Metachromatic leukodystrophy (MLD) is an inherited disease caused by a deficiency of the enzyme arylsulfatase A (ARSA). Lentivirus-modified autologous hematopoietic stem cell gene therapy (HSCGT) has recently been approved for clinical use in pre and early symptomatic children with MLD to increase ARSA activity. Unfortunately, this advanced therapy is not available for most patients with MLD who have progressed to more advanced symptomatic stages at diagnosis. Patients with late-onset juvenile MLD typically present with a slower neurological progression of symptoms and represent a significant burden to the economy and healthcare system, whereas those with early onset infantile MLD die within a few years of symptom onset. We conducted a pilot study to determine the safety and benefit of HSCGT in patients with postsymptomatic juvenile MLD and report preliminary results. The safety profile of HSCGT was favorable in this long-term follow-up over 9 years. The most common adverse events (AEs) within 2 months of HSCGT were related to busulfan conditioning, and all AEs resolved. No HSCGT-related AEs and no evidence of distorted hematopoietic differentiation during long-term follow-up for up to 9.6 years. Importantly, to date, patients have maintained remarkably improved ARSA activity with a stable disease state, including increased Functional Independence Measure (FIM) score and decreased magnetic resonance imaging (MRI) lesion score. This long-term follow-up pilot study suggests that HSCGT is safe and provides clinical benefit to patients with postsymptomatic juvenile MLD.
Humans ; Leukodystrophy, Metachromatic/genetics* ; Pilot Projects ; Genetic Therapy/methods* ; Hematopoietic Stem Cell Transplantation ; Male ; Follow-Up Studies ; Female ; Lentivirus/genetics* ; Child ; Child, Preschool ; Hematopoietic Stem Cells/metabolism* ; Cerebroside-Sulfatase/metabolism* ; Adolescent

OBJECTIVE: To observe the characteristics of changes in non-invasive hemodynamic parameters in neonates with septic shock so as to provide clinical reference for diagnosis and treatment. METHODS: A observational study was conducted. The neonates with sepsis complicated with septic shock or not admitted to neonatal intensive care unit (NICU) of the First Affiliated Hospital of Shantou University Medical College were enrolled as the study subjects, who were divided into preterm infant (< 37 weeks) and full-term infant (≥ 37 weeks) according to the gestational age. Healthy full-term infants and hemodynamically stable preterm infants transferring to NICU after birth were enrolled as controls. Electronic cardiometry (EC) was used to measure hemodynamic parameters, including heart rate (HR), mean arterial pressure (MAP), stroke volume (SV), stroke volume index (SVI), cardiac output (CO), cardiac index (CI), systemic vascular resistance (SVR) and systemic vascular resistance index (SVRI), before treatment in the septic shock group, at the time of diagnosis of sepsis in the sepsis without shock group, and before the discharge from the obstetric department or on the day of transferring to NICU in the control group. RESULTS: Finally, 113 neonates with complete data and parental consent for non-invasive hemodynamic monitoring were enrolled, including 32 cases in the septic shock group, 25 cases in the sepsis without shock group and 56 cases in the control group. In the septic shock group, there were 17 cases at the compensated stage and 15 cases at the decompensated stage. There were 21 full-term infants (20 cured or improved and 1 died) and 11 premature infants (7 cured or improved and 4 died), with the mortality of 15.62% (5/32). There were 18 full-term infants and 7 premature infants in the sepsis without shock group and all cured or improved without death. The control group included 28 full-term infants and 28 premature infants transferring to NICU after birth. Non-invasive hemodynamic parameter analysis showed that SV, SVI, CO and CI of full-term infants in the septic shock group were significantly lower than those in the sepsis without shock group and control group [SV (mL): 3.52±0.99 vs. 5.79±1.32, 5.22±1.02, SVI (mL/m²): 16.80 (15.05, 19.65) vs. 27.00 (22.00, 32.00), 27.00 (23.00, 29.75), CO (L/min): 0.52±0.17 vs. 0.80±0.14, 