Objective:This study aimed to investigate the role of bone morphogenetic protein 2 (BMP2)/Smad8 signaling pathway in T-2 toxin-induced apoptosis of rat articular chondrocytes.Methods:Primary chondrocytes from SD rats were cultured in vitro and exposed to varying concentrations of T-2 toxin (0.00, 0.32, 1.60, 8.00 ng/ml). The changes in chondrocytes survival rate were determined by CCK8, and the apoptosis changes of chondrocytes were determined by TUNEL assay kit. Using a group design, chondrocytes were cultured in complete culture media and culture media containing T-2 toxin (1.60 ng/ml), BMP2 cytokine (500 ng/ml), or T-2 toxin (1.60 ng/ml) + BMP2 cytokine (500 ng/ml), referred to as the control group, T-2 toxin group, BMP2 group, and T-2 toxin + BMP2 group, respectively. The survival rate and apoptosis changes of chondrocytes in each group were determined. The expression levels of Caspase-3, BMP2, BMP receptor Ⅱ (BMP-R Ⅱ), and Smad1/4/5/8 were determined by quantitative real-time PCR. Results:Compared with the 0.00 ng/ml of T-2 toxin group [(100.00 ± 0.00)%, (4.33 ± 0.32)%], the chondrocyte survival rates [(85.77 ± 2.96)%, (72.79 ± 2.31)%, (48.87 ± 1.83)%] of the 0.32, 1.60, and 8.00 ng/ml of T-2 toxin groups were significantly lower ( P < 0.05), and the apoptosis rates [(5.43 ± 0.32)%, (6.17 ± 0.15)%, (5.07 ± 0.13)%] were significantly higher ( P < 0.05). Compared with the control group, the T-2 toxin group had a lower survival rate and a higher apoptosis rate of chondrocytes ( P < 0.05). Compared with the T-2 toxin group, the T-2 toxin + BMP2 group had a higher survival rate and lower apoptosis rate of chondrocytes ( P < 0.05). Compared with the control group, the T-2 toxin group showed higher expression level of Caspase-3 mRNA in chondrocytes, while the expression levels of BMP2, BMP-R Ⅱ, and Smad1/4/8 mRNA were lower ( P < 0.05). Compared with the T-2 toxin group, the expression level of Caspase-3 mRNA was lower in the T-2 toxin + BMP2 group, while the expression levels of BMP2 and Smad8 mRNA were higher ( P < 0.05). Conclusion:BMP2 may partially block the apoptosis of chondrocytes caused by T-2 toxin by regulating the BMP2/Smad8 signaling pathway.

Objective:To investigate the effect of alendronate sodium (ALN) on osteoporosis in postmenopausal mice and the expression of growth differentiation factor-15 (GDF-15).Methods:C57BL/6J female mice (7 - 8 weeks old, weight 18 - 20 g) were selected for ovarian removal surgery (OVX) to establish a postmenopausal osteoporosis animal model. According to the random number table method, they were divided into three groups: control (sham surgery) group, OVX group, and OVX + ALN (0.5 mg·kg -1·d -1, administered orally one week before surgery) group, with 10 mice in each group. After 8 weeks of surgery, femurs and tibias were collected, and bone mineral density (BMD), bone volume fraction (BV/TV), mineral content (BMC), number of trabeculae (Tb.N), thickness of trabeculae (Tb.Th), and degree of trabecular separation (Tb. Sp) were measured in each group of mice using micro computed tomography(Micro-CT) technology. Enzyme linked immunosorbent assay and real-time quantitative PCR (qRT-PCR) were used to detect changes in serum GDF-15 levels and bone tissue GDF-15 mRNA expression levels in mice, respectively. Results:The Micro-CT examination results showed that the OVX group had a decrease of trabeculae, an increase in spacing, and sparse arrangement compared to the control group. The OVX + ALN group had an increase of bone trabeculae and a decrease in spacing compared to the OVX group. Quantitative analysis showed that the levels of BMD, BV/TV, BMC, Tb.N, and Tb.Th of the femur in the OVX group were lower than those in the control group, while the level of Tb.Sp was higher than that in the control group ( P < 0.05). The BMD, BV/TV, BMC, and Tb.N levels of the femur in the OVX + ALN group were higher than those in the OVX group, while the Tb.SP level was lower than that in the OVX group ( P < 0.05). Compared with the control group (1.86 ± 0.08, 1.02 ± 0.21), the serum GDF-15 levels and bone tissue GDF-15 mRNA expression levels in the OVX group (2.80 ± 0.45, 6.04 ± 0.42) were higher ( P < 0.05). Compared with OVX group, the serum GDF-15 levels and bone tissue GDF-15 mRNA expression levels in the OVX + ALN group (1.99 ± 0.15, 0.99 ± 0.13) were lower ( P < 0.05). Conclusion:ALN can prevent osteoporosis in postmenopausal mice, and GDF-15 is involved in its regulatory process.

