[Current status and progress of detection methods for common clinical toxicants].

With the progress of science and technology and the development of society, more and more chemical substances have been discovered and countless chemicals have been artificially synthesized, and the risk of exposure to some toxic chemicals by human beings has been greatly increased, resulting in the increasing incidence of acute poisoning, which has seriously endangered the public's physical health and life safety. As the poisoned patients are unconscious or refuse treatment when they are admitted to the hospital, it is difficult to understand the drug exposure history by asking the medical history, so the toxicity detection has become the key to the clinical diagnosis and treatment, and this paper briefly introduces some common toxicity detection methods in the clinic in the hope that it will bring help to the clinical doctors.

[A long-term evaluation of periodontal phenotypes before and after the periodontal-orthodontic-orthognathic combined treatment of lower anterior teeth in patients with skeletal Angle class â ¢ malocclusion].

OBJECTIVE: To evaluate the changes of periodontal phenotype (width of keratinized gingiva, thickness and height of alveolar bone) of lower anterior teeth in patients with skeletal class Ⅲ malocclusion before and after the periodontal-orthodontic-orthognathic combined treatment. METHODS: In the study, 20 patients with skeletal class Ⅲ malocclusion (6 males and 14 females) completed the periodontal-orthodontic-orthognathic combined treatment were included from March 2017 to June 2022, with 39 central incisors, 40 lateral incisors and 40 canines. The mean age was (25.40±4.27) years (20-34 years). The mean follow-up time was (3.70±1.05) years from the beginning of periodontal corticotomy regenerative surgery (PCRS) to the end of the combined treatment. Cone-beam computed tomography (CBCT) was used to measure the thickness, area and height of alveolar bone by the same researcher, taken before the PCRS (T0), 6 months after the PCRS (T1), 12 months after the PCRS (T2), before the orthognathic surgery (T3), and after the periodontal-orthodontic-orthognathic combined treatment (T4). The periodontal clinical parameters were used to evaluate changes in the soft tissue by another researcher, measured before the PCRS (T0) and after the combined treatment (T4). Changes of soft and hard tissue were evaluated by the periodontal phenotype. RESULTS: The width of keratinized gingiva increased significantly (all P < 0.001) in lower anterior teeth, the central incisors, lateral incisors and canines increased by (1.82±1.57) mm, (2.03±1.48) mm and (2.05±1.27) mm, respectively. The proportion of thick periodontal biotype in the central and lateral incisors increased significantly (all P < 0.001), while the changes of periodontal biotypes in the lower canines were not obvious. The thickness of labial alveolar bone of lower anterior teeth all increased significantly after periodontal corticotomy regenerative surgery and the combined treatment (all P < 0.001). The area of labial alveolar bone of lower anterior teeth also increased significantly after the combined treatment (all P < 0.001). The whole area of labial and lingual alveolar bone of central and lateral incisors increased (P < 0.001), while the whole area of canines remained the same. All The height of the alveolar bone increased (all P < 0.001) on the labial side after the treatment. CONCLUSION: The periodontal phenotypes of lower anterior teeth were significantly improved after the periodontal-orthodontic-orthognathic combined treatment in patients with skeletal Angle class Ⅲ malocclusion. The improvement was long-termly stable, and the periodontal risk was reduced.

[Effects of specimen preservation and transportation on blood paraquat concentration in rats].

Objective: To investigate the effects of duration, temperature and shake on paraquat (PQ) concentration in the blood of PQ-exposed rats during the specinen preservation and transportation. Methods: In March 2021, 60 SD male rats of Specific Pathogen Free class were randomly divided into low-dose group (10 mg/kg PQ) and high-dose group (80 mg/kg PQ). Each group was divided into 5 subgroups (normal temperature group, cold storage group, 37 ℃ storage group, shaking on normal temperature group and shaking on 37 ℃ group), six rats in each subgroup. The rats were given intraperitoneal injection of PQ, 1 h after exposure, the blood samples were obtained by cardiac extraction. After different interventions, the concentrations of PQ were detected and compared before and after the intervention in each subgroup. Results: In the shaking on 37 ℃ group, the results of PQ concentrations in PQ-exposed rats were significantly lower than those before the intervention (P<0.05). In the other subgroups, the results were not significantly different compared with before intervention (P>0.05) . Conclusion: The concentration of PQ in the blood of rats exposed to PQ was decreased by shaking for 4 hours at 37 ℃.

[Identification and preservation of arm lymphatics in axillary lymph node dissection to prevent arm lymphedema: a single center randomized controlled trial].

