Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.

Objective:To preliminarily explore the relationship between intraventricular pressure gradients (IVPG) measured by ultrasound hemodynamic analysis and left ventricular cardiotoxicity after anthracycline chemotherapy.Methods:This was a retrospective cohort study. Patients with diffuse large B-cell lymphoma (DLBCL) who completed 6 cycles of R-CHOP chemotherapy at Fudan University Shanghai Cancer Center from 2014 to 2015 were included. Echocardiography was performed at baseline (T0), after 2 cycles of chemotherapy (T1), after 4 cycles of chemotherapy (T2), and after all chemotherapy cycles (T3). Left ventricular global longitudinal strain (LVGLS), left ventricular global circumferential strain (LVGCS), and left ventricular ejection fraction (LVEF) were analyzed using speckle-tracking imaging technology, and IVPG was measured using hemodynamic analysis technology, including IVPG of long-axis (IVPG-LA) and IVPG of short-axis. The change rate of each index from T0 to T2 was marked as Δ. Left ventricular cardiotoxicity was defined as a decrease in LVEF of ≥10% from the baseline level or LVEF ≤50%. Univariate logistic regression analysis was used to explore the related factors of left ventricular myocardial toxicity, and the receiver operating characteristic curve was drawn to analyze their evaluation efficiency for left ventricular myocardial toxicity.Results:A total of 55 patients were included, including 28 males (51%), aged (46.5±11.7) years. Twelve patients (22%) developed left ventricular cardiotoxicity. Compared with T0, IVPG-LA decreased at T1 ((10.73±2.51)% vs. (11.52±3.62)%, P=0.037); while LVGLS, LVGCS, and LVEF only decreased at T3 (all P<0.05). Univariate logistic regression analysis showed that ΔIVPG-LA and ΔLVGLS were related factors for left ventricular myocardial toxicity in patients with DLBCL receiving chemotherapy (all P<0.05). The receiver operating characteristic curve showed that the area under the curve of ΔLVGLS was 0.702, with an optimal cut-off value of 13.15% (sensitivity 66.7%, specificity 62.8%); the area under the curve of ΔIVPG-LA was 0.812, with an optimal cut-off value of 20.74% (sensitivity 75.0%, specificity 90.7%). Conclusions:Hemodynamic analysis technology shows promise clinical application value in evaluating subclinical changes in left ventricular function in tumor patients after anthracycline chemotherapy; the change rate of IVPG-LA could be used as an early indicator of left ventricular toxicity after anthracycline chemotherapy.

Objective Explored the clinical efficacy and mechanism of improved CO2 fractional laser on treatment for refractory wounds.Methods A total of 122 patients with refractory wounds who were admitted to the hospital from May 2022 to May 2024 were selected and randomly divided into a conventional group(conventional dressing change treatment+photodynamic therapy+conventional CO2 fractional laser treatment)and an improved group(conventional dressing change treatment+photodynamic therapy+improved CO2 fractional laser treatment)by lottery,with 61 cases in each group.The serum C-reactive protein(CRP)and erythrocyte sedimentation rate(ESR)levels,the number of wound bacteria before treatment and after 1,2,and 3 weeks of treatment were compared between the two groups.The wound healing rates after 1,2,and 3 weeks of treatment were compared between the two groups.The clinical efficacy of the two groups was compared.The levels of wound exudate basic fibroblast growth factor(bFGF)and angiopoietin-1(Ang-1)before treatment and after 1,2,and 3 weeks of treat-ment were compared.The adverse reactions of the two groups were compared.Results Repeated measures analy-sis of variance showed that there were an interaction effect between groups and over time in the serum CRP and ESR levels,the number of wound bacteria in both groups(P<0.05).There were no significant differences in the serum CRP and ESR levels and the number of wound bacteria between the two groups before treatment(P>0.05).The serum CRP and ESR levels and the number of wound bacteria in the improved group were lower than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).Repeated measures analysis of variance showed that there was an interaction effect between groups and over time in the wound healing rates of both groups(P<0.05).The wound healing rates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).The total clinical effective rate of the improved group was higher than that of the conventional group(P<0.05).Repeated measures analysis of variance showed that there was an inter-action effect between groups and over time in the levels of bFGF and Ang-1 in wound exudates of both groups(P<0.05).There was no significant difference in the levels of bFGF and Ang-1 in wound exudates between the two groups before treatment(P>0.05).The levels of bFGF and Ang-1 in wound exudates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).There were no signifi-cant differences in the incidence of pain,pruritus and rash between the two groups(P>0.05).Conclusions In the treatment of refractory wounds,the combination of improved CO2 fractional laser and photodynamic therapy could promote the secretion of wound exudate growth factors,inhibit wound bacterial growth and inflammatory response,accelerate wound healing,and improve clinical efficacy,with high safety.The mechanism of action is related to the promotion of wound exudate growth factor secretion.

