Fangji Dihuang formulation ameliorated DNCB-induced atopic dermatitis-like skin lesions by IL-17 signaling pathway: integrating network analysis and experimental validation.

Background: The Fangji Dihuang formulation (FJDHF) is a widely recognized Traditional Chinese Medicine (TCM) formula that consists of five plant drugs: Stephaniae Tetrandrae Radix, Cinnamomi Ramulus, Rehmanniae Radix, Saposhnikoviae Radix, and Glycyrrhiza Urensis Fisch. This formulation has been known to exhibit clinical therapeutic effects in the treatment of inflammatory skin diseases. However, there is a lack of pharmacological research on its anti-atopic dermatitis (AD) activity. Methods: To investigate the potential anti-AD activity of FJDHF, DNCB was used to induce AD-like skin inflammation in the back of mice. Following successful modeling, the mice were administered FJDHF orally. The extent of the inflammatory skin lesions was recorded at day 4, 7, 14 and 28. UHPLC-Q-Exactive Orbitrap MS was used to identify and match the compounds present in FJDHF with ITCM, TCMIP and TCMSID. In silico predictions of potential target proteins of the identified compounds were obtained from SwishTargetPrediction, ITCM and TargetNet databases. AD-related genes were identified from GSE32924 data set, and FJDHF anti-AD hub genes were identified by MCODE algorithm. ClueGo enrichment analysis was employed to identify the core pathway of FJDHF's anti-AD effect. To further investigate the anti-AD effect of FJDHF, single-cell RNA sequencing data set (GSE148196) from AD patients was analyzed to determine the target cells and signaling pathways of FJDHF in AD. Finally, rt-PCR, flow cytometry, and mouse back skin RNA sequencing were utilized to validate our findings. Results: FJDHF was found to be effective in improving the degree of the AD-like lesions in the mice. Network pharmacological analysis revealed the core pathway of FJDHF to be the IL-17 signaling pathway, which is interactively associated with cytokines. Single-cell RNA sequencing analysis suggested that FJDHF may play an anti-AD role by influencing dendritic cells. Flow cytometry and rt-PCR results showed that FJDHF can reduce the influence of AD sample of IL-4, IFN-γ and the expression of IL-17. The RNA sequencing of mouse back skin also confirmed our conclusion. Conclusion: FJDHF may inhibit DNCB-induced AD-like skin inflammation in mice by inhibiting the IL-17 signaling pathway. Thus, FJDHF can be considered as a potential therapeutic agent for AD.

[Nonradiological methods for implant surgical guide accuracy measurement].

PURPOSE: To evaluate the accuracy of static computer-navigated implantation with surgical guides, based on a non-radiological method. Traditional measurements with a second cone-beam CT (CBCT) were applied to verify the accuracy. METHODS: A total of thirty template-guided implantations were designed and performed on 15 resin models. Two paralleled bone-level implants were planned in the edentulous space of each model, between which the distance was 4 mm. Postoperative implant positions were detected with both CAD/CAM-based measurements applying an intraoral scanner (3Shape TRIOS) and traditional ways via CBCT. Both methods were conducted with a CAD quality-control, reverse engineering software, Geomagic Studio 2013, comparing the positions with the virtual ones. Statistical analysis was processed with SPSS 23.0 software package. RESULTS: Measurements using CBCT (control group) showed a trend toward greater deviations when the results were directly compared(P<0.05). In the CAD/CAM-based evaluation of the 30 samples, the mean deviation of the insertion axis from the planned implant axis was 1.134°. The mean deviations of the implant shoulders in the horizontal direction and at the implant apices were 0.447 mm and 0.557 mm, respectively. No significant difference was observed when measuring distance deviation with the two assessment ways. CONCLUSIONS: Compared with evaluation based on radiology, CAD/CAM based evaluation system is able to evaluate implant accuracy precisely, effectively reduce radiological exposure of patients, being suitable for clinical evaluation.

[Study on cushioning and energy absorption of sports mouthguard].