Objective: To evaluate the feasibility of identification and preservation of arm lymphatics (DEPART) in axillary lymph node dissection (ALND) for breast cancer to prevent arm lymphedema. Methods: A randomized controlled study method was used. Two hundred and sixty-five patients who underwent breast cancer surgery at the Department of Thyroid and Breast Surgery, Zhongnan Hospital of Wuhan University from November 2017 to June 2018 were included, and the patients were randomly divided into ALND+ DEPART group (132 patients) and standard ALND group (133 patients) by random number table method. In the ALND+ DEPART group, indocyanine green and methylene blue were injected as tracers before surgery, and the arm sentinel nodes was visualized by staged tracing during intraoperative dissection of axillary lymph nodes. Partial frozen sections were made of arm lymph nodes >1 cm in length and hard and suspicious of metastasis, and arm lymph nodes and lymphatic vessels were selectively preserved. Patients in the standard ALND group underwent standard ALND. Objective and subjective indexes of arm lymphedema were evaluated by 5-point circumference measurement and Norman questionnaire. Results: Among 132 breast cancer patients in the ALND+ DEPART group, 121 (91.7%) completed DEPART. There were no statistically significant differences in age, body mass index, pathological type, dissection number of axillary lymph node, N stage, TNM stage, molecular typing, and regional radiotherapy between the ALND+ DEPART and standard ALND groups (P>0.05). At a median follow-up of 24 months, assessment by the 5-point circumference measurement showed that the incidence rates of lymphedema in the ALND+ DEPART and standard ALND groups were 5.0% (6/121) and 15.8% (21/133), respectively, with statistically significant differences (P=0.005). Assessment by the Norman questionnaire showed that the incidence rates of lymphedema in the ALND+ DEPART and standard ALND groups were 5.8% (7/121) and 21.8% (29/133), respectively, with a statistically significant difference (P<0.001). No local regional recurrence was observed in either group during the follow-up period. Conclusion: For breast cancer patients with positive axillary lymph nodes, the administration of DEPART during ALND can reduce or avoid the occurrence of arm lymphedema without compromising oncology safety.

[Expression and distribution of calprotectin in healthy and inflamed periodontal tissues].

OBJECTIVE: Calprotectin, the heterdimer of S100A8 and S100A9, is the major cytoplasmic protein of neutrophils, which is also expressed or induced in gingival epithelial cells, activated mononuclear macrophages and vascular endothelial cells. Calprotectin is intimately associated with the initiation and progression of periodontitis, but the in vivo expression patterns of calprotectin in healthy and inflamed periodontal tissue are not fully understood. To observe the expression, distribution and cellular localization of calprotectin in the samples of healthy periodontal tissues and experimental periodontitis tissues of Beagles and to explore their relationship with periodontal inflammation and possible effect. METHODS: Experimental periodontitis model was established by ligation around the mandibular second molar of the Beagle dogs, while the contralateral teeth were healthy controls. Induction duration was 12 weeks, before the dogs were executed. Tissue specimens were demineralized and serial sections were made conventionally. The in vivo expression of calprotectin in the healthy and inflamed periodontal tissues were examined by immunohistochemistry. The in vitro expression of calprotectin in human primary gingival fibroblasts (GFs) and periodontal ligament (PDL) cells were detected by immunocytochemistry. RESULTS: Immunohistochemistry analysis indicated that calprotectin was expressed in gingival epithelial cells and infiltrated neutrophils in the healthy periodontium within the gingival epithelium, S100A8/A9 was most strongly expressed in the junctional epithelium, followed by surface epithelium, and least expressed in the sulcular epithelium. The S100A8/A9 expression levels were sharply defined at the junction between the junctional epithelium and the sulcular epithelium. In periodontal inflammatory lesions, the expression level of calprotectin in sulcular epithelium and junctional epithelium was up-regulated than that in the healthy gingival epithelium. Calprotectin was inducibly expressed in fibroblast-like cells in gingival connective tissue and periodontal ligament tissue, microvascular endothelial cells (ECs) and bone marrow fibroblasts under inflammatory conditions. Additionally, the expression of calprotectin in primary human GFs and PDL cells was confirmed by immunnocytochemistry staining. CONCLUSION: Constitutively expressed in neutrophils and gingival epithelial cells, and calprotectin might maintain the homeostasis and integrity of periodontium. Inflammation-induced expression of calprotectin in GFs, PDL cells, microvascular ECs and bone marrow fibroblasts might process anti-microbial function and promote leukocytes transmigration to defend the host against the microorganisms.

[Functional axillary dissection based on lymphatic drainage for breast cancer: a single center randomized clinical trial].

