BACKGROUND:Inflammation affects the osteogenic differentiation of periodontal ligament stem cells,and the osteogenic ability and autophagy level of periodontal ligament stem cells are closely related.However,there are no relevant reports on whether inflammation affects the osteogenic ability and autophagy level of periodontal ligament stem cells at different stages of osteogenic differentiation. OBJECTIVE:To explore alkaline phosphatase expression and autophagy periodontal ligament stem cells levels in periodontitis and normal conditions. METHODS:Periodontal ligament stem cells from normal and periodontitis patients were isolated and cultured,and underwent Vimentin,pan-CK,and Stro-1 fluorescence staining.At 3,7,and 14 days of osteogenic differentiation,western blot assay was used to detect the protein expression levels of alkaline phosphatase,LC3B,Beclin1,and ATG5 in normal and inflammatory periodontal ligament stem cells.The mRNA expression levels of alkaline phosphatase,bone sialoprotein,osteocalcin,Runx2,LC3B,Beclin1,and ATG5 were detected by real-time PCR. RESULTS AND CONCLUSION:(1)Stro-1 was positive,Vimentin was positive,and pan CK was negative in periodontal ligament stem cells.(2)At 3,7,and 14 days after osteogenic differentiation,compared with normal periodontal ligament stem cells,the mineralization nodules formed by periodontal ligament stem cells from inflammatory sources were significantly reduced(P<0.01);the expression of alkaline phosphatase protein and mRNA was significantly lower(P<0.05);the mRNA expression levels of bone sialoprotein,osteocalcin,and Runx2 were significantly decreased(P<0.05).(3)At 7 and 14 days after osteogenic differentiation,compared with normal periodontal ligament stem cells,the expression levels of ATG5,LC3B,and Beclin1 proteins and mRNA of periodontal ligament stem cells were downregulated(P<0.05).These findings suggest that inflammation reduces the activity of periodontal ligament stem cells in mineralizing nodule formation and the expression of alkaline phosphatase and weakens the autophagy potential of periodontal ligament stem cells at 7 and 14 days after osteogenic differentiation.

Tyrosine kinase inhibitors （TKIs） represent a class of small-molecule targeted drugs that improve the survival time of patients with gastrointestinal stromal tumor （GIST）. Imatinib， sunitinib， regorafenib， ripretinib， and avapritinib are commonly used TKIs in the clinical treatment of various types of GIST. This article provides a comprehensive review of the pharmacokinetics and therapeutic drug monitoring （TDM） of these five drugs， finding that there is significant individual variability in the pharmacokinetics of these drugs. Among them， the absorption of imatinib， regorafenib， and avapritinib are influenced by food intake. Imatinib should be taken with meals and 200 mL of water， regorafenib is taken with a low-fat meal， while avapritinib is taken on an empty stomach. TKIs are mainly metabolized by cytochrome P450 3A4 （CYP3A4）， and when used in combination with CYP3A4 inducers or inhibitors， drug exposure levels will significantly change； apart from metabolic enzymes， the exposure levels of TKIs are also influenced by interactions with the transporter proteins P-glycoprotein and breast cancer resistance protein. Currently， research on TDM for TKIs is still in the exploratory stage， with a substantial amount of literature reporting the effective concentrations of imatinib， sunitinib and regorafenib. However， the precise relationship between exposure levels and efficacy/ toxicity needs further exploration. Currently， there is a lack of research on the correlation between exposure levels and efficacy/ toxicity of ripretinib and avapritinib. It is recommended to implement TDM in patients taking these drugs and explore their therapeutic window in combination with pharmacokinetic models. The commonly used methods for clinical TDM of TKIs include immunoassay， chromatography， and surface-enhanced Raman spectroscopy， providing a technical basis for clarifying the therapeutic window of TKIs.

