Objective To establish the reference interval of urine Alzheimer-associated neuronal thread protein(AD7c-NTP)for healthy adults in Mianyang area using the indirect method.Methods The detection results of urine AD7c-NTP from 5 093 healthy in-dividuals were collected from the information management database of Medical Laboratory Department of Sichuan Science City Hospital from March 2017 to March 2022.Skewness-kurtosis and Kolmogorov-Smirnov tests were used to determine whether the data followed a normal distribution.After removing outliers using the Box Plots method,the enrolled subjects were grouped by gender and age.The Mann-Whitney U or Kruska-Wallis H tests were used to analyze the between-group differences of urine AD7c-NTP in healthy individu-als with different genders and ages.The adjacent age groups without statistically significant difference(P＞0.05)were combined,and the indirect method(non-parametric test method)was used to calculate the reference intervals for different gender and age groups.Results Skewness-kurtosis and Kolmogorov-Smirnov tests showed that the data followed a non-normal distribution.After removing 293 outliers using the Box Plots method,a total of 4 800 subjects,including 3 199 males and 1 601 females,were enrolled.The enrolled subjects were grouped by gender and age,and the non-parametric test method were used to establish the reference intervals of urine AD7c-NTP in healthy populations with different genders.The Mann-Whitney U test confirmed that urine AD7c-NTP levels existed gen-der differences(Z=14.09,P＜0.01),and the reference intervals for males and females were≤1.10 ng/mL and≤1.40 ng/mL,re-spectively.There were also statistical differences in urine AD7c-NTP levels among different age groups of the same gender.After combi-ning adjacent age groups without statistically significant difference(P＞0.05),the reference intervals of urine AD7c-NTP in healthy populations with different genders and ages were established by the non-parametric test method,which were≤1.00 ng/mL for male 20-39 years old group,≤1.10 ng/mL for male 40-79 years old group,≤1.60 ng/mL for male≥80 years old group,≤1.30 ng/mL for female 20-69 years old group,and≤1.60 ng/mL for female≥70 years old group,respectively.The established reference intervals of urine AD7c-NTP were further verified by healthy individuals,and the results met the standards.Conclusion The reference intervals of urine AD7c-NTP in healthy populations with different genders and ages in Mianyang area are established successfully using the indi-rect method,which may help to predict the risk of Alzheimer's disease in clinical practice and provide support for the diagnosis and treatment of the disease.

OBJECTIVE: To observe the effects of acupuncture on podocyte autophagy and long non-coding RNA SOX2 overlapping transcript (LncRNA SOX2OT)/mammalian target of rapamycin C1 (mTORC1)/Unc-51-like kinase 1 (ULK1) pathway in rats with diabetic kidney disease (DKD), and to explore the mechanism by which acupuncture reduces urinary protein. METHODS: A total of 40 SPF-grade male Sprague-Dawley rats were randomly divided into a control group (n=10) and a modeling group (n=30). The DKD model was established by feeding a high-fat, high-sugar diet combined with intraperitoneal injection of streptozotocin (STZ) in the modeling group. Twenty rats with successful DKD model were randomly divided into a model group (n=10) and an acupuncture group (n=10). The acupuncture group received "spleen and stomach-regulating" acupuncture at bilateral "Zusanli" (ST36), "Fenglong" (ST40), "Yinlingquan" (SP9), and "Zhongwan" (CV12), 30 min per session, once daily, five times per week, for four weeks. The general condition, fasting blood glucose (FBG), 2-hour postprandial glucose (2hPG), serum creatinine (SCr), blood urea nitrogen (BUN), 24-hour urinary protein quantification, and urine albumin-to-creatinine ratio (UACR) were compared before and after the intervention. After intervention, urinary podocyte injury marker SPON2 was measured by ELISA. Podocyte autophagosomes and glomerular basement membrane ultrastructure in renal tissue were observed via transmission electron microscopy. Podocyte apoptosis was assessed by TUNEL staining. The protein expression of microtubule-associated protein 1 light chain 3Ⅱ (LC3-Ⅱ), mTORC1, ULK1, Beclin-1, and p62 in renal tissue was detected by Western