ObjectiveTranscranial magneto-acoustic stimulation (TMAS) is an emerging non-invasive neuromodulation technique that may provide a novel non-pharmacological intervention strategy for Parkinson's disease (PD). PD is characterized by the progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc), leading to motor impairments such as bradykinesia, tremor, and rigidity. Increasing evidence indicates that mitochondrial dysfunction and impaired mitochondrial quality control are central mechanisms underlying dopaminergic neuronal loss. In particular, abnormalities in mitophagy and mitochondrial fission-fusion balance contribute substantially to oxidative stress, energy metabolic failure, and neuronal injury. At present, most clinical treatments for PD mainly alleviate symptoms but do not effectively halt disease progression. Therefore, exploring new interventions targeting the core pathological mechanisms is of considerable significance. This study aims to investigate whether TMAS can improve neural damage and motor dysfunction in PD mice by regulating mitophagy and the fission/fusion dynamic balance, thereby providing theoretical and experimental support for its application in PD treatment. MethodsMale C57BL/6 mice were used in this study. A PD model was established by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 7 consecutive days. After model induction, mice in the intervention group received TMAS once daily for 14 consecutive days, whereas the corresponding control group received sham stimulation. The stimulation target was positioned over the primary motor cortex (M1). Motor performance was evaluated using the pole test and the open-field test. To verify the activation effect of TMAS on the target cortical region, c-Fos immunohistochemistry was performed in the M1. To assess nigral dopaminergic neuronal injury, tyrosine hydroxylase (TH) immunohistochemistry was used to quantify TH-positive neurons in the SNc. Mitochondrial function was evaluated by measuring reactive oxygen species (ROS) levels and adenosine triphosphate (ATP) content in the SNc. Western blot was further performed to determine the expression of mitophagy-related proteins, including PINK1, Parkin, LC3-II, and p62, as well as mitochondrial dynamics-related proteins, including Drp1 and Opa1. ResultsTMAS significantly increased the number of c-Fos-positive cells in M1 (P<0.000 1), indicating effective activation of neurons in the targeted cortical region. Compared with the control group, MPTP-treated mice exhibited marked motor dysfunction, including a significant reduction in total distance traveled in the open-field test (P<0.000 1) and mean speed (P=0.000 1), as well as significant prolongation of turn time and total climbing time in the pole test (P<0.000 1). These behavioral impairments were accompanied by a substantial loss of TH-positive dopaminergic neurons in the SNc, whereas TMAS significantly increased TH-positive neuron survival (P<0.000 1). In parallel, MPTP induced a pronounced increase in ROS levels and a significant reduction in ATP content, indicating severe mitochondrial dysfunction and energy metabolism impairment (P<0.01). TMAS treatment significantly improved motor performance, as reflected by the reversal of MPTP-induced impairment in the open-field and pole tests, and significantly reduced ROS accumulation (P<0.01) while restoring ATP production (P<0.001). At the molecular level, MPTP markedly downregulated PINK1 and Parkin, decreased p62 expression, increased LC3-II accumulation, elevated Drp1 expression, and reduced Opa1 expression, whereas TMAS significantly reversed these abnormalities, suggesting restoration of mitophagy-related mitochondrial quality control and re-establishment of mitochondrial fission-fusion balance. Collectively, these findings indicate that TMAS ameliorates MPTP-induced neurotoxicity and restores mitochondrial homeostasis and energy metabolism. ConclusionTMAS effectively attenuates neural damage and improves motor dysfunction in MPTP-induced PD mice. Its neuroprotective effects are closely associated with multidimensional regulation of the mitochondrial quality control system, including restoration of PINK1/Parkin-mediated mitophagy and rebalancing of Drp1/Opa1-related mitochondrial dynamics. Rather than acting only as a symptomatic neuromodulatory intervention, TMAS may influence a key pathological axis of PD by improving mitochondrial homeostasis in SNc and protecting nigral dopaminergic neurons. These findings provide experimental evidence supporting TMAS as a promising non-invasive physical intervention for PD.

