein which is also an apoptosis-related factor RPS3 decreased. Taken together, our study revealed that aflatoxin B1 impairs intracellular membrane system ER, Golgi apparatus, lysosome and mitochondria function to affect porcine oocyte maturation quality.

Single-cell transcriptome analysis reveals development atlas of mouse molar pulp cells / 中华口腔医学杂志

Objective: Single-cell RNA sequencing (scRNA-seq) was used to analyze the developing mouse molars, in order to construct a spatiotemporal development atlas of pulp cells, and further to reveal the developmental process and regulatory mechanism of tooth development. Methods: Ten mandibular first molars from C57BL/6 mice in postnatal day (PN) 0 and 3 were respectively dissected and digested to obtain single-cell suspensions. scRNA-seq was performed on 10× Genomics platform. PN 7 mouse molar scRNA-seq data were obtained from our previous study. PN 0, 3, and 7 scRNA-seq data were integrated for following analysis. The initial quality control, mapping and single cell expression matrix construction were performed by Cell Ranger. Quality control, standardization, dimensional reduction and cluster analysis were performed by using Seurat. Monocle was used to generate the pseudotime trajectory. Scillus was used to perform gene ontology analysis. In order to detect the spatiotemporal change of different population of pulp cells, the marker genes of each cluster were demonstrated by RNAscope in situ hybridization. Results: There were twenty-six cell clusters within mouse molars, which were identified as eight different cell types, including dental pulp cells, dental follicle cells, epithelial cells, immune cells, endothelial cells, perivascular cells, glial cells and erythrocytes. We further re-clustered and analyzed dental pulp cells. Cluster 0 were mature pulp cells, which located at the upper portion of crown. The main functions of cluster 0 were osteogenesis and extracellular structure organization. Cluster 1 were apical papilla cells, which located at the apical part of roots, whose main functions were extracellular structure organization and organ development. Cluster 2 were cycling cells, which were actively proliferated, resided in the lower portion of the crown. Cluster 3 and 4 were preodontoblasts and odontoblasts, respectively. Their functions were closely related to biomineralization. The proportion of mature pulp cells increased with the development process, while the proportion of cycling cells and odontoblast lineage decreased. According to the expression pattern of marker genes of each cluster, we constructed a cell atlas of dental pulp. Pseudotime trajectory analysis found there were two development trajectories within dental pulp. They both started from SPARC related modular calcium binding 2 (Smoc2)+ dental papilla cells, then went through DNA topoisomerase Ⅱ alpha (Top2a)+ cycling cells, and finally divided into coxsackie virus and adenovirus receptor (Cxadr)+ mature pulp cells or dentin sialophosphoprotein (Dspp)+ odontoblasts two lineages. Conclusions: scRNA-seq could fully discover the intercellular heterogeneity of cells on transcriptome level, which provides a powerful tool to study the process and regulatory mechanism of organ development.

Nonylphenol exposure-induced oocyte quality deterioration could be reversed by melatonin supplementation in mice.

Nonylphenol (NP) belongs to the metabolites of commercial detergents, which acts as an environmental endocrine disruptor. NP is reported to have multiple toxicity including reproductive toxicity. In present study, we reported the protective effects of melatonin on the NP-exposed oocyte quality. We set up a mouse in vivo model of NP exposure (500 µg/L), by daily drinking and continued feeding for 4 weeks; and we gave a daily dose of melatonin (30 mg/kg) to the NP-exposed mice. Melatonin supplementation restores the development ability of oocytes exposed to NP, and this was due to the reduction of ROS level and DNA damage by melatonin. Melatonin could rescue aberrant mitochondria distribution, mitochondria membrane potential, which also was reflected by ATP content and mtDNA copy number. Moreover, melatonin could restore the RPS3 expression to ensure the ribosome function for protein synthesis, and reduced GRP78 protein level to protect against ER stress and ER distribution defects. We also found that vesicle protein Rab11 from Golgi apparatus was protected by melatonin at the spindle periphery of oocytes of NP-exposed mice, which further moderated LAMP2 for lysosome function. Our results indicate that melatonin protects oocytes from NP exposure through its effects on the reduction of oxidative stress and DNA damage, which might be through its amelioration on the organelles in mice.