Objective Explored the clinical efficacy and mechanism of improved CO2 fractional laser on treatment for refractory wounds.Methods A total of 122 patients with refractory wounds who were admitted to the hospital from May 2022 to May 2024 were selected and randomly divided into a conventional group(conventional dressing change treatment+photodynamic therapy+conventional CO2 fractional laser treatment)and an improved group(conventional dressing change treatment+photodynamic therapy+improved CO2 fractional laser treatment)by lottery,with 61 cases in each group.The serum C-reactive protein(CRP)and erythrocyte sedimentation rate(ESR)levels,the number of wound bacteria before treatment and after 1,2,and 3 weeks of treatment were compared between the two groups.The wound healing rates after 1,2,and 3 weeks of treatment were compared between the two groups.The clinical efficacy of the two groups was compared.The levels of wound exudate basic fibroblast growth factor(bFGF)and angiopoietin-1(Ang-1)before treatment and after 1,2,and 3 weeks of treat-ment were compared.The adverse reactions of the two groups were compared.Results Repeated measures analy-sis of variance showed that there were an interaction effect between groups and over time in the serum CRP and ESR levels,the number of wound bacteria in both groups(P<0.05).There were no significant differences in the serum CRP and ESR levels and the number of wound bacteria between the two groups before treatment(P>0.05).The serum CRP and ESR levels and the number of wound bacteria in the improved group were lower than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).Repeated measures analysis of variance showed that there was an interaction effect between groups and over time in the wound healing rates of both groups(P<0.05).The wound healing rates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).The total clinical effective rate of the improved group was higher than that of the conventional group(P<0.05).Repeated measures analysis of variance showed that there was an inter-action effect between groups and over time in the levels of bFGF and Ang-1 in wound exudates of both groups(P<0.05).There was no significant difference in the levels of bFGF and Ang-1 in wound exudates between the two groups before treatment(P>0.05).The levels of bFGF and Ang-1 in wound exudates in the improved group were higher than those in the conventional group after 1,2,and 3 weeks of treatment(P<0.05).There were no signifi-cant differences in the incidence of pain,pruritus and rash between the two groups(P>0.05).Conclusions In the treatment of refractory wounds,the combination of improved CO2 fractional laser and photodynamic therapy could promote the secretion of wound exudate growth factors,inhibit wound bacterial growth and inflammatory response,accelerate wound healing,and improve clinical efficacy,with high safety.The mechanism of action is related to the promotion of wound exudate growth factor secretion.

Objective:To evaluate the application of metagenomic next-generation sequencing（mNGS）in the identification of non-tuberculous mycobacteria（NTM）.Methods:A retrospective analysis was conducted on mNGS results of 358 bronchoalveolar lavage fluid（BALF）samples positive for NTM collected at the First Affiliated Hospital of Zhejiang University School of Medicine from February 2021 to January 2024. The analysis included the distribution of NTM species，the detection of mixed pathogens，and the performance of conventional mycobacterial detection methods.Results:The results showed that 362 strains of 15 NTM species were identified from 350 specimens，8 specimens were not precise to the species level. The most frequently detected species were Mycobacterium intracellulare（37.3%，135/362）， Mycobacterium abscessus（26.8%，97/362），followed by Mycobacterium avium（11.0%，40/362）， Mycobacterium kansasii（8.0%，29/362）and Mycobacterium chelonae（7.7%，28/362）. Single NTM species were detected in 339 specimens，while two or three NTM species were simultaneously detected in 11 specimens（3.1%，11/358）. Non-NTM microorganisms co-infected were detected in 53.4%（191/358）of NTM-positive BALF samples，including common pathogens such as Pseudomonas aeruginosa， Staphylococcus aureus，and Aspergillus fumigatus；and difficult-to-identify pathogens such as Legionella pneumophila and Talaromyces marneffei. In NTM-positive patients detected by mNGS，the results supported the diagnosis of NTM infection in 298 cases（298/358，83.2%）and 105 cases（105/358，29.3%）initiated anti-NTM treatment accordingly；while in 60 cases（60/358，16.8%）the positive results were considered as colonization or unrelated to clinical infection. For samples tested with acid-fast staining，mycobacterial liquid culture，and DNA microarray，the positivity rates for NTM were 31.5%（73/232），48.7%（57/117），and 43.0%（46/107），respectively. Conclusions:mNGS demonstrates advantages in identification of NTM. However，the test may detect multiple microorganisms，in that case，the interpretation with clinical and radiological results is requried to determine the main pathogens.