Objective: To evaluate the effectiveness and safety of functional axillary dissection based on lymphatic drainage (FUND) in decreasing breast cancer-related lymphedema (BCRL) events. Methods: A total of 168 eligible patients in Zhongnan Hospital of Wuhan University from July 2018 to February 2019 were randomly assigned to the FUND group or axillary lymph node dissection (ALND) group using random number table generated by SPSS. In the FUND group, methylene blue (MB) was adopted to reveal the sentinel lymph node (SLN) for all patients; 0.1 ml MB was injected into the SLNs before resection to reveal the efferent lymphatic channels and subsequent-echelon lymph node. The blue-stained lymphatic channels were mapped by bluntly dissecting along the lymphatic drainage channels from the breast to the axilla. Then, the SLNs were removed and pathologically analyzed by immediate frozen sectioning (FS); if the SLNs were positive, the blue-stained bALNs in breast lymphatic level (BLL) Ⅱ were removed and sent for immediate FS; if the blue-stained ALNs in BLL Ⅱ were confirmed negative, the tissues in BLL Ⅱ were removed'en bloc'. Clinicopathologic information for all the patients in the two groups were collected. The fixed-point circumference volume measurement method and the Norman questionnaire scoring method were used to evaluate the arm lymphedema between the two groups. Clinicopathological characteristics, incidences of arm lymphedema, locoregional recurrence, and distant metastasis between the two groups were compared. Results: The mean age were (50.3±8.0) in the FUND group and (51.1±9.0) in the ALND group. Seventy-four cases (88.1%) in the FUND group successfully underwent FUND surgery, and patients whose breast lymphatics failed to be stained blue underwent standard ALND. There was no statistically significant difference in terms of age, BMI, histological types, surgical approaches and adjunct therapy between the FUND group (n=74) and ALND group (n=84) (P>0.05). The average operation time of the FUND group and the stand ALND group were (169±15) and (123±12) min respectively (range: 145-198, 103-146 min) (P<0.001), and the number of lymph nodes removed [M (Q1, Q3)] were 8.3 (6, 15) and 12.9 (7, 18) (P=0.019). The cumulative BCRL rate, within a median follow-up of 24 months and 23 months respectively for FUND and ALND group, were 10.8% (8/74) vs 23.8% (20/84) (P=0.033) measured by fixed-point circumference volume measurement method, and was 12.2% (9/74) vs 27.4% (23/84) by Norman questionnaire (P=0.018). There were no local regional recurrence events during the follow-up period between the two groups. Conclusion: For breast cancer patients with clinically node-positive axilla or positive SLN, FUND based on lymphatic drainage was a less radical axillary surgery, with which eliminating the risk of BCRL might be achieved.

[Factors associated with identification of lymph node detected by axillary reverse mapping for breast cancer].

Objective: To investigate the different tracer materials in identifying the axillary reverse mapping(ARM) lymph nodes. Methods: A retrospective analysis of clinical and pathological data of 478 breast cancer female patients(mean age: 50.5±8.0) under axillary lymph node dissection(ALND) with ARM technique was conducted between March 2019 and November 2020 in Wuhan University Zhongnan Hospital. Of the 478 patients, methylene blue was applied in 147 patients, indocyanine green in 119, and indocyanine green plus methylene blue in 212 patients. Wilcoxon rank-sum test, Chi-squire test or Fisher test, and binary logistic regression were carried out to identify the factors associated with identifying ARM lymph nodes. Results: The recognition rates of ARM lymph nodes were 73.5%, 79.0%, and 83.0%(P=0.091), and the recognition rate of ARM lymphatic vessels was 62.6%, 92.4%, 89.6%(P<0.001), respectively. The coincidence rate of ARM lymph node and SLN was 8.1%(12/148), and the metastasis rate was 16.1%(61/378). Supplemental injection of 1 ml of methylene blue or indocyanine green can improve the identification of ARM lymph nodes. The larger BMI and the performance of neoadjuvant therapy were associated with the lower recognition rate of ARM lymph nodes. Neoadjuvant therapy was an independent factor for the identification rate of ARM lymph nodes. Conclusions: Indocyanine green combined with methylene blue can improve the recognition rate of ARM lymph nodes. Obese patients have a lower recognition rate of ARM lymph nodes, and the supplemental injection tracer can be injected to improve the recognition rate. In breast cancer patients whose ARM lymph nodes are not successfully identified during operation, it may be that the ARM lymph nodes are not located in the axilla.

[Lesion removal plus whole breast exploration and washing plus micro-plastic procedures in the treatment of granulomatous lobular mastitis: a randomized controlled study].

