[Single-cell transcriptome analysis reveals development atlas of mouse molar pulp cells].

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.

[To reconstruct facial defects after removing skin lesions by using local flap].

Objective:The aim of this study is to investigate the method and effect of reconstruction of facial skin defects after removing the lesions by applying local flap. Method:Fifty-three patients （30 males and 23 females） with facial skin lesions. Complete surgical resection of the lesion; malignant tumor resection should be strictly non-tumor principle; intraoperative frozen section pathological examination confirmed negative margin; benign lesions can be cut off the lesion. According to the defect site, the appropriate local flap was used to repair the defect: 13 cases of modified VY propulsion flap, 8 cases of nasolabial flap, 8 cases of A/O-T shaped flap, 6 cases of rotating flap, 5 cases of direct sliding flap, multi-type There were 6 cases with flap, 2 cases with double leaf, 2 cases with prismatic flap and 3 cases with free flap. Result:Patients with malignant tumor were followed for 12-36 months postoperatively while followed for 10-12 months in the benign. Two patients with malignant tumor developed local recurrence and removed again. At half a year after first resection. Distal partial necrosis occurred in 5 cases while the wound dehiscence in1case, others were well developed. No others major complications occurred. Conclusion:There are various types of local skin flap for repairing facial skin defects. It is very important to excise the primary lesion radically before reconstruction,the satisfactory curative effect can be obtained through reasonable design of the flap.

[Epidemiology of allergic rhinitis in children in grassland of Inner mongolia].

Objective: To investigate the self-reported prevalence, clinical characteristics, complications of allergic rhinitis (AR) and the sensitization of outdoor air pollen allergens in children in the Inner mongolia grassland region. Methods: A multistage, stratified and random clustered sampling with a face-to-face interview survey study in children from 0 to 17 years old was performed together with 10 common allergen skin prick tests (SPT) and measurements of the daily pollen count in 6 regions in the Inner mongolia grassland region from May to August of 2015. SAS 9.4 software was used for data analysis. Results: A total of 2 443 subjects completed the study. The self-reported prevalence of AR was 26.6%. The prevalence of boys was higher than that of girls (28.8% vs 24.3%, χ(2)=6.157, P<0.05). Subjects from urban areas showed higher prevalence than rural areas (34.7% vs 18.8%, χ(2)=79.107, P<0.05). There was significant regional difference in the prevalence of AR among the six areas investigated (χ(2)=221.416, P<0.05). The main clinical symptoms of AR were sneezing (88.2%) and nasal congestion (78.6%). Among combined diseases, asthma accounted for 16.5% (107/650), rhinoconjunctivitis accounted for 47.9% (311/650). The peak season of AR was April and July, with the top SPT positive allergens of Artemisia species and chenopodium in this area. Conclusions: The prevalence AR in children in the Inner mongolia grassland region is extremely high. Sneezing is the main clinical symptom. Rhinoconjunctivitis is the most common combined disease. High summer and autumn pollen exposure is the main cause of AR.

[Observation on pollinosis with vegetable food allergy].

Objective: To investigate the incidence of plant food allergy in patients with pollinosis and the effect of food allergens on the clinical symptoms of pollinosis patients. Method: A total of 40 patients with pollinosis and food allergy attended to the allergy Department of Duolun People's Hospital of Inner Mongolia were accepted skin prick test of inhaled allergens while the rhino conjunctivitis quality of life questionnaire(RQLQ) were also completed. The patients were divided into A and B groups randomly. Patients in group A were required for avoiding allergic plant foods intake but not them in group B. The two groups of patients with RQLQ, VAS and clinical symptom scores were statistically analyzed by P<0.05, the difference was statistically significant. Result: Forty cases were allergic to Artemisia. The most common allergic plant foods was peach, which accounted for 47.5%. Twenty-four patients were allergic to multiple foods simultaneously. Seventeen cases of pollinosis were preceded by food allergy, and 23 cases of food allergy were preceded by pollinosis. The mean values of RQLQ, VAS and symptom scores in group A were 81.44±14.31, 6.02±1.39, 10.60±3.68, respectively. The mean values of group B patients after 1 years were 100.73±21.66, 8.30±1.00, 13.45±3.51, the difference was statistically significant (P<0.01). The patients in group A complained that the symptoms were better than before. The mean values of RQLQ, VAS and symptom scores before intervention were 105.2±26.69, 7.00±1.71, 14.83±3.66, with significant difference (P<0.01). There was no significant improvement in the symptoms of Group B patients (P>0.05). Conclusion: Patients with pollinosis are often associated with food allergies. Reducing the intake of allergic plant foods should help alleviate symptoms.