Objective:To preliminarily explore the relationship between intraventricular pressure gradients (IVPG) measured by ultrasound hemodynamic analysis and left ventricular cardiotoxicity after anthracycline chemotherapy.Methods:This was a retrospective cohort study. Patients with diffuse large B-cell lymphoma (DLBCL) who completed 6 cycles of R-CHOP chemotherapy at Fudan University Shanghai Cancer Center from 2014 to 2015 were included. Echocardiography was performed at baseline (T0), after 2 cycles of chemotherapy (T1), after 4 cycles of chemotherapy (T2), and after all chemotherapy cycles (T3). Left ventricular global longitudinal strain (LVGLS), left ventricular global circumferential strain (LVGCS), and left ventricular ejection fraction (LVEF) were analyzed using speckle-tracking imaging technology, and IVPG was measured using hemodynamic analysis technology, including IVPG of long-axis (IVPG-LA) and IVPG of short-axis. The change rate of each index from T0 to T2 was marked as Δ. Left ventricular cardiotoxicity was defined as a decrease in LVEF of ≥10% from the baseline level or LVEF ≤50%. Univariate logistic regression analysis was used to explore the related factors of left ventricular myocardial toxicity, and the receiver operating characteristic curve was drawn to analyze their evaluation efficiency for left ventricular myocardial toxicity.Results:A total of 55 patients were included, including 28 males (51%), aged (46.5±11.7) years. Twelve patients (22%) developed left ventricular cardiotoxicity. Compared with T0, IVPG-LA decreased at T1 ((10.73±2.51)% vs. (11.52±3.62)%, P=0.037); while LVGLS, LVGCS, and LVEF only decreased at T3 (all P<0.05). Univariate logistic regression analysis showed that ΔIVPG-LA and ΔLVGLS were related factors for left ventricular myocardial toxicity in patients with DLBCL receiving chemotherapy (all P<0.05). The receiver operating characteristic curve showed that the area under the curve of ΔLVGLS was 0.702, with an optimal cut-off value of 13.15% (sensitivity 66.7%, specificity 62.8%); the area under the curve of ΔIVPG-LA was 0.812, with an optimal cut-off value of 20.74% (sensitivity 75.0%, specificity 90.7%). Conclusions:Hemodynamic analysis technology shows promise clinical application value in evaluating subclinical changes in left ventricular function in tumor patients after anthracycline chemotherapy; the change rate of IVPG-LA could be used as an early indicator of left ventricular toxicity after anthracycline chemotherapy.