Objective:To investigate the clinical characteristics of sepsis-induced myocardial injury and microbial distribution.Methods:It was a retrospective observational study conducted from Jan 2023 to Dec 2023 in the Department of Emergency Intensive Care Medicine, Changshu Hospital Affiliated to Soochow University. Patients meeting the sepsis 3.0 criteria were included, excluding those with underlying cardiovascular diseases or incomplete data. Patients were categorized into myocardial injury (SIMI) and non-myocardial injury (Non-SIMI) groups based on troponin levels. General patient information, laboratory results, microbial findings, and prognostic indicators were collected. Differences in clinical parameters between the two groups were compared. Factors showing statistical differences in univariate analysis were further analyzed using multivariable logistic regression to identify risk factors for SIMI. Conduct propensity score matching among Pulmonary infection patients who underwent bronchoalveolar lavage high-throughput sequencing to compare microbial distribution between groups. Bracken was used to estimate species-level abundance from Kraken2 results, and α and β diversity analyses were conducted on the metagenomic samples.Results:A total of 179 patients were included in the study, with 98 (54.4%) in the Non-SIMI group and 81 (45.5%) in the SIMI group. There were 69 deaths overall (38.5%), with 23 (23.7%) in the Non-SIMI group and 46 (56.8%) in the SIMI group (χ 2=20.347, P<0.01). The 28-day survival curve indicated survival rates in the SIMI group were significantly lower compared to the Non-SIMI group (Log Rank χ 2=21.270, P<0.01). Univariate analysis revealed that fungal infection rate ( P=0.007), C-reactive protein ( P=0.021), procalcitonin, blood urea nitrogen, creatinine, alanine transaminase, and lactate levels were higher in the SIMI group compared to the Non-SIMI group (all P<0.01), prothrombin time was prolonger ( P<0.01) and APACHEⅡ scores were higher ( P<0.01), while serum albumin, base excess, and platelet levels were lower (all P<0.01). Multivariable logistic regression analysis indicated that fungal infection ( OR=3.441, P=0.015) was a risk factor for SIMI, whereas base excess and platelets were protective factors ( OR=0.845, 0.988, both P<0.01). Comparison of bronchoalveolar lavage high-throughput sequencing results in the pulmonary infection subgroup showed the relative abundance of Haemophilus paraininfluenzae in Non-SIMI group was higher than SIMI group among the top 20 species ( P=0.013). There were no statistically significant differences in microbial αand β-diversity between the two groups. Conclusions:The incidence of SIMI is relatively highamong sepsis patients and it affects their prognosis. Risk factors for SIMI include fungal infection, decreased platelet count, and reduced base excess levels. Among patients with pulmonary infections, there is a lower risk of SIMI associated with Haemophilus influenzae infection.