ObjectiveTranscranial magneto-acoustic stimulation (TMAS) is an emerging non-invasive neuromodulation technique that may provide a novel non-pharmacological intervention strategy for Parkinson's disease (PD). PD is characterized by the progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc), leading to motor impairments such as bradykinesia, tremor, and rigidity. Increasing evidence indicates that mitochondrial dysfunction and impaired mitochondrial quality control are central mechanisms underlying dopaminergic neuronal loss. In particular, abnormalities in mitophagy and mitochondrial fission-fusion balance contribute substantially to oxidative stress, energy metabolic failure, and neuronal injury. At present, most clinical treatments for PD mainly alleviate symptoms but do not effectively halt disease progression. Therefore, exploring new interventions targeting the core pathological mechanisms is of considerable significance. This study aims to investigate whether TMAS can improve neural damage and motor dysfunction in PD mice by regulating mitophagy and the fission/fusion dynamic balance, thereby providing theoretical and experimental support for its application in PD treatment. MethodsMale C57BL/6 mice were used in this study. A PD model was established by intraperitoneal injection of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) for 7 consecutive days. After model induction, mice in the intervention group received TMAS once daily for 14 consecutive days, whereas the corresponding control group received sham stimulation. The stimulation target was positioned over the primary motor cortex (M1). Motor performance was evaluated using the pole test and the open-field test. To verify the activation effect of TMAS on the target cortical region, c-Fos immunohistochemistry was performed in the M1. To assess nigral dopaminergic neuronal injury, tyrosine hydroxylase (TH) immunohistochemistry was used to quantify TH-positive neurons in the SNc. Mitochondrial function was evaluated by measuring reactive oxygen species (ROS) levels and adenosine triphosphate (ATP) content in the SNc. Western blot was further performed to determine the expression of mitophagy-related proteins, including PINK1, Parkin, LC3-II, and p62, as well as mitochondrial dynamics-related proteins, including Drp1 and Opa1. ResultsTMAS significantly increased the number of c-Fos-positive cells in M1 (P<0.000 1), indicating effective activation of neurons in the targeted cortical region. Compared with the control group, MPTP-treated mice exhibited marked motor dysfunction, including a significant reduction in total distance traveled in the open-field test (P<0.000 1) and mean speed (P=0.000 1), as well as significant prolongation of turn time and total climbing time in the pole test (P<0.000 1). These behavioral impairments were accompanied by a substantial loss of TH-positive dopaminergic neurons in the SNc, whereas TMAS significantly increased TH-positive neuron survival (P<0.000 1). In parallel, MPTP induced a pronounced increase in ROS levels and a significant reduction in ATP content, indicating severe mitochondrial dysfunction and energy metabolism impairment (P<0.01). TMAS treatment significantly improved motor performance, as reflected by the reversal of MPTP-induced impairment in the open-field and pole tests, and significantly reduced ROS accumulation (P<0.01) while restoring ATP production (P<0.001). At the molecular level, MPTP markedly downregulated PINK1 and Parkin, decreased p62 expression, increased LC3-II accumulation, elevated Drp1 expression, and reduced Opa1 expression, whereas TMAS significantly reversed these abnormalities, suggesting restoration of mitophagy-related mitochondrial quality control and re-establishment of mitochondrial fission-fusion balance. Collectively, these findings indicate that TMAS ameliorates MPTP-induced neurotoxicity and restores mitochondrial homeostasis and energy metabolism. ConclusionTMAS effectively attenuates neural damage and improves motor dysfunction in MPTP-induced PD mice. Its neuroprotective effects are closely associated with multidimensional regulation of the mitochondrial quality control system, including restoration of PINK1/Parkin-mediated mitophagy and rebalancing of Drp1/Opa1-related mitochondrial dynamics. Rather than acting only as a symptomatic neuromodulatory intervention, TMAS may influence a key pathological axis of PD by improving mitochondrial homeostasis in SNc and protecting nigral dopaminergic neurons. These findings provide experimental evidence supporting TMAS as a promising non-invasive physical intervention for PD.