Background Salidroside (SAL) has a protective effect on multiple organ systems. Exposure to fine particulate matter (PM_2.5) in the atmosphere may lead to disruptions in gut microbiota and impact intestinal health. The regulatory effect of SAL on the gut microbiota of mice exposed to PM_2.5 requires further investigation. Objective To evaluate gut microbiota disruption in mice after being exposed to PM_2.5 and the potential effect of SAL. Methods Forty male C57BL/6 mice, aged 6 to 8 weeks, were randomly divided into four groups: a control group, an SAL group, a PM_2.5 group, and an SAL+PM_2.5 group, each containing 10 mice. In the SAL group and the SAL+PM_2.5 group, the mice were administered SAL (60 mg·kg⁻¹) by gavage, while in the control group and the PM_2.5 group, sterile saline (10 mL·kg⁻¹) was administered by gavage. In the PM_2.5 group and the SAL+PM_2.5 group, PM_2.5 suspension (8 mg·kg⁻¹) was intratracheally instilled, and in the control group and SAL group, sterile saline (1.5 mL·kg⁻¹) was intratracheally administered. Each experiment cycle spanned 2 d, with a total of 10 cycles conducted over 20 d. Histopathological changes in the ileum tissue of the mice were observed after HE staining. Colon contents were collected for gut microbiota sequencing and short-chain fatty acids (SCFAs) measurements. Results The PM_2.5 group showed infiltration of inflammatory cells in the ileum tissue, while the SAL+PM_2.5 group exhibited only a small amount of inflammatory cell infiltration. Compared to the control group, the PM_2.5 group showed decreased Shannon index (P<0.05) and increased Simpson index (P<0.05), indicating that the diversity of gut microbiota in this group was decreased; the SAL+PM_2.5 group showed increased Shannon index compared to the PM_2.5 group (P<0.05) and decreased Simpson index (P<0.05), indicating that the diversity of gut microbiota in mice intervened with SAL was increased. The principal coordinates analysis (PCoA) revealed a significant separation between the PM_2.5 group and the control group, while the separation trend was less evident among the control group, the SAL group, and the SAL+PM_2.5 group. The unweighted pair-group method with arithmetic means (UPGMA) clustering tree results showed that the control group and the SAL group clustered together first, followed by clustering with the SAL+PM_2.5 group, and finally, the three groups clustered with the PM_2.5 group. The PCoA and UPGMA clustering results indicated that the uniformity and similarity of the microbiota in the PM_2.5 group were significantly decreased. Compared to the control group, the PM_2.5 group showed decreased abundance of phylum Bacteroidetes and Candidatus_Saccharimonas (P<0.05) and increased abundance of phylum Proteobacteria, genus Escherichia, genus Bacteroides, genus Prevotella, genus Enterococcus, and genus Proteus (P<0.05). Compared to the PM_2.5 group, the SAL+PM_2.5 group showed decreased abundance of phylum Proteobacteria, phylum Actinobacteria, genus Prevotella, and genus Proteus (P<0.05), and increased abundance of Candidatus_Saccharimonas (P<0.05). The PM_2.5 group showed reduced levels of propionic acid, valeric acid, and hexanoic acid compared to the control group (P<0.05), while the SAL+PM_2.5 group showed increased levels of propionic acid, isobutyric acid, butyric acid, valeric acid, and hexanoic acid compared to the PM_2.5 group (P<0.05). Conclusion Exposure to PM_2.5 can cause pathological alterations, microbial dysbiosis, and disturbing production of SCFAs in intestinal tissue in mice. However, SAL can provide a certain degree of protective effect against these changes.

Objective To analyze the changes in left ventricular deformation function in patients with diffuse large B-cell lymphoma(DLBCL)treated with different doses of anthracycline chemotherapy using 3D speckle-tracking imaging(3D-STI).Methods 66 DLBCL patients receiving anthracycline chemotherapy were enrolled.Based on the cumulative dose of anthracycline received,the patients were divided into a high-dose group(＞360 mg/m2,n=39)and a low-dose group(≤360 mg/m2,n=27).Patients underwent transthoracic echocardiography before chemotherapy and within one week after completion of the entire chemotherapy cycle.Left ventricular global longitudinal strain(LVGLS),left ventricular global circumferential strain(LVGCS),and other indices were analyzed using 3D-STI to assess changes in left ventricular deformation indices after chemotherapy and between two groups.Results Compared to baseline,DLBCL patients showed significant reductions in LVGLS,LVGCS and left atrial global longitudinal strain(LAGLS)after treatment completion(P＜0.001).When comparing the high-dose group with the low-dose group,there was a significant increase in relative LVGCS change rate at the end of chemotherapy(21.12[6.52,35.37]vs 5.49[-14.73,27.01];P=0.03).However,there were no statistically significant differences in relative left ventricular ejection fraction(LVEF),LVGLS,LVGCS,LVEF change rate,or LVGLS change rate between the two groups.Conclusions 3D-STI can be a potential method to identify the sub-clinical deterioration of left ventricular systolic function in patients received anthracycline chemotherapy,the difference of change rate of LVGCS may predict the variation of sub-clinical deterioration of left ventricular function between patients received high and low doses of anthracycline chemotherapy.