Exposure to nonylphenol impairs oocyte quality via the induction of organelle defects in mice.

Nonylphenol (NP) is an environmental endocrine disruptor, which is mainly used in the production of surfactants, lubricants, additives, pesticides, and emulsifiers. NP is widely found in sewage and sludge, which has neurotoxicity, immunotoxicity, metabolic toxicity and reproductive toxicity. In this study, we investigated the effects of NP exposure on mammalian oocyte quality from organelle aspects with mouse in vivo model. The results showed that the ovarian weight of mice exposed to 500 µg/L NP for 4 weeks increased and the development ability of oocytes decreased, showing with lower rate of polar body extrusion. Further analysis indicated that exposure to NP caused the abnormal distribution of mitochondria, following with altered membrane potential drop. NP exposure disrupted the spindle periphery localization of ER, and affected the expression of GRP78 for the induction of ER stress. Moreover, Golgi apparatus fragment in the oocytes was observed, and Rab11-based vesicle transport was disturbed. We also found that the protein degradation might be affected since LAMP2 expression increased and LC3 decreased, indicating the lysosome and autophagy dysfunction. Taken together, our findings suggested that the exposure of NP to mice in vivo affected oocyte quality through its effects on the distribution and function of organelles.

Mapping of dwarfing QTL of Ari1327, a semi-dwarf mutant of upland cotton.

BACKGROUND: Upland Cotton (Gossypium hirsutum L.) has few cotton varieties suitable for mechanical harvesting. The plant height of the cultivar is one of the key features that need to modify. Hence, this study was planned to locate the QTL for plant height in a 60Co γ treated upland cotton semi-dwarf mutant Ari1327. RESULTS: Interestingly, bulk segregant analysis (BSA) and genotyping by sequencing (GBS) methods exhibited that candidate QTL was co-located in the region of 5.80-9.66 Mb at D01 chromosome in two F2 populations. Using three InDel markers to genotype a population of 1241 individuals confirmed that the offspring's phenotype is consistent with the genotype. Comparative analysis of RNA-seq between the mutant and wild variety exhibited that Gh_D01G0592 was identified as the source of dwarfness from 200 genes. In addition, it was also revealed that the appropriate use of partial separation markers in QTL mapping can escalate linkage information. CONCLUSIONS: Overwhelmingly, the results will provide the basis to reveal the function of candidate genes and the utilization of excellent dwarf genetic resources in the future.

The association between the two more common genetic causes of spermatogenic failure: a 7-year retrospective study.

Chromosomal abnormalities and Y chromosome microdeletions are considered to be the two more common genetic causes of spermatogenic failure. However, the relationship between chromosomal aberrations and Y chromosome microdeletions is still unclear. This study was to investigate the incidence and characteristics of chromosomal aberrations and Y chromosome microdeletions in infertile men, and to explore whether there was a correlation between the two genetic defects of spermatogenic failure. A 7-year retrospective study was conducted on 5465 infertile men with nonobstructive azoospermia or oligozoospermia. Karyotype analysis of peripheral blood lymphocytes was performed by standard G-banding techniques. Y chromosome microdeletions were screened by multiplex PCR amplification with six specific sequence-tagged site (STS) markers. Among the 5465 infertile men analyzed, 371 (6.8%) had Y chromosome microdeletions and the prevalence of microdeletions in azoospermia was 10.5% (259/2474) and in severe oligozoospermia was 6.3% (107/1705). A total of 4003 (73.2%) infertile men underwent karyotyping; 370 (9.2%) had chromosomal abnormalities and 222 (5.5%) had chromosomal polymorphisms. Karyotype analysis was performed on 272 (73.3%) patients with Y chromosome microdeletions and 77 (28.3%) had chromosomal aberrations, all of which involved sex chromosomes but not autosomes. There was a significant difference in the frequency of chromosomal abnormalities between men with and without Y chromosome microdeletions (P< 0.05).

Comprehensive genetic diagnosis of patients with Duchenne/Becker muscular dystrophy (DMD/BMD) and pathogenicity analysis of splice site variants in the DMD gene.

Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are caused by mutations in the DMD gene. The aim of this study is to identify pathogenic DMD variants in probands and reduce the risk of recurrence of the disease in affected families. Variations in 100 unrelated DMD/BMD patients were detected by multiplex ligation-dependent probe amplification (MLPA) and next-generation sequencing (NGS). Pathogenic variants in DMD were successfully identified in all cases, and 11 of them were novel. The most common mutations were intragenic deletions (69%), with two hotspots located in the 5' end (exons 2-19) and the central of the DMD gene (exons 45-55), while point mutations were observed in 22% patients. Further, c.1149+1G>A and c.1150-2A>G were confirmed by hybrid minigene splicing assay (HMSA). This two splice site mutations would lead to two aberrant DMD isoforms which give rise to severely truncated protein. Therefore, the clinical use of MLPA, NGS, and HMSA is an effective strategy to identify variants. Importantly, eight embryos were terminated pregnancies according to prenatal diagnosis and a healthy boy was successfully delivered by preimplantation genetic diagnosis (PGD). Early and accurate genetic diagnosis is essential for prenatal diagnosis/PGD to reduce the risk of recurrence of DMD in affected families.

Preimplantation genetic diagnosis and screening (PGD/S) using a semiconductor sequencing platform.

BACKGROUND: Recent advances in semiconductor sequencing platform (SSP) have provided new methods for preimplantation genetic diagnosis/screening (PGD/S). The present study aimed to evaluate the applicability and efficiency of SSP in PGD/S. METHODS: The artificial positive single-cell-like DNAs and normal single-cell samples were chosen to test our semiconductor sequencing platform for preimplantation genetic diagnosis/screening (SSP-PGD/S) method with two widely used whole-genome amplification (WGA) kits. A total of 557 single blastomeres were collected from in vitro fertilization (IVF) couples, and their WGA products were processed and analyzed by our SSP-PGD/S method in comparison with array comparative genomic hybridization (array-CGH). RESULTS: Our SSP-PGD/S method indicated high compatibilities with two commercial WGA kits. For 557 single blastomeres, our method with four million reads in average could detect 24-chromosome aneuploidies as well as microdeletion/microduplication of the size over 4 Mb, providing 100% consistent conclusion with array-CGH method in the classification of whether it was transplantable. CONCLUSIONS: Our studies suggested that SSP-PGD/S represents a valuable alternative to array-CGH and brought PGD/S into a new era of more rapid, accurate, and economic.

Detection of fetal subchromosomal aberration with cell-free DNA screening led to diagnosis of parental translocation: Review of 11344 consecutive cases in a university hospital.

BACKGROUND: Fetal chromosome aberrations and sub-chromosomal copy number variations (CNVs) are not rare. There are several ways to detect duplications and deletions; cell-free DNA screening (cfDNA screening) is nowadays an accurate and safe detection method. The objective of this study is to report the feasibility of cfDNA screening as an indicator of parental balanced chromosome translocation. RESULTS: From February 2015 to March 2016, cfDNA screening was offered to 11344 pregnant women. 137 out of 11344 individuals tested positive for aneuploidies using cfDNA screening were confirmed by karyotyping. 6 additional cases also tested positive for other deletion/duplication were confirmed by chromosomal microarray analysis (CMA). 11201 patients tested negative and 10342 of them were confirmed through interviews after delivery. Among the 137 cases that were screened positive in cfDNA screening, 91 were common trisomies (63 cases of trisomy 21, 25 cases of trisomy 18 and 3 cases of trisomy 13) and 46 cases were positive for sex-chromosomal abnormalities. In addition, 6 cases were positive for other deletion/duplication in which 2 were identified as terminal duplication and deletion on different chromosomes. The cfDNA screening findings were confirmed by CMA or karyotyping, and the origins of CNVs were validated afterward by karyotyping or fluorescence in situ hybridization (FISH) using parental blood samples. CONCLUSION: CfDNA screening may help identify deletions and duplications in fetus, which in some cases may indicate risk of a parent being a balanced rearrangement carrier, and that the diagnostic follow-up testing is necessary.

Comparative study of differentiation potential of mesenchymal stem cells derived from orofacial system into vascular endothelial cells / 北京大学学报(医学版)

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.

A feasible diagnostic approach for the translocation carrier from the indication of products of conception.