Objective:To explore the clinical effectiveness of a self-designed robot reduction system for femoral intertrochanteric fractures.Methods:A retrospective study was conducted to analyze the 57 patients with intertrochanteric fracture who had been treated at Department of Orthopedics, The Fourth Affiliated Central Hospital of Tianjin Medical University from June 2022 to February 2023. The patients were divided into a robot group (using the self-designed robot reduction system to assist intramedullary nailing) and a traction bed group (using a traction bed to assist intramedullary nailing) based on their fracture reduction method. The robot group: 31 patients, 11 males and 20 females, with an age of (78.7±9.3) years; 16 left and 15 right sides; 17 cases of type 31-A1, 12 cases of type 31-A2 and 2 cases of type 31-A3 by the AO/OTA classification. The traction bed group: 26 patients, 12 males and 14 females, with an age of (78.7±7.7) years; 13 left and 13 right sides; 16 cases of type 31-A1, 9 cases of type 31-A2 and 1 cases of type 31-A3 by the AO/OTA classification. The 2 groups were compared in terms of reduction and operation time, intraoperative blood loss, fluoroscopy frequency, reduction quality, and VAS and Harris score at preoperation, 1 week and 6 months postoperation.Results:The 2 groups were comparable due to insignificant differences in their preoperative general data ( P>0.05). The robot group was significantly better than the traction bed group in reduction time [(4.4±2.2) min versus (9.4±3.2) min], operation time [(29.0±13.5) min versus (49.3±13.3) min], intraoperative blood loss [(76.5±30.5) mL versus (115.0±38.4) mL], fluoroscopy frequency [(10.2±2.6) times versus (14.8±3.2) times], and good/excellent rate of reduction [80.6% (25/31) versus 50.0% (13/26)] ( P<0.05). All patients were followed up for (6.8±0.3) months. Respectively, the VAS scores at preoperation and 6 months postoperation was (6.2±1.3) and (2.4±0.8) points for the robot group, and (6.3±1.3) and (2.7±0.8) points for the traction bed group, showing no statistically significant differences between the 2 groups ( P>0.05). However, the VAS score was (3.3±1.2) points for the robotic group and (4.8±1.5) points for the traction bed group at 1 week postoperation, showing a statistically significant difference between the 2 groups ( P<0.001). Respectively, the Harris scores at preoperation and 6 months postoperation were (35.3±3.0) and (88.7±3.4) points for the robot group, and (35.6±2.9) and (87.2±3.5) points for the traction bed group, showing no statistically significant differences between the 2 groups ( P>0.05). However, the Harris score was (57.3±3.7) points for the robotic group and (46.7±2.8) points for the traction bed group at 1 week postoperation, showing a statistically significant difference between the 2 groups ( P<0.05). The patient satisfaction rates in the robot and traction bed groups were 96.8% (30/31) and 92.3% (24/26), respectively, showing no statistically significant difference ( P>0.05). Conclusion:Our self-designed robot reduction for femoral intertrochanteric fractures can effectively shorten reduction and operation time, reduce bleeding and fluoroscopy frequency, and enhance anatomical reduction.

Objectives:To explore the causal relationship between body mass index,uric acid and congestive heart failure(CHF),and provide genetic evidence to support the association between uric acid-mediated body mass index and the risk of CHF. Methods:Using the published data set of genome-wide association studies(GWAS)in East Asia,inverse variance weighting(IVW)method was used as the main analysis method,and MR-Egger method,weighted median model(WME),simple model and weighted model were used to analyze the causal relationship between body mass index,uric acid and CHF.MR-Egger regression was used to detect pleiotropy,Cochran Q test was used to detect heterogeneity,leave-one-out sensitivity was used to detect bias,funnel plot was drawn to detect bias,and MR-PRESSO package was used to remove outlier single nucleotide polymorphism(SNP).After Mendelian analysis,the mediating effect ratio was calculated,and the reverse Mendelian randomization study results between body mass index,uric acid and CHF were analyzed. Results:IVW method showed that body mass index(OR=1.685,95%CI:1.417-2.003,P<0.001)and uric acid(OR=1.225,95%CI:1.087-1.380,P<0.001)were risk factors of CHF in two-sample Mendelian analysis.Body mass index(OR=1.204,95%CI:1.139-1.273,P<0.001)was a risk factor for uric acid.The mediating effect of uric acid was 7.23%.The P values of MR-Egger regression intercept terms were all>0.05,that is,there was no pleiotropy of the selected SNP,and the causal inference method was valid.The Cochran Q test P values of body mass index and uric acid,and body mass index and CHF were<0.01,indicating heterogeneity.IVW analysis of CHF and body mass index in reverse Mendelian analysis was OR=0.977,95%CI:0.947-1.008,P>0.05 and CHF and uric acid was OR=1.000,95%CI:0.963-1.038,P>0.05,so the reverse causal inference was not valid.However,the analysis of uric acid and body mass index showed pleiotropy,so the causal inference method was invalid. Conclusions:There is a causal and positive correlation between body mass index and CHF.There is a causal relationship and positive correlation between uric acid and CHF.However,uric acid is an incomplete mediator between body mass index and CHF.