0.72±0.12, CI (mL×s^-1×m^-2): 40.00 (36.67, 49.18) vs. 62.51 (56.34, 70.85), 60.01 (53.34, 69.68), all P < 0.05], while SVR and SVRI were significantly higher than those in the sepsis without shock group and control group [SVR (kPa×s×L^-1): 773.46±291.96 vs. 524.17±84.76, 549.38±72.36, SVRI (kPa×s×L^-1×m^-2): 149.27±51.76 vs. 108.12±12.66, 107.81±11.87, all P < 0.05]. MAP, SV, SVI, CO and CI of preterm infants in the septic shock group were significantly lower than those in the control group [MAP (mmHg, 1 mmHg ≈ 0.133 kPa): 38.55±10.48 vs. 47.46±2.85, SV (mL): 2.45 (1.36, 3.58) vs. 3.96 (3.56, 4.49), SVI (mL/m²): 17.60 (14.20, 25.00) vs. 25.50 (24.00, 29.00), CO (L/min): 0.32 (0.24, 0.63) vs. 0.56 (0.49, 0.63), CI (mL×s^-1×m^-2): 40.01 (33.34, 53.34) vs. 61.68 (56.68, 63.35), all P < 0.05], while SVR and SVRI were similar to the control group [SVR (kPa×s×L^-1): 1 082.88±689.39 vs. 656.63±118.83, SVRI (kPa×s×L^-1×m^-2): 126.00±61.50 vs. 102.37±11.68, both P > 0.05]. Further analysis showed that SV, SVI and CI of neonates at the compensation stage in the septic shock group were significantly lower than those in the control group [SV (mL): 3.60±1.29 vs. 4.73±1.15, SVI (mL/m²): 19.20±8.33 vs. 26.34±3.91, CI (mL×s^-1×m^-2): 46.51±20.34 vs. 61.01±7.67, all P < 0.05], while MAP, SVR and SVRI were significantly higher than those in the control group [MAP (mmHg): 52.06±8.61 vs. 48.54±3.21, SVR (kPa×s×L^-1): 874.95±318.70 vs. 603.01±111.49, SVRI (kPa×s×L^-1×m^-2): 165.07±54.90 vs. 105.09±11.99, all P < 0.05]; MAP, SV, SVI, CO and CI of neonates at the decompensated stage in the septic shock group were significantly lower than those in the control group [MAP (mmHg): 35.13±6.08 vs. 48.54±3.21, SV (mL): 2.89±1.17 vs. 4.73±1.15, SVI (mL/m²): 18.50±4.99 vs. 26.34±3.91, CO (L/min): 0.41±0.19 vs. 0.65±0.15, CI (mL×s^-1×m^-2): 43.34±14.17 vs. 61.01±7.67, all P < 0.05], while SVR and SVRI were similar to the control group [SVR (kPa×s×L^-1): 885.49±628.04 vs. 603.01±111.49, SVRI (kPa×s×L^-1×m^-2): 114.29±43.54 vs. 105.09±11.99, both P > 0.05]. CONCLUSIONS Full-term infant with septic shock exhibit a low cardiac output, high vascular resistance hemodynamic pattern, while preterm infant with septic shock show low cardiac output and normal vascular resistance. At the compensated stage the hemodynamic change is low output and high resistance type, while at the decompensated stage it is low output and normal resistance type. Non-invasive hemodynamic monitoring can assist in the identification of neonatal septic shock and provide basis for clinical diagnosis and treatment.
Humans ; Shock, Septic/physiopathology* ; Infant, Newborn ; Hemodynamics ; Female ; Male ; Case-Control Studies ; Infant, Premature

Objective:To construct a preoperative prehabilitation program for prostate cancer based on evidence-based and Delphi method, so as to provide theoretical basis for medical staff to carry out preoperative prehabilitation research for prostate cancer patients.Methods:From June 2023 to March 2024, the first draft of the prehabilitation plan for prostate cancer before surgery was formed through evidence summary, semi-structured interviews and expert meetings. The experts in related fields were selected for two rounds of Delphi expert consultation. The items were revised according to the expert consultation opinions to establish the final plan.Results:Totally 16 experts were included, aged (43.19 ± 7.57) years. Five were males and 11 were females. The response rates of the two rounds of expert consultation were both 16/16, and the authority coefficients of the two rounds expert consultation were both 0.85. The Kendall coordination coefficients of the importance and feasibility of the items in the second round of consultation were 0.213 and 0.224, both P<0.05. In the second round of consultation, the value of importance assignment of items at all levels was 4.19-4.94 points, and the full score rate was 43.75%-93.75%. The final scheme included 5 first-level items, 13 second-level items, and 34 third-level items. Conclusions:The preoperative prehabilitation program for prostate cancer is scientific, importanceand applicable, which provides a theoretical basis for clinical preoperative prehabilitation for prostate cancer patients.