In this study, Saccharomyces cerevisiae R0 was used as the chassis cell to synthesize oleanolic acid from scratch through the heterologous expression of β-amyrin synthase(β-AS) from Glycyrrhiza uralensis, cytochrome P450 enzyme CYP716A154 from Catharanthus roseus, and cytochrome P450 reductase AtCPR from Arabidopsis thaliana. The engineered strain R1 achieved shake flask titres of 5.19 mg·L~(-1). By overexpressing enzymes in the pentose phosphate pathway(PPP)(ZWF1, GND1, TKL1, and TAL), the NADH kinase gene in the mitochondrial matrix(POS5), truncated 3-hydroxy-3-methylglutaryl-CoA reductase(tPgHMGR1) from Panax ginseng, and farnesyl diphosphate synthase gene(SmFPS) from Salvia miltiorrhiza, the precursor supply and intracellular reduced nicotinamide adenine dinucleotide phosphate(NADPH) supply were enhanced, resulting in an 11.4-fold increase in squalene yield and a 3.6-fold increase in oleanolic acid yield. Subsequently, increasing the copy number of the heterologous genes tPgHMGR1, β-AS, CYP716A154, and AtCPR promoted the metabolic flow towards the final product, oleanolic acid, and increased the yield by three times. Shake flask fermentation data showed that, by increasing the copy number, precursor supply, and intracellular NADPH supply, the final engineered strain R3 could achieve an oleanolic acid yield of 53.96 mg·L~(-1), which was 10 times higher than that of the control strain R1. This study not only laid the foundation for the green biosynthesis of oleanolic acid but also provided a reference for metabolic engineering research on other pentacyclic triterpenoids in S. cerevisiae.
Oleanolic Acid/biosynthesis* ; Saccharomyces cerevisiae/metabolism* ; Industrial Microbiology ; Microorganisms, Genetically-Modified/metabolism* ; Plants/enzymology* ; Fermentation ; Metabolic Engineering

Many triterpenoid compounds have been successfully heterologously synthesized in Saccharomyces cerevisiae. To increase the yield of triterpenoids, various metabolic engineering strategies have been developed. One commonly applied strategy is to enhance the supply of precursors, which has been widely used by researchers. Squalene, as a precursor to triterpenoid biosynthesis, plays a crucial role in the synthesis of these compounds. This study primarily investigates the effect of different squalene levels in chassis strains on the synthesis of triterpenoids(oleanolic acid and ursolic acid), and the underlying mechanisms are further explored using real-time quantitative PCR(qPCR) analysis. The results demonstrate that the chassis strain CB-9-5, which produces high levels of squalene, inhibits the synthesis of oleanolic acid and ursolic acid. In contrast, chassis strains with moderate to low squalene production, such as Y8-1 and CNPK, are more conducive to the synthesis of oleanolic acid and ursolic acid. The qPCR analysis reveals that the expression levels of ERG1, βAS, and CrCYP716A154 in the oleanolic acid-producing strain CB-OA are significantly lower than those in the control strains C-OA and Y-OA, suggesting that high squalene production in the chassis strains suppresses the transcription of certain genes, leading to a reduced yield of triterpenoids. Our findings indicate that when constructing S. cerevisiae strains for triterpenoid production, chassis strains with high squalene content may suppress the expression of certain genes, ultimately lowering their production, whereas chassis strains with moderate squalene levels are more favorable for triterpenoid biosynthesis.
Squalene/analysis* ; Saccharomyces cerevisiae/genetics* ; Triterpenes/metabolism* ; Metabolic Engineering ; Oleanolic Acid/biosynthesis* ; Ursolic Acid

Chemical investigation of the stems of Fritillaria unibracteata P.K. Hsiao & K.C. Hsia resulted in the isolation of nine compounds, by means of silica gel column chromatography, and preparative HPLC. Based on spectroscopic and chemical evidence, these compounds were identified as: 27-hydroxychlorogenone (1), sieboldogenin (2), (3β, 25S)-spirost-5-ene-3,17,27-triol (3), laxogenin (4), tigogenone (5), cerevisterol (6), ergosterol peroxide (7), stigmaterol (8), and β-sitosterol (9). Compound 1 was a new compound, and compounds 2-9 were isolated from the stems of Fritillaria unibracteata for the first time. The inhibitory effects of compounds 1−9 on A549 cells were determined using the MTT method. The results show that compound 6 exhibits moderate inhibitory activity with an IC50 value of (14.16 ± 1.11) μmol/L.

