OBJECTIVES: To investigate the regulatory role of astrocytes in the dorsomedial hypothalamus (DMH) during sevoflurane anesthesia emergence. METHODS: Forty-two male C57BL/6 mice were randomized into 6 groups (n=7) for assessing astrocyte activation in the dorsomedial hypothalamus (DMH) under sevoflurane anesthesia. Two groups of mice received microinjection of agfaABC1D promoter-driven AAV2 vector into the DMH for GCaMP6 overexpression, and the changes in astrocyte activity during sevoflurane or air inhalation were recorded using calcium imaging. For assessing optogenetic activation of astrocytes, another two groups of mice received microinjection of an optogenetic virus or a control vector into the DMH with optic fiber implantation, and sevoflurane anesthesia emergence was compared using behavioral experiments. In the remaining two groups, electroencephalogram (EEG) recording during sevoflurane anesthesia emergence was conducted after injection of the hChR2-expressing and control vectors. Anesthesia induction and recovery were assessed by observing the righting reflex. EEG data were recorded under 2.0% sevoflurane to calculate the burst suppression ratio (BSR) and under 1.5% sevoflurane for power spectrum analysis. Immunofluorescence staining was performed to visualize the colocalization of GFAP-positive astrocytes with viral protein signals. RESULTS: Astrocyte activity in the DMH decreased progressively as sevoflurane concentration increased. During 2.0% sevoflurane anesthesia, the mice injected with the ChR2-expressing virus exhibited a significantly shortened wake-up time (P<0.05), and optogenetic activation of the DMH astrocytes led to a marked reduction in BSR (P<0.001). Under 1.5% sevoflurane anesthesia, optogenetic activation resulted in a significant increase in EEG gamma power and a significant decrease in delta power in ChR2 group (P<0.01). CONCLUSIONS Optogenetic activation of DMH astrocytes facilitates sevoflurane anesthesia emergence but does not significantly influence anesthesia induction. These findings offer new insights into the mechanisms underlying anesthesia emergence and may provide a potential target for accelerating postoperative recovery and managing anesthesia-related complications.
Animals ; Astrocytes/physiology* ; Sevoflurane ; Mice, Inbred C57BL ; Mice ; Male ; Electroencephalography ; Anesthetics, Inhalation/pharmacology* ; Hypothalamus/cytology* ; Anesthesia Recovery Period ; Methyl Ethers/pharmacology*

ObjectiveTo explore the impact of intrauterine growth discordance on physical growth and development during infancy and early childhood in twins, and to provide an evidence-based basis for early childhood healthcare and clinical intervention. MethodsA retrospective cohort study design was adopted, with data sourced from the Yiwu Maternal and Child Health Information Platform, Zhejiang Province from January 2016 to July 2023. A total of 439 pairs of twins were included in the study. Generalized estimating equations (GEE) were used to analyze the association between twin growth discordance and weight, length, and head circumference within 36 months of age. ResultsAmong the 439 pairs of twins, 51 pairs (11.6%) exhibited intrauterine growth discordance. GEE analyses showed that the percentage difference of intrauterine growth discordance was negatively associated with weight (β=-0.013, P<0.001), length (β=-0.033, P<0.001), and head circumference (β=-0.015, P<0.001) within 36 months of age. Among the discordant pairs, the twin with the lower birth weight had a significantly higher risk of being classified as growth-retarded for weight (OR=6.057, 95%CI: 3.956‒9.274), length (OR=5.233, 95%CI: 3.499‒7.827), and head circumference (OR=3.476, 95%CI: 2.004‒6.031) during early childhood. ConclusionAmong twins with intrauterine growth discordance, the twin born with lower birth weight is a risk factor for growth retardation in weight, length, and head circumference during infancy and early childhood. It is suggested that an early identification mechanism should be established for such high-risk groups, dynamic monitoring should be implemented in clinical diagnosis and treatment as well as child health care services, and comprehensive measures such as personalized nutritional interventions should be taken as early as possible.

