Apical microsurgery is accurate and minimally invasive, produces few complications, and has a success rate of more than 90%. However, due to the lack of awareness and understanding of apical microsurgery by dental general practitioners and even endodontists, many clinical problems remain to be overcome. The consensus has gathered well-known domestic experts to hold a series of special discussions and reached the consensus. This document specifies the indications, contraindications, preoperative preparations, operational procedures, complication prevention measures, and efficacy evaluation of apical microsurgery and is applicable to dentists who perform apical microsurgery after systematic training.
Microsurgery/standards* ; Humans ; Apicoectomy ; Contraindications, Procedure ; Tooth Apex/diagnostic imaging* ; Postoperative Complications/prevention & control* ; Consensus ; Treatment Outcome

Approximately 20% to 30% of the global workforce is engaged in shift work. As a significant cause of circadian disruption, shift work is closely associated with an increased risk for periodontitis. Nevertheless, how shift work-related circadian disruption functions in periodontitis remains unknown. Herein, we employed a simulated shift work model constructed by controlling the environmental light-dark cycles and revealed that shift work-related circadian disruption exacerbated the progression of experimental periodontitis. RNA sequencing and in vitro experiments indicated that downregulation of the core circadian protein brain and muscle ARNT-like protein 1 (BMAL1) and activation of the Gasdermin D (GSDMD)-mediated pyroptosis were involved in the pathogenesis of that. Mechanically, BMAL1 regulated GSDMD-mediated pyroptosis by suppressing NOD-like receptor protein 3 (NLRP3) inflammasome signaling through modulating nuclear receptor subfamily 1 group D member 1 (NR1D1), and inhibiting Gsdmd transcription via directly binding to the E-box elements in its promoter. GSDMD-mediated pyroptosis accelerated periodontitis progression, whereas downregulated BMAL1 under circadian disruption further aggravated periodontal destruction by increasing GSDMD activity. And restoring the level of BMAL1 by circadian recovery and SR8278 injection alleviated simulated shift work-exacerbated periodontitis via lessening GSDMD-mediated pyroptosis. These findings provide new evidence and potential interventional targets for circadian disruption-accelerated periodontitis.
Pyroptosis/physiology* ; ARNTL Transcription Factors/metabolism* ; Animals ; Periodontitis/etiology* ; Mice ; Phosphate-Binding Proteins/metabolism* ; Shift Work Schedule/adverse effects* ; Intracellular Signaling Peptides and Proteins/metabolism* ; Mice, Inbred C57BL ; Male ; Disease Models, Animal ; Gasdermins