Objective: To evaluate the effects of dietary intervention on blood pressure and renal function in patients with hypertensive nephropathy, so as to provide dietary and nutritional guidances for this population. Methods: Hypertensive nephropathy patients who were treated at Zhucheng People's Hospital from March 2023 to February 2024 were selected as the study subjects and randomly divided into the intervention group and the control group. The control group received routine antihypertensive treatment and health lifestyle guidance. On the basis of the treatment and guidance received by the control group, the intervention group implemented dietary intervention in accordance with the Clinical Practice Guidelines for Nutritional Therapy of Chronic Kidney Disease in China (2021 edition) for a period of 3 months. Systolic blood pressure (SBP), diastolic blood pressure (DBP) were measured before and after the intervention, and serum creatinine (Scr), blood urea nitrogen (BUN), uric acid (UA), cystatin and β2-microglobulin were detected. Differences of indicators before and after intervention between the two groups were compared using generalized estimation equation. Results: A total of 83 patients with hypertensive nephropathy were followed up, including 43 cases in the intervention group and 40 cases in the control group. There were no statistically significant differences in gender, age, body mass index, duration of hypertension, family history of hypertension, hypertension grade, physical activity index, or smoking status between the two groups (all P>0.05). The differences in SBP, DBP, Scr, BUN, and UA between the two groups, as well as the differences before and after the intervention, were statistically significant, and there was an interaction between the groups and the intervention time (all P<0.05). After intervention, the levels of SBP, DBP, Scr, BUN, and UA in the intervention group were lower than those in the control group (all P<0.05). The differences in cystatin and β2-microglobulin between the two groups and before and after the intervention were not statistically significant, and there was no interaction between the groups and the intervention time (all P>0.05). Conclusion Dietary intervention has a certain effect on reducing blood pressure and improving renal function indicators in patients with hypertensive nephropathy.

AIM:To compare the effect of AT TORBI 709M and Tecnis ZMT intraocular lenses on astigmatism correction in patients with corneal astigmatism at 3 mo after operation based on the standard astigmatism vector analysis.METHODS: This was a retrospective case-control study. The clinical data of 69 patients(69 eyes)with corneal astigmatism who underwent phacoemulsification and implantation of toric intraocular lens(IOL)from June 2021 to December 2021 in Day Surgery Center of Xi'an No.1 Hospital was analyzed. The patients were divided into two groups. In group one, 38 cases(38 eyes)were implanted with AT TORBI 709M, and 31 patients(31 eyes)with Tecnis ZMT in group two. The axial length, preoperative astigmatism and axis, and the degree of intraocular lens were recorded. The uncorrected distance visual acuity(UCDVA), best corrected distance visual acuity(BCDVA), diopter, residual astigmatism and axis were recorded preoperatively and at 1 wk, 1 and 3 mo postoperatively. The postoperative surgical indicators, including spherical equivalent(SE), target induced astigmatism vector(TIA), surgically induced astigmatism vector(SIA), magnitude of error(ME), absolute value of angle of error(|AE|), absolute value of difference vector(|DV|), correction index(CI), and index of success(IOS)were evaluated by the standard astigmatism vector analysis.RESULTS:Postoperative UCDVA and BCDVA were significantly improved(all P<0.001), and there were statistically significant differences compared to preoperative UCDVA and BCDVA(all P<0.001). While, there was no significant difference in UCDVA and BCDVA between the two groups(P=0.275, 0.124). The standard astigmatism vector analysis showed that a good astigmatism correction was achieved in both AT TORBI 709M group and Tecnis ZMT group, and both |DV| and IOS were close to 0(P=0.329, 0.288). The CI of the AT TORBI 709M group was closer to 1, indicating a better astigmatism correction, while the CI of the Tecnis ZMT group was higher than 1, suggesting an overcorrection of astigmatism. However, the difference between the two groups was not statistically significant(P=0.193). The mean residual astigmatism at 3 mo postoperatively was -0.11±0.91 D in the AT TORBI 709M group and -0.46±0.76 D in the Tecnis ZMT group, respectively, showing no statistically significance difference(t=1.732, P=0.088).CONCLUSION:Both the flat loop AT TORBI 709M and the double C-loop Tecnis ZMT intraocular lenses can effectively improve postoperative visual acuity in patients with regular corneal astigmatism, showing good rotational stability and comparable correction abilities for both astigmatism with the rule and against-the-rule astigmatism.