Objective:To observe the effects of Yiyuan moxibustion on bladder function and antioxidant level of spinal cord tissues in rats with urinary retention after spinal cord injury (SCI); To explore the mechanism of inhibition of ferroptosis in spinal cord neurons after SCI by Yiyuan moxibustion.Methods:Wistar female rats were divided into sham-operation group, model group, and Yiyuan moxibustion group according to random number table method, with 12 rats in each group. The modified Allen′s vertical percussion method was used to construct the model of urinary retention after SCI in T10 segment. The rats in the Yiyuan moxibustion group were moxibued at the Zhongji acupoint, Guanyuan acupoint, and Shenque acupoint for 20 min per day, and the intervention was continued for 2 weeks. Urodynamic test was used to observe the degree of urinary retention in rats; HE staining was used to observe the morphological changes of the injured spinal cord tissues; transmission electron microscopy was used to observe the mitochondrial ultrastructure of the spinal cord tissues; ferric ion kit was used to detect the ferric ion content of the spinal cord tissues; ELISA was used to detect the GSH and MDA contents of the spinal cord tissues of the rats; Western blot was used to measure the relative expressions of glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (SLC7A11) proteins in rat spinal cord tissue.Results:Compared with the model group, the basal and leakage point pressures of the bladder, and bladder compliance were significantly reduced in the Yiyuan moxibustion group ( P<0.05); the spinal cord tissue structure was restored and mitochondrial morphology improved; the levels of iron ions and MDA in spinal cord tissue decreased ( P<0.05), while the level of GSH increased ( P<0.05), and the relative expressions of GPX4 and SLC7A11 proteins increased ( P<0.05). Conclusion:Yiyuan moxibustion can improve bladder function in rats with urinary retention after SCI, and the mechanism may involve the initiation of antioxidant defense and reduction of lipid peroxidation in spinal cord neuronal cells, thus preventing the occurrence of ferroptosis and achieving the protection of neuronal cells.

Objective:To understand the molecular evolutionary characteristics of the recombinant strain GII.3[P12] of norovirus acute gastroenteritis outbreaks and aggregated outbreaks in China from 2022 to 2023.Methods:Epidemiological information, case information, clinical samples, as well as detection and genotyping information of norovirus outbreaks and aggregated outbreaks from 2022 to 2023 were collected; positive samples of the GII.3[P12] recombinant strain were subjected to nucleic acid extraction, whole-genome amplification, and sequence analysis; and homology simulation method were used to construct a three-dimensional structure and predict antigenic epitopes.Results:From January 2022 to December 2023, a total of 1 136 norovirus outbreaks and aggregated outbreaks were reported in China′s norovirus outbreak surveillance network, and genotyping result were successfully obtained for 942 outbreaks, with GII dominating, accounting for 76.0% (716/942), and the proportion of GI and mixed genotypes being 15.8% (149/942) and 8.2% (77/942). Norovirus outbreaks caused by GII were dominated by GII.3[P12] (22.5%, 161/716), while other major genotypes included GII.17[P17] (18.7%, 134/716), GII.4_Sydney 2012[P16] (11.6%, 83/716) and GII.6[P7] ( 11.3%, 81/716). 2022-2023 Outbreaks caused by GII.3[P12] were concentrated in February-March (54.0%, 87/161), with the main outbreaks occurring in nursery and primary schools (87.5%), the mode of transmission was mainly human-to-human (68.9%), and the main susceptible population was children aged 3-7 years (93.3%). In this study, the genome sequences of 25 GII.3[P12] recombinant strains were obtained, and according to the phylogenetic analysis, it was shown that the GII.3[P12] recombinant strains in China in 2022-2023 belonged to the Cluster IV cluster of sublineage b (7 strains) and sublineage c (18 strains). A total of 11 linear and 8 conformational epitopes were predicted by epitope prediction analysis, and the predicted linear and conformational epitopes had overlapping positions, and each conformational epitope was part of the predicted linear epitope with conserved potential antigen-binding and receptor-binding sites.Conclusions:The recombinant strain GII.3[P12] is one of the epidemic strains that will cause outbreaks and clusters of norovirus in China in 2022-2023, and its genome did not undergo significant mutation.

