Objective:Drug safety assessments based on real-world data are often challenged by both treatment switching and rare events. In this study, we used statistical simulations to investigate the effects of switching rates and treatment effects on the statistical performance of commonly used analytical strategies and methods under overlapping scenarios of treatment switching and rare events.Methods:The simulation scenario was set up as a bidirectional treatment switching (allowing the control group to switch to the treatment group and the treatment group to switch to the control group), and the event rates were set at approximately 2%, 5%, and 20%. Different simulation scenarios were generated with sufficient sample size to consider switching rate and relative treatment effect. The simulated datasets were analyzed using three types of analysis strategy, i.e. intention to treat (ITT), per protocol (PP), and as treated (AT). The performance of five indicators, namely percentage bias, mean square error, empirical standard error, coverage, and rejection rate, were compared among the different methods in different scenarios, and recommendations for method selection were given.Results:In terms of analytical strategies and methods, AT analysis were relatively optimal in terms of percentage bias and accuracy, followed by PP analysis and ITT analysis. When the relative treatment effects converged (e.g. HR=1.0), both the ITT analysis and the time-dependent AT approaches (marginal structural model, time-dependent Cox regression model or time-dependent propensity score matching) performed well; when the relative treatment effects were small (e.g. HR=0.8), the marginal structural model was the most optimal; when the relative treatment effects were large (e.g. HR=0.6 or 0.4), the approaches of using a censored treatment for switchers in the AT analysis were more accurate. In addition, the time-dependent AT approaches had the highest rejection rate when there was a difference in treatment effect between the two groups, and the ITT analysis had the lowest rejection rate. Conclusions:For the dual challenges of bidirectional switching and rare events in real-world drug safety evaluations, adequate sample size is a prerequisite for accurate estimation of treatment effects, while switching rates and effect sizes of switched drugs might also affect estimation accuracy. Appropriate strategies and methods should be selected for the analysis. It is necessary to consider whether the event is rare or not, the switching rate and the expected treatment effect size of the two types of treatment to select appropriate analysis strategies and methods.

Tooth pulpitis is a prevalent oral disorder. Understanding the underlying mechanisms of pulpitis and developing effective treatment strategies hold great significance. Ferroptosis has recently emerged as a new form of cell death, but the role of ferroptosis in pulpitis remains largely unknown. In our study, single-cell RNA sequencing (scRNA-seq) was used to identify cellular heterogeneity between 3 pulpitis tissue and 3 healthy pulp tissue, and explored ferroptosis occurrence in pulpitis tissue and inflamed dental pulp cells (DPCs). In scRNA-seq, 40 231 cells (Pulpitis: 17 814; Healthy pulp: 22 417) were captured, and visualized into 12 distinct cell clusters. Differentially expressed ferroptosis-related genes (DE-FRGs) were almost presented in each cluster in pulpitis vs healthy pulp. ROS and Fe²⁺ levels significantly rose, and immunohistochemistry showed low expression of GPX4 and high expression of PTGS2 in pulpitis. In LPS-stimulated DPCs, thymosin α1 increased the expression of GPX4 and FTL, and decreased expression of TNF-α, IL-1β, IL-6, and Fe²⁺ levels. In rat pulpitis models, both prothymosin α (PTMA, precursor of thymosin α1) gelatin sponge placed at the hole of pulp (LPS-P(gs)) and PTMA injection in pulp (LPS-P(i)) significantly reduced infiltration of inflammatory cells and expression of PTGS2, and increased the expression of GPX4. In RNA sequencing, the expression of DE-FRGs were reversed when thymosin α1 were added in LPS-stimulated DPCs. Collectively, single-cell atlas reveals cellular heterogeneity between pulpitis and healthy pulp, and ferroptosis occurrence in pulpitis. Thymosin α1 may reduce ferroptosis in DPCs to alleviate pulpitis and thus potentially has the ability to treat pulpitis.
Ferroptosis/drug effects* ; Dental Pulp/drug effects* ; Animals ; Pulpitis/pathology* ; Rats ; Thymalfasin/pharmacology* ; Humans ; Male ; Thymosin/pharmacology* ; Disease Models, Animal ; Rats, Sprague-Dawley