[The clinical features and surgical treatment of facial basal cell carcinoma].

Objective:To investigate the clinical features and reasonable surgical treatment of facial basal cell carcinoma (BCC). Method:The clinical features of primary facial BCC from 37 patients were analyzed.All patients were cured by standard surgical excision with intraoperative frozen section examination and ensuring margins negative,the defects were reconstructed by using adaptive flaps after resection. Result:The patients were followed up for 12 to 36 months after operation.Two cases of 37(2/37,5.41%) with unclear margin developed local recurrence during 4-6 months after resection,the cure rate was 94.59% (35/37). Except 4 cases were suffered with distal skin flap necrosis, the others were well developed. Conclusion:The clinical feature of primary facial basal cell carcinoma was various. The complete resection of primary nidus with clear margin is the key to a successful treatment of facial basal cell carcinoma, and satisfactory therapeutic effects can be obtained after repairing defects by using adaptive flaps after resection.

Relation between blood pressure variability and early renal damage in hypertensive patients.

OBJECTIVE: The objective of the present study was to observe the relation between blood pressure variability (BPV) and early renal damage in hypertensive patients. PATIENTS AND METHODS: A total of 118 hypertensive patients were consecutively selected. General parameters including sex, age, duration, and grade of hypertension, antihypertensive drugs taken, smoking status, blood sugar, blood lipid level, body mass index, indexes of 24-h ambulatory blood pressure monitoring and renal function including cystatin C (CysC), serum creatinine (SCr), angiotensin II (Ang II), microalbuminuria (mALb), and urine creatinine (UCr) were measured. Glomerular filtration rate (eGFR), endogenous creatinine clearance rate (Ccr), and urine albumin/creatinine ratio (UACR) were calculated by CysC level, SCr level, and mALb and UCr level respectively. The 24-h ambulatory blood pressure monitoring indexes included 24-h mean systolic blood pressure variability (24h-SBPV), 24-h mean diastolic blood pressure variability (24h-DBPV), day mean systolic blood pressure variability (d-SBPV), day mean diastolic blood pressure variability (d-DBPV), night mean systolic blood pressure variability (n-SBPV), and night mean diastolic blood pressure variability (n-DBPV). RESULTS: Sixty-four hypertensive patients (54.24%) were non-dipper, and the baseline data of the two groups were comparable. The 24h-SBPV, 24h-DBPV, d-DBPV, n-SBPV and SCr, eGFR, and Ccr of the two groups showed no significant differences. The d-SBPV, n-DBPV, CysC, and Ang II of the non-dipper group were significantly higher than those of the dipper group (p<0.05). The mALb in both groups increased and was more obvious in the non-dipper group. UACR of the non-dipper group was significantly higher than that of dipper group (p<0.05), while UCr showed no difference. By Pearson correlation, d-SBPV and n-DBPV correlated positively (p<0.05) with CysC, Ang II, mALb, and UACR. CONCLUSIONS: BPV of hypersensitive patients, especially the d-SBPV and n-DBPV, was closely related to indexes of early renal damage including CysC, Ang II, mALb, and UACR.

RUNX2 Mutation Impairs 1α,25-Dihydroxyvitamin D3 mediated Osteoclastogenesis in Dental Follicle Cells.

