Objective: To investigate the efficacy of magnesium-strontium co-doped hydroxyapatite mineralized collagen (MSHA/Col) in improving the bone repair microenvironment and enhancing bone regeneration capacity, providing a strategy to address the insufficient biomimetic composition and limited bioactivity of traditional hydroxyapatite mineralized collagen (HA/Col) scaffolds. Methods: A high-molecular-weight polyacrylic acid-stabilized amorphous calcium magnesium strontium phosphate precursor (HPAA/ACMSP) was prepared. Its morphology and elemental distribution were characterized by high-resolution transmission electron microscopy (TEM) and energy-dispersive spectroscopy. Recombinant collagen sponge blocks were immersed in the HPAA/ACMSP mineralization solution. Magnesium-strontium co-doped hydroxyapatite was induced to deposit within collagen fibers (experimental group: MSHA/Col; control group: HA/Col). The morphological characteristics of MSHA/Col were observed using scanning electron microscopy (SEM). Its crystal structure and chemical composition were analyzed by X-ray diffraction and Fourier transform infrared spectroscopy, respectively. The mineral phase content was evaluated by thermogravimetric analysis. The scaffold's porosity, ion release, and in vitro degradation performance were also determined. For cytological experiments, CCK-8 assay, live/dead cell staining, alkaline phosphatase staining, alizarin red S staining, RT-qPCR, and western blotting were used to evaluate the effects of the MSHA/Col scaffold on the proliferation, viability, early osteogenic differentiation activity, late mineralization capacity, and gene and protein expression levels of key osteogenic markers [runt-related transcription factor 2 (Runx2), collagen type Ⅰ (Col-Ⅰ), osteopontin (Opn), and osteocalcin (Ocn)] in mouse embryonic osteoblast precursor cells (MC3T3-E1). Results: HPAA/ACMSP appeared as amorphous spherical nanoparticles under TEM, with energy spectrum analysis showing uniform distribution of carbon, oxygen, calcium, phosphorus, magnesium, and strontium elements. SEM results of MSHA/Col indicated successful complete intrafibrillar mineralization. Elemental analysis showed the mass fractions of magnesium and strontium were 0.72% (matching the magnesium content in natural bone) and 2.89%, respectively. X-ray diffraction revealed characteristic peaks of hydroxyapatite crystals (25.86°, 31°-34°). Infrared spectroscopy results showed characteristic absorption peaks for both collagen and hydroxyapatite. Thermogravimetric analysis indicated a mineral phase content of 78.29% in the material. The scaffold porosity was 91.6% ± 1.1%, close to the level of natural bone tissue. Ion release curves demonstrated sustained release behavior for both magnesium and strontium ions. The in vitro degradation rate matched the ingrowth rate of new bone tissue. Cytological experiments showed that MSHA/Col significantly promoted MC3T3-E1 cell proliferation (130% increase in activity at 72 h, P < 0.001). MSHA/Col exhibited excellent efficacy in promoting osteogenic differentiation, significantly upregulating the expression of osteogenesis-related genes and proteins (Runx2, Col-Ⅰ, Opn, Ocn) (P < 0.01). Conclusion The MSHA/Col scaffold achieves dual biomimicry of natural bone in both composition and structure, and effectively promotes osteogenic differentiation at the genetic and protein levels, breaking through the functional limitations of pure hydroxyapatite mineralized collagen. This provides a new strategy for the development of functional bone repair materials