Objective To explore the mechanism of Jiawei Jisheng Shenqi Decoction in improving the hypoxic microenvironment and antagonizing liver cirrhosis.Methods In vivo experiments were conducted using a rat model of carbon tetrachloride(CCL4)-induced liver cirrhosis.Rats were divided into normal,model,colchicine,JWJSSQ low-dose,JWJSSQ medium-dose,and JWJSSQ high-dose group.Pathological changes in liver tissues in each group were examined by hematoxylin and eosin(HE)and Masson staining,changes in serum liver function were detected using test kits,levels of hyaluronic acid(HA),laminin(LN),procollagen Ⅲ(PC Ⅲ),and collagen typeⅣ(COL4)were detected by enzyme-linked immunosorbent assay(ELISA),and protein expression levels of hypoxia-inducible factor-1α(HIF-1α),Notch1,Jagged1,and α-smooth muscle actin(α-SMA)were detected by Western blot.In vitro experiments were conducted in HSC-T6 cells,and the optimal concentration of CoCl2(100 μ mol/L,200μmol/L,400 μmol/L,600 μmol/L and 800 μmol/L)in the cultured cells and the optimal concentration of drug-containing serum(5％,10％,15％,20％)were determined by Cell Counting Kit-8(CCK-8)assay.The migration ability of cells in each group was detected by scratch testing,and changes in the apoptosis rates were determined by flow cytometry.Protein expression levels of HIF-1α,Notch1,Jagged1,α-SMA,matrix metallopeptidase 9(MMP9),and tissue inhibitor of metalloproteinases 1(TIMP-1)were detected by Western blot.Results In the in vivo experiments,liver swelling,inflammatory cell infiltration,collagen deposition,and the appearance of pseudolobules were significantly increased in the model group compared with those in the normal group.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),HA,LN,PCⅢ,and COL4 were significantly increased and albumin(ALB)was significantly decreased in the model group,while liver levels of HIF-1α,Notch1,Jagged1,and α-SMA proteins were significantly increased(P＜0.01).Liver swelling,inflammatory cell infiltration,and collagen deposition were significantly reduced in each treatment group compared with those in the model group,and the degree of fibrosis was reduced.Serum ALT,AST,HA,LN,PCⅢ,and COL4 were significantly decreased and ALB was significantly increased,while liver levels of HIF-1α,Notch1,Jagged1,and α-SMA proteins were also significantly decreased to varying degrees(P＜0.05).In the in vitro experiments,hypoxia promoted HSC-T6 migration and reduced apoptosis,increased the protein expression levels of HIF-1α,Notch1,Jagged1,α-SMA,and TIMP-1,and reduced the expression levels of MMP9(P＜0.01).Serum containing Jiawei Jisheng Shenqi Decoction inhibited HSC-T6 migration,promoted HSC-T6 apoptosis,lowered the expression of HIF-1α,Notch1,Jagged1,α-SMA,and TIMP-1 proteins,and enhanced the expression of MMP9 protein(P＜0.01).The inhibitory effect of Jiawei Jisheng Shenqi on HSC-T6 cell activation was reversed by the HIF-1α agonist dimethyloxalylglycine.Conclusions Jiawei Jisheng Shenqi Decoction can improve the hypoxic microenvironment via the HIF-1α/Notch pathway,thereby exerting an anti-liver cirrhosis effect.