Objective: To examine the effect of"lesion removal plus whole breast exploration and washing plus micro-plastic surgery"in granulomatous lobular mastitis. Methods: A single-center prospective randomized controlled study method was used to enroll patients diagnosed with granulomatous lobular mastitis for whom surgical procedures were projected from March 2017 to September 2019 at Department of Thyroid and Breast Surgery, Zhongnan Hospital of Wuhan University. The sample size is determined by the superiority test. Based on the literatures and the previous work, the two groups require 97 cases. Fifty-two patients underwent"lesion removal+whole breast exploration and washing plus micro-plastic surgery"(observation group). Forty-five cases underwent"empirical breast lesion resection plus fascia tissue flap plasty plus nipple and areola correction"(control group). The primary study endpoint is the recurrence rate, and the secondary study endpoints include surgical complications, incision healing time, and postoperative patient satisfaction. Independent sample t test, Wilcoxon rank-sum test, χ² test and Fisher exact test were used for comparison between groups. Results: All procedures were completed successful, with no severe complications. All patients were followed up for (15.2±1.9) months (range: 12 to 24 months). There were no significant differences in incidence of postoperative complications (7.7% (4/52) vs. 6.7%(3/45), P=1), drainage time ((8.6±0.6) days vs. (8.4±0.8) days, t=1.921, P=0.053) and hospital stay ((7.7±0.6) days vs. (7.6±0.5) days, t=1.633, P=0.102) between the two groups. The recurrence rate of the observation group was lower significantly than that of the control group (3.8% (2/52) vs. 24.4%(11/45), χ²=8.819, P=0.003). The observation group had better cosmetic effects (Z=-2.657, P=0.008) and patient satisfaction than control group (Z=-5.730, P=0.000). Conclusion: The "lesion removal plus whole breast exploration and flushing plus micro-plastic surgery" has a good therapeutic effect and cosmetic value for patients with refractory granulomatous lobular mastitis.

[Comparison of residual cement between CAD/CAM customized abutments and stock abutments via digital measurement in vitro].

OBJECTIVE: To compare the residual cement between computer aided design/computer aided manufacturing customized abutments (CCA) and stock abutments (SA), and to evaluate the feasibility of digital measurement for residual cement volume by three-dimensional scanning. METHODS: Twenty master models needed in this study were all taken from one 47-year-old patient with arrested periodontitis, who had already had an implant placed at his right upper central incisor site in the Department of Periodonto-logy, Peking University School and Hospital of Stomatology. After 4 weeks of soft tissue conditioning by means of customized healing abutment, the height of peri-implant soft tissue was measured, from the implant platform to mucosal margin, as 5 mm. Using customized impression coping, the impression was taken and twenty models were fabricated and allocated to 4 groups according to the type of abutments: CCA1 (5 mm transmucosal height CCA, with margin at tissue level), CCA2 (4 mm transmucosal height CCA, with 1 mm submucosal margin), SA1 (3 mm transmucosal height SA, with 2 mm submucosal margin) and SA2 (1 mm transmucosal height SA, with 4 mm submucosal margin). Crowns were cemented to the abutments, which were seated on the working models. Excess cement was removed by a prosthodontic specialist. Thereafter, the volume of residual cement was evaluated by using three-dimensional scanning technique. The area proportion of residual cement was calculated on photographs taken by a single lens reflex camera. The weight of residual cement was weighed by an analytical balance. And the correlation of residual cement volume data with residual cement area proportion or weight of residual cement acquired by traditional methods was analyzed. RESULTS: Residual cement was observed on all the experiment samples. The residual cement volume of CCA was significantly less than that of SA [(0.635 3±0.535 4) mm3 vs. (2.293 8±0.943 8) mm3, P < 0.001]. Consistently, CCA had less residual cement area proportion and weight than those of SA [area proportion: 7.57%±2.99% vs. 22.68%±10.06%, P < 0.001; weight: (0.001 5±0.001 0) g vs. (0.003 7±0.001 4) g, P < 0.001]. The residual cement volume was strongly correlated with the residual cement area proportion and residual cement weight (r>0.75, P < 0.001). CONCLUSION: These in vitro results suggest that CCA minimized the residual cement more effectively than SA. The method to digitally evaluate the residual cement volume is feasible, but its validity and reliability need to be further studied.

[Effect of initial periodontal therapy on blood parameters related to erythrocyte and platelet in patients with type 2 diabetes mellitus and chronic periodontitis].