Objective To investigate the effect of the mutant of adenovirus-mediated HIF-1α triple mutant(Ad-HIF-1α-Trip)on the apoptosis of H9c2 cardiomyocytes under high glucose and hypoxia conditions.Methods H9c2 cardiomyocytes were cultured under hypoxic conditions in vitro and randomly divided into four groups based on glucose concentration and viral transfection status:low oxygen+normal glucose concentration group(LO+NG group),low oxygen+high glucose concentration group(LO+HG group),low oxygen+high glucose+adenoviral null vector group(LO+HG+Ad-Null group),and low oxygen+high glucose+adenovirus HIF-1α triple mutant group(LO+HG+Ad-HIF-1α-Trip group).After 12 hours of hypoxia,HIF-1α,PI3K,and Akt expression were measured via qRT-PCR and Western blot,and apoptosis of H9c2 cardiomyocytes was assessed by flow cytometry.Results HIF-1α,PI3K,and Akt expression decreased in cardiomyocytes under high glucose and hypoxia,while Ad-HIF-1 α-Trip enhanced their expression.Flow cytometry revealed increased apoptosis under high glucose and hypoxia,which was reduced by Ad-HIF-1α-Trip.Conclusion Ad-HIF-1α-Trip upregulates HIF-1α,PI3K,and Akt in H9c2 cardiomyocytes under high glucose and hypoxia,likely reducing apoptosis via PI3K-Akt pathway activation.

Objective:Drug safety assessments based on real-world data are often challenged by both treatment switching and rare events. In this study, we used statistical simulations to investigate the effects of switching rates and treatment effects on the statistical performance of commonly used analytical strategies and methods under overlapping scenarios of treatment switching and rare events.Methods:The simulation scenario was set up as a bidirectional treatment switching (allowing the control group to switch to the treatment group and the treatment group to switch to the control group), and the event rates were set at approximately 2%, 5%, and 20%. Different simulation scenarios were generated with sufficient sample size to consider switching rate and relative treatment effect. The simulated datasets were analyzed using three types of analysis strategy, i.e. intention to treat (ITT), per protocol (PP), and as treated (AT). The performance of five indicators, namely percentage bias, mean square error, empirical standard error, coverage, and rejection rate, were compared among the different methods in different scenarios, and recommendations for method selection were given.Results:In terms of analytical strategies and methods, AT analysis were relatively optimal in terms of percentage bias and accuracy, followed by PP analysis and ITT analysis. When the relative treatment effects converged (e.g. HR=1.0), both the ITT analysis and the time-dependent AT approaches (marginal structural model, time-dependent Cox regression model or time-dependent propensity score matching) performed well; when the relative treatment effects were small (e.g. HR=0.8), the marginal structural model was the most optimal; when the relative treatment effects were large (e.g. HR=0.6 or 0.4), the approaches of using a censored treatment for switchers in the AT analysis were more accurate. In addition, the time-dependent AT approaches had the highest rejection rate when there was a difference in treatment effect between the two groups, and the ITT analysis had the lowest rejection rate. Conclusions:For the dual challenges of bidirectional switching and rare events in real-world drug safety evaluations, adequate sample size is a prerequisite for accurate estimation of treatment effects, while switching rates and effect sizes of switched drugs might also affect estimation accuracy. Appropriate strategies and methods should be selected for the analysis. It is necessary to consider whether the event is rare or not, the switching rate and the expected treatment effect size of the two types of treatment to select appropriate analysis strategies and methods.