Objective:To understand the molecular evolutionary characteristics of the recombinant strain GII.3[P12] of norovirus acute gastroenteritis outbreaks and aggregated outbreaks in China from 2022 to 2023.Methods:Epidemiological information, case information, clinical samples, as well as detection and genotyping information of norovirus outbreaks and aggregated outbreaks from 2022 to 2023 were collected; positive samples of the GII.3[P12] recombinant strain were subjected to nucleic acid extraction, whole-genome amplification, and sequence analysis; and homology simulation method were used to construct a three-dimensional structure and predict antigenic epitopes.Results:From January 2022 to December 2023, a total of 1 136 norovirus outbreaks and aggregated outbreaks were reported in China′s norovirus outbreak surveillance network, and genotyping result were successfully obtained for 942 outbreaks, with GII dominating, accounting for 76.0% (716/942), and the proportion of GI and mixed genotypes being 15.8% (149/942) and 8.2% (77/942). Norovirus outbreaks caused by GII were dominated by GII.3[P12] (22.5%, 161/716), while other major genotypes included GII.17[P17] (18.7%, 134/716), GII.4_Sydney 2012[P16] (11.6%, 83/716) and GII.6[P7] ( 11.3%, 81/716). 2022-2023 Outbreaks caused by GII.3[P12] were concentrated in February-March (54.0%, 87/161), with the main outbreaks occurring in nursery and primary schools (87.5%), the mode of transmission was mainly human-to-human (68.9%), and the main susceptible population was children aged 3-7 years (93.3%). In this study, the genome sequences of 25 GII.3[P12] recombinant strains were obtained, and according to the phylogenetic analysis, it was shown that the GII.3[P12] recombinant strains in China in 2022-2023 belonged to the Cluster IV cluster of sublineage b (7 strains) and sublineage c (18 strains). A total of 11 linear and 8 conformational epitopes were predicted by epitope prediction analysis, and the predicted linear and conformational epitopes had overlapping positions, and each conformational epitope was part of the predicted linear epitope with conserved potential antigen-binding and receptor-binding sites.Conclusions:The recombinant strain GII.3[P12] is one of the epidemic strains that will cause outbreaks and clusters of norovirus in China in 2022-2023, and its genome did not undergo significant mutation.

Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Objective To investigate the role of glutamatergic neurons in the dorsomedial periaqueductal grey(dmPAG)in regulating excessive defensive behaviors in mice with post-traumatic stress disorder(PTSD).Methods Eight-week-old male C57BL/6 mice were subjected to stereotactic injections of different recombinant adeno-associated viral vectors(rAAV2/9-CaMKⅡ-mCherry,rAAV2/9-CaMKⅡ-hM3Dq-mCherry and rAAV2/9-CaMKⅡ-hM4Di-mCherry)into the bilateral dmPAG for chemogenetic activation or inhibition of the glutamatergic neurons,followed 2 weeks later by PTSD modeling by single prolonged stress.The looming test,response to whisker stimulation test and contextual fear conditioning(CFC)test were used to observe changes in defensive behaviors of the PTSD mice.The activity of glutamatergic neurons in the dmPAG were observed using immunofluorescence staining.Results Compared with the control mice,the mouse models of PTSD showed a shortened latency of flights with increased time spent in the nest,response scores of defensive behaviors and freezing time(all P<0.01).Immunofluorescence staining revealed significantly increased c-fos-positive glutamatergic neurons in the dmPAG of PTSD mice with defensive behaviors.Activation of the glutamatergic neurons in the dmPAG(in PTSD hM3Dq group)did not cause significant changes in the latency of flights or time in nest but obviously increased response scores of defensive behaviors and freezing time of the mice,whereas inhibiting the glutamatergic neurons in the dmPAG(in PTSD hM4Di group)caused the reverse changes and obviously alleviated defensive behaviors in the PTSD mice(P<0.05 or 0.01).Conclusion Inhibiting the activity of glutamatergic neurons in the dmPAG can alleviate defensive behaviors in mice with PTSD.

