OBJECTIVEThis study measures the glutaredoxin (Grx) gene and protein expression in umbilical vein endothelial cells upon exposure to Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS). The involvement of the Akt-signaling pathway is also determined.

METHODSEA-hy926 cells were pretreated with 1,000 ng · mL⁻¹ P. gingivalis LPS for 4, 12, 18, and 24 h, and then real-time reverse transcription polymerase chain reaction was employed to detect Grx1 expression. The effect of Grx on Akt activity was investigated using Western blot for the control, LPS (1,000 ng · mL⁻¹ LPS), and carmus- tine (BCNU) groups (1,000 ng · mL⁻¹ LPS, and the EA-hy926 cells were pretreated with 25 μmol · ml⁻¹ BCNU for 30 min).

RESULTSGene expression of Grx1 significantly increased in LPS group compared with that in the control group. The Grx1 expression reached the peak level in 12 h, and the variation between the expression in 4 and 12 h was significant (P < 0.05). After 12 h, the protein levels of Grx and phosphorylated-Akt (p-Akt) significantly increased in the LPS group (P < 0.05), whereas the BCNU group showed a considerable decrease in both Grx and p-Akt expression levels (P < 0.05). Moreover, a slight difference was observed in the total Akt protein levels in the three groups (P > 0.05).

CONCLUSIONGrx expression increased upon exposure of EA-hy926 cells to the LPS. Akt activity could be inhibited by BCNU (a Grx inhibitor), which indicated that Akt might act as a downstream regulator of Grx.

Endothelial Cells ; Glutaredoxins ; genetics ; Humans ; Lipopolysaccharides ; pharmacology ; Oxidative Stress ; drug effects ; Phosphorylation ; Porphyromonas gingivalis ; pathogenicity ; Proto-Oncogene Proteins c-akt ; drug effects ; Signal Transduction ; drug effects ; Umbilical Veins

Objective We aimed to (i) observe the therapeutic effect of Yiqi Wenyang Prescription (YQWYP, which consists of milkvetch root, tangshen, dried ginger, cassig twig, ephedra, biond magnolia flower-bud, Chinese magnoliavine fruit, earthworm, and liquorice root) on an allergic rhinitis (AR) murine model and the protective effect on the tight junction (TJ) key proteins zonula occludens-1 (ZO-1) and occludin and (ii) elucidate whether YQWYP exerts these effects by inhibiting autophagy in nasal mucosal cells.Methods Thirty C57BL/6 mice were randomly divided into six groups with 5 mice per group: (i) the blank group, (ii) the model group, (iii-v) the YQWYP low-dose, mid-dose, and high-dose groups(7, 14, 28 g/kg), and (vi) the cetirizine group(1.667 mg/kg). The model was established by intraperitoneal injection and nasal stimulation of ovalbumin. 1 hour before the nasal stimulation, mice in the blank group or in the model group were given normal saline by gavage, mice in the YQWYP groups and those in the cetirizine group were given the corresponding medicines solution by gavage. After 7 days, mice behavior was scored. After that, the mice were sacrificed and the nasal mucosa tissues were harvested. The inflammatory reaction of the nasal mucosa was observed by HE staining, the proliferation of goblet cells in the nasal mucosa was observed by PAS staining, the IgE content in nasal lavage fluid was determined by ELISA, the expression levels of ZO-1 and occludin were analyzed by immunohistochemistry, and the expression levels of LC3B, P62, and Beclin1 in the nasal mucosa of the blank group, model group, and YQWYP high-dose group were analyzed by Western blotting.Results After the successful establishment of the AR model, compared with the blank group, the symptom scores of nose scratching, sneezing, and runny nose in the model group were higher, while the total scores was significantly higher(P<0.01). In the model group, the nasal mucosal epithelium was disrupted and destroyed, a large number of inflammatory cells were infiltrated, goblet cells showed hyperplasia, mucus production was increased, mucosal swelling was obvious, the amount of IgE in nasal lavage fluid was increased (P<0.01). The protein expression levels of ZO-1 and occludin were decreased (P<0.05), the protein expression levels of LC3BⅡ/Ⅰ and Beclin1 were increased (P<0.05), P62 protein expression was decreased (P<0.05). After medication intervention, compared with the model group, in the YQWYP high-dose group and the cetirizine group, the total score were reduced (P<0.01), the swelling of the nasal mucosa was reduced, the infiltration of inflammatory cells was inhibited, the number of goblet cells was decreased, the amount of IgE in nasal lavage fluid was reduced(P<0.01), the protein expression of ZO-1 and occludin were increased (P<0.05), the protein levels of LC3BII/I and Beclin1 were decreased (P<0.05), P62 protein expression was increased (P<0.05). Conclusion YQWYP has a good therapeutic effect on AR mice, and its mechanism may be related to the promotion of nasal mucosal repair by inhibiting autophagy of nasal mucosal cells.

Gingival pigmentation is a nonplaque gum disease. Patients are often afraid to communicate with others because of gum color problems, which affect the social and mental health of patients. The commonly used treatment methods for gingival pigmentation include scalpel excision, gingival grinding, laser therapy, cryosurgery and electrosurgery. In this paper, the progress of gingival pigmentation treatment was reviewed in terms of bleeding, pain, tissue healing and recoloring. The results showed that the clinical effect of laser treatment was better. Among them, the semiconductor laser had more advantages in reducing bleeding, pain and the restaining rate, while the Er:Cr:YSSG/Er:YAG laser performed better for promoting tissue healing. Clinicians can choose the best kind of laser to use according to the actual situation. For patients with thin gingival biotypes, floating gingival transplantation or substitute materials can be selected to restore the gingival morphology. With the in-depth study of melanin regulation mechanisms, various drugs, such as ascorbic acid, natural peptides, synthetic peptides and derivatives, may be the main research direction for the treatment of gingival pigmentation in the future.