blot. LncRNA SOX2OT expression in renal tissue was measured by real-time PCR. RESULTS: After the intervention, compared with the control group, the model group exhibited increased food and water intake, increased urine output, weight loss, and loose stools; compared with the model group, the food and water intake, urine volume, and loose stools were improved in the acupuncture group. Compared with the control group, FBG, 2hPG, SCr, BUN, 24-hour urinary protein quantification, UACR, and urinary SPON2 were all higher in the model group (P<0.01); compared with the model group, the FBG, 2hPG, SCr, BUN, 24-hour urinary protein quantification, UACR, and urinary SPON2 were all lower in the acupuncture group (P<0.01). Compared with the control group, the model group showed reduced podocyte autophagosomes and thickened glomerular basement membrane; compared with the model group, the acupuncture group had increased podocyte autophagosomes and less thickened basement membrane. Compared with the control group, the podocyte apoptosis index (AI) was higher in the model group (P<0.01); compared with the model group, the AI was lower in the acupuncture group (P<0.01). Compared with the control group, the expression of ULK1, Beclin-1, and LC3-Ⅱ proteins was lower, and the expression of mTORC1 and p62 proteins was higher in the model group (P<0.01). Compared with the model group, the expression of ULK1, Beclin-1, and LC3-Ⅱ proteins was higher, and the expression of mTORC1 and p62 proteins was lower in the acupuncture group (P<0.01). Compared with the control group, the LncRNA SOX2OT expression was lower in the model group (P<0.01). Compared with the model group, LncRNA SOX2OT expression was higher in the acupuncture group (P<0.01). CONCLUSION The "spleen and stomach-regulating" acupuncture method could improve renal function in DKD rats, reduce blood glucose and urinary protein excretion, alleviate podocyte injury, and enhance podocyte autophagy. The mechanism may be related to modulation of the renal LncRNA SOX2OT/mTORC1/ULK1 pathway.
Animals ; Podocytes/cytology* ; Diabetic Nephropathies/physiopathology* ; Rats, Sprague-Dawley ; Male ; Rats ; Mechanistic Target of Rapamycin Complex 1/genetics* ; Autophagy ; Acupuncture Therapy ; Autophagy-Related Protein-1 Homolog/genetics* ; RNA, Long Noncoding/metabolism* ; Humans ; Signal Transduction

Objective:This study aims to investigate the behavioral changes, gut microbiota alterations, and short-chain fatty acid levels in mice with chronic dizocilpine (MK-801) administration and to evaluate the correlation between gut microbiota and its metabolites with schizophrenia-like behaviors.Methods:This study was conducted from March to September 2024, involving 24 male C57BL/6 mice aged 6 to 8 weeks, purchased from the Guangdong Provincial Medical Laboratory Animal Center. The mice were randomly divided into a control group and a model group using a random number table method. The model group mice were intraperitoneally injected with MK-801 (0.6 mg/kg) for 14 days, while the control group mice were intraperitoneally injected with an equal amount of normal saline for 14 days. After modeling, behavioral performance was assessed using the open field test (OFT), novel object recognition test (NOR), forced swimming test (FST), tail suspension test (TST), elevated plus maze test (EPM), and prepulse inhibition test (PPI). Additionally, mouse feces were collected for 16S rRNA sequencing to analyze the composition of gut microbiota. The gas chromatography-tandem mass spectrometry (GC-MS/MS) was applied to detect the levels of short-chain fatty acids quantitatively. Between-group comparisons of behavioral data were performed using an independent samples t-test or repeated measures ANOVA with adjusted Bonferroni or corrected Greenhouse-Geisser. Between-group comparisons of gut microbiota composition were conducted using an independent samples t-test or the Mann-Whitney U test. Between-group comparisons of short-chain fatty acids were performed using an independent samples t-test. The correlations among behavioral indicators, gut microbiota composition, and short-chain fatty acids were analyzed using Spearman′s rank correlation. Results:(1) Behavioral experiments showed that the model group mice exhibited significantly prolonged immobility times in the FST and TST ( t=-4.84, -2.54; P<0.05), significantly