ObjectiveTo investigate the effects of Tongxinluo capsules on traditional Chinese medicine （TCM） syndrome elements and major adverse cardiovascular events （MACEs） in patients with chronic coronary syndrome of Qi deficiency and blood stasis type. MethodsA multicenter and prospective cohort study was conducted. The intervention of Tongxinluo Capsules was used as the exposure factor， and the patients were divided into an exposure group （integrated traditional Chinese and western medicine treatment group） and a non-exposure group （western medicine treatment group）. The patients were followed up for one year. The TCM syndrome element scores were assessed by using a syndrome element diagnosis scale on the day of enrollment and in the third， sixth， and twelfth months， and the incidence of MACE within one year was recorded. ResultsA total of 186 patients were included， with 128 patients in the exposure group and 58 patients in the non-exposure group. There was no significant difference in baseline data between the two groups. Compared with those in the pretreatment period for each group， the Qi deficiency and blood stasis syndrome scores in the treatment and follow-up period were significantly improved （P<0.05）. Compared with the non-exposure group， the exposure group exhibited significantly decreased Qi deficiency syndrome scores in the treatment and follow-up period （P<0.01） and significantly reduced blood stasis syndrome scores in the sixth month （P<0.05）. In the remaining follow-up period， there was no statistically significant difference between the two groups. Compared with that of the non-exposure group， during the treatment period （the third month）， the difference in Qi deficiency and blood stasis syndrome scores of the exposure group was statistically significant （P<0.05， P<0.01）. At the end of the follow-up period， patients in the non-exposure group had a MACE probability of 6.90% （4/58）， higher than 3.13% in the exposure group （4/58）. Compared with patients with angina pectoris who used conventional medicine， patients administered with Tongxinluo Capsules had a relative risk（RR） of 0.45 ［95%confidence interval（95%CI） 0.12-1.75， P=0.26］. There was no significant difference in the incidence of MACE within one year between the two groups. ConclusionTongxinluo capsules can improve the degree of Qi deficiency in patients with chronic coronary syndrome in the short term， and the improvement effect of blood stasis syndrome appears in the medium and long term. They can better improve the Qi deficiency syndrome in the long term. Within one year， the incidence of MACE in the exposure group was lower than that in the non-exposure group.

ObjectiveTo investigate the effects of Tongxinluo capsules on traditional Chinese medicine （TCM） syndrome elements and major adverse cardiovascular events （MACEs） in patients with chronic coronary syndrome of Qi deficiency and blood stasis type. MethodsA multicenter and prospective cohort study was conducted. The intervention of Tongxinluo Capsules was used as the exposure factor， and the patients were divided into an exposure group （integrated traditional Chinese and western medicine treatment group） and a non-exposure group （western medicine treatment group）. The patients were followed up for one year. The TCM syndrome element scores were assessed by using a syndrome element diagnosis scale on the day of enrollment and in the third， sixth， and twelfth months， and the incidence of MACE within one year was recorded. ResultsA total of 186 patients were included， with 128 patients in the exposure group and 58 patients in the non-exposure group. There was no significant difference in baseline data between the two groups. Compared with those in the pretreatment period for each group， the Qi deficiency and blood stasis syndrome scores in the treatment and follow-up period were significantly improved （P<0.05）. Compared with the non-exposure group， the exposure group exhibited significantly decreased Qi deficiency syndrome scores in the treatment and follow-up period （P<0.01） and significantly reduced blood stasis syndrome scores in the sixth month （P<0.05）. In the remaining follow-up period， there was no statistically significant difference between the two groups. Compared with that of the non-exposure group， during the treatment period （the third month）， the difference in Qi deficiency and blood stasis syndrome scores of the exposure group was statistically significant （P<0.05， P<0.01）. At the end of the follow-up period， patients in the non-exposure group had a MACE probability of 6.90% （4/58）， higher than 3.13% in the exposure group （4/58）. Compared with patients with angina pectoris who used conventional medicine， patients administered with Tongxinluo Capsules had a relative risk（RR） of 0.45 ［95%confidence interval（95%CI） 0.12-1.75， P=0.26］. There was no significant difference in the incidence of MACE within one year between the two groups. ConclusionTongxinluo capsules can improve the degree of Qi deficiency in patients with chronic coronary syndrome in the short term， and the improvement effect of blood stasis syndrome appears in the medium and long term. They can better improve the Qi deficiency syndrome in the long term. Within one year， the incidence of MACE in the exposure group was lower than that in the non-exposure group.