Objective: To analyze the epidemiological characteristics of acute paraquat(PQ)poisoning in children in southwest Shandong, and the risk factors for pulmonary interstitial fibrosis. Methods: This retrospective study was performed on the clinical data of children with acute PQ poisoning admitted from January 2013 to December 2017 in 12 hospitals in southwest Shandong.All participants were divided into pulmonary interstitial fibrosis group and no pulmonary interstitial fibrosis group on the basis of the chest CT 14 days after poisoning.The epidemiological characteristics and risk factors of pulmonary interstitial fibrosis were analyzed. Results: During the study period, a total of 307 children with acute PQ poisoning were admitted to 12 hospitals, of which 61 (19.87%) were suffering from acute PQ poisoning.Forty-nine cases with complete clinical data were analyzed, including 26 male and 23 female patients poisoned by oral.The age distribution ranged from 8 months to 14 years.Poisoning mainly occured from July to September of each year.The mortality of acute PQ poisoning was 8.2%(4/49), and the incidence of pulmonary interstitial fibrosis in survival patients was 44.4%(20/45). Statistical differences (P<0.05) were found between the pulmonary interstitial fibrosis and no pulmonary interstitial fibrosis, with regard to the times of blood purification, the time from poison exposure to blood purification, the application rate of glucocorticoids, the concentration of PQ in urine, the pediatric critical illness score, the time from poison exposure to gastric lavage, the white blood count at admission, serum creatinine, arterial blood lactate, PaO₂, PaCO₂, and PaO₂/FiO₂; however, there was no significant difference in the proportion of blood purification treatment, the mode of blood purification treatment, alanine aminotransferase, aspartate aminotransferase, urea nitrogen, creatine kinase and troponin.Stepwise logistic regression analysis showed that the time from exposure to poison to gastric lavage(OR=0.683, 95%CI 0.210-2.222)and to blood purification(OR=0.0133, 95%CI 0.004-0.042), the times of blood purification(OR=2.862, 95%CI 1.450-5.648), concentration of PQ in urine(OR=1.435, 95%CI 1.085-1.898), and the use of glucocorticoids(OR=0.190, 95%CI 0.048-0.757) were the risk factors for pulmonary interstitial fibrosis(P<0.05). Conclusion Early gastric lavage and blood purification, increasing the frequence of adminitrating purification appropriately, using low-dose glucocorticoids can reduce the incidence of pulmonary interstitial fibrosis of children with acute PQ poisoning.

Objective To observe the effects of olprinone on ischemia/reperfusion (I/R) induced myocardial injury in male (Sprague-Dawley, SD rats) and explore its mechanisms. Methods Rats were subjected to a 30-min coronary arterial occlusion followed by 24-hour reperfusion. The survival rats were randomly divided into sham group (n=6), ischemia reperfusion group (I/R group, n=9), ischemia reperfusion+low dose of olprinone group(IR+olprinone-L group, n=6), ischemia reperfusion+medium dose of olprinone group (IR+olprinone-M group, n=6),ischemia reperfusion +high dose of olprinone group (IR+olprinone-H group, n=6). A MAP heart function analysis system was used to measure hemodynamic parameters; TTC staining method was used to detect the myocardial infarct size;24-hour mortality of SD rats was recorded; western blot was used to detect the levels of Caspase-3, Bax,Bcl-2, LC3B/LC3A,Beclin-1. Results Cardiac function in I/R group was lower than that in sham group, which was significantly improved by pretreatment with olprinone (P＜0.01),but systolic arterial pressure (SAP) diastolic arterial pressure (DAP) mean arterial pressure (MAP) mean pressure developed in left ventricle (Pmean) had no significant difference (P＞0.05). The percentage of myocardial infarct size in olprinone-M and olprinone-H group was lower than that in I/R group (P＜0.05).There was no significant difference in mortality among groups within 24 hours. Compared with sham group, the expressions of Caspase-3 and Bax were obviously up-regulated in I/R group (P＜0.01), whereas caspase-3 was down-regulated in olprinone-M group (P＜0.05) and Bax was inhibited by different doses of olprinone (P＜0.05), but the expression of Bcl-2 increased (P＜0.05); furthermore, the ratio of Bcl-2/Bax decreased in I/R group (P＜0.01) and increased with different degrees in different doses of olprinone (P＜0.05). Meanwhile, compared with sham group, the expression of Beclin-1 was up-regulated in I/R group(P＜0.05),and also increased in olprinone-L and olprinone-M groups(P＜0.05), but the ratio of Bcl-2 /Beclin-1 decreased in different doses of olprinone making statistically significant difference only in olprinone-M group (P＜0.05). Moreover, different doses of olprinone elevated the different ratios of LC3B/LC3A (P＜0.05), and this elevated ratio in olprinone-M group at median among groups. Conclusions Olprinone can strengthen the cardiac function after myocardial ischemia/reperfusion injury, without leading to disorders in hemodynamics; by regulating autophagy with anti-apoptotic protein, olprinone can make autophagy to an appropriate level using the mechanism of autophagy to preventing the myocardium from injury.