ObjectiveTo investigate the protective effect of genistein （Gen） on etoposide-induced chondrocyte senescence and its underlying mechanism. MethodsThe C28/I2 cell line was treated with different concentrations of Gen and etoposide， and the cell viability was detected by the CCK-8 assay. The senescence model of C28/I2 chondrocytes was induced by etoposide， with Gen intervention. Senescence-associated β-galactosidase （SA-β-gal） staining was performed to detect the senescence-positive rate and staining characteristics of chondrocytes. The expressions of peroxiredoxin 6 （Prdx6）， cyclin-dependent kinaseto clarify the functional necessity of Prdx6. ResultsCompared with the etoposide group， the C28/I2 chondrocyte viability significantly increased （P<0.01）， the expression ofsenescence-associated proteins p21 and p16 decreased （P<0.01， P<0.05）， the expression of senescence-associated genes p21 and p16 reduced （both P<0.01）， the fluorescence intensity of senescence-associated proteins p21 and p16 was diminished （P<0.05， P<0.01）， and the proportion of SA-β-gal-positive cells decreased （P<0.01） in the Gen+etoposide group. Compared with the Control group， the expression of Prdx6 was downregulated in the etoposide group （P<0.05）. Compared with the etoposide group， the expression of Prdx6 was upregulated in the Gen+etoposide group （P<0.01）. Compared with the Control group， the GPx activity significantly decreased in the si-Prdx6 group （P<0.01）. Furthermore， compared with the si-Prdx6 group， the GPx activity increased in the si-Prdx6+Gen group （P<0.05）. Molecular docking results revealed that Gen formed hydrogen bond interactions with the active site of Prdx6. After Prdx6 knockdown， the expression of senescence-associated genes p21 and p16 and the fluorescence intensity of senescence-associated proteins p21 and p16 both increased in the Gen+etoposide+si-Prdx6 group （both P<0.01）. ConclusionGen can inhibit etoposide-induced senescence of C28/I2 chondrocytes by upregulating the expression of Prdx6. This study provides potential drug targets and experimental basis for the prevention and treatment of chondrocyte senescence-related diseases.

Objective To develop an assessment scale of oropharyngeal candidiasis risks in HIV-infected patients,and to test its reliability and validity.Methods The draft of the assessment scale was constructed by the literature analysis method,Delphi consultations and analytic hierarchy process.Totally 231 HIV-infected patients in a tertiary A hospital of Hangzhou were recruited for questionnaire survey in October 2023 to January 2024.Results The scale includes 3 dimensions,including general factors,HIV related factors and oral related factors,12 first-level items and 21 second-level items.The Cronbach's α coefficient of the scale was 0.813;the content validity index was 0.98;the correlation of each dimension was 0.574～0.733(P＜0.05).Confirmatory factor analysis showed that the 3 factor models fitted the data well.The critical value(30.5 points)of the scale was determined by ROC curve;the area under the curve was 0.909;the sensitivity of index system was 0.962,and specificity was 0.697.Conclusion The assessment scale of oropharyngeal candidiasis risks in HIV-infected patients has good reliability and validity,and it can be used as an auxiliary tool for effective identification and prediction of oropharyngeal candidiasis risk.