Objective:To observe whether endothelial cells undergo pyroptosis in the inflammatory periodontal environment by using a model in vivo and in vitro, providing an experimental basis for indepth understanding of the underlying pathogenesis of periodontitis. Methods:According to the classification of periodontal diseases of 2018, gingival tissues were collected from periodontally healthy subjects and patients with stage Ⅲ-Ⅳ, grade C periodontitis, who presented Department of Oral and Maxillofacial Surgery and Department of Periodontology, School of Stomatology, The Fourth Military Medical University from April to May 2022. Immunohistochemical staining was performed to detect the expression level and distribution of gasdermin D (GSDMD), a hallmark protein of cell pyroptosis, in gingival tissues. Periodontitis models were established in each group by ligating the maxillary second molar teeth of three mice for 2 weeks (ligation group). The alveolar bone resorption was determined by micro-CT (mice without ligation treatment were used as the control group), and the colocalization of GSDMD and CD31 were quantitatively analyzed by immunofluorescence staining in gingival tissues of healthy and inflammatory mice. Human umbilical vein endothelial cells (HUVECs) were cultured in vitro and treated with lipopolysaccharide (LPS) of Porphyromonas gingivalis (Pg) combined with adenosine triphosphate (ATP) at various concentrations of 0.5, 1.0, 2.5, 5.0, and 10.0 mg/L, respectively, and the 0 mg/L group was set as the control group at the same time. Scanning electron microscopy was used to observe the morphology of HUVECs. Western blotting was used to detect the expression of gasdermin D-N terminal domains (GSDMD-N) protein and immunofluorescence cell staining was used to detect the expression and distribution of GSDMD. Cell counting kit-8 (CCK-8) was used to detect the proliferative ability of HUVECs, and propidium iodide (PI) staining was used to detect the integrity of cell membrane of HUVECs. Results:Immunohistochemistry showed that GSDMD in gingival tissues of periodontitis was mainly distributed around blood vessels and its expression level was higher than that in healthy tissues. Micro-CT showed that alveolar bone resorption around the maxillary second molar significantly increased in ligation group mice compared with control subjects ( t=8.88, P<0.001). Immunofluorescence staining showed significant colocalization of GSDMD with CD31 in the gingival vascular endothelial cells in mice of ligation group. The results of scanning electron microscopy showed that there were pores of different sizes, the typical morphology of pyroptosis, on HUVECs cell membranes in the inflammatory environment simulated by ATP combined with different concentrations of LPS, and 2.5 mg/L group showed the most dilated and fused pores on cell membranes, with the cells tended to lyse and die. Western blotting showed that the expression of GSDMD-N, the hallmark protein of cell pyroptosis, was significantly higher in 2.5 and 5.0 mg/L groups than that in the control group ( F=3.86, P<0.01). Immunofluorescence cell staining showed that the average fluorescence intensity of GSDMD in 2.5 mg/L group elevated the most significantly in comparison with that in the control group ( F=35.25, P<0.001). The CCK-8 proliferation assay showed that compared to the control group (1.00±0.02), 0.5 mg/L (0.52±0.07), 1.0 mg/L (0.57±0.10), 2.5 mg/L (0.58±0.04), 5.0 mg/L (0.55±0.04), 10.0 mg/L (0.61±0.03) groups inhibited cell proliferation ( F=39.95, P<0.001). PI staining showed that the proportion of positive stained cells was highest [(56.07±3.22)%] in 2.5 mg/L group ( F=88.24, P<0.001). Conclusions:Endothelial cells undergo significant pyroptosis in both in vivo and in vitro periodontal inflammatory environments, suggesting that endothelial cell pyroptosis may be an important pathogenic factor contributing to the pathogenesis of periodontitis.

Objective To evaluate the effect of preoperative catheterization on the success rate of ureteroscopy placement.Methods A total of 400 patients with ureteric stones admitted from October 2020 to September 2023 were selected as the study subjects,and all underwent ureteroscopic holmium laser lithotripsy.According to the preoperative catheterization,the patients were divided into two groups:200 in the observation group(catheterization group)and 200 in the control group(non-catheterization group).The number of successful cases,time of lens surgery and complication rate were recorded and analyzed.Results The placement time in the observation group was(126.35±1.55)s,which was significantly lower than that in the control group(241.33±1.50)s(P＜0.01).The intraoperative complication rate(0.5% )was significantly lower than that in the control group(5.5% )(P＜0.05).The difference was not statistically significant(P＞0.05).Conclusion Preoperative catheterization can shorten the ime of ureteroscopy,and the success rate and safety are higher,which is worthy of clinical promotion.