PURPOSE: Using impact test to study cushioning and energy absorption of sports mouthguard of different thickness, material and combination. METHODS: According to the thickness, material and combination, the mouthguards were divided into 7 groups with 6 testing pieces in each group. A pendulum device was used to apply impact energy to the mouthguard and dentition model, and the magnitude of the impact energy was measured by changing the release height of the pendulum. The force sensor and the laser vibrometer were respectively connected to the data acquisition and signal analyzer to obtain the incident speed, rebound speed, maximum collision force, collision start time and collision end time of the hammer head. Collision time, energy absorption and energy absorption ratio of the mouthguard were calculated, which reflected different mouthguard cushioning and energy absorption characteristics. One-way analysis of variance and t test in SPSS 22.0 software package was used to analyze the difference between different groups. RESULTS: No matter what type of mouthguard was worn by the dentition model, the impact force was significantly reduced than without mouthguards. At the same height, there was significant difference between the average maximum collision force and the average collision time among soft mouthguards with thicknesses of 2, 3, 4, and 5 mm(P<0.05); among which the average maximum collision force of the 5 mm was the smallest and the average collision time was the shortest. There was no significant difference in the average energy absorption and the average energy absorption ratio among these 4 groups. There was significant difference in the average maximum collision force, average collision time, average energy absorption and average energy absorption ratio between the two groups of mouthguards with the same thickness of splints and different materials(P<0.05). Among them, the average maximum impact force of the 4 mm soft mouthguard was smaller and the average collision time was longer. CONCLUSIONS: Wearing sports mouthguard can reduce the risk of tooth fracture during collision. Soft mouthguard has the best cushioning performance and it is positively related to thickness. Locally strengthening hard materials can enhance energy absorption and provide better protection.

The occurrence and treatment of hemophagocytic lymphohistiocytosis caused by multiple factors: a case report and literature review.

Hemophagocytic lymphohistiocytosis (HLH) is a high-fatality disease caused by hereditary or acquired immune dysfunction, and is characterized by pathological inflammatory response. Primary HLH (pHLH) has hereditary genetic defects, and secondary HLH (sHLH) is caused by a variety of underlying diseases. Here, we report the case of a patient with aggressive natural killer cell leukemia and HLH-related gene defects who achieved long-term survival after treatment. A 20-year-old man presented to our hospital with symptoms of fever and fatigue. Investigations revealed splenomegaly, cytopenia, hyperferritinemia, hypofibrinogenemia, elevated levels of soluble CD25 (sCD25), and hemophagocytosis in bone marrow. Bone marrow flow cytometry showed 23.4% abnormal natural killer cells, the cells were CD2, CD7, CD16, CD94, NKG2A positive, met the diagnosis of aggressive NK-Cell leukemia. Investigation of the patient's pedigree revealed that mutations of pHLH-related genes (LYST and UNC13D) were inherited from his father and mother, but neither of the parents had the disease. The patient received hematopoietic stem-cell transplantation (HSCT), after which he achieved complete remission. As of 2020-10-19, the patient's survival has exceeded 3 years, and he has returned to his normal life. A variety of factors contribute to the onset of HLH, and this case gives greater insight into the etiology of HLH. Allogeneic HSCT is a key treatment for HLH patients with underlying genetic mutations.

Association between TERT gene polymorphisms and acute myeloid leukemia susceptibility in a Chinese population: a case-control study.

BACKGROUND: The aim of this study was to investigate the association between telomerase reverse transcriptase (TERT) gene polymorphisms and acute myeloid leukemia (AML) susceptibility in a Chinese Han population. METHODS: A total of 102 AML patients and 108 healthy controls were enrolled in this case-control study. TERT gene rs2853669 and rs2736100 polymorphisms were genotyped via polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Chi-square test was applied to compare polymorphism distributions between case and control groups. The strength of the association between TERT gene polymorphisms and AML susceptibility was evaluated utilizing odds ratio (OR) with corresponding 95% confidence interval (CI). RESULTS: CC genotype and C allele of rs2736100 polymorphism were more frequent in AML patients (P < 0.05), and individuals carrying CC genotype showed higher risk of suffering from AML (OR = 2.632, 95% CI 1.129-6.133). But for rs2853669 polymorphism, no significant differences were detected in either genotype or allele distributions between groups (P > 0.05). CONCLUSIONS: This study suggested a positive association between TERT gene rs2736100 polymorphism and AML susceptibility in Chinese Han population.

[Study of the timing of tooth movement after repair of alveolar bone defects by rabbit BMSCs combined with beta-TCP].