Objective To investigate the effect of the mutant of adenovirus-mediated HIF-1α triple mutant(Ad-HIF-1α-Trip)on the apoptosis of H9c2 cardiomyocytes under high glucose and hypoxia conditions.Methods H9c2 cardiomyocytes were cultured under hypoxic conditions in vitro and randomly divided into four groups based on glucose concentration and viral transfection status:low oxygen+normal glucose concentration group(LO+NG group),low oxygen+high glucose concentration group(LO+HG group),low oxygen+high glucose+adenoviral null vector group(LO+HG+Ad-Null group),and low oxygen+high glucose+adenovirus HIF-1α triple mutant group(LO+HG+Ad-HIF-1α-Trip group).After 12 hours of hypoxia,HIF-1α,PI3K,and Akt expression were measured via qRT-PCR and Western blot,and apoptosis of H9c2 cardiomyocytes was assessed by flow cytometry.Results HIF-1α,PI3K,and Akt expression decreased in cardiomyocytes under high glucose and hypoxia,while Ad-HIF-1 α-Trip enhanced their expression.Flow cytometry revealed increased apoptosis under high glucose and hypoxia,which was reduced by Ad-HIF-1α-Trip.Conclusion Ad-HIF-1α-Trip upregulates HIF-1α,PI3K,and Akt in H9c2 cardiomyocytes under high glucose and hypoxia,likely reducing apoptosis via PI3K-Akt pathway activation.

Objective·To explore the efficacy and safety of compound amino acid capsules in the treatment of malnutrition and calcium and phosphorus metabolism disorders in maintenance hemodialysis patients.Methods·In this prospective,randomized,controlled,single-center study,forty maintenance hemodialysis patients from Renji Hospital,Shanghai Jiao Tong University School of Medicine were randomly divided into two groups,the treatment group(n=21)and the control group(n=19).The treatment group was given oral compound amino acid capsules on the basis of regular hemodialysis treatment,while the control group received no special nutritional intervention.Serum albumin,prealbumin,hemoglobin,ferritin,calcium,phosphorus,1,25-(OH)2-D3 and intact parathyroid hormone levels were analyzed every 3 months,and the incidence of adverse events including death,cardio-cerebrovascular accidents and vascular access failure was recorded.The total follow-up period was 9 months.Results·Serum albumin and prealbumin in the treatment group at 6-month and 9-month were significantly higher than the baseline parameters(albumin,t=3.574,5.599,both P<0.05;prealbumin,t/Z=-2.485,2.921,both P<0.05).Albumin in the control group increased at 9-month with a lower amplification compared to the treatment group(t=3.877,P=0.001),while the difference of prealbumin showed no statistical significance during follow-up.Hemoglobin and serum ferritin in the treatment group started to increase at 3-month(hemoglobin,t=2.192;ferritin,t=2.994;both P<0.05).Phosphorus in treatment group decreased at 3-month and 9-month(t/Z=-2.743,-2.103,both P<0.05),while phosphorus in the control group remained relatively stable during the first 6 months and increased at 9-month(Z=-2.178,P=0.029).Calcium and 1,25-(OH)2-D3 in the treatment group at 3-month and 6-month were significantly higher than the baseline parameters(calcium,t=4.581,4.922,both P=0.000;1,25-(OH)2-D3 t/Z=4.504,-2.374,both P<0.05),while the increase in blood calcium in the control group was significantly smaller than that in the treatment group during the same period.1,25-(OH)2-D3 in the control group showed no significant improvement.There was no significant difference in intact parathyroid hormone level,incidence of adverse events and other laboratory examination results between the two groups.Conclusion·Compound amino acid capsules can ameliorate the nutrition status and regulate calcium and phosphorus metabolism effectively and safely in maintenance hemodialysis patients.