reduced exploration frequency and time in the open arms of the EPM ( t=3.31, 2.48; P<0.05), significantly lower PPI at 76 dB, 79 dB, and 85 dB ( F=23.28, 10.65, 17.82; P<0.05), and a significantly reduced NOR index ( t=2.90, P<0.05) compared to the control group, indicating successful modeling. (2)16S rRNA sequencing revealed that, compared with the control group, the model group exhibited significantly lower relative abundances of Actinobacteriota, Coriobacteriia, Coriobacteriales, and Ileibacterium( Z=-3.10--2.04, all P<0.05). Conversely, the relative abundances of Tannerellaceae, Rikenellaceae, Alistipes, Bacteroides, Intestinimonas, Parabacteroides, Unclassified_f_Prevotellaceae, and unclassified_f_Erysipelotrichaceae significantly increased ( Z=-3.78--2.04; all P<0.05). (3)GC-MS/MS analysis showed that the concentrations of acetic acid and butyric acid in the feces of the model group were significantly lower compared to the control group ( t=2.66, 2.10; P<0.05). (4)Spearman correlation analysis with FDR correction (Benjamini-Hochberg method) revealed that Actinobacteriota significantly positively correlated with the open-arm exploration frequency in the EPM ( r=0.69, Q<0.05), whereas unclassified_f_Erysipelotrichaceae significantly negatively correlated with this measure ( r=-0.66, Q<0.05). Additionally, Bacteroides and Intestinimonas were significantly negatively correlated with PPI at 85 dB ( r=-0.71, -0.63; Q<0.05).Conversely, Ileibacterium demonstrated a significant positive correlation with PPI at 85 dB ( r=0.64, Q<0.05). Conclusion:Alterations in gut microbiota and SCAFs may be associated with MK-801-induced schizophrenia-like behaviors.

Objective To establish the reference interval of urine Alzheimer-associated neuronal thread protein(AD7c-NTP)for healthy adults in Mianyang area using the indirect method.Methods The detection results of urine AD7c-NTP from 5 093 healthy in-dividuals were collected from the information management database of Medical Laboratory Department of Sichuan Science City Hospital from March 2017 to March 2022.Skewness-kurtosis and Kolmogorov-Smirnov tests were used to determine whether the data followed a normal distribution.After removing outliers using the Box Plots method,the enrolled subjects were grouped by gender and age.The Mann-Whitney U or Kruska-Wallis H tests were used to analyze the between-group differences of urine AD7c-NTP in healthy individu-als with different genders and ages.The adjacent age groups without statistically significant difference(P＞0.05)were combined,and the indirect method(non-parametric test method)was used to calculate the reference intervals for different gender and age groups.Results Skewness-kurtosis and Kolmogorov-Smirnov tests showed that the data followed a non-normal distribution.After removing 293 outliers using the Box Plots method,a total of 4 800 subjects,including 3 199 males and 1 601 females,were enrolled.The enrolled subjects were grouped by gender and age,and the non-parametric test method were used to establish the reference intervals of urine AD7c-NTP in healthy populations with different genders.The Mann-Whitney U test confirmed that urine AD7c-NTP levels existed gen-der differences(Z=14.09,P＜0.01),and the reference intervals for males and females were≤1.10 ng/mL and≤1.40 ng/mL,re-spectively.There were also statistical differences in urine AD7c-NTP levels among different age groups of the same gender.After combi-ning adjacent age groups without statistically significant difference(P＞0.05),the reference intervals of urine AD7c-NTP in healthy populations with different genders and ages were established by the non-parametric test method,which were≤1.00 ng/mL for male 20-39 years old group,≤1.10 ng/mL for male 40-79 years old group,≤1.60 ng/mL for male≥80 years old group,≤1.30 ng/mL for female 20-69 years old group,and≤1.60 ng/mL for female≥70 years old group,respectively.The established reference intervals of urine AD7c-NTP were further verified by healthy individuals,and the results met the standards.Conclusion The reference intervals of urine AD7c-NTP in healthy populations with different genders and ages in Mianyang area are established successfully using the indi-rect method,which may help to predict the risk of Alzheimer's disease in clinical practice and provide support for the diagnosis and treatment of the disease.