PURPOSE: To investigate the effect of tooth movement at different time after repair of alveolar bone defects with tissue engineering bone constructed by rabbit bone marrow mesenchymal stem cells (BMSCs) and beta-tricalcium phosphate (ß-TCP). METHODS: Alveolar bone defect (6 mm×4 mm×8 mm) was made on the right side of 40 New Zealand rabbits, which was filled with tissue engineering bone constructed by BMSCs and ß-TCP as experimental sides. Tooth extraction on the other side was performed as control. The mandibular second molars in both sides were moved mesially 2, 4, 8 and 12 weeks after surgery. The specimens were taken 4 weeks after exertion. The distance of mandibular second molar moved was measured. The periodontal tissues were observed after H-E staining. TRAP staining was performed and osteoclasts were counted in the periodontal tissues on the pressure side. BMP-2 immunohistochemical staining was used to observe the average optical density of periodontal tissue on tension side. The results were analyzed with SAS 8.0 software package. RESULTS: The distance of tooth movement, the amount of TRAP positive cells and the optical density of BMP-2 in the experimental side of the 2-week and 4-week groups were all significantly lower than those in the control side, while there was no significant difference between the 8-week and 12-week groups. CONCLUSIONS: Eight weeks after repair of alveolar bone defect with rabbit BMSCs and ß-TCP is an appropriate time for orthodontic tooth movement.

Effect of miRNA-19a antisense oligonucleotide and Ara-C on the proliferation and apoptosis of HL60 cells.

BACKGROUND: This study aimed to investigate the effects of microRNA19a (miR-19a) antisense oligonucleotide (ASODN) on the proliferation and apoptosis of acute myeloid leukemia cells (HL60). METHODS: In experiment 1, HL60 cells were divided into the blank control group, the blank transfection group, the scrambled oligonucleotide (SODN) group and the ASODN group. MiR-19a ASODN and SODN were independently transferred into HL60 cells. The miR-19a expression was detected by real-time quantitative RT-PCR (qRT-PCR) after 48-h and 72-h transfection; CCK8 assay was used to detect the proliferation inhibition rate at 48 and 72 h; Hoechst 33258 staining was performed to examine apoptotic cells at 48 h; the apoptosis rate was detected by flow cytometry after AnnexinV/PI staining at 48 and 72 h; the protein expression of E2F1 and Bim was detected by Western blotting at 48 h. In experiment 2, cells were divided into the Ara-C group, the SODN + Ara-C group and the ASODN + Ara-C group. The cell proliferation inhibition rate at 48 and 72 h and apoptosis rate at 72 h were assessed as mentioned above. RESULTS: MiR-19a expression in the miR-19a ASODN group was lower than in the SODN group and the blank control group (P<0.05). MiR-19a ASODN significantly inhibited the growth of HL60 cells (P<0.05) and increased their apoptosis, and the apoptosis rate peaked at 48 h. The protein expression of E2F1 and Bim in the ASODN group was higher than in the blank control group, blank transfection group and SODN group. In addition, Ara-C further inhibited the growth and induced the apoptosis of miR-19a ASODN-transfected cells (P<0.05) in a time dependent manner. The growth inhibition rate and apoptosis rate in the ASODN + Ara-C group were higher than the sum of those in both Ara-C group and ASODN group. CONCLUSIONS: miRNA-19a ASODN can inhibit the proliferation and induce apoptosis of HL60 cells and may exert synergistic effects with Ara-C on HL60 cells.

[Study on energy absorption of mouthguard splints by impact test].

PURPOSE: Using impact test to study energy absorption ability of mouthguard splints of different thickness and materials. METHODS: In this experiment, group 1 was BIOPLAST 5 mm splint, group 2 was BIOPLAST 4 mm splint, group 3 was BIOPLAST 3 mm splint，group 4 was BIOPLAST 2 mm splint and group 7 was BIOPLAST 1 mm splint. Group 5 was ERKOLOC-PRO 2 mm splint, group 6 was DURAN 2 mm splint. Mouthguard splints were clashed by impact head from different heights. Digital Image Correlation System was used to record the process of the test. Images were transferred to data processing software, to analyze incident speed, incident depth and rebound height of the impact head. Absorbed energy and energy absorption ratio were calculated to analyze energy absorption ability of mouthguard splints of different thickness and materials. The data were analyzed using SPSS 22.0 software package for one-way analysis of variance and LSD. RESULTS: The higher impact head fell, the larger energy absorption ratio was. There was significant difference among group 1, 2, 3, 4, 7 in average energy absorption ratio（P<0.05）, and group 7 was the largest. There was significant difference among group 4, 5, 6 in average energy absorption ratio（P<0.05）, among which group 4 and group 5 were larger. CONCLUSIONS: 3 mm splint is good enough to be used to make mouthguard, which is also thinner and more comfortable. Splint of soft material is more suitable for mouthguard than splint of hard material.