Objective:To explore the clinical efficacy of pelvic floor biofeedback combined with electroacupuncture therapy in the treatment of chronic pelvic floor pain syndrome (CPPS).Methods:Ninety CPPS patients admitted to the Dongguan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine from August 2022 to July 2023 were selected prospectively, and they were divided into two groups by random number table method, each group with 45 cases. The control group received pelvic floor biofeedback treatment alone, while the observation group received pelvic floor biofeedback combined with electroacupuncture treatment. The two groups were treated once a day, 10 times as a course of treatment. After two courses of treatment, the clinical efficacy, pain degree before and after treatment, inflammatory factor levels, SF-36 health questionnaire (SF-36) scores, pelvic floor surface myoelectric parameters, and incidence of adverse reactions were compared between the two groups.Results:The total effective rate in the observation group after treatment was higher than that in the control group: 95.56% (43/45) vs. 75.56% (34/45), there was statistical difference ( χ2 = 7.28, P<0.05). After treatment, the visual analogue scale (VAS) scores, levels of procalcitonin (PCT), interleukin-6 (IL-6) and C-reactive protein (CRP) in the two groups were decreased, and SF-36 scores were increased, there were statistical differences ( P<0.05); the VAS scores, PCT, IL-6 and CRP levels in the observation group were lower than those in the control group, and SF-36 scores was higher than that in the control group: (2.11 ± 0.24) scores vs. (2.87 ± 0.34) scores, (0.92 ± 0.08) ng/L vs. (1.26 ± 0.09) ng/L, (24.08 ± 2.52) μg/L vs. (28.49 ± 3.10) μg/L, (4.55 ± 0.51) mg/L vs. (6.06 ± 0.63) mg/L, (74.55 ± 7.29) scores vs. (70.18 ± 7.80) scores, there were statistical differences ( P<0.05). After treatment, the surface muscle potential of pelvic floor muscle were increased in the front resting stage, rapid contraction stage, tension contraction stage, endurance contraction stage and post-resting stage, and the above parameters in the observation group were higher than those in the control group: (3.56 ± 0.34)μV vs.(3.20 ± 0.37) μV, (35.26 ± 3.05) μV vs. (31.47 ± 3.08) μV, (34.22 ± 3.25) μV vs. (31.15 ± 3.01)μV, (29.77 ± 3.17) μV vs. (27.04 ± 2.68) μV, (3.21 ± 0.27) μV vs. (3.00 ± 0.34) μV, there were statistical differences ( P<0.05). The incidence rate of adverse reactions in the two groups had no statistical difference ( P>0.05). Conclusions:Pelvic floor biofeedback combined with electroacupuncture in the treatment of CPPS has good clinical effect, and can relief pain response, regulate the inflammatory response and pelvic floor function, and have high safety.

Objective:To establish a method for the determination of 2-amino-5-chloro-N, 3-dimethylbenzamide in blood by ultra-performance liquid chromatography-quadrupole-orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) .Methods:In October 2022, 200 μl of blood samples were taken and 800 μl of acetonitrile was added. The samples were centrifuged at 3500 r/min (with a centrifugation radius of 6.6 cm) for 15 minutes. The supernatant was detected in the positive ion target tandem mass spectrometry (Target MS 2) mode and quantified by the external standard method. Methodological validation was also carried out for the established method. Results:The working curve of 2-amino-5-chloro-N, 3-dimethylbenzamide showed good linearity in the concentration range of 2.07-165.44 μg/L, with a correlation coefficient ( r) of 0.9994. The intra-day and inter-day relative standard deviations ( RSD) for the detection of 2-amino-5-chloro-N, 3-dimethylbenzamide were 2.6%-3.0% and 3.1%-3.5%, respectively. Analytical accuracy of 2-amino-5-chloro-N, 3-dimethylbenzamide quantification ranged from 101.0 % to 108.4 %, and the limit of detection and limit of quantification were 0.11 μg/L and 0.36 μg/L, respectively. The blood samples were able to be stored at -20 ℃ for at least 7 days. Conclusion:The UPLC-Q-Orbitrap HRMS determination method is sensitive, efficient and accurate, and can be used for the determination of 2-amino-5-chloro-N, 3-dimethylbenzamide in blood.

Objective: To explore the association between maternal lipid levels during pregnancy and the risk of overweight and obesity in offspring at 3 years of age, providing scientific evidences for the prevention and control of childhood obesity. Methods: A total of 2 432 mother-child pairs with maternal lipid tests during pregnancy and offspring s physical growth data at 3 years of age were included from the Borin in Guangzhou Cohort Study up to September 2021. Lipid indicators, including high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol(LDL-C), triglycerides (TG), and total cholesterol (TC), were measured at 13-19 +6 weeks (mid pregnancy) and 32-39 +6 weeks (late pregnancy). Children s body mass index (BMI) Z score were calculated according to the World Health Organization s growth standards for children under 5 years old. The lipid Z score were divided into four quartiles: Q 1, Q 2, Q 3 and Q 4. Linear regression was used to analyze the relationship between maternal lipid levels during pregnancy and offspring’s BMI Z score at 3 years of age. Poisson regression with a robust error variance was employed to evaluate the association between maternal lipid levels during pregnancy and the at risk of overweight and obesity in offspring at 3 years of age, after adjusting for maternal age at conception, education level, parity, pre pregnancy BMI and gestational diabetes mellitus. Results: There was a statistically significnt difference in the detection rate of overweight and obesity risk among children with different mothers s pre pregnancy BMI ( χ 2=22.85, P <0.05). Linear regression analysis showed that TG levels in late pregnancy were positively related to BMI Z score ( β=0.10, 95%CI=0.02-0.18, P <0.05). Poisson regression with a robust error variance indicated that, compared with the Q 1 group of TC, the Q 4 group of TC in mid pregnancy was associated with an increased risk of overweight and obesity in offspring at 3 years of age ( RR=1.59, 95%CI =1.04-2.44); compared with the Q 1 group of TG, the Q 4 group of TG during late pregnancy increased the risk of overweight and obesity in offspring at 3 years of age ( RR=1.79, 95%CI =1.02-3.12) (both P <0.05). Conclusions Maternal serum TC level during mid pregnancy can increase the risk of overweight and obesity in offspring at 3 years of age. Maternal serum TG levels during late pregnancy is positively correlated with BMI and the risk of overweight and obesity in offspring at 3 years of age.