Objective:This study aims to investigate the behavioral changes, gut microbiota alterations, and short-chain fatty acid levels in mice with chronic dizocilpine (MK-801) administration and to evaluate the correlation between gut microbiota and its metabolites with schizophrenia-like behaviors.Methods:This study was conducted from March to September 2024, involving 24 male C57BL/6 mice aged 6 to 8 weeks, purchased from the Guangdong Provincial Medical Laboratory Animal Center. The mice were randomly divided into a control group and a model group using a random number table method. The model group mice were intraperitoneally injected with MK-801 (0.6 mg/kg) for 14 days, while the control group mice were intraperitoneally injected with an equal amount of normal saline for 14 days. After modeling, behavioral performance was assessed using the open field test (OFT), novel object recognition test (NOR), forced swimming test (FST), tail suspension test (TST), elevated plus maze test (EPM), and prepulse inhibition test (PPI). Additionally, mouse feces were collected for 16S rRNA sequencing to analyze the composition of gut microbiota. The gas chromatography-tandem mass spectrometry (GC-MS/MS) was applied to detect the levels of short-chain fatty acids quantitatively. Between-group comparisons of behavioral data were performed using an independent samples t-test or repeated measures ANOVA with adjusted Bonferroni or corrected Greenhouse-Geisser. Between-group comparisons of gut microbiota composition were conducted using an independent samples t-test or the Mann-Whitney U test. Between-group comparisons of short-chain fatty acids were performed using an independent samples t-test. The correlations among behavioral indicators, gut microbiota composition, and short-chain fatty acids were analyzed using Spearman′s rank correlation. Results:(1) Behavioral experiments showed that the model group mice exhibited significantly prolonged immobility times in the FST and TST ( t=-4.84, -2.54; P<0.05), significantly reduced exploration frequency and time in the open arms of the EPM ( t=3.31, 2.48; P<0.05), significantly lower PPI at 76 dB, 79 dB, and 85 dB ( F=23.28, 10.65, 17.82; P<0.05), and a significantly reduced NOR index ( t=2.90, P<0.05) compared to the control group, indicating successful modeling. (2)16S rRNA sequencing revealed that, compared with the control group, the model group exhibited significantly lower relative abundances of Actinobacteriota, Coriobacteriia, Coriobacteriales, and Ileibacterium( Z=-3.10--2.04, all P<0.05). Conversely, the relative abundances of Tannerellaceae, Rikenellaceae, Alistipes, Bacteroides, Intestinimonas, Parabacteroides, Unclassified_f_Prevotellaceae, and unclassified_f_Erysipelotrichaceae significantly increased ( Z=-3.78--2.04; all P<0.05). (3)GC-MS/MS analysis showed that the concentrations of acetic acid and butyric acid in the feces of the model group were significantly lower compared to the control group ( t=2.66, 2.10; P<0.05). (4)Spearman correlation analysis with FDR correction (Benjamini-Hochberg method) revealed that Actinobacteriota significantly positively correlated with the open-arm exploration frequency in the EPM ( r=0.69, Q<0.05), whereas unclassified_f_Erysipelotrichaceae significantly negatively correlated with this measure ( r=-0.66, Q<0.05). Additionally, Bacteroides and Intestinimonas were significantly negatively correlated with PPI at 85 dB ( r=-0.71, -0.63; Q<0.05).Conversely, Ileibacterium demonstrated a significant positive correlation with PPI at 85 dB ( r=0.64, Q<0.05). Conclusion:Alterations in gut microbiota and SCAFs may be associated with MK-801-induced schizophrenia-like behaviors.