The incidence of psycho-cardiological diseases， i.e.， cardiovascular diseases combined with psychological disorders， is increasing year by year. Brain-derived neurotrophic factor （BDNF） plays a role in the pathogenesis of such diseases. According to the theory of collateral diseases， our team innovates the concept of regulating mental activity by dredging collaterals in the treatment of psycho-cardiological diseases and summarizes the concepts of "heart of Qi and collaterals" and "heart of vessels and collaterals". We believe that obstructed collaterals and disturbed mental activity run through the whole course of psycho-cardiological diseases， being the core pathogenesis. BDNF closely related to the core pathogenesis can regulate nerve and vascular inflammation， alleviate oxidative stress， and mediate a variety of signaling pathways， thereby promoting the survival and repair of nerve cells and vascular endothelial cells to regulate emotion and protect the heart. Therefore， BDNF is one of the potential biomarkers for clinical treatment of psycho-cardiological diseases. Collateral obstruction caused by blood stasis is specifically manifested as collateral deficiency， blood stasis， and Qi stagnation in collaterals. It can easily lead to inflammation， free radical generation， and antioxidant system changes in the patients with psycho-cardiological diseases， which can cause oxidative stress damage， affect the BDNF level， and result in mental disorders， such as anxiety and depression. Disturbed mental activity is mainly caused by the disturbance in the heart of Qi and collaterals， which is specifically manifested as the disturbance of the mind and liver soul. It is prone to cause anxiety or depression symptoms， which is closely related to the BDNF-mediated abnormal activation of neural circuits， nerve injury， and inflammation. This article elaborates on the theoretical connotation and pathological mechanism of regulating mental activity by dredging collaterals in the treatment of psycho-cardiological diseases from the perspective of BDNF， aiming to provide new ideas for the prevention and treatment of psycho-cardiological diseases and collateral diseases.

This article systematically summarizes the theoretical framework, training content, training methods, and evaluation tools of TeamSTEPPS curriculum, and explores its application status and future research directions in nursing clinical training and college teaching, in order to provide a basis for further promoting this curriculum in nursing clinical training and college teaching in China.

Objective To establish a fast,stable,and sensitive zebrafish model of osteoporosis(OP)using different method.Methods OP models were induced by iron overload or prednisolone(Pred),and bone formation and mortality were observed.The groups were divided into:Control group,model group(include FAC group and Pred group),and positive control group(AC group).Ammonium ferric citrate was used as the model drug in the iron-overload induction method.For the Pred induction models,the modeling time for the Pred-3 days post-fertilization(dpf)method was 3～9 dpf,the modeling time for the Pred-5 dpf method was 5～10 dpf,and Pred was administered from 3 dpf and removed from 7～9 dpf for the Pred withdrawal method.To compare the anti-osteoporosis(OP)effects of commonly used drugs such as Alfacalcidol(AC),Calcitriol(CA),and Alendronate(AL),it's important to select a stable and sensitive positive control drug and to further optimize different staining methods and conditions.Results There was no significant effect of ammonium ferric citrate 500 μg/mL on bone formation.Bone formation and the length of the first vertebra were significantly decreased in the Pred group induced by Pred-3 dpf compared with those in the control group(P＜0.01,P＜0.05),but zebrafish mortality was higher.There was no significant difference between the Pred-5 dpf method,but bone formation was significantly reduced in the Pred withdrawal group(P＜0.01),with no mortality.Alfacalcidol,calcitriol,and alendronate all had anti-OP effects,with CA having the most sensitive and stable anti-OP effect.Alizarin red staining showed that the optimal dye parameters were 0.02%concentration for dyeing 2 h,with washing in 0.5%KOH and glycerol under the conditions of a 3∶1 ratio for 3 h followed by a 1∶1 ratio for 14 h.The result of staining showed that calcein was more sensitive for staining bone nodes and ARS staining was more sensitive for staining the first vertebra.Conclusions The Pred withdrawal method can be used to establish a rapid,stable,and sensitive OP model in zebrafish as a reliable model for studying OP.