OBJECTIVE: To compare the blood parameters related to erythrocyte and platelet between baseline and 3 months after initial periodontal therapy in patients with both type 2 diabetes mellitus and chronic periodontitis (DM-P). METHODS: According to the International Symposium on Classification of Periodontal Diseases and Conditions in 1999 and the diagnostic criteria of type 2 diabetes mellitus proposed by the World Health Organization in 1999, 35 patients with DM-P were recruited. All the participants received initial periodontal therapy, including oral hygiene instruction, scaling, and root planning provided by one senior periodontist. Original diet, exercise, and medication for blood glucose control were unchanged for all the participants. At baseline and 3 months after initial periodontal therapy, the clinical periodontal parameters, including probing depth (PD), bleeding index (BI) and clinical attachment loss (CAL); erythrocyte-related indexes, including red blood cell (RBC) count, hemoglobin (HGB), hematocrit (HCT), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), RBC volume distribution width (RDW); platelet-related indexes, including platelet (PLT) count, mean platelet volume (MPV), platelet distribution width (PDW), plateletocrit (PCT) were measured and compared. RESULTS: Compared with baseline, the periodontal parameters, including PD [(3.370±0.601) mm vs. (2.729±0.431) mm], BI [2.160 (1.550~3.410) vs. 1.420 (1.000~2.970)] and CAL [(3.307±1.577) mm vs. (2.990±1.587) mm], were significantly reduced (P < 0.001) three months after the initial periodontal therapy; the erythrocyte-related indexes, including RBC count [(4.727±0.392)×1012/L vs. (4.825±0.394)×1012/L, P=0.010], HGB [(145.886±11.792) g/L vs. (149.200±12.979) g/L, P=0.007] and HCT [43.40% (37.50%~48.50%) vs. 43.80% (38.50%~53.20%), P=0.003], were significantly increased three months after the initial periodontal therapy; PLT count [(216.714±61.900)×109/L vs. (205.886±62.051)×109/L, P=0.016] was significantly reduced 3 months after the initial periodontal therapy. CONCLUSIONS: The initial periodontal therapy can significantly improve blood parameters related to RBC and PLT, which might decrease the risk of vascular complications in DM-P patients.

[Alveolar crest and relevant analysis of labial side of anterior teeth on skeletal Angle class III patients].

OBJECTIVE: To measure the distance from cemento-enamel junction (CEJ) to alveolar crest (AC) of labial side of anterior teeth on skeletal Angle class III patients under direct vision during periodontal bone augmentation surgery and to make relevant analysis to find the relevant factors. METHODS: In the study, 46 skeletal Angle class III patients (10 males and 36 females) received periodontal bone augmentation surgery of anterior teeth were included, with 67 jaws (27 maxillae and 40 mandibles) and 400 anterior teeth (161 maxillary anterior teeth and 239 mandibular anterior teeth). The mean age was 23.65 years. Maxillary anterior teeth consisted of 54 central incisors, 53 lateral incisors and 54 canines. Mandibular anterior teeth consisted of 79 central incisors, 80 lateral incisors and 80 canines. CEJ-AC was measured in three sites (mesial sites, central sites and distal sites) by Williams periodontal probes during periodontal bone augmentation surgery under direct vision by the same researcher. RESULTS: The average CEJ-AC of 400 anterior teeth was (2.21±1.48) mm. The average CEJ-AC of maxillary anterior teeth was (1.72±1.13) mm, more than (2.54±1.60) mm of mandibular anterior teeth (P<0.05). The average CEJ-AC of canines was (2.42±1.78) mm, more than (2.06±1.27) mm of central incisors or (2.16±1.32) mm of lateral incisors (P<0.05). The average CEJ-AC of central sites was (3.04±2.01) mm, more than (1.79±0.86) mm of mesial sites or (1.81±0.89) mm of distal sites (P<0.05). CEJ-AC of 233 anterior teeth was more than 2 mm, accounting for 58.25%, and 117 anterior teeth with dehiscence were found, accounting for 29.25%. Multilevel and multivariate Logistic regression showed age, jaw, tooth and site were the relevant factors to the position of alveolar crest. CONCLUSION: The position of alveolar crest of skeletal Angle class III patients who received periodontal bone augmentation surgery was lower than that of the general population, causing periodontal risks during decompensation orthodontics therapy before orthognathic surgery. The position of alveolar crest was lower in older patients than in younger patients, in mandibular teeth than in maxillary teeth, in canines than in central incisors or lateral incisors, and in central sites than in mesial sites or distal sites of labial side, which showed much higher risk.

[Clinical outcomes of ultrasonic subgingival debridement combined with manual root planing in severe periodontitis].