Objective:To investigate whether bone marrow mesenchymal stem cells (BMMSCs) derived apoptotic extracellular vesicles (ApoEVs) could regulate the polarization of mouse macrophage cell line RAW264.7 and whether BMMSCs derived ApoEVs could attenuate pro-inflammatory condition of RAW264.7 induced by Porphyromonas gingivalis lipopolysaccharide (Pg-LPS), so as to provide experimental evidence and theoretical basis for using BMMSCs derived ApoEVs as a method to treat periodontitis. Methods:The Operetta CLS high-content analysis system was used to observe the time-dependent apoptosis process of BMMSCs. Besides, field emission scanning electron microscopy (FESEM), dynamic light scattering technology and streaming potential method were used to measure the surface characteristics of BMMSCs derived ApoEVs. The Operetta CLS high-content analysis system was used to observe the process of RAW264.7 phagocyting 5-carboxy-tetramethylrhodamine, succinimidyl ester (5-TAMRA-SE) labeled ApoEVs. Real-time quantitative PCR was used to detect the mRNA expression of arginase-1 (Arg-1). Cell immunofluorescence and Western blotting were used to detect the number of inducible nitric oxide synthase (iNOS)(+) macrophages and iNOS protein expression level in each experiment group. Enzyme linked immunosorbent assay was used to detect tumor necrosis factro-α (TNF-α) level in the Pg-LPS induced pro-inflammatory macrophage culture supernatant in each experiment group.Results:After treating with 0.5 μmol/L staurosporine for 12 hours, mouse BMMSCs underwent shrinking with obvious vesicles structure around. The FESEM showed the ApoEVs were in spherical shapes. The size range of ApoEVs was about 100-1 000 nm and the average Zeta potential was -16.6 mV. The Operetta CLS high-content analysis system showed RAW264.7 could phagocytose 5-TAMRA-SE labeled ApoEVs by pseudopodia. The relative mRNA expression of Arg-1 was significantly increased in RAW 264.7 after being treated with interleukin 4 (IL-4) and ApoEVs (261.97±15.91) compared to that with IL-4 alone (115.29±15.42) ( P<0.01). Cell immunofluorescence showed that ApoEVs could reduce the number of iNOS(+) macrophages induced by Pg-LPS (39.33±4.70) comparing to those without ApoEVs (95.33±4.70) ( P=0.007). In the meanwhile, ApoEVs could also down-regulate the iNOS protein level of macrophages induced by Pg-LPS (5.84±1.05) comparing to those without ApoEVs (14.91±3.87) ( P<0.01). Besides, ApoEVs could also reduce the TNF-α secretion in the culture supernatant of pro-inflammatory macrophages induced by Pg-LPS [(21 899.71±409.73) ng/L] comparing to those without ApoEVs [(71 296.50±2 344.22) ng/L] ( P=0.003). Conclusions:BMMSCs derived ApoEVs could regulate the polarization of macrophages and could also attenuate the pro-inflammatory condition of macrophages induced by Pg-LPS.

Objective:To study factors influencing postoperative pancreatic fistula rates with a view to prevent postoperation pancreatic fistula from happening.Methods:This is a retrospective study on 281 patients who underwent distal pancreatectomy at the First Affiliated Hospital of China Medical University from March 2011 to April 2018. There were 89 males and 192 females, with the age of (51.01±13.65) years. Univariate and multivariate logistic regression analyses were used to analyze the following factors on the occurrence of pancreatic fistula after operation: gender, age, body mass index(BMI), tumor characteristics, preoperative fasting blood glucose, blood biochemistry, liver function and surgical indications.Results:Of the 281 patients who underwent distal pancreatectomy in this study, 245 (87.2%) did not develop pancreatic fistula / biochemical leakage, while 36(12.8%) patients developed clinically significant pancreatic fistula (B/C grade). Univariate analysis showed the factors which affected the incidence of pancreatic fistula after surgery to include: BMI, preoperative fasting blood glucose, and whether the main pancreatic duct was ligated (all P<0.05). Multivariate logistic regression analysis showed that the independent factors affecting pancreatic fistula incidence after surgery were BMI≥25 kg/m 2 ( OR=2.354, 95% CI: 1.137-4.873, P<0.05), and main pancreatic duct was not ligated ( OR=4.067, 95% CI: 1.191-13.885, P<0.05). Conclusions:A high BMI increased the risk of postoperative pancreatic fistula, while ligation of main pancreatic duct during surgery reduced the risk.