BACKGROUND:The research of dental stem cells in the fields of regenerative medicine and tissue engineering has been deepening,bringing hope for the repair of tooth-related tissues and the treatment of systemic diseases.However,there is a lack of systematic research and analysis on the biological characteristics of dental stem cells in different age groups. OBJECTIVE:To explore the biological characteristics of the human deciduous tooth and permanent tooth pulp stem cells cultured in umbilical cord blood platelet lysate to provide a reliable basis for human platelet lysates to replace fetal bovine serum. METHODS:The pulp tissues of deciduous teeth,juvenile permanent teeth and adult permanent teeth were taken out and cultured in DMEM/F-12 medium supplemented with 10%fetal bovine serum or different concentrations(5%,10%and 15%)of human platelet lysates.Cell proliferation in the four groups was detected by cytometry.The optimal concentration of human platelet lysates was selected for subsequent experiments.Under the optimal concentration of human platelet lysates,human deciduous tooth and juvenile and adult permanent tooth pulp stem cells were cultured in vitro.The cell growth status was observed under the microscope.The specific antigen on the cell surface was detected by flow cytometry.The cell proliferation ability was tested by the cell counting method and CCK-8 assay.The cell differentiation ability in vitro was observed by a three-line differentiation assay. RESULTS AND CONCLUSION:(1)The cell proliferation rate of the 10%human platelet lysate group was the highest.(2)In all three groups,fusiform fibrous cells grew and expanded from around the tissue block.There was no significant difference between deciduous teeth and juvenile permanent tooth cells,but the adult permanent tooth cells were larger than the deciduous and juvenile permanent tooth cells of the same generation.(3)The results of flow cytometry showed that deciduous teeth,juvenile permanent teeth and adult permanent teeth conformed to the phenotypic characteristics of mesenchymal stem cells.(4)The proliferative capacity of adult permanent dental pulp stem cells was significantly lower than those of deciduous teeth and juvenile permanent dental pulp stem cells(P<0.01).(5)mRNA expressions of osteoblast-related genes alkaline phosphatase and bone morphogenetic protein 2,lipoprotein lipase and peroxisome proliferator-activated receptor γ2,mRNA expressions of chondroblast related gene type II collagen α1 and cartilage oligomeric matrix protein in adult pulp stem cells of permanent teeth were significantly lower than those of deciduous teeth and juvenile permanent teeth pulp stem cells(P<0.01).(6)Compared with adult dental pulp stem cells,human deciduous teeth and juvenile permanent teeth dental pulp stem cells have the stronger proliferative capacity and multidirectional differentiation potential,and are more suitable for clinical research and disease treatment.

The high incidence of complications in preterm infants has a serious negative impact on neonatal health.Therefore,the discovery of predictors of complications in preterm infants has a positive clinical value.Umbilical cord blood is rich in various biomarkers and has the advantages of easy access and no harm to the infants,making it an ideal specimen for testing.A number of studies had explored the predictive value of de-tecting markers in umbilical cord blood,either alone or in combination with other assays,for various types of preterm complications,in order to facilitate earlier intervention for preterm complications and to improve the prognosis of the disease.This paper reviews the recent development of cord blood testing for predicting com-plications in various types of preterm infants.