Objective To investigate the protective effect and mechanism of astragaloside Ⅳ(AS-Ⅳ)on the pulmonary arterial hypertension(PAH)model induced by monocrotaline(MCT)/monocrotaline pyrrole(MCTP)in SD rats/human pulmonary artery endothelial cell(HPAEC).Methods In vivo experiment,60 male SD rats were randomly assigned to control group,PAH model group,AS-Ⅳ low-dose(20 mg/kg)group,AS-Ⅳ medium-dose(40 mg/kg)group,AS-Ⅳ high-dose(80 mg/kg)group,or sildenafil(Sil,100 mg/kg)group,with 10 rats in each group;except for the control group,PAH rat models were established by single intraperitoneal injection of MCT(60 mg/kg)in other groups.In vitro experiment,HPAECs were randomly assigned to control group,PAH model group,AS-Ⅳ low-dose(10 μmol/L)group,AS-Ⅳ medium-dose(20 μmol/L)group,MCTP+AS-Ⅳ high-dose(40 μmnol/L)group,or p38 mitogen-activated protein kinase(MAPK)signaling pathway inhibitor(SB203580,5 μmol/L)group;except for the control group,in vitro PAH cell models were established by MCTP(60 μg/mL)induction for 24 h in other groups.In vivo experiments,after 4 weeks of drug intervention,the right ventricular systolic pressure(RVSP)and mean pulmonary artery pressure(mPAP)of rats were measured by hemodynamic methods,the right ventricle hypertrophy index was measured by weighing methods,the percentage of pulmonary arteriole wall thickness to outer diameter(WT％)and percentage of the wall area to total vascular area(WA％)were observed by hematoxylin-eosin staining,the expression of cysteine aspartic acid specific protease 3(caspase 3)protein in lung tissue was observed by immunohistochemistry(IHC),and the apoptosis of lung tissue cells was detected by TUNEL assay.In vitro experiments,JC-1 staining was used to detect the mitochondrial membrane potential in cells,and immunofluorescence was used to detect caspase 3 protein expression.In vitro and in vivo experiments,Western blotting was used to detect the expression of caspase 3,B-cell lymphoma gene 2(Bcl-2),Bcl-2 associated X protein(Bax),p38 MAPK,and phosphorylated p38 MAPK proteins in lung tissue and HPAECs.Results In vivo experiments,the RVSP,mPAP,and right ventricle hypertrophy index were decreased in the Sil group and each dose group of AS-Ⅳ(all P＜0.01);the WA％and WT％of each dose group of AS-Ⅳ were decreased(all P＜0.01),the expression of caspase 3 protein in lung tissue was decreased(all P＜0.01),and the apoptosis of lung tissue cells was decreased(all P＜0.01).In vitro experiments showed that after intervention with each dose of AS-Ⅳ and SB203580,the mitochondrial membrane potential of HPAEC was increased(all P＜0.01)and the expression of caspase 3 was decreased(all P＜0.01).In vivo and in vitro experiments,each dose of AS-Ⅳand SB203580 reduced the expression of Bax and phosphorylated p38 MAPK proteins,and increased the expression of Bcl-2 protein(all P＜0.01).Conclusion AS-Ⅳ reduces apoptosis by inhibiting p38 MAPK signaling pathway,improving PAH in SD rats.