Objective:To establish a colloidal gold technique assay for the rapid detection of immunoglobulin(Ig)M and IgG antibodies against 2019 novel coronavirus (2019-nCoV) and to evaluate its clinical performance.Methods:A total of 278 patients who were respectively treated at Wuhan Hankou Hospital and the People′s Hospital of Honghu from February 12, 2020 to February 20, 2020 were collected. According to the diagnostic criteria, 89 patients were confirmed with positive 2019-nCoV nucleic acid, and 189 were 2019-nCoV nucleic acid-negative suspected patients. A total of 273 medical examiners from Nanfang Hospital, Southern Medical University from 2015 to 2018 were selected as controls. The serum samples of patients were collected. 2019-nCoV nucleic proteins were obtained from prokaryotic expression vectors. Indirect IgM and IgG colloidal gold techniques were established by using recombinant nuclear protein. 2019-nCoV nucleic acid detection by reverse transcription-polymerase chain reaction (RT-PCR) was used as control. Serum specimens were tested for 2019-nCoV IgM and IgG. The specificity and sensitivity of colloidal gold assay were analyzed.Results:The positive rates of IgM and IgG with the colloidal gold detection in confirmed patients with positive 2019-nCoV nucleic acid were 78.7%(70/89) and 73.0%(65/89), respectively. The positive rates of IgM and IgG in medical examiners were 1.8%(5/273) and 0.7%(2/273), respectively. The sensitivity and specificity of IgM detection reagents were 78.7% and 98.2%, respectively, those of IgG detection reagents were 73.0% and 99.3%, respectively, and those of IgM combined with IgG detection were 87.6% and 98.2%, respectively. For suspected patients with negative 2019-nCoV nucleic acid, the positive rates of IgM and IgG were 59.8%(113/189) and 52.9%(100/189), respectively, and the positive rate of IgM combined with IgG detection was 66.1%(125/189).Conclusion:This reagent of 2019-nCoV antibodies detection (colloidal gold technique) fulfills the requirement for clinical application with high specificity and sensitivity, which can be served as a supplementary detection method for 2019-nCoV nucleic acid detection by RT-PCR.

Objective: To establish a colloidal gold technique assay for the rapid detection of immunoglobulin(Ig) M and IgG antibodies against 2019 novel coronavirus (2019-nCoV) and to evaluate its clinical performance. Methods: A total of 278 patients who were treated at Wuhan Hankou Hospital and the People's Hospital of Honghu from February 12, 2020 to February 20, 2020 were collected. According to the diagnostic criteria, 89 patients were confirmed with 2019-nCoV nucleic acid positive diagnosis, and 189 were 2019-nCoV nucleic acid-negative suspected patients. A total of 273 medical examiners from Nanfang Hospital, Southern Medical University from 2015 to 2018 were selected as controls. The serum samples of patients were collected. 2019-nCoV nucleic proteins were obtained from prokaryotic expression vectors. Indirect IgM and IgG colloidal gold techniques were established by using recombinant N protein. 2019-nCoV nucleic acid detection by reverse transcription-polymerase chain reaction (RT-PCR) was used as control. Serum specimens were tested for 2019-nCoV IgM and IgG. The specificity and sensitivity of colloidal gold assay were analyzed. Results: The sensitivity and specificity of IgM detection reagents were 78.7% and 98.2%, respectively, those of IgG detection reagents were 73.0% and 99.3%, respectively, and those of IgM combined with IgG detection were 87.6% and 98.2%, respectively. For suspected patients with negative 2019-nCoV nucleic acid, the positive rates of IgM and IgG were 59.8% (113/189) and 52.9% (100/189), respectively, and the positive rate of IgM combined with IgG detection was 66.1% (125/189). Conclusion This reagent of 2019-nCoV antibodies detection (colloidal gold technique) fulfills the requirement for clinical application with high specificity and sensitivity, which can be served as a supplementary detection method for 2019-nCoV nucleic acid detection by RT-PCR.