Objective:To prepare novel targeted lipid nanobubbles（MEB-NBs）that can be loaded with exosomes（Exo），and test the property and explore the specific invitro target-seeking ability of MEB-NBs. Methods:The core lipid nanobubbles（NBs）were prepared using mechanical oscillation methods，and Exo was connected to the NBs through membrane fusion. The target antibody MYH6 was linked to the NBs using a phospholipid coupling covalent method，resulting in the preparation of targeted lipid nanobubbles MEB-NBs loaded with Exo. The morphology，particle size，and surface potential of the prepared lipid nanobubbles MEB-NBs were observed，and the co-loading of Exo and NBs was verified using laser confocal and fluorescence resonance energy transfer techniques. The in vitro imaging capability and biosafety of MEB-NBs were evaluated，and the loading rate and drug content of Exo carried by MEB-NBs were measured. The ability of MEB-NBs to specifically target hiPSC-CMs cells was observed using flow cytometry，small animal in vivo imaging systems，and laser confocal microscopy. Results:The average particle size of the prepared MEB-NBs was（597.10 ± 47.70）nm，the surface potential was（-11.70 ± 0.21）mV，and the concentration was（2.43 ± 0.06）×10 7/ml. Laser confocal microscopy and FRET results confirmed that the Exo was effectively connected with the NBs；When the Exo concentration was 250 μg/ml，the Exo loading rate and drug loading of MEB-NBs were both 73.8%，and the results of laser confocal microscopy showed that MEB-NBs could be effectively targeted to hiPSC-CMs cells. MEB-NBs were well developed in vitro and had good biological safety. Conclusions:This study constructs novel Exo-loaded targeted lipid nanobubbles（MEB-NBs）that demonstrate stable physicochemical properties and ultrasound imaging capabilities. The developed MEB-NBs precisely target hiPSC-CMs cells，providing an innovative drug delivery system with enhanced therapeutic efficacy for Exo-based therapies.

Objective:To prepare novel targeted lipid nanobubbles（MEB-NBs）that can be loaded with exosomes（Exo），and test the property and explore the specific invitro target-seeking ability of MEB-NBs. Methods:The core lipid nanobubbles（NBs）were prepared using mechanical oscillation methods，and Exo was connected to the NBs through membrane fusion. The target antibody MYH6 was linked to the NBs using a phospholipid coupling covalent method，resulting in the preparation of targeted lipid nanobubbles MEB-NBs loaded with Exo. The morphology，particle size，and surface potential of the prepared lipid nanobubbles MEB-NBs were observed，and the co-loading of Exo and NBs was verified using laser confocal and fluorescence resonance energy transfer techniques. The in vitro imaging capability and biosafety of MEB-NBs were evaluated，and the loading rate and drug content of Exo carried by MEB-NBs were measured. The ability of MEB-NBs to specifically target hiPSC-CMs cells was observed using flow cytometry，small animal in vivo imaging systems，and laser confocal microscopy. Results:The average particle size of the prepared MEB-NBs was（597.10 ± 47.70）nm，the surface potential was（-11.70 ± 0.21）mV，and the concentration was（2.43 ± 0.06）×10 7/ml. Laser confocal microscopy and FRET results confirmed that the Exo was effectively connected with the NBs；When the Exo concentration was 250 μg/ml，the Exo loading rate and drug loading of MEB-NBs were both 73.8%，and the results of laser confocal microscopy showed that MEB-NBs could be effectively targeted to hiPSC-CMs cells. MEB-NBs were well developed in vitro and had good biological safety. Conclusions:This study constructs novel Exo-loaded targeted lipid nanobubbles（MEB-NBs）that demonstrate stable physicochemical properties and ultrasound imaging capabilities. The developed MEB-NBs precisely target hiPSC-CMs cells，providing an innovative drug delivery system with enhanced therapeutic efficacy for Exo-based therapies.

Objective To explore the correlation between sarcopenia and body fat percentage(BF%)in elderly male patients with type 2 diabetes mellitus(T2DM).Methods A total of 291 elderly male T2DM patients who were hospitalized in the Endocrinology Department of Xuanwu Hospital,Capital Medical University were enrolled in this study from December 2018 to September 2019.All the patients were divided into sarcopenia group(n=35)and non sarcopenia group(n=256).BF%,skeletal muscle mass index(SMI),grip strength,gait speed,HbA1c,hemoglobin(Hb),blood uric acid(SUA),albumin and vitamin D(Vit D)were measured in all the participants.Results Compared with the non sarcopenia group,age,HbA1c and BF%were increased,while BMI,Vit D,SMI,Hb,SUA,ALB,grip strength,and gait speedwere decreased in sarcopenia group(P＜0.05 or P＜0.01).Pearson correlation analysis showed that SMI was positively correlated with BMI,Vit D,Hb,and SUA(P＜0.05 or P＜0.01),and negatively correlated with HbA1c(P＜0.01).Grip strength and gait speed are positively correlated with Vit D and Hb(P＜0.01),and negatively correlated with BF%(P＜0.01).Logistic regression analysis showed that age and BF%were risk factors for sarcopenia,while BMI and Vit D were protective factors for sarcopenia.Conclusions High BF%significantly increases the risk of sarcopenia in elderly male T2DM patients.