Objective To establish a drosophila reproductive dysfunction model and observe whether Danggui Shaoyao San(DSS)has an improving effect.Methods Tripterygium glycosides were used to establish the drosophila reproductive dysfunction model.The number of offspring pupae,the parental superoxide dismutase(SOD)and catalase(CAT)activities,the parental reproductive-related gene expression,the F1 generation body weight,and the F1 generation development-related genes were measured to evaluate the efficacy of DSS.Results Tripterygium glycosides in a dose of 20 mg·mL-1 significantly inhibited the quantity of offspring(P<0.01)and were suitable for establishing the drosophila reproductive dysfunction model.Compared with the model control group,medium(10 mg·mL-1)and high(20 mg·mL-1)dose DSS significantly increased the number of offspring pupae(P<0.01,P<0.05).High-dose DSS significantly increased the SOD and CAT activities of the parent drosophila(P<0.01).Medium and high doses of DSS promoted the expression of drosophila parental reproduction-related genes,among which the medium dose DSS increased the mRNA expression of DEAD-Box helicase 4(VASA)and factor in the germline alpha(Figla)(P<0.05,P<0.01),and the high dose DSS increased the mRNA expression of VASA,Figla and forkhead box O(FoxO)(P<0.01).The medium dose of DSS significantly increased the body weight of F1-generation female drosophila(P<0.05),and the high dose of DSS significantly increased the body weight of both F1-generation female and male drosophila(P<0.05,P<0.01).Both the medium and high dose DSS increased the mRNA expression of the F1 generation development-related gene mammalian target protein of rapamycin(mTOR)(P<0.01).Conclusions DSS improves the reproductive function injury of drosophila induced by tripterygium glycosides.

OBJECTIVE To establish a method for concentration determination of caffeine and its three metabolites， theophylline， paraxanthine and theobromine in urine， and apply it in clinical practice. METHODS Using caffeine-13C3-d3 as internal standard （IS）， and the urine samples were protein precipitated with acetonitrile； HPLC-MS/MS method was adopted to determine the concentrations of caffeine and its three metabolites. The determination was performed on Waters ACQUITY UPLC® BEH HILIC column with mobile phase consisting of 60 mmol/L ammonium acetate （A）-acetonitrile （B） （gradient elution） at the flow rate of 0.5 mL/min. The column temperature was set at 38 ℃ ， and the sample size was 2 μL. The electrospray ionization detection was operated in a positive mode by multiple reaction monitoring. The detection ions for quantitative analysis were m/z 195.1→110.0 for caffeine， m/z 181.1→124.0 for theophylline， m/z 181.1→124.0 for paraxanthine， m/z 181.1→138.0 for theobromine， and m/z 198.1→ 140.1 for IS. The above method was used to determine the concentrations of caffeine and its three metabolites in the urine of 19 infants with apnea of prematurity （AOP）. RESULTS The linear ranges of mass concentration of caffeine， theophylline， paraxanthin and theobromine were 0.200-200， 0.050-50.0，0.050 0-50.0， and 0.100-100 μg/mL， respectively. The lower limits of quantification were 0.200， 0.050， 0.050 and 0.100 μg/mL （r＞0.990）， respectively. RSDs of intra-day and intra- day precision were not above 10.37%， and matrix factors were 85.68%-109.90%； extraction recoveries were 93.53%-109.40% （RSD≤15%）， and RSDs of stability tests were all lower than 15%. The concentrations of caffeine and its three metabolites in the urine of 19 cases were （27.346±7.951）， （0.351±0.223）， （0.428±0.395） and （0.472±0.374） μg/mL， respectively. CONCLUSIONS The established HPLC-MS/MS method is simple， sensitive and can be used for the determination of caffeine and its three metabolites in urine samples of AOP.

Objective:To evaluate the feasibility of indocyanine green fluorescence imaging technology applied in thyroid surgery to identify parathyroid gland.Methods:From Oct 2021 to May 2022, data of 42 patients undergoing thyroidectomy via gasless unilateral axillary approach in Zhejiang Provincial Hospital of Traditional Chinese Medicine were retrospectively analyzed. Cases using intraoperative fluorescence imaging technology(42 cases) were compared with conventional laparocopic approach.Results:The number of parathyroid glands dissected in the study group 1.57±0.61 was higher than that in the control group 0.56± 0.59 ( t=-5.472, P<0.05). The PTH value of the study group was (2.88±1.23)pmol/L on the first day after operation, which was higher than that of the control group (2.16±0.10)pmol/L ( t=-1.844, P<0.05). The blood parathyroid hormone value on the third day(3.22±1.31)pmol/L was higher than that of the control group (2.55±0.81) pmol/L ( t=-2.041, P<0.05). There were 2 cases of hypoparathyroidism in the study group, less than 5 cases in the control group, but there was no significant difference between the two groups( χ2=0.942, P>0.05). There was 1 case of hypocalcemia in the study group and 3 cases in the control group ( χ2=0.731, P>0.05). Conclusion:Using indocyanine green fluorescence imaging technology to identify parathyroid gland is feasible, simple, fast, safe and effective.