Objective To analyze the expression changes of transforming growth factor-β (TGF-β) and vascular endothelial growth factor (VEGF) in malignant brain gliomas and their values in tumor resectability evaluation.Methods One hundred and eighty-eight patients with malignant brain gliomas received surgery in our hospital from January 2010 to January 2017 were chosen in our study as patient group;during the surgery,the glioma tissues and peritumoral tissues were collected;Westem blotting was employed to detect the VEGF and TGF-β expressions.Fifty healthy controls accepted healthy examinations at the same time-period were enrolled.Peripheral blood 24 h before surgery of the two groups was collected.ELISA was used to test the serum VEGF and TGF-β levels.The values of serum VEGF and TGF-β levels in tumor resectability were analyzed by receiver operating characteristic (ROC) curve.Results In the patient group,significantly higher levels of VEGF and TGF-β in the tumor tissues were noted as compared with those in the para-tumor tissues (P＜0.05).Significantly higher levels of VEGF and TGF-β in the patient group were noted as compared with those in the control group (P＜0.05).There were 126 patients received total tumor resection,while 62 just had partial tumor resection;one year after surgery,22 patients with total tumor resection (17.46％) died,and 20 patients with partial tumor resection (32.26％) died,which indicated that the cumulative mortality rate one year after surgery in patients with total tumor resection was significantly lower than that in patients with partial tumor resection (P＜0.05).With cut-off point value of 588.0 pg/mL,ROC analysis showed that the sensitivity of VEGF in prediction of total tumor resection was 80.65％,and specificity 80.16％,and area under curve 0.82,which were better than those of TGF-β.Conclusion For malignant brain glioma patients,the levels of VEGF and TGF-β increase,and VEGF shows an ideal value in tumor resectability evaluation.

OBJECTIVE:To explore clinical efficacy and safety of Ulinastatin injection combined with Xingnaojing injec-tion in the treament of severe craniocerebral injury(CCI). METHODS:A total of 120 severe CCI patients selected from our hospital during Sept. 2014-Nov. 2015 were divided into ulinastatin group,Xingnaojing group and combination group according to therapy plan,with 40 cases in each group. Three groups were given routine treatment timely after admission. On the basis of routine treatment,Ulinastatin group additionally received Ulinastatin injection 200 000 U,ivgtt,bid;Xingnaojing group addi-tionally received Xingnaojing injection 20 mL,ivgtt,qd;combination group additionally received Ulinastatin injection com-bined with Xingnaojing injection,same usage as above(with 1 h intervals). Three groups received therapy for consecutive 14 d. Serum inflammatory factors(CRP,IL-1,IL-6,TNF-α),serologic indexes of craniocerebral injury [neuron specific enolase (NSE),myelin basic protein(MBP),S100B protein(S100B)] and GCS scores before and after treatment as well as GOS scores after treatment were all observed in 3 groups. The occurrence of ADR was recorded during treatment. RESULTS:Before treatment,there was no statistical significance in serum inflammatory factors,serologic indexes of craniocerebral injury or GCS scores among 3 groups(P>0.05). Compared to before treatment,inflammatory factors of 3 groups were decreased signifi-cantly after treatment,the ulinastatin group was significantly lower than the Xingnaojing group,combination group was signifi-cantly lower than two single drug groups,with statistical significance(P<0.05). Levels of serologic indexes of craniocerebral injury and GCS scores of 3 groups were improved significantly,and the combination group was significantly better than the two single drug groups,with statistical significance(P<0.05). There was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). Six months after treatment,GOS score of combination group(4.17±0.81)was significantly better than those of ulinastatin group(3.05±0.97)and Xing-naojing group(2.97 ± 0.89),with statistical significance (P<0.05);there was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). During treatment,the incidence of ADR in combination group(27.50%)was significantly lower than ulinastatin group(50.00%)and Xingnaojing group(42.50%),with statistical significance(P<0.05);there was no statistical significance between ulinastatin group and Xingnaojing group(P>0.05). CONCLUSIONS:Ulinastatin injection combined with Xingnaojing injection can sig-nificantly decrease serum inflammatory factor levels,relieve craniocerebral injury,protect cerebral tissue and improve short-term prognosis with good safety.