OBJECTIVE: To compare the clinical effects of ultrasonic subgingival debridement and ultrasonic subgingival debridement combined with manual root planing on severe periodontitis and then to investigate the necessity and significance of manual root planing. METHODS: Twenty-three patients with severe periodontitis participated in this split-mouth randomized-controlled clinical trial. Baseline examination and randomization were performed after supragingival scaling: each of the upper and lower jaws had a quadrant as the test group treated with ultrasonic subgingival debridement combined with manual root planing, whereas the other two quadrants were the control group treated with ultrasonic subgingival debridement. Treatment of each patient was at intervals of one week and completed in two visits. Clinical indicators concerning probing depth (PD), clinical attachment loss (CAL) and bleeding index (BI) were recorded at baseline and 1 month, 3 months, 6 months after treatment. RESULTS: There was no significant difference of periodontal indicators between the test group and the control group at baseline. Both the test group and control group resulted in significant improvement of PD, CAL and BI. One and three months after treatment, reduction of PD in the test group was higher than that in the control group [1 month: (2.13±1.31) mm vs. (1.79±1.33) mm, P<0.01; 3 months: (2.46±1.33) mm vs. (2.17±1.38) mm, P<0.01] and reduction of CAL in the test group was higher than that in the control group [1 month: (1.89±2.03) mm vs. (1.65±1.93) mm, P<0.01; 3 months: (2.03±2.05) mm vs. (1.83±1.97) mm, P<0.05]. Six months after treatment, PD in the test group and the control group decreased by (2.52±1.40) mm and (2.35±1.37) mm respectively, and the improvement in the test group was significantly better than that in the control group (P<0.01). CAL in the test group and the control group decreased by (1.89±2.14) mm and (1.77±2.00) mm respectively, and there was no statistical difference between the groups. There was no significant difference in the changes of BI between the two groups 1, 3 and 6 months after treatment. CONCLUSION: Ultrasonic subgingival debridement combined with manual root planing has more reduction in PD and CAL compared with ultrasonic subgingival debridement. Therefore, it is still necessary to use manual instruments for root planing following ultrasonic subgingival debridement.

[Influence of vitamin D receptor FokI polymorphism on expression of CYP24A1 in periodontal cells].

OBJECTIVE: There is asingle nucleotide polymorphism (SNP) in the exon 2 of the vitamin D receptor (VDR) gene that can be distinguished using the restriction endonuclease FokI, and accordingly divided into three genotypes: FF, Ff and ff. VDR-FokI polymorphism was the only known SNP that could alter the protein structure of VDR. CYP24A1 is the gene encoding vitamin D 24 hydroxylase and is a vitamin D responsive gene. The influence of rs2228570 on transcriptional activation by VDR in human gingival fibroblasts (hGF) and periodontal ligament cells (hPDLC) was investigated in this study. METHODS: hGF and hPDLC of 12 donors' were primarily cultured and genomic DNA was extracted. A part of genomic DNA with the length of 267 bp was obtained using PCR, which contained the SNP. VDR-Fok I genotypes were determined according to the results of restriction fragment length polymorphism. hGF and hPDLC were stimulated with 10 nmol/L 1α,25 dihydroxy vitamin D3 (1,25OH2D3) or 1 000 nmol/L 25 hydroxy vitamin D3 (25OHD3) for 48 h before RNA was extracted. Then VDR antagonist ZK159222 was used or not used during 1,25OH2D3 or 25OHD3 stimulation with hGF and hPDLC. After 1,25OH2D3 stimulation for 48 h, the proteins in hGF and hPDLC were also collected. The protein expressions of CYP24A1 and VDR were detected using Western blot. RESULTS: Among the 12 donors' cell cultures, the number of FF, ff and Ff genotypes was 4, 3 and 5, respectively.After stimulation with 1,25OH2D3 or 25OHD3 for 48 h,CYP24A1 mRNA levels in FF-hGF were significantly higher than those in other hGF genotypes(1,25OH2D3: F=31.147, P<0.01; 25OHD3: F= 32.061,P <0.01), as was in FF-hPDLC (1,25OH2D3: F=23.347, P<0.01; 25OHD3: F=32.569,P<0.01). When ZK159222 was used before 1,25OH2D3 stimulation, this statistically significant difference disappeared (hGF: F=0.246, P=0.787; hPDLC: F=0.574, P=0.583). When ZK159222 was used before 25OHD3 stimulation, the trend was similar (hGF: F=1.636, P=0.248; hPDLC: F=0.582, P=0.578).After stimulation with 1,25OH2D3 for 48 h, CYP24A1 protein levels in FF-hGF were significantly higher than those in the other hGF genotypes (F=12.368, P <0.01), as was in FF-hPDLC (F=15.749, P <0.01). In hGF and hPDLC, the mRNA or protein expression of VDR of different genotypes was not significantly different under different stimulation conditions.The paired comparison showed that there was no statistically significant difference between the expression of CYP24A1 in hGF and that in hPDLC under all the stimulation conditions, as was the expression of VDR. CONCLUSION: In hGF and hPDLC, the FF-VDR genotype is associated with the more remarkable up-regulation of CYP24A1than the other genotypes, indicating that transcriptional activation of FF-VDR might be higher than those of other vitamin D receptors.