Objective:This study aimed to investigate the role of bone morphogenetic protein 2 (BMP2)/Smad8 signaling pathway in T-2 toxin-induced apoptosis of rat articular chondrocytes.Methods:Primary chondrocytes from SD rats were cultured in vitro and exposed to varying concentrations of T-2 toxin (0.00, 0.32, 1.60, 8.00 ng/ml). The changes in chondrocytes survival rate were determined by CCK8, and the apoptosis changes of chondrocytes were determined by TUNEL assay kit. Using a group design, chondrocytes were cultured in complete culture media and culture media containing T-2 toxin (1.60 ng/ml), BMP2 cytokine (500 ng/ml), or T-2 toxin (1.60 ng/ml) + BMP2 cytokine (500 ng/ml), referred to as the control group, T-2 toxin group, BMP2 group, and T-2 toxin + BMP2 group, respectively. The survival rate and apoptosis changes of chondrocytes in each group were determined. The expression levels of Caspase-3, BMP2, BMP receptor Ⅱ (BMP-R Ⅱ), and Smad1/4/5/8 were determined by quantitative real-time PCR. Results:Compared with the 0.00 ng/ml of T-2 toxin group [(100.00 ± 0.00)%, (4.33 ± 0.32)%], the chondrocyte survival rates [(85.77 ± 2.96)%, (72.79 ± 2.31)%, (48.87 ± 1.83)%] of the 0.32, 1.60, and 8.00 ng/ml of T-2 toxin groups were significantly lower ( P < 0.05), and the apoptosis rates [(5.43 ± 0.32)%, (6.17 ± 0.15)%, (5.07 ± 0.13)%] were significantly higher ( P < 0.05). Compared with the control group, the T-2 toxin group had a lower survival rate and a higher apoptosis rate of chondrocytes ( P < 0.05). Compared with the T-2 toxin group, the T-2 toxin + BMP2 group had a higher survival rate and lower apoptosis rate of chondrocytes ( P < 0.05). Compared with the control group, the T-2 toxin group showed higher expression level of Caspase-3 mRNA in chondrocytes, while the expression levels of BMP2, BMP-R Ⅱ, and Smad1/4/8 mRNA were lower ( P < 0.05). Compared with the T-2 toxin group, the expression level of Caspase-3 mRNA was lower in the T-2 toxin + BMP2 group, while the expression levels of BMP2 and Smad8 mRNA were higher ( P < 0.05). Conclusion:BMP2 may partially block the apoptosis of chondrocytes caused by T-2 toxin by regulating the BMP2/Smad8 signaling pathway.

Objective:To investigate the effect of alendronate sodium (ALN) on osteoporosis in postmenopausal mice and the expression of growth differentiation factor-15 (GDF-15).Methods:C57BL/6J female mice (7 - 8 weeks old, weight 18 - 20 g) were selected for ovarian removal surgery (OVX) to establish a postmenopausal osteoporosis animal model. According to the random number table method, they were divided into three groups: control (sham surgery) group, OVX group, and OVX + ALN (0.5 mg·kg -1·d -1, administered orally one week before surgery) group, with 10 mice in each group. After 8 weeks of surgery, femurs and tibias were collected, and bone mineral density (BMD), bone volume fraction (BV/TV), mineral content (BMC), number of trabeculae (Tb.N), thickness of trabeculae (Tb.Th), and degree of trabecular separation (Tb. Sp) were measured in each group of mice using micro computed tomography(Micro-CT) technology. Enzyme linked immunosorbent assay and real-time quantitative PCR (qRT-PCR) were used to detect changes in serum GDF-15 levels and bone tissue GDF-15 mRNA expression levels in mice, respectively. Results:The Micro-CT examination results showed that the OVX group had a decrease of trabeculae, an increase in spacing, and sparse arrangement compared to the control group. The OVX + ALN group had an increase of bone trabeculae and a decrease in spacing compared to the OVX group. Quantitative analysis showed that the levels of BMD, BV/TV, BMC, Tb.N, and Tb.Th of the femur in the OVX group were lower than those in the control group, while the level of Tb.Sp was higher than that in the control group ( P < 0.05). The BMD, BV/TV, BMC, and Tb.N levels of the femur in the OVX + ALN group were higher than those in the OVX group, while the Tb.SP level was lower than that in the OVX group ( P < 0.05). Compared with the control group (1.86 ± 0.08, 1.02 ± 0.21), the serum GDF-15 levels and bone tissue GDF-15 mRNA expression levels in the OVX group (2.80 ± 0.45, 6.04 ± 0.42) were higher ( P < 0.05). Compared with OVX group, the serum GDF-15 levels and bone tissue GDF-15 mRNA expression levels in the OVX + ALN group (1.99 ± 0.15, 0.99 ± 0.13) were lower ( P < 0.05). Conclusion:ALN can prevent osteoporosis in postmenopausal mice, and GDF-15 is involved in its regulatory process.