Objective To establish the energy expenditure(EE)prediction models of health Qigong Wuqinxi based on heart rate combined with accelerometer counts in different body parts,so to provide a reference for monitoring EE of Wuqinxi.Methods Seventy-four healthy college students aged 18-30 were selected as the research objects.They were divided into a skilled group of 39(21 males and 18 females)and a primary group of 35(17 males and 18 females)according to their level of practice,with 55 in the model group and 19 in the validation group.When performing a set of Wuqinxi,all subjects were recorded heart rate(HR),accelerometer counts[the average counts of X-axis,Y-axis,Z-axis and vector magnitude(VM)],and EE-related indicators by wearing the Polar heart rate moni-tor,ActiGraph-GT3X+accelerometers(9 parts:waist,both arms,both wrists,both thighs,and both ankles)and CORTEX Meta Max3B-R2 portable indirect calorimeter.Based on the accurate EE mea-sured by the gas metabolism meter,linear regression models of only HR,only accelerometer counts,or HR combined accelerometer counts were established,and their accuracy was analyzed and com-pared.Results The EE of Wuqinxi was significantly correlated with gender,height,weight,muscle mass,HR-related indicators,and accelerometer counts in different parts(X-axis counts of the waist,VM values of the left thigh and counts of the X-axis and Y-axis,VM values of the right thigh and the Y-axis,Z-axis count value,left ankle VM value and Y-axis count value,right ankle VM value and Y-axis,Z-axis count value)(P<0.05).Moreover,the adjusted R2 of the prediction model of only HR,only accelerometer counts,and the both were 0.582,0.508 and 0.678,respectively(P<0.05).The correlation between the predicted and measured values ranged between 0.706 and 0.817.Accord-ing to the Bland-Altman analysis,for each model,all except one error value fell outside the 95%confidence interval.That is,all models had an excellent fitting effect and high accuracy.Among them,the combined model was of the best prediction effect:EE(kcal)=-20.089+0.279×body weight(kg)+0.243×ΔHR(exercise HR-quiet HR,bpm)+0.001×Right thigh Y-axis count value+0.181×exer-cise HR(bpm)-4.202×gender(male=0,female=1).Conclusion The EE prediction model of Wuqinxi es-tablished on HR combined with accelerometer counts has the best effect and can calculate the EE more accurately.

Ferroptosis is a form of regulated cell death that depends on iron and reactive oxygen spe-cies.Different from apoptosis,necrosis,and autophagy,ferroptosis is characterized by the accumulation of lipid peroxides in cells.Studies have discovered that ferroptosis is closely associated with the occurrence and develop-ment of tumors and inducing ferroptosis in tumor cells can enhance the therapeutic effects of drugs on tumors.This article summarizes the latest research progress in ferroptosis regarding its mechanisms and associations with tumors,aiming to provide a reference for further understanding the interaction mechanisms between ferroptosis and tumors and offering new insights and targets for the treatment of tumors.

Objective To investigate the distribution of cell membrane tension in a gradient fluid shear stress(FSS)field.Methods A gradient plate flow chamber model was constructed.Fluid-solid coupling numerical simulations were conducted to analyze the distribution of membrane tension with different FSS gradients and FSS amplitudes under varying hydrostatic pressures.Results With an increase in the flow rate at the inlet of the flow chamber,the FSS gradient exhibited a proportionally positive increase.Under the gradient FSS field,the cell membrane tension initially decreased and then increased from the bottom to the top of the cell.Under normal blood pressure,higher hydrostatic pressure was correlated with increased membrane tension.Larger FSS amplitudes result ed in higher membrane tension.When the FSS amplitude was constant,the average difference in membrane tension between the high-and low-FSS regions increased with the FSS gradient.Similarly,with a constant FSS gradient,the average difference in membrane tension between the high-and low-FSS regions increased with the FSS amplitude.Conclusions Local variation in cell membrane tension induced by gradient FSS is a crucial factor influencing the directional migration of osteoclast precursors in a gradient FSS field.