Fibronectin chorused cohesion between endothelial progenitor cells and mesenchymal stem cells of mouse bone marrow.

Endothelial progenitor cells (EPCs) could function as niche cells to promote self—renewal of mesenchymal stem cells (MSCs) in the mouse bone marrow. Cohesion was the basis of the two cells to display their biological functions to each other. In this study, we investigated the mechanism of cohesion between MSCs and EPCs. And demonstrated that fibronectin (FN) in EPCs activated the integrin α5β1 of MSCs and further mediated cell-cell cohesion. Integrin α5β1 and its FN ligand played critical roles not only in single—cell line adhesion, but also in adhesion between stem and niche cells. This novel finding is important to understand the cross—talk between MSCs and their niche cells.

Two missense mutations in exon 9 of caprine PRLR gene were associated with litter size.

Guanzhong (n = 321) and Boer (n = 191) goat breeds were used to detect single nucleotide polymorphisms (SNPs) in the coding regions of the prolactin receptor (PRLR) gene by DNA sequencing and PCR-RFLP. Two SNPs (c.1457G>A and c.1645G>A) were identified that caused amino acid variations p.Ser485Asn and p.Val548Met respectively. Statistical results indicated that the c.1457G>A and c.1645G>A SNPs were significantly associated with litter size in Boer and Guanzhong goat breeds. Further analysis revealed that combined genotype C4 (GGGG) and haplotype G-G were better than the others for litter size in both goat breeds. These results might contribute to goat genetic resources and breeding.

Two mutations in the 5'-flanking region of the KITLG gene are associated with litter size of dairy goats.

In this study, Xinong Saanen (SN) and Guanzhong (GZ) dairy goat breeds were used to detect single nucleotide polymorphisms (SNPs) in the 5'-flanking region of the KITLG gene by DNA sequencing and primer-introduced restriction analysis-polymerase chain reaction. Two novel SNPs (g.13090G>T and g.13664C>A) were identified (GenBank Accession no. KM658964). Furthermore, g.13090G>T and g.13664C>A loci were closely linked in SN and GZ breeds (r(2) > 0.33). Association analysis results showed that g.13090G>T and g.13664C>A SNPs significantly affected litter size (P < 0.05). The litter size of individuals with the combined genotype GG/CC from both dairy goat breeds was greater than that of individuals with TT/AA in average parity (P < 0.05). Known biochemical and physiological functions, along with our results, indicated that GG/CC could be used in marker-assisted selection to choose individuals with greater litter size from both breeds. These results extend the spectrum of genetic variation in the caprine KITLG gene and may contribute to genetic resources and breeding of goats.

Polymorphisms of PRLR and FOLR1 genes and association with milk production traits in goats.

We investigated the polymorphisms of PRLR and FOLR1 genes in Xinong Saanen, Guanzhong, and Boer goat breeds by DNA sequencing and PCR-RFLP. Two novel SNPs were identified: KC109741: g.62130C>T in the 3ꞌ-UTR of goat gene PRLR, and KC136296: g.7884A>C in exon 3 of goat gene FOLR1. In the three goat breeds, the polymorphism information content was 0.20-0.27 at the g.62130C>T locus. At the g.7884A>C locus, it was 0.36 in Boer goats. The three goat breeds were in Hardy-Weinberg disequilibrium at the g.62130C>T locus. The g.62130C>T SNP was found to be significantly associated with milk production traits in Xinong Saanen and Guanzhong breeds. These results are consistent with the regulatory function of PRLR in mammary gland development, milk secretion, and expression of milk protein genes; they extend the spectrum of genetic variation of the goat PRLR gene, which could be useful for breeding programs.

Polymorphism identification in goat GNRH1 and GDF9 genes and their association analysis with litter size.

This study investigated the polymorphisms of GNRH1 and GDF9 genes in 641 goats of three breeds: Xinong Saanen, Guanzhong and Boer. Two allelic variants were identified in the GNRH1 gene (JN645280:g.3548A>G and JN645281:g.3699G>A) and one allelic variant was found in the GDF9 gene (JN655693:g.4093G>A). Furthermore, g.4093G>A was a missense mutation (p.Val397Ile of GDF9). Results of an association analysis indicated that SNPs g.3548A>G and g.4093G>A had significant effects on litter size (P < 0.05). The combination genotypes of SNPs g.3548A>G, g.3699G>A and g.4093G>A also affected litter size with the C5 genotype having the highest litter size in the first, third, fourth and average parity. Hence, the biochemical and physiological functions, together with the results obtained in our investigation, suggest that the GNRH1 and GDF9 genes could serve as genetic markers for litter size in goat breeding.