Objective To develop an assessment scale of oropharyngeal candidiasis risks in HIV-infected patients,and to test its reliability and validity.Methods The draft of the assessment scale was constructed by the literature analysis method,Delphi consultations and analytic hierarchy process.Totally 231 HIV-infected patients in a tertiary A hospital of Hangzhou were recruited for questionnaire survey in October 2023 to January 2024.Results The scale includes 3 dimensions,including general factors,HIV related factors and oral related factors,12 first-level items and 21 second-level items.The Cronbach's α coefficient of the scale was 0.813;the content validity index was 0.98;the correlation of each dimension was 0.574～0.733(P＜0.05).Confirmatory factor analysis showed that the 3 factor models fitted the data well.The critical value(30.5 points)of the scale was determined by ROC curve;the area under the curve was 0.909;the sensitivity of index system was 0.962,and specificity was 0.697.Conclusion The assessment scale of oropharyngeal candidiasis risks in HIV-infected patients has good reliability and validity,and it can be used as an auxiliary tool for effective identification and prediction of oropharyngeal candidiasis risk.

Objective:To explore the clinical efficacy of pelvic floor biofeedback combined with electroacupuncture therapy in the treatment of chronic pelvic floor pain syndrome (CPPS).Methods:Ninety CPPS patients admitted to the Dongguan Hospital of Traditional Chinese Medicine Affiliated to Guangzhou University of Traditional Chinese Medicine from August 2022 to July 2023 were selected prospectively, and they were divided into two groups by random number table method, each group with 45 cases. The control group received pelvic floor biofeedback treatment alone, while the observation group received pelvic floor biofeedback combined with electroacupuncture treatment. The two groups were treated once a day, 10 times as a course of treatment. After two courses of treatment, the clinical efficacy, pain degree before and after treatment, inflammatory factor levels, SF-36 health questionnaire (SF-36) scores, pelvic floor surface myoelectric parameters, and incidence of adverse reactions were compared between the two groups.Results:The total effective rate in the observation group after treatment was higher than that in the control group: 95.56% (43/45) vs. 75.56% (34/45), there was statistical difference ( χ2 = 7.28, P<0.05). After treatment, the visual analogue scale (VAS) scores, levels of procalcitonin (PCT), interleukin-6 (IL-6) and C-reactive protein (CRP) in the two groups were decreased, and SF-36 scores were increased, there were statistical differences ( P<0.05); the VAS scores, PCT, IL-6 and CRP levels in the observation group were lower than those in the control group, and SF-36 scores was higher than that in the control group: (2.11 ± 0.24) scores vs. (2.87 ± 0.34) scores, (0.92 ± 0.08) ng/L vs. (1.26 ± 0.09) ng/L, (24.08 ± 2.52) μg/L vs. (28.49 ± 3.10) μg/L, (4.55 ± 0.51) mg/L vs. (6.06 ± 0.63) mg/L, (74.55 ± 7.29) scores vs. (70.18 ± 7.80) scores, there were statistical differences ( P<0.05). After treatment, the surface muscle potential of pelvic floor muscle were increased in the front resting stage, rapid contraction stage, tension contraction stage, endurance contraction stage and post-resting stage, and the above parameters in the observation group were higher than those in the control group: (3.56 ± 0.34)μV vs.(3.20 ± 0.37) μV, (35.26 ± 3.05) μV vs. (31.47 ± 3.08) μV, (34.22 ± 3.25) μV vs. (31.15 ± 3.01)μV, (29.77 ± 3.17) μV vs. (27.04 ± 2.68) μV, (3.21 ± 0.27) μV vs. (3.00 ± 0.34) μV, there were statistical differences ( P<0.05). The incidence rate of adverse reactions in the two groups had no statistical difference ( P>0.05). Conclusions:Pelvic floor biofeedback combined with electroacupuncture in the treatment of CPPS has good clinical effect, and can relief pain response, regulate the inflammatory response and pelvic floor function, and have high safety.