Objective To investigate the effect of bone marrow mesenchymal stem cells(BMSC)transplantation on rats with painful diabetes peripheral neuropathy(PDPN).Methods Fourteen SPF grade male GK rats were randomly divided into model group and treat-ment group,with 7 rats in each group.Seven Wistar rats of the same-sex and same-age were used as the normal group.The treatment group received intraperitoneal injection of BMSC for 6 weeks of treatment.Detection of thermal pain threshold;take the sciatic nerve and observe the pathological morphological changes of the sciatic nerve using hematoxylin-eosin(HE)staining and Masson staining;Western blot was used to detect the expression of JAK2/STAT3 signaling pathway related proteins p-JAK2 and p-STAT3.Results After treat-ment with BMSC,the thermal pain threshold of rats increased slightly;the sciatic nerve injury was repaired,the infiltration of inflammato-ry cells was alleviated,and the phenomenon of demyelination was improved;the expression of pathway related proteins p-JAK2 and p-STAT3 decreased.Conclusion BMSC transplantation can alleviate inflammatory response,repair sciatic nerve,and alleviate pain by in-hibiting the JAK2/STAT3 signaling pathway.

Objective To explore the effect of 12-week simplified Tai Chi training on rehabilitation and neuromuscular control in individuals with chronic ankle instability(CAI).Methods Thirty-four partic-ipants with CAI were randomly divided into an experimental group and a control group,each of 17.The control group received 12-week health education,while the experimental group underwent simpli-fied Tai Chi training for the same length.Before and after the intervention,both groups were evaluat-ed their self-reported instability feeling,ankle muscle strength,proprioception and dynamic postural stability.Results After intervention,a significant increase was observed in the average Cumberland An-kle Instability Tool(CAIT),the maximum extension distance in the posteromedial and posterolateral di-rections of the modified Star Excursion Balance Test(mSEBT)in,as well as the ankle peak torque in plantarflexion,inversion and eversion(P<0.05)in the experimental.Moreover,the ankle passive motion proprioceptive threshold in plantarflexion and inversion decreased significantly in the experimental group after intervention(P<0.05).Meanwhile,after intervention,the average CAIT score,maximum ex-tension distances in the posteromedial and posterolateral directions in the mSEBT,and the ankle peak torques in plantarflexion,inversion and eversion of the experimental group were significantly higher than the control group(P<0.05),while the ankle passive motion proprioceptive threshold in plantarflex-ion was significantly lower than the latter(P<0.05).Conclusion Twelve-week simplified Tai Chi train-ing can improve the clinical subjective instability of CAI participants,maybe related to the improve-ment of ankle muscle strength,proprioception and dynamic postural stability.Therefore,it is suggest-ed that simplified Tai Chi should be one of the rehabilitation methods for CAI patients.

Aristolochic acid (AA), extracted from Aristolochiaceae plants, plays an essential role in traditional herbal medicines and is used for different diseases. However, AA has been found to be nephrotoxic and is known to cause aristolochic acid nephropathy (AAN).AA-induced acute kidney injury (AKI) is a syndrome in AAN with a high morbidity that manifests mitochondrial damage as a key part of its pathological progression. Melatonin primarily serves as a mitochondria-targeted antioxidant. However, its mitochondrial protective role in AA-induced AKI is barely reported. In this study, mice were administrated 2.5 mg/kg AA to induce AKI. Melatonin reduced the increase in Upro and Scr and attenuated the necrosis and atrophy of renal proximal tubules in mice exposed to AA. Melatonin suppressed ROS generation, MDA levels and iNOS expression and increased SOD activities in vivo and in vitro. Intriguingly, the in vivo study revealed that melatonin decreased mitochondrial fragmentation in renal proximal tubular cells and increased ATP levels in kidney tissues in response to AA. In vitro, melatonin restored the mitochondrial membrane potential (MMP) in NRK-52E and HK-2 cells and led to an elevation in ATP levels. Confocal immunofluorescence data showed that puncta containing Mito-tracker and GFP-LC3A/B were reduced, thereby impeding the mitophagy of tubular epithelial cells. Furthermore, melatonin decreased LC3A/B-II expression and increased p62 expression. The apoptosis of tubular epithelial cells induced by AA was decreased. Therefore, our findings revealed that melatonin could prevent AA-induced AKI by attenuating mitochondrial damage, which may provide a potential therapeutic method for renal AA toxicity.