Objective:To prepare dual-targeted lipid ultrasound microbubbles loaded with ANM33 (HA-PANBs) and evaluate its feasibility in targeting foam cells by stages and achieving precise intracellular drug release in vitro. Methods:The dual-targeteded lipid ultrasound microbubbles were designed with nanobubbles (NBs) as the microbubble core, hyaluronic acid (HA) as the first-stage targeting ligand for damaged endothelial cells, aptamer PM1 as the second-stage targeting moiety for foam cells, and ANM33 as the therapeutic factor. Simultaneously with the characterization of the lipid bubbles, the stability and in vitro contrast-enhanced ultrasound imaging capability were detected. Then a co-culture model of damaged human umbilical vein endothelial cells (HUVEC) and macrophages (RAW264.7, MΦ) was established, combined with flow cytometry, oil red O staining and small animal in vivo imaging to evaluate the ability of HA-PANBs in targeting foam cells precisely and releasing ANM33. Results:The HA-PANBs exhibited regular morphology and good structural stability, with a particle size of (1 357.53±140.20)nm and a surface potential of (-5.61±0.73)mV. HA, PM1 and ANM33 were effectively connected. In the damaged HUVEC/MΦ co-culture system, the HA-PANBs group demonstrated the best targeting effect on foam cells, with an effective uptake of (80.65±2.12)%, which was 56.74% higher than that of the NBs group. Oil red O staining revealed that the cholesterol efflux capacity of foam cells in the HA-PANBs group was significantly better than that in the NBs group, the results were statistically different [(629.80±21.99) a.u.vs (1 071.00±55.49)a.u., P<0.05]. Conclusions:The dual-targeted lipid ultrasound microbubbles (HA-PANBs) can precisely target foam cells and significantly enhance their cholesterol efflux, providing a new strategy for the early non-invasive diagnosis and treatment of atherosclerosis.

Objective:To evaluate the safety and effectiveness of dual-plane breast augmentation via endoscopic assisted axillary approach.Methods:A retrospective analysis was conducted on 215 female patients who underwent dual-plane breast augmentation via endoscopic assisted axillary approach from March 2019 to March 2021 (15 cases at Longhua Maternal and Child Health Hospital and 200 cases at Chongqing Huamei Hospital). The patient′s age was 22-45 (32.32±5.67) years. 42 cases underwent dual-plane breast augmentation via endoscopic assisted axillary approach, by using a combination of blunt and sharp methods to separate the cavity; while 173 cases used sharp methods to separate cavities. The complications related to breast augmentation were evaluated during a follow-up period of 6 to 18 months.Results:None of the 215 patients experienced wound infection, postoperative hematoma formation, or skin burns. The long-term complications included 1 case (0.46%) of grade Ⅰ capsule contracture, 1 case (0.46%) of grade Ⅲ capsule contracture, and 12 cases (5.58%) of nipple areola sensory impairment or reduction. A patient with grade Ⅲ capsule contracture underwent right capsulotomy and replacement of the prosthesis.Conclusions:The endoscope-assisted transaxillary dual-plane breast augmentation surgery has higher safety and satisfactory, worthy of clinical promotion and application.