Objective:To assess medication adherence and identify independent associated factors among elderly patients with multimorbidity attending the Department of General Practice at Zhoukou Central Hospital.Methods:This cross-sectional study enrolled 380 elderly patients with multimorbidity through convenience and snowball sampling between March 2023 and March 2024. Face-to-face interviews were conducted with the subjects through community general practice clinics and resident group discussions. The survey covered the subjects′ general clinical information and medication adherence. Medication adherence was assessed using the Chinese version of the 8-item Morisky Medication Adherence Scale (MMAS-8). Multivariate logistic regression analyzed independent factors of medication adherence in elderly patients with multimorbidity.Results:A total of 380 patients were included in the analysis, with an mean age of (71.57±7.58) years, and 211 (55.5%) were males. Only 50 patients (13.2%) demonstrated good adherence. Multivariate logistic regression revealed lower adherence in ages 60-69 vs.≥80 ( OR=4.595, 95% CI:1.730-12.207, P=0.002), higher adherence in high school and above education vs. primary/illiterate ( OR=0.278, 95% CI:0.082-0.944, P=0.040), reduced adherence with increased household responsibility ( OR=3.684, 95% CI:1.799-7.543, P<0.001). Conclusions:Medication adherence was suboptimal among elderly patients with multimorbidity attending the Department of General Practice at Zhoukou Central Hospital. Age, education, and household responsibility burden were independent predictors for medication adherence.

Objective:To explore the correlation between abnormal vital signs (e.g., heart rate, oxygen saturation, and body temperature) and acute exacerbations in patients with chronic obstructive pulmonary disease (COPD), as well as to evaluate the clinical value of continuous monitoring via wearable devices for the early warning and intervention.Methods:A multicenter cross-sectional study enrolled 335 patients with stable chronic obstructive pulmonary disease (COPD) from 12 community health centers in Beijing and Chengdu between June 2023 and May 2024. General demographic and clinical data were collected, and each participant underwent continuous monitoring of resting heart rate, oxygen saturation, and body temperature using wearable devices for 21 days. Based on whether participants had experienced acute exacerbations requiring outpatient, emergency, or inpatient treatment within the previous year, they were categorized into the acute exacerbation group and the non-exacerbation group. Differences in physiological parameters between the acute exacerbation group and non-exacerbation group were analyzed, and contributing factors were assessed using logistic regression analysis.Results:A total of 335 patients with stable COPD were enrolled, including 252 cases (75.22%) in the acute exacerbation group and 83 cases (24.78%) in the non-acute exacerbation group. There were no statistically significant differences in age, sex distribution, comorbidities, or baseline lung function between the two groups (all P>0.05). Compared with the non-acute exacerbation group, patients in the acute exacerbation group had a faster resting heart rate((76.01 ± 7.78) beats/min vs. (72.72 ± 7.35) beats/min, t=3.126, P=0.002), a higher proportion of patients with decreased oxygen saturation (1.75% (0.97%, 3.03%) vs. 0.86% (0.44%, 1.65%), Z=11.086, P=0.001), and a higher proportion of patients with elevated body temperature (0.60% (0.39%, 1.03%) vs. 0.31% (0.17%, 0.54%), Z=7.314, P=0.007). Logistic regression analysis showed that advanced age ( OR=1.051, 95% CI: 1.023-1.080), increased heart rate ( OR=1.055, 95% CI:1.013-1.098), decreased oxygen saturation ( OR=1.197, 95% CI:1.023-1.400), and elevated body temperature ( OR=1.777, 95% CI:1.148-2.752) were positively associated factors for acute exacerbation of COPD. Conclusions:Abnormalities in physiological indicators such as heart rate, oxygen saturation, and body temperature are associated with acute exacerbations in COPD patients. Continuous monitoring using wearable devices may provide a new method for early warning and timely intervention in COPD exacerbations.