BACKGROUND: Chromosome translocations are rare but frequently associated with infertility. The objective of this study is to investigate the feasibility of using chromosomal microarray analysis (CMA) on products of conception (POC) samples as an indicator of parental balanced translocation. From January 2011 to December 2016, CMA using Affymetrix Cytoscan™750K array was performed on 1294 POC samples in our hospital. Karyotyping and fluorescence in situ hybridization (FISH) using parental blood samples were performed to validate the origin of subchromosomal copy number variations (CNVs). RESULTS: In the 1294 cases of POCs, we detected CNVs of terminal duplication and deletion that imply unbalanced translocation derivatives in 16 cases, and accurate diagnosis with the parental study was made in all the cases by karyotyping and/or FISH. In 10/16 (62.5%) of these cases, CNVs were inherited from one carrier parent of balanced translocation (Cases 1 to 10), while 6/16 (37.5%) cases occurred de novo (Cases 11 to 16). CONCLUSION: This study clearly illustrated the importance of the utilization of CMA on POC, followed by parental karyotyping and FISH to better characterize CNVs. This approach is especially useful for couples in whom one partner carries a cryptic/submicroscopic balanced translocation but has an apparently normal karyotype.

Three-dimensional analysis of the early correction of anterior crossbite using eruption guidance appliance / 北京大学学报(医学版)

OBJECTIVE@#To investigate the clinical effect of eruption guidance appliance in the treatment of anterior cross bite in mixed-dentition children.@*METHODS@#In the study,10 mixed-dentition children with anterior cross bite, totally 12 incisors, were selected. Alginate was used to take upper and lower dentition impression and make a hard plaster model,which served as the eruption guidance appliance for treatment. The pre- and post-operative dental casts were digitized with SmartOptics Activity 880 scanner,and the three-dimensional overlapping models were obtained by reverse engineering software,Geomagic Studio 2012,then the three-dimensional movements of the upper and lower incisors were analyzed using Imageware 13.2 software. The overbite and overjet were analyzed using the same methods. Measurement with copper wire was used to analyze the upper and lower arch length. Space analysis was the result by the sum of crown width minus the arch length. The crown width of unerupted permanent teeth was according to X-ray method to predict. The SPSS 17.0 software was used to analyze the pre- and post-operative measurements of the same child. The normality test of the measured data showed that it conformed to the normal distribution. Therefore,the t test and double side test were used,and the significance level was 0.05.@*RESULTS@#The course of treatment was (5.6±2.7) months. During orthodontic treatment, the upper incisors moved mainly labially (P<0.001) in three-dimensional displacement, and the lower incisors moved mainly the same direction (P=0.025). During the treatment of eruption guidance appliance,the average overbite decreased (1.01±0.9) mm (t=-3.531, P=0.006), and the difference was statistically significant. There was no statistically significant difference between the pre- and post-operative average overjet (t=0.771, P=0.460). The severity of crowding in upper arch decreased (1.9±0.99) mm (t=-6.042, P<0.001),and that in lower arch decreased (1.9±0.74) mm (t=-8.143, P<0.001), both of the differences were statistically significant.@*CONCLUSION@#The anterior cross bite in mixed dentition could be corrected by eruption guidance appliance, and at the same time, the normal overjet and overbite were established, and the teeth were aligned.

Comparison of the properties of CD146 positive and CD146 negative subpopulations of stem cells from human exfoliated deciduous teeth / 北京大学学报(医学版)

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.

Analysis of dental agenesis patterns of the oligodontia patients using the method of tooth agenesis code / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To analyze the common dental agenesis patterns of the oligodontia patients.</p><p><b>METHODS</b>The information of 64 oligodontia patients was collected, including the histories, oral examinations and panoramic radiographs. The Tooth Agenesis Code (TAC) procedure was used to analyze the agenesis pattern of each quadrant.</p><p><b>RESULTS</b>In the maxilla, 63% (40/64) (right side) and 58% (37/64) (left side) could be described using eight different patterns. The most common pattern was agenesis of the maxillary lateral incisor, canine and both premolars.In the mandible, 52% (33/64) (right side) and 53% (34/64) (left side) of the patients could be described using only five different patterns, the most common pattern was agenesis of both mandibular premolars.</p><p><b>CONCLUSIONS</b>Common patterns of tooth agenesis were successfully identified in non-syndromic oligodontia patients.</p>