Objective:To study the effect of continuous subcutaneous insulin infusion(CSII)on the emotional disorder of children and adolescents with type 1 diabetes mellitus(T1DM)and their parents.Methods:A total of 72 children and adolescents with T1DM were divided into CSII group( n=40)and multiple daily injection(MDI)group( n=32).There were 58 healthy children and adolescents with their parents selected as control group.The emotional condition of children and adolescents in T1DM group and control group was evaluated by Depression Self-rating Scale for Children(DSRS)and the Screen for Child Anxiety Related Emotional Disorders(SCARED)respectively, and Symptom Checklist-90(SCL-90)was used for evaluating the mental health of all parents. Results:The average glycated hemoglobin A1c(HbA1c)of T1DM group was at the optimal level(7.406±1.294)%.The average HbA1c of CSII group was significantly lower than that of MDI group[(7.040±1.082)% vs(7.863±1.404)%, t=2.728, P=0.008].The depression rate of children and adolescents in T1DM group increased significantly than that of control group(31.9% vs 15.5%, χ2 =4.671, P=0.031).There were statistically significant differences among CSII group, MDI group and control group(20.0% vs 46.9% vs 15.5%, χ2 =11.591, P=0.003).The depression rate of children and adolescents in MDI group increased significantly than that of CSII group and control group(all P<0.05).CSII group showed similar results as compared with control group( P>0.05).Concerning the anxiety in children and adolescents, there was no significant difference between T1DM group and control group(19.4% vs 13.8%, χ2=0.730, P=0.393), and there were no significant differences among CSII group, MDI group and control group(15.0% vs 25.0% vs 13.8%, χ2=1.994, P=0.369).The emotional disorder prevalence of parents in T1DM group was remarkably higher than that of the control group(31.9% vs 5.2%, χ2=52.927, P<0.01).The factor scores of obsessive-compulsive symptoms, interpersonal sensitivity, depression, anxiety, hostility, paranoia and psychotic symptoms in T1DM group were higher than that of control group( P<0.05).There were statistically significant differences among CSII group, MDI group and control group(17.5% vs 50.0% vs 5.2%, χ2=26.126, P<0.01).The emotional disorder prevalence of parents in MDI group increased significantly than that of CSII group and control group(all P<0.05).But CSII group was same as that of control group( P>0.05). Conclusion:The children and adolescents with T1DM and their parents were high-risk population of emotional disorder.CSII can reduce not only the depression in the children and adolescents with T1DM, but also emotional disorder of their parents, thus CSII can improve the mental health in families suffering from T1DM.

Objective To investigate the gene expression of sigma factors in vivo, and to explore the sigma factors that may be closely related to the virulence of pathogenic Mycobacterium tuberculosis Methods Tuberculosis (TB) patients diagnosed in the outpatient department of Tianjin Tuberculosis Control Center from January to December 2018 were selected, and 20 sputum-positive specimens were randomly selected from TB patients confirmed with Xpert-positive for the present study. Two immediate sputum specimens were collected from each case of pulmonary tuberculosis before treatment, one for RNA extraction and one for in vitro culture. In vitro cultured strains in the logarithmic phase of growth were harvested for RNA extraction. The specific primers for 13 sigma factors were designed. The differential expression of the 13 sigma factors between sputum isolates and in vitro cultured strains was analyzed by fluorescence quantitative PCR. Taking ribosomal 16s as the reference gene, the transcription level of sigma factors was analyzed by 2^ΔCt. Using the stably expressed sigA as the control reference, the expression differences of other sigma factors were analyzed by one-way ANOVA. Results Within 0 days, stress-associated sigma factors have a different expression profile in clinical isolate strains vs H37Rv or in vitro. All the sigma factors induced up regulation in sputum ,while no difference transcription between clinical isolate strains vs H37Rv（P＞0.05）. When compared to in vitro culture ,only sigM transcript highest in sputum(P＜0.05). Conclusion SigM plays an important role in the initial stages of bacterial infection, but its exact role is unclear.We assumed it could have a role in the interplay between the host immune defenses and the bacterial escape mechanisms.