Objective:To analyze the correlation between paraspinal muscle parameters and the angle of degenerative thoracolumbar segmental kyphosis.Methods:From November 2021 to April 2023, a total of 90 female patients with lumbar degenerative disease who underwent surgical treatment in the Department of Spine Surgery, Beijing Jishuitan Hospital were retrospectively analyzed. The average age was 67.62±4.98 years (range, 60-80 years), and the average height was 1.58±0.05 m (range, 1.48-1.70 m). Weight 63.79±9.13 kg (range, 47-90 kg), body mass index 25.48±3.35 kg/m 2 (range, 18.37-36.05 kg/m 2). The angle of kyphosis of the thoracolumbar segments was 6.65°±10.38° (range, -17.34° to 9.34°). Disease diagnosis: 32 cases of lumbar disc herniation and 58 cases of lumbar spinal stenosis. Frontal and lateral radiographs of the thoracolumbar segments in the standing position were taken to measure the angle of kyphosis of the thoracolumbar segments; quantitative CT of the thoracolumbar segments and Osirix software were used to measure the parameters of the paravertebral muscles at the levels of T 12, L 3, and L 5, including paravertebral muscle cross-sectional area, skeletal muscle area, and tissue density, the proportion of fat infiltration, and the height-corrected skeletal muscle area index. The correlation between paraspinal muscle parameters and the angle of thoracolumbar segmental kyphosis was analyzed, and the factors affecting the angle of thoracolumbar segmental kyphosis were analyzed using multiple linear regression. Results:Correlation analysis showed a negative correlation between the angle of thoracolumbar segmental kyphosis and the tissue density of T 12 skeletal muscle ( r=-0.303, P=0.004) and L 5 skeletal muscle ( r=-0.219, P=0.038). Age was negatively correlated with T 12 skeletal muscle tissue density ( r=-0.263, P=0.012), T 12 height-corrected skeletal muscle area index ( r=-0.221, P=0.036), T 12 paravertebral muscle cross-sectional area ( r=-0.280, P=0.007), L 3 skeletal muscle tissue density ( r=-0.266, P=0.011) and L 5 skeletal muscle tissue density ( r=-0.290, P=0.006). There was no correlation between bone mineral density and paravertebral muscle parameters ( P>0.05). Multiple linear regression showed that T 12 skeletal muscle tissue density ( β=-1.125, P<0.001), T 12 fat-infiltrated proportion ( β=-0.849, P=0.001), L 3 skeletal muscle tissue density ( β=0.996, P<0.001), and L 3 fat-infiltrated proportion ( β=0.496, P=0.020) were independent factors influencing the angle of thoracolumbar segmental kyphosis . Conclusion:T 12 and L 3 paraspinal muscle density and fat-infiltrated proportion are independent factors affecting the angle of thoracolumbar kyphosis. The smaller the density and fat-infiltrated proportion of T 12 paraspinal muscle, and the larger the density and fat-infiltrated proportion of L 3 paraspinal muscle, the larger the angle of thoracolumbar kyphosis.

Objective : To observe the clearance of smear layer on the root canal wall in different action time by scanning electron microscope (SEM), and to determine the optimal amount of time using sonically activated irrigation to wash root canal in clinic. Methods: Fifty-six ex vivo human anterior teeth with single straight root canal were selected. After routine mechanical preparation, they were divided into two experimental groups according to different irrigating agents: saline group and EDTA group. Each group was assisted by VDW sonic activation EDDY. The saline group was divided into three subgroups according to the irrigating time: 5 s, 30 s and 50 s; EDTA group was divided into six subgroups according to the irrigating time: 5 s, 10 s, 20 s, 30 s, 40 s and 50 s. The control group did not undergo root canal irrigation. After irrigation, the root was cut longitudinally. The smear layer of crown, middle and apical of root canal wall was observed by SEM. Results: After irrigating for 30 seconds, there was a significant difference between the normal saline group and the control group and the 5 second group (P<0.05), and there was no difference in the middle and apical part (P>0.05). After 50 seconds, there was a significant difference in the score of the smear layer between the apical area and the other groups (P<0.05). After irrigating for 5 seconds or 10 seconds in EDTA group, there was a significant difference between the scores of the crown and middle area of the root canal and the control group (P<0.05), and there was no significant difference in the apical area (P>0.05). There was a significant difference between the 20-40 second group and the first two groups (P<0.05). There was a significant difference between the 50 second group and the other groups (P<0.05). Comparing the cleaning effect on the smear layer after 50 seconds of irrigating between the two experimental groups, the whole root canal showed significant statistical difference (P<0.05). Conclusion The EDTA-assisted sonic activated device used for 50 seconds has the best cleaning effect.