Objective:To establish a method for the determination of 2-amino-5-chloro-N, 3-dimethylbenzamide in blood by ultra-performance liquid chromatography-quadrupole-orbitrap high resolution mass spectrometry (UPLC-Q-Orbitrap HRMS) .Methods:In October 2022, 200 μl of blood samples were taken and 800 μl of acetonitrile was added. The samples were centrifuged at 3500 r/min (with a centrifugation radius of 6.6 cm) for 15 minutes. The supernatant was detected in the positive ion target tandem mass spectrometry (Target MS 2) mode and quantified by the external standard method. Methodological validation was also carried out for the established method. Results:The working curve of 2-amino-5-chloro-N, 3-dimethylbenzamide showed good linearity in the concentration range of 2.07-165.44 μg/L, with a correlation coefficient ( r) of 0.9994. The intra-day and inter-day relative standard deviations ( RSD) for the detection of 2-amino-5-chloro-N, 3-dimethylbenzamide were 2.6%-3.0% and 3.1%-3.5%, respectively. Analytical accuracy of 2-amino-5-chloro-N, 3-dimethylbenzamide quantification ranged from 101.0 % to 108.4 %, and the limit of detection and limit of quantification were 0.11 μg/L and 0.36 μg/L, respectively. The blood samples were able to be stored at -20 ℃ for at least 7 days. Conclusion:The UPLC-Q-Orbitrap HRMS determination method is sensitive, efficient and accurate, and can be used for the determination of 2-amino-5-chloro-N, 3-dimethylbenzamide in blood.

Autism Spectrum Disorders (ASDs) are reported as a group of neurodevelopmental disorders. The structural changes of brain regions including the hippocampus were widely reported in autistic patients and mouse models with dysfunction of ASD risk genes, but the underlying mechanisms are not fully understood. Here, we report that deletion of Trio, a high-susceptibility gene of ASDs, causes a postnatal dentate gyrus (DG) hypoplasia with a zigzagged suprapyramidal blade, and the Trio-deficient mice display autism-like behaviors. The impaired morphogenesis of DG is mainly caused by disturbing the postnatal distribution of postmitotic granule cells (GCs), which further results in a migration deficit of neural progenitors. Furthermore, we reveal that Trio plays different roles in various excitatory neural cells by spatial transcriptomic sequencing, especially the role of regulating the migration of postmitotic GCs. In summary, our findings provide evidence of cellular mechanisms that Trio is involved in postnatal DG morphogenesis.
Animals ; Dentate Gyrus/metabolism* ; Mice ; Morphogenesis/physiology* ; Neurons/pathology* ; Cell Movement ; Mice, Inbred C57BL ; Autism Spectrum Disorder/pathology* ; Mice, Knockout ; Neural Stem Cells ; Male ; Neurogenesis ; Autistic Disorder/genetics*

Objective To compare the anti-inflammatory,antitumor and anti-bacterial effects of the single extract(in granules)and the prepared drug in pieces of Forsythia Suspense(Lianqiao,a traditional Chinese herbal medicine).Methods In zebrafish embryo models of CuSO4 exposure,tail transection and LPS microinjection-induced inflammation,the anti-inflammatory effects of 10 μg/mL DEX,single extract of Forsythia Suspense,and the water extract of the prepared drug(400,600,and 800 μg/mL)were evaluated by observing neutrophil counts,RT-qPCR,HE staining and survival analysis.Zebrafish embryo models bearing different human tumor cell xenografts were used to assess the anti-tumor effect of the drugs in different dosage forms by fluorescence staining and HE staining.The microbroth dilution method was used to evaluate the antibacterial efficacy of the drugs.Results In the zebrafish embryo models of inflammation,both of the two dosage forms of Forsythia Suspense significantly inhibited neutrophil aggregation,reduced the mRNA expressions of TNF-α,IL-6,P38,Jnk,Erk and P65,and increased the survival rate of zebrafish.They both showed obvious inhibitory effects against xenografts of different human cancer cells including colon cancer cells(HCT116),pancreas adenocarcinoma cells(PANC-1),lung cancer cells(A549),liver cancer cells(Hep3B)and cervical carcinoma cells(Hela)in zebrafish embryos,and exhibited strong anti-bacterial effects at the concentration of 15.63 mg/mL.Conclusion The two dosage forms of Forsythia Suspense have similar anti-inflammatory,antitumor and antibacterial effects,but their effects for inhibiting IL-6,P65,and Jnk mRNA expressions and HCT116 cell proliferation differ significantly at low doses in zebrafish.