Molecular cloning and characterization of KISS1 promoter and effect of KISS1 gene mutations on litter size in the goat.

Kisspeptins, the product of the KISS1 gene, play an essential role in the regulation of reproductive functions, acting primarily at the hypothalamic level of the gonadotropic axis. We detected polymorphisms of the goat KISS1 gene in 723 individuals from three goat breeds (Xinong Saanen, Guanzhong, and Boer) by DNA pooling, PCR-RFLP, and DNA sequencing methods. We cloned the promoter sequence of this gene and found it to share high similarity with that of the bovine KISS1 promoter. Six TATA boxes were found in the goat KISS1 promoter region. Two novel SNPs (g.2124T>A and g.2270C>T) were identified in the intron 1 of the KISS1 gene of all three goat breeds. The three goat breeds were in Hardy-Weinberg disequilibrium at g.2124T>A and g.2270C>T loci. The g.2124T>A and g.2270C>T loci were closely linked in the three goat breeds (r2 > 0.33). The g.2124T>A and g.2270C>T SNPs were significantly associated with litter size, and the C1 female goats had a larger litter size than did those with the other genotypes. These results extend the spectrum of genetic variation of the goat KISS1 gene, which contributes to our knowledge of goat genetic resources for breeding programs.

[Clinical and radiographic effect of simultaneously labial and lingual augmented corticotomy: a prospective case series study].

Objective: To investigate soft-and hard-tissue changes after simultaneously labial and lingual augmented corticotomy in patients with insufficient alveolar bone thickness of lower anterior teeth both in labial and lingual side during orthodontic treatment. Methods: From January 2021 to June 2022, 10 patients [2 males and 8 females, (26.2±3.1) years old] who received orthodontic and orthognathic combined treatment from the Fourth Clinical Division, Peking University School and Hospital of Stomatology were selected. The alveolar bone thickness of lower anterior teeth both in labial and lingual side in these patients was less than 0.5 mm according to cone-beam CT examination before or during treatment, and 60 lower anterior teeth were included. The 10 patients were treated with simultaneously labial and lingual augmented corticotomy. The differences in gingival recession, papilla index and the differences in labial and lingual alveolar bone thickness of lower anterior teeth were compared. Results: Six months after surgery, the alveolar bone thicknesses at the 4 mm under cemento-enamel junction (CEJ), 8 mm under CEJ and at the apical level [labial side: (1.02±0.39), (2.22±0.89) and (4.87±1.35) mm; lingual side: (1.07±0.46), (2.31±1.04) and (3.91±1.29) mm] were significantly higher than that before surgery [labial side: (0.02±0.09), (0.06±0.21) and (2.71±1.33) mm]; lingual side: (0.14±0.29), (0.40±0.52) and (2.13±1.02) mm] (P<0.001), respectively. The increases in alveolar bone thickness of central incisors [apical level on labial side: (2.53±1.20) mm, 8 mm under CEJ on lingual side: (2.27±1.24) mm, apical level on lingual side: (2.66±1.49) mm] and lateral incisors [apical level on labial side: (2.42±1.30) mm, 8 mm under CEJ on lingual side: (2.28±0.92) mm, apical level on lingual side: (1.94±1.15) mm] were significantly higher than that of canines [apical level on labial side: (1.52±1.47) mm, 8 mm under CEJ on lingual side: (1.17±1.09) mm,apical level on lingual side: (0.74±1.37) mm] (P<0.01). There were no significant differences in the degree of gingival recession [labial side before surgery: (0.72±0.88) mm, lingual side before surgery: (0.80±1.09) mm; labial side 6 months after surgery: (0.72±0.81) mm,lingual side 6 months after surgery: (0.89±0.21) mm] and gingival papilla index [before surgery: 1.00(0.75, 2.00); 6 months after surgery: 1.00(1.00, 2.00) ] between pre-operation and 6 months after surgery (P>0.05). No serious complications occurred. Conclusions: The method used in this article for simultaneously labial and lingual augmented corticotomy was safe and feasible. This surgery has positive clinical significance for the stability of the periodontal tissue in orthodontic treatment for patients with alveolar bone thickness less than 0.5 mm of lower anterior teeth both in labial and lingual side.