Objective:To prepare a nano-barium titanate@gold Schottky junction (nano-BaTiO 3@Au) coating and investigate its effects on the adhesion, proliferation, and osteogenic differentiation of bone marrow stem cells (BMSCs), aiming to explore a titanium surface modification strategy with superior osteogenic activity. Methods:Pure titanium specimens served as the control group (Ti group). Titanium dioxide coatings were prepared on their surfaces via anodic oxidation. Nano-barium titanate (nBTO group) was further synthesized using the hydrothermal method. Gold nanoparticles were grown in situ on the nano-BaTiO 3 via high-temperature reduction of chloroauric acid using sodium citrate, yielding the nano-barium titanate@gold Schottky junction coating (nBTO@Au group). Surface morphology was observed by scanning electron microscopy (SEM). Elemental composition was analyzed using X-ray energy dispersive spectrum (EDS) and X-ray photoelectron spectroscopy (XPS). Crystal structure was analyzed using X-ray diffraction (XRD) and Raman spectroscopy. Hydrophilicity was assessed via water contact angle measurement. Specimens were co-cultured with BMSCs to evaluate biocompatibility and osteogenic properties. Cell proliferation on days 1, 3, 5, and 7 was assessed using the cell counting kit-8 (CCK-8) assay. Cytotoxicity towards BMSCs was assessed using live/dead cell staining. Cell morphology and adhesion were observed using cytoskeleton staining. Alkaline phosphatase (ALP) expression in BMSCs after 7 days was quantified using an ALP activity assay and ALP staining. Extracellular matrix mineralization after 7 days was evaluated using alizarin red staining and quantification assay. Each experiment was performed using three specimens per group. Results:Scanning electron microscopy revealed that gold nanoparticles with the diameter of(14.838±0.718) nm, uniform in size and homogeneously distributed, were successfully grown in situ on the surface of the nBTO coating. EDS and XPS confirmed the presence of Ba, Ti, O, and Au elements in the nBTO@Au composite coating. XRD and Raman spectroscopy analysis indicated that the nanostructured barium titanate (nBTO) coating was synthesized via a hydrothermal method.Water contact angle measurements showed that the contact angle was 66.8°± 0.45° for the control group, 22.55°±0.42° for the nBTO group, and 26.78°±1.15° for the nBTO@Au group, indicating good hydrophilicity of both nBTO and nBTO@Au coatings. On day 1 and day 3 of culture, the cell proliferation in the nBTO group was significantly lower than that in the control group ( P<0.05). In contrast, no significant differences were observed between the nBTO@Au group and either the control group or the nBTO group (all P>0.05). By day 5, the cell proliferation of nBTO@Au groups was significantly lower than that of the control group ( P<0.05), and the cell proliferation of nBTO group was significantly lower than that of the control group and that of the nBTO@Au group ( P<0.05). By day 7, there were no statistically significant differences in cell proliferation among all experimental groups ( F=1.62, P>0.05).Live/dead cell staining demonstrated that the cell survival rate exceeded 90% in all groups, with normal morphology and few dead cells, indicating good biocompatibility of the nBTO@Au coating. Compared to the control group, both nBTO and nBTO@Au groups promoted cell adhesion and spreading, although no significant difference in cell morphology was noted between the two modified groups. ALP staining revealed a larger stained area and deeper coloration in the nBTO@Au group. Quantitative results showed that ALP activity in the nBTO@Au group was significantly higher than that in both the nBTO and control groups ( P<0.05), and the nBTO group also exhibited significantly higher activity than the control group( P<0.05). Alizarin red staining indicated the deepest coloration in the nBTO@Au group, followed by the nBTO group, and the lightest in the control group. Quantitative analysis further confirmed that the amount of calcium nodule deposition in the nBTO@Au group was significantly greater than that in the other two groups ( P<0.05), and the nBTO group also showed significantly more deposition than the control group( P<0.05). Conclusions:This study successfully prepared an nBTO@Au coating possessing good biocompatibility and enhanced osteogenic properties.