Objective:To analyze the relationship between blood electrolytes and the prognosis of patients with severe coronavirus disease 2019 (COVID-19) and to provide assistance for clinical decision-making.Methods:The clinical data of patients with severe COVID-19 admitted to intensive care unit (ICU) of the Wuhan Third Hospital by the Shanghai aid-Hubei medical team from January 21 to March 4, 2020 were collected. Excluding ineligible patients, 110 patients were finally enrolled. The patients' gender, age, temperature, heart rate, systolic and diastolic blood pressure, clinical symptoms at admission, time of symptom onset, duration of fever, and relevant indicators at admission to ICU (including blood potassium, chloride, sodium, calcium, phosphorus, and magnesium, etc.) and prognosis were analyzed. The patients were grouped by blood potassium or calcium levels or blood potassium/calcium ratio. The Kaplan-Meier survival curves were used to analyze the survival of patients in each group. The relationship between the potassium/calcium ratio and the prognosis was analyzed using restricted cubic spline plots. The relationship between each index in the different models and the prognosis was analyzed using Cox regression models.Results:Among 110 severe COVID-19 patients, 78 cases survived, and 32 cases died. Compared with the surviving group, patients in the death group had higher blood potassium levels [mmol/L: 4.25 (3.80, 4.65) vs. 3.90 (3.60, 4.20), P < 0.05] and lower blood calcium levels (mmol/L: 2.00±0.14 vs. 2.19±0.18, P < 0.05). The Kaplan-Meier survival curves showed that patients in the potassium > 4.2 mmol/L group had a worse prognosis than the potassium < 3.8 mmol/L group and the potassium 3.8-4.2 mmol/L group ( P = 0.011), patients in the calcium > 2.23 mmol/L group had a better prognosis than the calcium < 2.03 mmol/L group and the calcium 2.03-2.23 mmol/L group, and the lower calcium group had a worse prognosis ( P = 0.000 15). Cox regression analysis showed that the hazard ratio ( HR) of blood potassium and calcium were 2.08 and 0.01, respectively, in model 1 (single blood potassium or calcium) and in model 2 (model 1 plus age and gender), the HR of blood potassium and calcium were 1.98 and 0.01 respectively, which were significantly associated with patient prognosis (all P < 0.05). Patients in the group with the potassium/calcium ratio > 1.9 had higher blood potassium levels and a higher proportion of mechanical ventilation, lower calcium levels and lower proportion of survival, and longer time of ICU admission compared with the groups with the potassium/calcium ratio < 1.7 and 1.7-1.9. The Kaplan-Meier survival curves showed that the survival rate of the potassium/calcium ratio > 1.9 group was the lowest ( P < 0.000 1), and there was no statistically significant difference in survival between the potassium/calcium ratio < 1.7 group and the potassium/calcium ratio 1.7-1.9 group. A restricted cubic spline plot corrected for age and gender showed that patients in the potassium/calcium ratio > 1.8 group had HR values > 1. Cox regression analysis corrected for other indicators showed that the potassium/calcium ratio was still associated with patient prognosis ( HR = 4.85, P = 0.033). Conclusions:Blood potassium, calcium, and the potassium/calcium ratio at ICU admission are related to the prognosis of patients with severe COVID-19, and the potassium/calcium ratio is an independent risk factor for the death of patients. The higher the potassium/calcium ratio, the worse the prognosis of patients.

Subgingival microorganisms can orchestrate the microbial community under the influence of environmental factors, thus causing the initiation and progression of periodontitis. Keystone taxa play an important role in the dynamic changes of microbial community. The co-occurrence network analysis may pave the way for screening periodontitis associated candidate keystone taxa in the periodontal microflora. These microorganisms, e.g. Porphyromonas gingivalis, might function via myriad of intermediary taxa or as the chokepoint community-remodeling drivers in response of environmental factors. Herein, we review and discuss the construction of microbial co-occurrence network, the way of screening and identifying keystone taxa using network analysis as well as the characteristic of candidate keystone taxa related to periodontitis for the purpose that keystone taxa and the role they played in dynamic succession of microflora could be well investigated.

Treponema denticola (Td) is a gram-negative anaerobic bacterium closely related to the occurrence and development of periodontal disease and it accounts for a considerable proportion of mature plaque. As a later colonizer of the subgingival plaque biofilm, Td may have complex interactions with earlier and concurrent colonists including symbiotic relationship as while as synergistic or antagonistic effects under the regulation of quorum sensing molecules. Adhesin and coaggregation, mediated by a series of surface molecules, are the basis of the interaction. These interactions are ultimately manifested as gene expression changes in metabolism and virulence, in which are mainly metabolism changes with up- or down-regulation of multiple enzymes related to amino acid metabolism. This article reviews the related researches on the interaction between Td and microorganisms of subgingival plaque.

Periodontitis is closely related to systemic health, especially cardiovascular disease. Periodontal pathogens from periodontal infection actively participate in the formation and development of coronary atherosclerosis. At present, accumulcted evidences show that periodontal pathogen infection is an independent risk factor for cardiovascular events, which promotes the deepening of atherosclerosis through a variety of immune inflammation and metabolism-related molecular mechanisms. The present article reviews multipal aspects of the correlation between periodontitis and periodontal pathogens and cardiovascular disease, and the mechanism of periodontal pathogens affecting the occurrence and development of atherosclerosis.