Cleidocranial dysplasia (CCD), a skeletal disorder characterized by delayed permanent tooth eruption and other dental abnormalities, is caused by heterozygous RUNX2 mutations. As an osteoblast-specific transcription factor, RUNX2 plays a role in bone remodeling, tooth formation and tooth eruption. To investigate the crosstalk between RUNX2 and 1α,25-dihydroxyvitamin D3 (1α,25-(OH)2D3) in human dental follicle cells (hDFCs) during osteoclast formation, we established a co-culture system of hDFCs from CCD patient and healthy donors with peripheral blood mononuclear cells (PBMCs). Expression of the osteoclast-associated genes and the number of TRAP(+) cells were reduced in CCD hDFCs, indicating its suppressed osteoclast-inductive ability, which was reflected by the downregulated RANKL/OPG ratio. In addition, 1α,25-(OH)2D3-stimulation elevated the expression of osteoclast-related genes, as well as RANKL mRNA levels and RANKL/OPG ratios in control hDFCs. Conversely, RUNX2 mutation abolished this 1α,25-(OH)2D3-induced RANKL gene activation and osteoclast formation in CCD hDFCs. Therefore, RUNX2 haploinsufficiency impairs dental follicle-induced osteoclast formation capacity through RANKL/OPG signaling, which may be partially responsible for delayed permanent tooth eruption in CCD patients. Furthermore, this abnormality was not rescued by 1α,25-(OH)2D3 application because 1α,25-(OH)2D3-induced RANKL activation in hDFCs is mediated principally via the RUNX2-dependent pathway.

Valve replacement for valvular heart disease with giant left ventricle.

OBJECTIVE: To discuss early efficacy of the cardiac patient with giant left ventricle underwent valve replacement. PATIENTS AND METHODS: Fifty-five patients, who are suffering cardiac valve disease with giant left ventricle, underwent valve replacement. Among them, sixteen patients underwent aortic valve replacement; thirty patients underwent mitral valve replacement; nine patients underwent double valve replacement. All of them use mechanical heart valve. RESULTS: The number of early death after operation was five. Two patients died of malignant arrhythmia; two died of intractable low cardiac output syndrome; the last one's mechanical valve lost its ability to function after operation and died of respiration-circulation failure after an emergency operation. The death rate was 9%. The remaining 55 patients were cured and their cardiac function was significantly improved. CONCLUSIONS: The definite effective myocardial protection and perfect and detailed preoperative treatment can reduce the possibility of operative complications and death rate of this kind of patients.

Abnormal differentiation of dental pulp cells in cleidocranial dysplasia.

Cleidocranial dysplasia (CCD) is a skeletal dysplasia caused by heterozygous mutations of RUNX2, a gene that is essential for the mineralization of bone and tooth. We isolated primary dental pulp cells from a 10-y-old patient and tested their proliferative capacity, alkaline phosphatase activity, and ability to form mineralized nodules, in comparison with those from 7 healthy children. All these measures were reduced in primary dental pulp cells from the CCD patient. The expression of the osteoblast/odontoblast-associated genes RUNX2, ALP, OCN, and DSPP was also found to be significantly decreased in the primary dental pulp cells of the CCD patient. The osteoclast-related markers TRAP, CTSK, CTR, and MMP9 were decreased in primary dental pulp cells cocultured with human peripheral blood mononuclear cells. Moreover, the expression of RANKL and the ratio of RANKL/OPG were both reduced in the cells from the CCD patient, indicating that the RUNX2 mutation interfered with the bone-remodeling pathway and decreased the capacity of primary dental pulp cells to support osteoclast differentiation. These effects may be partly responsible for the defects in tooth development and the retention of primary teeth that is typical of CCD.

A morphometric study of the lumbar spinous process in the Chinese population

Our goal was to analyze the anatomical parameters of the lumbar spine spinous process for an interspinous stabilization device designed for the Chinese population and to offer an anatomical basis for its clinical application. The posterior lumbar spines (T12-S1) of 52 adult cadavers were used for measuring the following: distance between two adjacent spinous processes (DB), distance across two adjacent spinous processes (DA), thickness of the central spinous processes (TC), thickness of the superior margin of the spinous processes (TS), thickness of the inferior margin of the spinous processes (TI), and height of the spinous processes (H). Variance and correlation analyses were conducted for these data, and the data met the normal distribution and homogeneity of variance. DB decreased gradually from L1-2 to L5-S1. DA increased from T12-L1 to L2-3 and then decreased from L2-3 to L4-5. The largest H in males was noted at L3 (25.45±5.96 mm), whereas for females the largest H was noted at L4 (18.71±4.50 mm). Usually, TS of the adjacent spinous process was lower than TI. Based on the anatomical parameters of the lumbar spinous processes obtained in this study, an “H”-shaped coronal plane (posterior view) was proposed as an interspinous stabilization device for the Chinese population. This study reports morphometric data of the lumbar spinous processes in the Chinese population, which provides an anatomical basis for future clinical applications.