Abstract： Objective　To analyze the species distribution of microorganisms in culture tubes reported positive byBACTEC™ MGIT960 (hereafter referred to as "MGIT960") after species identification using matrix-assisted laser desorption/ionization time of flight massspectrometry (MALDI-TOF MS) technique. Methods　From 2021 to 2022, a total of 2 662 positive tubes reported by the MGIT 960 instrument at Tuberculosis Reference Laboratory of Tianjin Center for Tuberculosis Control were collected. Liquid cultures were independently inoculated to blood plate and neutral L-J medium, and the resulting isolate strains were identified using the MALDI-TOF MS method. According to the MALDI-TOF MS results, the non-repetitive results of the same patient on the same culture medium were analyzed for the composition ratio of strain distribution. For the strains not identified by MALDI-TOF MS, 38 strains were selected for 16S rRNA gene sequencing. Results　A total of 605 isolates were obtained from blood plates, and 501 of those were analyzed. Among them, Mycobacterium accounted for 17.76% (89/501), predominant by Mycobacterium abscess 10.18% (51/501) and Mycobacterium fortuitum 3.19% (16/501). Bacteria other than Mycobacterium accounted for 68.06% (341/501), with the main ones being Nocardia farcinica 15.57% (78/501), Gordonia sputa 9.38% (47/501) and Gordonia bronchialis 7.58% (38/501). There were 71 unidentifiable strains, making up 14.17% (71/501). A total of 2 378 strains were isolated from neutral L-J mediums, 1 748 of which were used in the incoming analysis. Among these, 78.72% (1 376/1 748) were Mycobacterium, 60.53% (1 058/1 748) were Mycobacterium tuberculosis (MTB), 4.69% (82/1 748) were Mycobacterium chimaer intracellulare group and 3.55% (62/1 748) were Mycobacterium Lentiflavum. Bacteria other than Mycobacterium accounted for 17.11% (299/1 748) in neutral L-J medium isolates, with Nocardia farcinica 4.35% (76/1 748), Gordonia sputa 2.69% (47/1 748) and Gordonia bronchialis 2.12% (37/1 748) as the main species. There were 73 strains that couldn't be identified, comprising 4.18% (73/1 748). The 38 strains that not identified by MALDI-TOF MS were all found to be Actinobacteria and Firmicutes by sequencing. Conclusions　A variety of nontuberculous Mycobacterium and bacteria other than Mycobacterium were found in the positive culture tubes reported by the MGIT960 instrument, most of which could be quickly identified by mass spectrometry. Bacteria other than Mycobacterium are mainly Nocardia and Gordonia, which should be paid attention to in differential diagnosis.

Objective To investigate the distribution of mosquito species and their associated viruses, and identify Culex pipiens subspecies in Hami City, Xinjiang Uygur Autonomous Region. Methods Mosquitoes were captured using mosquito trapping lamps method in Yizhou District, Yiwu County, and Balikun County of Hami City in mi-July, 2019 and 2020. The species and subspecies of all captured mosquitoes were characterized. In addition, the flavivirus, alphavirus, bunyavirus, Japanese encephalitis virus, Liaoning virus, Tahyna virus, tick-borne encephalitis virus and West Nile virus were detected using reverse-transcription PCR assay in captured mosquitoes. Results A total of 1 496 mosquitoes were captured from Yizhou District, Yiwu County, and Balikun County of Hami City, belonging to 3 genus and 3 species. Cx. pipiens was the dominant mosquito species (986 mosquitoes, 65.91%), followed by Aedes caspius (457 mosquitoes, 30.55%), while Culiseta alaskaensis had the lowest number (53 mosquitoes, 3.54%). All captured Cx. pipiens mosquitoes were identified as Cx. pipiens pipiens based on the terminalia of male mosquitoes. RT-PCR assay tested negative for flavivirus, alphavirus, bunyavirus, Japanese encephalitis virus, Liaoning virus, Tahyna virus, tick-borne encephalitis or West Nile virus in captured Cx. pipiens mosquitoes. Conclusions There were 3 species of mosquitoes in Hami City from 2019 to 2020, including Cx. pipiens, Ae. Caspius and C. alaskaensis, with Cx. pipiens as the dominant mosquito species, and all captured Cx. pipiens mosquitoes were Cx. pipiens pipiens; however, no arboviruses were detected.