Chinese version of a face version of the modified child dental anxiety scale: transcultural adaptation and evaluation / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To develop the Chinese version of a face version of the modified child dental anxiety scale (MCDASf) and test the reliability and validity of MCDASf.</p><p><b>METHODS</b>The English version of MCDASf was translated and back-translated, as well as crosscultural adapted by the method of psychometrics to develop the Chinese version of MCDASf. Subsequently the Chinese version schedule was randomly investigated among 245 kindergarten children and school children aged greater than 4 and less than 12 years on two separate occasions 3 weeks apart to determine the reliability. A total of 248 children attending Pediatrics Dentistry of Peking University School and Hospital of Stomatology aged greater than 4 and less than 12 years old were selected and completed the Chinese version of MCDASf and the Chinese version of modified Children's fear survey schedule-dental subscale (CFSS-DS) before treatment to determine the validity. Then we rated the children's behavior during dental treatment by Venham's clinical anxiety rating scale and cooperative behavior rating scale to evaluate the relation between self-assessed dental anxiety scores and the behavioral reaction during the dental treatment.</p><p><b>RESULTS</b>In reliability study, 98.0% of 250 children completed the MCDASf. In validity study, 99.2% of 248 children completed the MCDASf. Cronbach's alpha coefficient of the translated scale was 0.814 and the test-retest reliability was 0.907. Principal component analysis of the translated scale confirmed that the scale consisted of a single unidimensional construct. The Chinese version of MCDASf significantly was correlated with the Chinese version of modified CFSS-DS (r = 0.843, P < 0.01) . It was also correlated with Venham's clinic anxiety rating scale and cooperative behavior rating scale (r = 0.675, P < 0.01).</p><p><b>CONCLUSIONS</b>The Chinese version of MCDASf demonstrated good reliability and validity and can be used as a simple self-report measurement of dental anxiety in Chinese children aged 4-11 years.</p>

Enrichment and purification of gardenia yellow from Gardenia jasminoides var. radicans Makino by column chromatography technique.

In present study, the performance and separation characteristics of nine macroporous resins for the enrichment and purification of gardenia yellow from Gardenia jasminoides var. radicans Makino have been evaluated. The adsorption and desorption properties of crude gardenia yellow solution on macroporous resins including HPD722, HPD100, HPD100A, HPD400, HPD400A, D101, AB-8, XAD-16, and NKA-9 have been compared. Then, HPD722 was chosen to purify gardenia yellow because of its strong adsorption and desorption abilities as well as high selectivity. Column packed with HPD722 resin was used to perform dynamic adsorption and desorption tests to optimize the separation process of gardenia yellow. The optimal conditions were as follows: The crude gardenia yellow solution with concentration of 15 mg/mL was loaded in column packed with HPD722 resin at the flow rate of 1.0 mL/min, and the adsorbate-laden column was washed with 800 mL water, 600 mL 15% ethanol water solution respectively at the speed of 2.5 mL/min, then desorbed with 200 mL 80% ethanol water solution at the speed of 3.5 mL/min. The colority of the product obtained were up to 300. The method developed in this study provides a new approach for scale-up separation and purification of gardenia yellow from G. jasminoides var. radicans Makino.

The decline of fertility in male uremic patients is correlated with low expression of the cystic fibrosis transmembrane conductance regulator protein (CFTR) in human sperm.

BACKGROUND: The present study was designed to investigate the possible association between infertility of male uremic patients and expression of the cystic fibrosis transmembrane conductance regulator (CFTR) protein in their sperm. METHODS: Semen was collected and analyzed. Serum levels of FSH, LH and testosterone were measured by radioimmunoassay. The sperm CFTR expressions of 21 uremic patients and 15 renal transplant patients were measured and compared with those of 32 healthy and 33 infertile men. RESULTS: Only 9 ± 5.9% of sperm from uremic patients expressed CFTR, significantly less than those of the renal transplant patients (29 ± 14.3%, P< 0.001), the infertile men (42 ± 20.7%, P< 0.001) and the healthy men (51 ± 20.5%, P< 0.001). Furthermore, significantly fewer sperm from renal transplant patients expressed CFTR than those of the infertile men (P< 0.05) and the healthy men (P< 0.01). LH levels in uremic patients were significantly higher than in all other groups, whereas FSH levels in uremic patients were only significantly higher than in infertile and healthy men. There was no significant difference in testosterone level among the four categories. CONCLUSIONS: Sperm CFTR expression is depressed in uremic patients but recovers to some degree after renal transplant along with some improvement in fertility, indicating a 'reversible' change. These results suggest that the CFTR expression rate in sperm is correlated with the decline of uremic patients' fertility, and may be considered as a potential marker to assess the fertility of male uremic patients.