Objective:Vascular cell adhesion molecule 1(VCAM1)is involved in a series of physiological and pathological processes,such as immune and inflammatory response,tumor cell metastasis and invasion.But under trau-matic brain injury(TBI),specific types of cells with VC AM1 expression and the related functions are not clear.In order to further explore the specific functions of VCAM1 involved in TBI,this study constructed reporter mice of VCAM1 to explore the response of VCAM1 to TBI in detail.Methods:VCAM1-Cre/ERT2::Ai14 reporter mice were constructed by gene targeting technology and Cre/loxP system,the labeled cell types and labeling efficiency were validated by im-munofluorescence staining and reporter mice hybridization.The stab wound model was used to simulate TBI to induce the changes of VCAM1 expression in cells,and the characteristics of VCAM1 positive astrocytes were detected by immu-nofluorescence staining and fluorescent probe labeling.Results:The labeling efficiency of VCAM1-Cre/ERT2::Ai14 re-porter mice was higher than that of VCAM1 antibody as was seen by labeling of more endothelial cells of blood vessels and unique astrocytes.The distribution of these astrocytes was specific,for example in the nucleus accumbens,amygda-la,hypothalamus,and paraventricular fiber systems.TBI could significantly induce the expression of VCAM1 in astro-cytes(P＜0.0001).These induced astrocytes developed reactive qualities,including somal hypertrophy,GFAP ex-pression and proliferative ability.Conclusion:VCAM1-Cre/ERT2::Ai14 reporter mice could label cells with VCAM1 expression more sensitively,so they were more effective tools for observing expression and function of VCAM1.The up-regulation of VCAM1 expression in astrocytes after TBI surgery suggested that VCAM1 was an inflammatory response molecule in astrocytes,we recommended it as a new molecular indicator of reactive astrocytes.

Objective:To explore effect of Yishen Paidu Formula on inflammatory response in chronic renal failure(CRF)rats and role of Calcineurin/nuclear factor of activated T cell(NFAT).Methods:CRF rat model was constructed,and randomly grouped into model group,low,medium and high doses Yishen Paidu Formula groups,with 10 rats in each group,another 10 rats were fed normally as a blank group;general situation of rats was recorded;urine protein quantification kit was applied to detect 24-hour urine protein level of CRF rats in each group;serum creatinine and urea nitrogen levels of CRF rats were detected;ELISA was applied to detect serum IL-1β,IL-6 and TNF-α levels in CRF rats;pathological changes of renal tissue were observed by HE staining;Western blot was applied to detect expressions of fibroblast growth factor 23(FGF23),Klotho protein,Calcineurin,NFAT protein in renal tissue of CRF rats in each group.Results:Compared with blank group,levels of creatinine,urea nitrogen in serum,24 h urine protein in urine,IL-1β,IL-6,TNF-α in serum,and protein levels of FGF23,Calcineurin,NFAT in kidney tissue were obviously increased in model group,level of Klotho protein was obviously decreased(P<0.05).Compared with model group,levels of creatinine,urea nitrogen in serum,24 h urine protein in urine,IL-1β,IL-6,TNF-α in serum,and protein levels of FGF23,Calcineurin,NFAT in kidney tissue were obviously decreased in low,medium and high doses Yishen Paidu Formula groups,level of Klotho protein was obviously increased,which were more significant with dosage increase(P<0.05).Conclusion:Yishen Paidu Formula may alleviate inflammatory response in CRF rats by inhibiting Calcineurin/NFAT signaling pathway.

A porous composite scaffold is prepared through freeze-drying method using sodium alginate as the matrix and calcium citrate double salt as the functional enhancer.To thoroughly investigate the structure and properties of the composite scaffold,advanced instruments including Fourier transform infrared spectrometer,X-ray diffractometer,scanning electron microscope,and thermogravimetric analyzer are utilized for comprehensive structural characterization.Meanwhile,key properties such as porosity,mechanical performance,swelling ratio,gel fraction,and in vitro degradation rate are systematically tested and analyzed.The results show that the composite of sodium alginate and calcium citrate double salt is a physical complex.The porosity of the scaffold mainly ranges from 200 to 600 μm,and as the addition of double salt increases,the pore size firstly increases and then slightly decreases,while the pore distribution become more uneven.In terms of mechanical properties,with the gradual increase of the content of double salt in the scaffold,the mechanical properties of the scaffold are significantly improved.However,its swelling ratio and gel fraction exhibit decreasing trends.Additionally,the degradation rate of the alginate scaffold is relatively fast,while the addition of calcium citrate double salt does not significantly improve the degradation rate.This study not only provides valuable insights into the preparation and property research of porous composite scaffold composed of sodium alginate/calcium citrate double salt,but also offers important reference for their potential applications in biomedicine.