Objective:To evaluate the systemic absorption and safety of multiple doses of topical tazarotene/betamethasone dipropionate cream in healthy subjects and patients with psoriasis.Methods:From September 2008 to April 2009, 12 healthy subjects collected from Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College were randomly and equally divided into tazarotene 0.15%/betamethasone dipropionate 0.15% cream group and tazarotene 0.2%/betamethasone dipropionate 0.2% cream group; these subjects were instructed to apply 0.03 g of the test drug per day on each of the 4 body sites, including the flexor aspects of bilateral forearms, waist and back, for 7 consecutive days, and venous blood samples were obtained before, and 1, 3, 5 and 7 days after the start of drug application. From October 2010 to August 2011, 60 patients with non-cephalic psoriasis collected from the Hospital for Skin Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College were randomly divided into 3 groups at a ratio of 3∶1∶1, i.e., tazarotene 0.05%/betamethasone dipropionate 0.05% cream group ( n = 36) and tazarotene 0.05% gel group ( n = 12) topically treated with a cream vehicle in the morning and the test drug at night, and betamethasone dipropionate 0.05% cream group ( n = 12) topically treated with the test drug twice a day (once in the morning and again in the evening) ; the treatment lasted 6 consecutive weeks, and venous blood samples were collected before, and 2, 4 and 6 weeks after drug application. Liquid chromatography-tandem mass spectrometry was performed to determine the concentrations of tazarotenic acid and betamethasone in plasma. During the trial, adverse events in the subjects were recorded, routine blood and urine examinations were carried out, and liver and kidney function were evaluated before and after treatment. Results:The plasma concentrations of tazarotenic acid and betamethasone in the 12 healthy subjects were below the lower limit of quantitation (0.04 μg/L) after 1-, 3-, 5- and 7-day treatment. After the consecutive treatment, tazarotenic acid and betamethasone were detected in 2 (5.56%) and 4 (11.11%) patients respectively at week 2, 4 or 6 in the tazarotene 0.05%/betamethasone dipropionate 0.05% cream group, and the highest plasma concentrations of tazarotenic acid and betamethasone were 0.112 and 0.201 μg/L respectively; in the betamethasone dipropionate 0.05% cream group, betamethasone was detected in 2 of 12 patients, and the highest plasma concentration of betamethasone was 0.112 μg/L. No test drug-related systemic adverse reactions or laboratory abnormalities were observed in any of the healthy subjects or patients.Conclusion:Multiple doses of topical tazarotene/betamethasone dipropionate cream has advantages of little systemic absorption, no long-term accumulation and good systemic safety.

Objective To investigate the relationship between serum secreted phospholipase A2 (sPLA2) and the severity of the disease in cobra bite patients. Methods Fifty-two cobra bite patients admitted to our hospital from January 2015 to December 2015 were selected according to the inclusion and exclusion criteria. According to the degree of disease, patients were divided into the light and heavy risk groups. According to the degree of swelling of the affected limbs, patients were divided into the mild swelling group and moderate-severe swelling group. According to the time difference between visits, patients were divided into: visit time <8 h group and visit time ≥ 8 h group. Twenty healthy adult volunteers served as the control group. The levels of serum sPLA2, C reactive protein (CRP), D-dimer (DD), lactic acid (LAC) and peripheral blood leukocyte count (WBC) were measured and compared in different groups. Quantitative data were analyzed using t test and rank sum test. Results The levels of serum sPLA2 in the light and heavy risk group were statistically different from those in the control group (P<0.05), and were statistically significant between the light and heavy risk groups (P<0.01) and between the mild swelling group and moderate-severe swelling group (P<0.05). The serum sPLA2 levels in group A and group B were also significantly different from those in the control group (P<0.05). The level of sPLA2 was positively correlated with the level of LAC, DD and WBC, and the correlation coefficients were 0.3142, 0.2752 and 0.6534, respectively. Conclusions The higher the level of serum sPLA2, the more serious the patient's poisoning symptoms. It is of certain clinical value to evaluate the condition of cobra.