Association of ERCC2 Gene Polymorphisms with Susceptibility to Diffuse Large B-Cell Lymphoma.

BACKGROUND The objective of this study was to detect the association between ERCC excision repair 2, TFIIH core complex helicase subunit (ERCC2) gene polymorphisms and diffuse large B-cell lymphoma (DLBCL) susceptibility. MATERIAL AND METHODS This study used a case-control design. ERCC2 gene rs1799793 (Asp312Asn) and rs13181 (Lys751Gln) polymorphisms were genotyped by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) both in DLBCL patients and healthy controls. The association between ERCC2 gene polymorphisms and DLBCL risk was assessed by χ² test. Odds ratios (ORs) with corresponding 95% confidence intervals (95% CIs) were used to address the association strength. Subgroup analyses were also performed to investigate the genetic effects of ERCC2 polymorphisms on clinical characteristics of DLBCL patients. RESULTS A significant association was discovered between the rs1799793 A allele and increased DLBCL risk (P=0.031, OR=1.928, 95% CI=1.052-3.534). The C allele of rs13181 was obviously associated with elevated DLBCL susceptibility (P=0.047, OR=1.820, 95% CI=1.002-3.305). The subgroup analysis demonstrated that rs1799793 and rs13181 polymorphisms had no relationship with serum lactate dehydrogenase level, nidus number, B-symptoms, Ann Arbor stages, or immunological types in DLBCL cases (P>0.05 for all). CONCLUSIONS Minor allele carriers of ERCC2 gene rs1799793 (Asp312Asn) and rs13181 (Lys751Gln) polymorphisms had higher susceptibility to DLBCL.

Symmetry of root anatomy and root canal morphology in maxillary premolars analyzed using cone-beam computed tomography.

OBJECTIVE: This study aimed to use cone-beam computed tomography (CBCT) to evaluate the root anatomy and canal morphology of maxillary premolars in a Chinese population and determine their degree of bilateral symmetry. DESIGN: 774 CBCT images were retrospectively analyzed, representing 1387 maxillary first premolars and 1403 second premolars. The number of roots and canals were recorded. The morphology of root canal systems was determined according to Vertucci's classification. The symmetry of root and canal anatomies between maxillary contralateral premolars was further evaluated. RESULTS: The most common anatomy of maxillary first and second premolars was one-rooted with two canals (58.0%) and one-rooted with one canal (50.3%), respectively. The typical canal morphology was type IV (42.7%) in maxillary first premolars and type I (50.3%) in maxillary second premolars. One-rooted maxillary premolars exhibited a higher variability in the canal morphology, compared to two-rooted or three-rooted teeth. Maxillary second premolars exhibited greater anatomic symmetry than first premolars. The root and canal numbers showed bilateral symmetry between 80.2% of maxillary first premolar pairs and 81.8% of second premolar pairs. Bilateral symmetry in both number and morphology of roots and canals was observed for 72.3% of maxillary first premolar pairs and 73.2% of second premolar pairs. CONCLUSION: The root anatomy and canal morphology of maxillary premolars in a Chinese population were quite diversified. Maxillary contralateral premolars demonstrated a high degree of symmetry in root and canal anatomies, which enables practitioners to better determine the nature of the root canal system during treatment of opposite homonymous teeth.

[Study on periodontal responses on the compression side during early tooth movement into alveolar defect regenerated by a tissue engineering bone].