Isolation and identification of Beagle dog dental pulp stem cells / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To identify the existence of the dental pulp stem cells in Beagle's pulp tissue by using the same methods of isolating and culturing the human dental pulp stem cells.</p><p><b>METHODS</b>Pulp tissue was extirpated from the crown and root of the Beagle's healthy permanent tooth, and digested by dispase for cell culture. Classical identification methods of mesenchymal stem cells including observation of biological characteristics, capacity of multilineage differentiation, and expression of specific markers associated with mesenchymal stem cells were applied to verify the existence of Beagle's dental pulp stem cells.</p><p><b>RESULTS</b>A clonogenic, rapidly proliferative population of cells were isolated from Beagle' pulp tissue. Under the same culture condition, the Beagle's dental pulp stem cells had a significant higher colony-forming unit-fibroblast (CFU-F) formation rate (150 colony/10(4) cells) than the dental pulp cells derived from the human pulp tissue (60 colony/10(4) cells). These cells also had the multilineage differentiation ability. They could be induced to form mineralized nodules, lipid droplets and chondrocytes. Furthermore these cells expressed the mesenchymal stem cell markers including STRO-1, CD146, alkaline phosphatase, nestin, vimentin and cytokeratin-18.</p><p><b>CONCLUSIONS</b>There are dental pulp stem cells in the Beagle's pulp tissue.</p>

An experimental study on the penetration abilities of resin infiltration into proximal caries lesions in primary molars / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To compare the penetration abilities of resin infiltration into proximal lesions in primary molars with those of adhesive in vitro.</p><p><b>METHODS</b>Thirty-two extracted or exfoliated primary molars showing proximal white spot lesions were selected. Roots of the teeth were removed, and the crowns were cut across the white spot lesions perpendicular to the surface. Cut surfaces were examined (by stereo microscopy) and classified with respect to histological lesion extension (C1-C4): lesions confined to the outer half on enamel (C1), lesions confined to the inner half on enamel (C2), lesions confined to the outer half on dentin (C3), lesions extending into the inner half of dentin (C4). Corresponding lesion halves were etched for 120 s with 15% hydrochloric acid gel and were subsequently treated with either adhesive or resin infiltration. Specimens were observed with laser scanning confocal microscope (LSCM) in dual fluorescence mode. In confocal microscopic images, lesion depth and penetration depth of the resin infiltration or the adhesive in corresponding halves were measured, and penetration percentages were calculated respectively. Differences of the data between two groups were analyzed by Wilcoxon signed rank test. Variations of histological caries extensions were detected with Kruskal-Wallis H test.</p><p><b>RESULTS</b>At the same grading level (C1-C3) in histological caries extension, penetration depths of the resin infiltration group and the adhesive group were 240 (230, 260) µm vs 190 (150, 210) µm, 405 (300, 523) µm vs 180 (160, 200) µm, and 590 (430, 640) µm vs 180 (160, 200) µm respectively. There was significant statistical difference in the data between two groups (P < 0.05). Statistically significant difference in penetration depths of the resin infiltration group (at C1-C3) were found (P < 0.01). At the same grading level (C1-C3) in histological caries extension, percentage penetrations of the resin infiltration group and the adhesive group were [100.0% (96.2%, 100.0%)], [99.1% (95.7%, 100.0%)], [82.0% (81.1%, 92.2%)] and [79.2% (68.4%, 87.5%)], [41.8% (29.1%, 74.5%)], [30.2% (29.2%, 39.6%)], respectively. The difference between the above data was also significant (P < 0.05). Percentage penetrations of the resin infiltration group at C1 and C2 level was higher than those at C3 level (P < 0.05).</p><p><b>CONCLUSIONS</b>The resin infiltration is capable of penetrating almost completely into proximal lesions in primary molars.</p>