Objective:To analyze the clinical and radiographic effectiveness of a calcium silicate-based bioactive ceramic iRoot BP Plus? pulpotomy of immature permanent teeth with complicated crown fracture and to evaluate the factors influencing its long-term success rate.Methods:The digital medical records of patients under 13 years old who had undergone iRoot BP Plus? pulpotomy in the Department of Oral Emergency or the First Clinical Division,Peking University School and Hospital of Stomatology from March 2017 to September 2022 due to complicated crown fracture of anterior teeth,and had taken at least one post-operation apical radiograph were reviewed.The clinical and radiographic information at the initial examination and follow-up period were obtained,including crown color,mobility,percussion,cold test(partial pulpotomy teeth),dental restoration,fistula,swelling or inflammation of the gingival tissue,the formation of apical foramen,pathologic radiolucency and calcification of pulp chamber or root canal obliteration.Data were tested by Fisher exact test and a multiple comparison.Results:In the study,64 patients including 37 males(57.8％)and 27 females(42.2％)with a mean age of 9.1 years were finally enrolled.The total number of permanent teeth that received pulpotomy was 75,and the average follow-up time was 19.3 months.The success rate was 93.1％with the time interval between dental injury and treatment in 24 h,while the success rate dropped to 88.2％with the time intervals beyond 24 h.The time intervals did not significantly affect the pulp survival rate(P=0.61)after pulpotomy(partial or co-ronal).The success rate 6 months after pulpotomy was 96.0％,and one-year success rate was 94.7％.A total of 23 cases were reviewed for more than 2 years after pulpotomy,and 6 cases failed.The mobility had no significant effect on the success rate(P=0.28).Pulp chamber calcification and pulp canal obli-teration were not observed in all the post-operative radiographs.Conclusion:The one year clinical and radiographic success rates obtained in this study indicate that iRoot BP Plus? is an appropriate pulp cap-ping material option for pulpotomy treatment of complicated crown fracture in immature permanent teeth without displacement injuries.This technique has broad promotional value.

Objective:To analyze the therapeutic effect and safety of simultaneous integrated boost intensity-modulated radiotherapy for lung cancer brain metastases.Methods:A total of 300 patients with lung cancer brain metastases admitted to the Department of Oncology, Qingdao Municipal Hospital from March 2021 to March 2023 were selected as the study objects. The patients were divided into control group ( n=150) and study group ( n=150) by random number table method. The control group received sequential three-dimensional conformal radiotherapy, while the study group received simultaneous integrated boost intensity-modulated radiotherapy. The short-term and medium-term efficacy, target dose, and adverse reactions were compared between the two groups. Results:The short-term and medium-term total effective rates of the study group were 73.33% (110/150) and 88.67% (133/150), respectively, which were higher than those of the control group [51.33% (77/150) and 71.33% (107/150) ] ( χ2=15.46, P<0.001; χ2=14.08, P<0.001). The D min in gross tumor planning target volume and whole brain clinical planning target volume of the study group were (23.78±1.11) and (58.46±0.55) Gy, respectively, which were higher than those in the control group [ (16.67±1.08) and (53.44±0.74) Gy], with statistically significant differences ( t=56.22, P<0.001; t=66.68, P<0.001). The D mean in gross tumor planning target volume and whole brain clinical planning target volume of the study group were (44.12±0.87) and (62.19±0.57) Gy, respectively, which were higher than those in the control group [ (37.55±0.89) and (57.78±0.82) Gy], with statistically significant differences ( t=64.65, P<0.001; t=54.08, P<0.001). The total incidence of adverse reactions was 30.67% (46/150) in the study group and 36.67% (55/150) in the control group, with no significantly significant difference ( χ2=1.20, P=0.271) . Conclusion:Compared with sequential three-dimensional conformal radiotherapy, simultaneous integrated boost intensity-modulated radiotherapy has better short-term and medium-term efficacy in patients with lung cancer brain metastases. Target dose can be increased without increasing adverse reactions.