Endometriosis is a common estrogen-dependent disease in women of childbearing age,often leading to chronic pelvic pain,infertility,ovarian cancer,and other serious complications,and jeopardizing the health of women.The pathogenesis of endometriosis is complex and involves the alteration of multiple signaling pathways mediated by hormones,immunity,genetics,and the environment,and their interactions.Wnt/β-catenin signaling is involved in the regulation of embryonic development and tissue homeostasis,and it has recently been implicated in the pathogenesis of endometriosis via multiple pathways.This review considers the biological characteristics of the Wnt/β-catenin signaling pathway and summarizes the main mechanisms and signaling pathways involved in the pathogenesis of endometriosis,as well as the curr-ent research status of the regulation of this signaling pathway by traditional Chinese medicine for the treatment of endometriosis.We aim to clarify how the Wnt/β-catenin signaling pathway affects the development of endometriosis,and suggest that future studies should focus on exploring its potential role as an indicator for the clinical prediction and early diagnosis of endometriosis,thus providing theoretical support for the early diagnosis of this condition and the development of targeted drugs.

Objective To explore the mechanism of Jiawei Jisheng Shenqi Decoction in improving the hypoxic microenvironment and antagonizing liver cirrhosis.Methods In vivo experiments were conducted using a rat model of carbon tetrachloride(CCL4)-induced liver cirrhosis.Rats were divided into normal,model,colchicine,JWJSSQ low-dose,JWJSSQ medium-dose,and JWJSSQ high-dose group.Pathological changes in liver tissues in each group were examined by hematoxylin and eosin(HE)and Masson staining,changes in serum liver function were detected using test kits,levels of hyaluronic acid(HA),laminin(LN),procollagen Ⅲ(PC Ⅲ),and collagen typeⅣ(COL4)were detected by enzyme-linked immunosorbent assay(ELISA),and protein expression levels of hypoxia-inducible factor-1α(HIF-1α),Notch1,Jagged1,and α-smooth muscle actin(α-SMA)were detected by Western blot.In vitro experiments were conducted in HSC-T6 cells,and the optimal concentration of CoCl2(100 μ mol/L,200μmol/L,400 μmol/L,600 μmol/L and 800 μmol/L)in the cultured cells and the optimal concentration of drug-containing serum(5％,10％,15％,20％)were determined by Cell Counting Kit-8(CCK-8)assay.The migration ability of cells in each group was detected by scratch testing,and changes in the apoptosis rates were determined by flow cytometry.Protein expression levels of HIF-1α,Notch1,Jagged1,α-SMA,matrix metallopeptidase 9(MMP9),and tissue inhibitor of metalloproteinases 1(TIMP-1)were detected by Western blot.Results In the in vivo experiments,liver swelling,inflammatory cell infiltration,collagen deposition,and the appearance of pseudolobules were significantly increased in the model group compared with those in the normal group.Serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),HA,LN,PCⅢ,and COL4 were significantly increased and albumin(ALB)was significantly decreased in the model group,while liver levels of HIF-1α,Notch1,Jagged1,and α-SMA proteins were significantly increased(P＜0.01).Liver swelling,inflammatory cell infiltration,and collagen deposition were significantly reduced in each treatment group compared with those in the model group,and the degree of fibrosis was reduced.Serum ALT,AST,HA,LN,PCⅢ,and COL4 were significantly decreased and ALB was significantly increased,while liver levels of HIF-1α,Notch1,Jagged1,and α-SMA proteins were also significantly decreased to varying degrees(P＜0.05).In the in vitro experiments,hypoxia promoted HSC-T6 migration and reduced apoptosis,increased the protein expression levels of HIF-1α,Notch1,Jagged1,α-SMA,and TIMP-1,and reduced the expression levels of MMP9(P＜0.01).Serum containing Jiawei Jisheng Shenqi Decoction inhibited HSC-T6 migration,promoted HSC-T6 apoptosis,lowered the expression of HIF-1α,Notch1,Jagged1,α-SMA,and TIMP-1 proteins,and enhanced the expression of MMP9 protein(P＜0.01).The inhibitory effect of Jiawei Jisheng Shenqi on HSC-T6 cell activation was reversed by the HIF-1α agonist dimethyloxalylglycine.Conclusions Jiawei Jisheng Shenqi Decoction can improve the hypoxic microenvironment via the HIF-1α/Notch pathway,thereby exerting an anti-liver cirrhosis effect.