PURPOSE: To explore periodontal responses on the compression side during early tooth movement into alveolar defect regenerated by bone marrow mesenchymal stem cells (BMSCs) and porous granulated beta-tricalcium phosphate（ß-TCP） scaffolds. METHODS: Thirty New Zealand rabbits were used to establish bilateral mandibular defective alveolar bone model by extracting the mandibular first molars and expanding the sockets. The right mandibular alveolar defects were filled with a construct of ß-TCP scaffolds combined with BMSCs as experimental group. The left alveolar defects were repaired by ß-TCP scaffolds alone as control group. Eight weeks later, 6 rabbits were sacrificed to evaluate osteogenesis effect. The other rabbits were loaded orthodontic force to move the bilateral second molars forward for 4 weeks. Six rabbits in each group were sacrificed at 1, 2, 3, and 4 week after orthodontic tooth movement (OTM). The distance of OTM was measured, and the status of periodontal tissues was observed by H-E staining. The number of osteoclasts on the compression side of tooth was counted by tartrate-resistant acid phosphatase histochemistry. The results were compared between groups using SPSS 19.0 software package. RESULTS: After 8 weeks of bone grafting, the osteogenesis effect of the experimental group was better than the control group. The OTM distance in the experimental area was higher than that in the control area. At 2, 3 and 4 week of OTM, the number of osteoclasts in the experimental group was significantly higher than that in the control group. CONCLUSIONS: A tissue-engineered complex with ß-TCP scaffolds and BMSCs could well repair the alveolar bone defect. When the adjacent tooth moved into regenerated area, the new periodontal tissue had an active response, promoting to accelerate tooth movement.

The Effects of Antisense miRNA-20a Alone or in Combination with Imatinib on K562 Cell Proliferation.

Objective: The effects of microRNA-20a (miR-20a) antisense oligonucleotides (ASODNs) on the proliferation and apoptosis of K562 cells were investigated, and the effects of these ASODNs in combination with imatinib on K562 cells were preliminarily observed. Methods: miR-20a ASODNs and scrambled oligonucleotides (SODNs) were chemically synthesized, and the later was used as the control. miR-20a ASODNs were transfected into K562 cells using Lipofectamine 2000 transfection reagent, and the expression of miR-20a was detected using real-time quantitative RT-PCR (qRT-PCR). The CCK8 assay was performed to detect the inhibition of the cell growth rate. The cells were stained by Hoechst 33258 to detect apoptotic cell morphology. Annexin V/PI double staining was used to detect the cell apoptosis rate using flow cytometry. The protein expression levels of E2F1, P21, and Bim in the K562 cell line were detected using western blotting. Results: The qRT-PCR results showed that the expression level of miR-20a in K562 cells transfected with miR-20a ASODNs was lower than those in the normal control, SODN and blank transfection groups (p < 0.05). miR-20a ASODNs significantly inhibited the growth of K562 cells as compared to the controls (p < 0.05). The Hoechst staining results showed morphological changes, suggesting apoptosis. The cell apoptosis rates in the ASODN group was (13.9 ± 1.5)%, which was significantly higher than that in the normal control group (1.84 ± 0.21)%, blank transfection group (3.21 ± 0.32)%, and SODN group (3.72 ± 0.44)% (p < 0.05). The protein expression of E2F1 and P21 in K562 cells transfected with miR-20a ASODNs were higher, while the level of Bim protein was significantly lower than that in the control groups. When miR-20a ASODNs were combined with imatinib, the growth of K562 cells was significantly inhibited as compared to the ASODN treatment alone, imatinib alone, and SODN+imatinib groups (p < 0.05). Conclusions: miR-20a ASODNs could induce apoptosis and inhibit the proliferation of K562 cells. In addition, imatinib combined with miR-20a ASODNs can increase the inhibitory effect on K562 cell proliferation.

Sex-specific parental care strategies via nestling age: females pay more attention to nestling demands than males do in the horned lark, Eremophila alpestris.

In many species, nestling demands vary continuously during early development and both parents have different parental care strategies at each nestling age. Sexual conflict arises when each parent expects its partner investing more in parental care. It is largely unknown how the two parents respond to the dynamics of nestling demands and resolve the sexual conflict during nestling period, especially on Qinghai-Tibetan Plateau. To address this question, we monitored parental care behaviors of horned larks (Eremophila alpestris) using video-recording systems. We found that male horned larks invested less in parental care, but had a larger body size than females, which is consistent with the parental investment hypothesis. Only the female brooded nestlings, but both parents contributed to feeding efforts. Feeding rates of males and females were negatively correlated, indicating that they used evolutionarily stable strategies. Strategies of parental care via nestling age were sex-specific. Females continuously adjusted care behaviors to follow the dynamics of nestling demands as nestling age increased, such as decreasing brood attentiveness and increasing feeding rate. By contrast, male feeding rate showed no significant correlation with nestling age, but increased with the synchrony feeding rate. We suggest the synchrony feeding behavior may act as a control measure for females to promote and assess the males' contribution. We consider low mating opportunities drive males to act as assistants for females, and correspondingly cause males to pay less attention to nestling demands than females.